I was wondering if anyone could tell me if addition of caffeine to the fixative affects antigenicity, generally speaking. I understand it's good for dealing with plant materials high in polyphenolic compounds. Does anyone know exactly what it's doing to the polyphenolics? Thanks, Karen
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dhaslam-at-kmslawyers.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, January 1, 2003 at 18:45:55 ---------------------------------------------------------------------------
Email: dhaslam-at-kmslawyers.com Name: donald haslam
Organization: kornfeld mackoff silber
Education: Undergraduate College
Location: vancouver bc canada
Question: I am an amateur botanist interested in studying rhododendron seeds for the purposes of identification of species and sub-species. I will be using a trinocular microscope of 10x to 40x magnification. If I use a digital camera, what is the recommended minimum number of pixels that I should insist on to usefully record images? Thank you. don p.s. How useful in such endevour is a zoom microscope?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
] FROM BRo/johnson junior IVORY COST WEST AFRICA PHONE225 07464760 A CRY FOR HELP Dear Sir,
It is my pleasure to write you after much consideration since telephone communication can not be suitable enough to communicate to you at first. Being the only son of my father, late Chief johmson SMITH from KWAZULU NATAL in Republic of South Africa (SA) I am 18 years of age. My father was limited liability Cocoa and Gold merchant in JOHANNESBURG South Africa before his untimely death. After his business trip to Abidjan -C?te d'Ivoire, to negotiate on a cocoa and gold business he wanted to invest in Abidjan - C?te d'Ivoire. A Week after he came back from Abidjan, he was attacked with my mother by unknown assassins, which my mother died instantly but my father died after five days in a private hospital on that faithful afternoon. I didn't know that my father was going to leave me after I had lost my mother. But before he gave up the ghost, it was as if he knew he was going to die. He my father, MAY HIS SOUL REST IN PERFECT PEACE he disclosed to me that he deposited the sum of $15,800,000,00 US Dollars (FIFTEEN MILLION EIGHT HUNDRED THOUSAND DOLLARS) in a bank here in Abidjan- C?te d'ivoire.That the money was meant for his cocoa and Gold company he wanted to establish in Abidjan - C?te d'Ivoire though, according to my father he deposited the money in a confident account of the bank and he handed to me all the relievant documents of the deposited fund and instructed me to seek for a reliable and trust worthy business partner for my life time investment abroad. Now I have succeeded in locating the bank here in Abidjan - C?te d'Ivoire. Now I am soliciting for your assistance to help me to transfer this money out from Abidjan to your safe account abroad so that we will invest it in any meaningful lucrative business in your country because this is my only hope in life. Awaiting anxiously to hear from you so that we can discuss the modalities of this transaction. Please kindly contact me through email immediately for more discussion or call me on225-07464760. Thanks for your kind attention.
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id KAA22730 for dist-Microscopy; Thu, 2 Jan 2003 10:24:49 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA22721 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Thu, 2 Jan 2003 10:24:18 -0600 (CST) Received: from novafrica.com ([193.251.130.55]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA22710 for {Microscopy-at-sparc5.Microscopy.Com} ; Thu, 2 Jan 2003 10:24:03 -0600 (CST) Received: (qmail 8412 invoked by uid 100); 2 Jan 2003 14:49:02 -0000
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FROM THE DESK OF;Engr CHRIST GESO, ABIDJAN COTE D,IVORIE WEST AFRICA PHONE 225 07464760 e-MAIL;christ.geso-at-ONDIKOI.COM
Dear sir,
On behalf of group of IVORY COST Senators i hereby propose the above to you.
I am Engr CHRIST.I.C.GESO Chairman Senate Committee on Contracts Review and foreign payments. There is presently available the sum of US$50m {Fifty Million United States Dollars} which members of this committee wish to transfer into your account, to be used for our re-election in 2003 general elections in IVORY COST
SOURCE OF FUND. This amount was realized from inflated or over-invoiced aspect of contracts executed by some foreign firms in 1999 when IVORUY COST hosted the world youth soccer Championship.ln the course of our duty we observed that there were some un-paid claims.lt is one of them that we intend to lodge into your bank account for our mutual benefit.
REMUNERATION. We have unanimously agreed that you will be entitled to 30% of the total sum, while 10% is set aside to offset expenses incurred during the transaction.
INVESTMENT. You will be required to place 60% of the total sum into any high yield investment facility in your country, for an initial duration of 8 months. During the 2003 elections, you will return 30% to us, but this time as a foreign loan for our election purposes. The accrued interest and the remaining 30% is what you will re-invest in the same process for a period of 4 years.
OPERATIONAL MODALITIES. We shall present you as one of the contractors awaiting payment for over-due contract payment for job executed for the Federal RepubliQU DE COTE,D,IVOREA in 1999.Application for claims, processing of approvals will be undertaken by my committee in conjunction with some highly placed officials, to ensure that fund is wired into your nominated bank account.
REQUIREMENTS. Your company details, banking details, as well as your confidential tel and fax numbers should be sent to me immediately to indicate your interest.
CONCLUSION. You do not stand any risk at all for being parts of this project, and your present line of business or profession is no hindrance. Honestly, our re-election in 2003 will be determined by this transaction and your ability to partner with us in all sincerity will enhance its success.
Note: That this transaction is expected to last between 7-14 days from the time we received this information, so i am waiting for your reply via my private e-mail address christ.geso-at-caramail.com MY PHONE NUMBER 225 07464760 Thanks for your co-operation and understanding while your urgent response is expected.
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id MAA23262 for dist-Microscopy; Thu, 2 Jan 2003 12:17:02 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id MAA23254 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Thu, 2 Jan 2003 12:16:31 -0600 (CST) Received: from fort-point-station.mit.edu (FORT-POINT-STATION.MIT.EDU [18.7.7.76]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id MAA23247 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 2 Jan 2003 12:16:19 -0600 (CST) Received: from grand-central-station.mit.edu (GRAND-CENTRAL-STATION.MIT.EDU [18.7.21.82]) by fort-point-station.mit.edu (8.9.2/8.9.2) with ESMTP id NAA06367; Thu, 2 Jan 2003 13:11:00 -0500 (EST) Received: from melbourne-city-street.mit.edu (MELBOURNE-CITY-STREET.MIT.EDU [18.7.21.86]) by grand-central-station.mit.edu (8.9.2/8.9.2) with ESMTP id NAA03292; Thu, 2 Jan 2003 13:10:46 -0500 (EST) Received: from emlab.mit.edu (EMLAB.MIT.EDU [18.82.0.121]) by melbourne-city-street.mit.edu (8.9.2/8.9.2) with ESMTP id NAA20435; Thu, 2 Jan 2003 13:10:46 -0500 (EST) Message-Id: {5.0.2.1.2.20030102111322.00ad1438-at-hesiod} X-Sender: tonygr-at-hesiod (Unverified) X-Mailer: QUALCOMM Windows Eudora Version 5.0.2
I have available two surplus X-ray analyzers (without detectors):
1: LINK/Oxford eX/L Mk. I, ca. 1988. Has been out of use for about 3 years, was in working order when last turned on. Has no video monitor. Has pulse processor, microscope scan control and electron image acquisition. Comes with SEM software suite.
2: LINK/Oxford Isis 300, ca. 1996. In use until recently. You provide the computer (works well with 300MHz Pentium, Win98 - it needs a computer with an ISA slot for the Translink card). Comes with Isis software suite on floppies, and key disks for TEM/STEM software suite (X-ray analysis, X-ray mapping, digital imaging, drift correction, etc). Has no work table.
Both have the original Link documentation.
These systems are offered on a strictly as-is basis, free to an academic user, you arrange the transport and take all responsibility. First-come, first-served. Easiest if you can visit Cambridge, MA and pick up. The Isis will fit into a family car. The eX/L might fit a car with a large trunk, but will certainly fit a station wagon or small pick-up.
I am not prepared to split these systems up for parts. If you want/need a specific board, then you take the whole thing and arrange disposal of the parts you do not need.
You can try calling with questions, but e-mail works well.
Tony Garratt-Reed.
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
Does anyone have a good protocol for doing SEM on replicas (casts) of human skin on a living, breathing person. I presume one would use a silicon rubber. Can that be imaged directly or would one make an epoxy cast from it? This is our first foray into cosmetology.
Greg Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program University of Florida P.O. Box 118525 Gainesville, FL 32611 352-392-1295
If you are looking to blot against naturally soluble proteins, then my first suggestion is to work with an assumption that during preparation of the tissue such proteins generally end up in the buffers, or the alcohols, etc. If not soluble, then the method that you must use to homogenize/subdivide any tissue to 'solubilize' portions of otherwise insoluble macromolecules should be sufficient to provide a useful specimen for blotting even if substantial crosslinking has occurred. By any standard, the blotting method that starts with fixed, processed and sectioned tissue (or vibratomed???) tissue should be compared at every step with fresh, unfixed and otherwise unprocessed tissue. IF the only difference between the section and the fresh tissue is HCHO, then that and its solvent constitute the only variables. You can compare fresh with HCHO-solvent, and solvent alone to see the effect of each. It would seem that blotting is an appropriate method for such determinations when assaying either Ab dilutions or epitope availability/presence.
Please remember the following. The anatomist looks at the bone and wonders where the molecule is. The biochemist looks at the molecules and wonders how they were assembled to form the bone. The histochemist looks at the molecule in the bone and wonders if that's where it was when the bone was still part of the living organism. For insoluble molecules the answers appear much more apparent (e.g., glycogen!).
Forgetting for the moment about "soluble or insoluble". If fixation was permitted to proceed for days beyond 24hr, then you are working with tissue in which there will be at least some frequency of methylene bridges linking adjacent molecules together. I am not very experienced with blotting in any case, but I have some familiarity with the methods. If the epitope for which you are going to assay has been tested on fixed tissue, with the same monoclonal you will use for blotting, without a requirement for retrieval, then the homogenized tissue sections (I assume that is what you will do with them.) should have the exposed epitope as well. If retrieval is necessary for immunohistochemistry, then there is no reason to expect that the homogenizing procedure alone will not also 'retrieve' a substantial amount of the epitope. Controlling fixed sections against unfixed tissue sections for normal consituent proteins is problematic but doable if you have a torsion balance with which to weigh the starting sectioned material.
Returning to the soluble and insoluble point again. If one compares identical amounts of fixed and fresh liver and calculates the amount of protein per cell, one should find that there is substantially more protein per cell in the fresh tissue and that the amount of protein per cell in fixed tissue falls with duration of fixation until no more diffusibile protein can escape the matrix or none remains.
Just for the sake of argument. Years ago a method by which one could ostensibly remove bound HCHO from a section was 1N NaOH. While this method might remove an -MeHO group, it probably isn't more likely to remove a so-called methylene bridge (a cross link) than any other hydrolytic method. In fact, if one turned to look, dreamily, out the window during such a procedure, the section might magically disappear.
} From my point of view, your question poses its own experimental answer. In modern immunohistochemistry each Ag/Ab pair places its own set of demands on the operator of the method.
Hope this helps,
Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: Beveridge, Mark J. [mailto:bevermj-at-peds.ufl.edu] Sent: Monday, December 30, 2002 9:56 AM To: 'Microscopy-at-MSA.Microscopy.Com'
i would like to know if there is any way to unfix tissue fixed in paraformaldehyde for immunoblotting?
Can anyone tell me of the fate of RMC, Inc.? I haven't seen anything from them in several years. We have an MT6000-XL ultramicrotome in our lab that's in need of some service. Any suggestions are welcome.
Richard Geissler Director, EM Laboratory Department of Pathology & Lab Medicine 859-257-5068 geissle-at-mail.uky.edu
Hi Donald, No zoom is desirable for specific photmicrography. Indeed, zooming is OK for photographing for show, but is disconcerting for quantitative characterizing. Zoom if you must, but insure that you capture images at the same magnification for your quantified studies. As to a; camera, I would wager that the new Nikon Coolpix 4500? would do the trick nicely. You can find good and reasonable adapters widespread on the new (the Nikon adapter is the highest priced of all). The most important determinant for a CCD camera is the purpose for which you purchase it. You must know its limits - there are few with film because the enlarging capacity is so great. Such is not the case with CCD images. You can zoom in 1-3X and see pixels. Scan a 2x2" 35mm transparency at 24-bits and you have a file of size 285MB. VERY BIG PRINTOUT!!! or WORTHLESS RESOLUTION WHEN THERE IS NO PRINTER FOR BILLBOARDS AVAILABLE. My Olympus CCD camera is a 1.3M-Pixel and provides great 5x7 color prints. If I use a CCD camera on a microscope, I have to be close to the publishing mag when I capture the image. Not so with film - though there are limits in this as well.
Here's a link: http://www.leubner.ch/index.html
Here's another to an encyclopedic source for seeds: http://hort.cabweb.org/SeedSci/Pdfs/ssr08075.pdf The above is for a PDF file so if you don't have the reader, you should get it free from Adobe.
I am not a botanist, so might I suggest you find one at a nearby college. Having some help in such an endeavor would likely lead to the collection of data that are publishable. Even a lawyer or a legal assistant should appreciate the prospect of starting with some notion of what work would be most productive.
Also, when you discover the best species for pie, please let me know so I can purchase a pack of the seed.
Hope this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: dhaslam-at-kmslawyers.com [mailto:dhaslam-at-kmslawyers.com] Sent: Thursday, January 02, 2003 12:54 AM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dhaslam-at-kmslawyers.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, January 1, 2003 at 18:45:55 ---------------------------------------------------------------------------
Email: dhaslam-at-kmslawyers.com Name: donald haslam
Organization: kornfeld mackoff silber
Education: Undergraduate College
Location: vancouver bc canada
Question: I am an amateur botanist interested in studying rhododendron seeds for the purposes of identification of species and sub-species. I will be using a trinocular microscope of 10x to 40x magnification. If I use a digital camera, what is the recommended minimum number of pixels that I should insist on to usefully record images? Thank you. don p.s. How useful in such endevour is a zoom microscope?
I followed the discussions concerning pregnancy and sem operation with interest in the past. Unfortunately, I did not save the replies as it seemed unlikely to occur in our lab. We currently have a pregnant occassional operator. At her request, the decision has been made to transfer sem work to another individual for the duration of the pregnancy.
When I mentioned that the list server has covered this topic in the past, I was asked if I the information was still available. I welcome any new infomation and hope that some one can pass along archived info.
Gregory W. Erdos wrote: ============================================================= Does anyone have a good protocol for doing SEM on replicas (casts) of human skin on a living, breathing person. I presume one would use a silicon rubber. Can that be imaged directly or would one make an epoxy cast from it? This is our first foray into cosmetology. ============================================================= I would reference my own (old) publications:
"Characterizing Cosmetic Effects and Skin Morphology by Scanning Electron Microscopy", presented at SCC Annual Educational Symposium, May 1975, St. Louis; J. Soc. Cosmet. Chemists, 27, 509-531 (November 1976).
If one does not have easy access to this journal, a "sample" of this type of work can be seen on the SPI Supplies website at URL http://www.2spi.com/catalog/spec_prep/a.html
While we were not the very first to ever replicate, in vivo, human skin, we were the first to publish a testing method for evaluating skin care products for the cosmetics and toiletries industries, and later for the pharmaceutical industry.
For "cosmetology" which I assumed is for the substantiation of cosmetics industry advertising claims, one must use a very rapidly curing resin for the original cast or "negative" replica, otherwise the cast itself, and the occlusiveness of the covering creates changes ("moisturing effects" if you will) that are greater than the effects of the products being studied. Hence the need for a replicating system that cures in literally seconds, instead of minutes or even hours. The system we developed is what has evolved over the years into the SPI Supplies Wet Replica Kit, see URL http://www.2spi.com/catalog/spec_prep/wet_rep_kits.shtml
Can it be imaged directly? Well, yes it can but it is not the preferred way . It is far less confusing to make a positive replica from the negative, and photograph the positive. That is why the replicating kit described above comes with a pololefinic powder that so far as we are concerned makes the very best positives with the least amount of shrinkage or distortion.
Does it pass the test of being "pretty good?" I think so since many of the leading laboratories in the cosmetics and toiletries industry use this kit for their work. It has been pretty extensively peer-reviewed by a number of technical persons working on skin care products and "moisturizer" products.
I would be happy to elaborate more on this technique if there were any questions, either on or off the listserver.
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 ######################## ============================================
Greg Dental silicone is excellent for replicating skin. The product I am familiar with is Kerr Reflect, though there are others no doubt. The setting time at body temperature is rapid, the material is comparatively non-toxic, releases very cleanly and the replica contains information with resolution approaching 100nm. Positive replicas can be made with epoxy, but I have had better results using hot-melt plastics such as polycarbonate. My method was fairly unrefined - the silicone negative replica was placed in an oven at just above the melting point of the plastic, a piece of 3mm plastic sheet placed on top and weighted with a brass weight. PTFE or silicone sheet was used to prevent adhesion between plastic and weight. The specimens were allowed to cool to room temperature with the weights in place before separating positive from negative. With care more than one copy can be made if necessary, though I daresay fine detail is gradually degraded by this. I think the main reason this has worked better for me than replicating with epoxy is that I use it for replicating leaves, which are covered with wax crystals. The silicone picks up these waxes, so epoxy only sees their undersides, especially when cold- polymerized. At high temperature, polycarbonate displaces the molten wax and can replicate the negative replica of the outer surface. This may not be an issue for you in replicating skin, but the combination of high temperature and pressure also helps displace air from features like hairs, which epoxy can be reluctant to enter. Vacuum infiltration of epoxy at high temperature may improve this though.
Hope this is of some use. Best wishes for the new year Chris
On 2 Jan 03, at 15:39, Greg Erdos wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Fellow scopesters } } Does anyone have a good protocol for doing SEM on replicas (casts) of } human skin on a living, breathing person. I presume one would use a } silicon rubber. Can that be imaged directly or would one make an } epoxy cast from it? This is our first foray into cosmetology. } } Greg } Gregory W. Erdos, Ph.D. } Assistant Director, Biotechnology Program } University of Florida } P.O. Box 118525 } Gainesville, FL 32611 } 352-392-1295 } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 /8669 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
Here is the table of contents for the January '03 issue of Microscopy Today:
Carmichael: Out with the Old, In with the New Boyes: Pros and Cons of Low Voltage SEM EDX Elemental Analysis Basgen: Basic Stereology Sedgewick: Segmentation Before Quantization By Using Photoshop: Darkfield Images Clarke: Rediscovery of Darkfield Dispersion Staining while Building a Universal Student Microscope Beanland: Rapid Cross-Section TEM Specimen Prep. of III-V Materials Harmsen: Scan Speed, Mag and Accuracy. That is the Question! Maleeff: Removing Very Fine Wrinkles from Half-Micron Epoxy Sections Killius: Problems with Hydrophobicity of Coated Grids Mascorro: The Versatile Family of Epoxy Resins: Designing Embedding Media with Specific Viscosity Properties Fandrich, et al.: Secondary Melt Ornamentation On Westwater, Utah: Microspherules: Evidence Of An Extraterrestrial Provenance
Some news about Microscopy Today: As of 12/31/02 we have 17,411 subscribers, making us the largest microscopy magazine in the USA.
I will close the subscription list for mailing the January issue on January 7th. You must subscribe before then to receive this issue.
All subscriptions/changes/unsubscribes must come via our website: www.microscopy-today.com
Note that if you live in the USA and received the November issue of MT, along with the M&M-2003 Call for Papers, you probably already have a subscription.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
You could use a dental impression material; they are usually silicone-based but can be other things, and all are completely harmless to the subject and the experimenter. It is possible to image the negative made this way, but there are problems with its behaviour under vacuum. The negative can be used to make a positive in epoxy resin which is virtually indistinguishable under SEM from the original.
Wieland M, Chehroudi B, Textor M, Brunette DM. Use of Ti-coated replicas to investigate the effects on fibroblast shape of surfaces with varying roughness and constant chemical composition. J Biomed Mater Res. 2002 Jun 5;60(3):434-44. PMID: 11920667 [PubMed - indexed for MEDLINE]
2: Chehroudi B, Brunette DM. Subcutaneous microfabricated surfaces inhibit epithelial recession and promote long-term survival of percutaneous implants. Biomaterials. 2002 Jan;23(1):229-37. PMID: 11762842 [PubMed - indexed for MEDLINE]
Hope this helps.
Lesley Weston.
on 02/01/2003 12:39 PM, Greg Erdos at gwe-at-biotech.ufl.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 } -----------------------------------------------------------------------. } } } Fellow scopesters } } Does anyone have a good protocol for doing SEM on replicas (casts) of human } skin on a living, breathing person. I presume one would use a silicon } rubber. Can that be imaged directly or would one make an epoxy cast from } it? This is our first foray into cosmetology. } } Greg } Gregory W. Erdos, Ph.D. } Assistant Director, Biotechnology Program } University of Florida } P.O. Box 118525 } Gainesville, FL 32611 } 352-392-1295 } }
As I was the instigator of this subject, I thought that I would give you a synopsis of what I learned.
1. It is against Federal Law to ask any employee if they are pregnant, plan to get pregnant, or are sexually active (both male and female). 2. The lab is legally responsible for any damage which may occur to the mother or fetus due to working in the lab. So, the lab is responsible, and therefore the management liable, but it is against the law to know what condition your workers are in vis a vis reproduction. 3. What I have gone to here is treating everyone as if they are reproductive. With the appropriate protocols this is possible and makes for a much safer work environment for everyone. 4. The use of modern microscopes is not dangerous from radiation. If there is radiation coming out of your scope , get it fixed or get rid of it. There are many good scopes out there for a pittance. If you are really paranoid, get one of those badges and have your heaviest user wear it, if it show radiation, see above. 5. With the use of appropriate hoods and safety equipment, anyone should be able to prepare samples. Do however, use the safer chemical and/or procedure when possible. Such as Hepes, not Cacodylate; microwave fixation in hood, not conventional. 6. Once you know that you have a pregnant worker, contact your safety people and have the worker prepare a personal hygiene plan and have it approved by the safety office and suggest that her doctor have a look at it to see if he/she has any concerns. I had two pregnant workers in the lab at the same time, there were no problems.
Just one personal aside, it is much easier to manage an employee if you know that she is pregnant, some have little to no problems, but others can have a very hard time. If you do not know what is going on, the possibility is there to misjudge the performance of that individual, but the law allows for the worker to privacy. Just one more thing to deal with in the modern world.
Bill
William McManus Supervisor Microscopy Facility Utah State University Logan UT 84322-5305
-----Original Message----- } From: Cochran [mailto:fisher-at-meganet.net] Sent: Thursday, January 02, 2003 6:45 PM To: microscopy-at-sparc5.microscopy.com
Hi All,
I followed the discussions concerning pregnancy and sem operation with interest in the past. Unfortunately, I did not save the replies as it seemed unlikely to occur in our lab. We currently have a pregnant occassional operator. At her request, the decision has been made to transfer sem work to another individual for the duration of the pregnancy.
When I mentioned that the list server has covered this topic in the past, I was asked if I the information was still available. I welcome any new infomation and hope that some one can pass along archived info.
} 6. Once you know that you have a pregnant worker, contact your safety } people and have the worker prepare a personal hygiene plan and have it } approved by the safety office and suggest that her doctor have a look at } it to see if he/she has any concerns. } I had two pregnant workers in the lab at the same time, there were no } problems. } :
How long is it going to take to go through each step of the procedure mentioned above? By the end of that procedure it might be too late for the fetus.
I believe that as soon as a lady learned or suspect she is pregnant she should stop working with radiation stuff. There is no safe dose of radiation. Risk might be small but it is always positive.
Kind regards, Ayten.
} Just one personal aside, it is much easier to manage an employee if you } know that she is pregnant, some have little to no problems, but others } can have a very hard time. If you do not know what is going on, the } possibility is there to misjudge the performance of that individual, but } the law allows for the worker to privacy. Just one more thing to deal } with in the modern world. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } -----Original Message----- } } From: Cochran [mailto:fisher-at-meganet.net] } Sent: Thursday, January 02, 2003 6:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Pregnancy & SEM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } } I followed the discussions concerning pregnancy and sem operation with } interest in the past. Unfortunately, I did not save the replies as it } seemed unlikely to occur in our lab. We currently have a pregnant } occassional operator. At her request, the decision has been made to } transfer sem work to another individual for the duration of the } pregnancy. } } When I mentioned that the list server has covered this topic in the } past, I was asked if I the information was still available. I welcome } any new infomation and hope that some one can pass along archived info. } } Thanx, } Ray Cochran } } } }
I'm fairly interested in this, too, would you please copy responses to me?
I don't know how others feel, but I'm always a bit disappointed when people ask for off-list replies, as it shuts me out of the loop, having given a tantalising initial glimpse of a thread. It seems to me that one of the wonderful things about this list is the discussions, and I would hate for it to be merely a bulletin board on which people posted their requests for information.
Commercial and personal privacy issues occasionally preclude open discussion, I understand that, but that's pretty rare.
Happy New Year
rtch
} } Hi, } } We have a researcher on campus who would like to examine the cell size } of 3000 year old wood. Any ideas on the sample preparation protocol to } preserve the cell structure without shrinking? } } Please direct your responss to Cheryl Jensen (jensenc-at-missouri.edu). } } Thanks in advance. } Lou Ross } -- } Senior Electron Microscope Specialist } Electron Microscopy Core Facility } W136 Veterinary Medicine } University of Missouri } Columbia, MO 65211-5120 } (573) 882-4777, fax 884=5414 } email: rosslm-at-missouri.edu } web: www.biotech.missouri.edu/emc }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Your pont is well taken, but the law does not let us take this kind of action. In the past, companies have used pragnancy as a tool to demean women, and even if meant in the best of intentions, a lab manager cannot get involved until it is probably too late, a catch 22. That is why, I have set up my lab so that all workers are treated the same, is if they are pregnant, even the men. An issue generally not discussed is, what are the ramifications to male gametes when exposed to chemical and radiation exposure? But again, let me repeat, if you have an EM that is leaking radiation that is bad, and should be dealt with, period. Fix it or pitch it. In reality, we are exposed to more teratigens and carcinogens in the grocery store than in a well run EM lab.
Bill
-----Original Message----- } From: celik ayten [mailto:celik-at-ews.uiuc.edu] Sent: Friday, January 03, 2003 1:20 PM To: William McManus Cc: fisher-at-meganet.net; microscopy-at-sparc5.microscopy.com
I was once told that the only possible way to get irradiated by an SEM was to have your body under the beam while it is operating. I have not tried it yet to see if it is true or not.
Ron L
-----Original Message----- } From: celik ayten [mailto:celik-at-ews.uiuc.edu] Sent: Friday, January 03, 2003 3:20 PM To: William McManus Cc: fisher-at-meganet.net; microscopy-at-sparc5.microscopy.com
On Fri, 3 Jan 2003, William McManus wrote:
[skip]
} 6. Once you know that you have a pregnant worker, contact your safety } people and have the worker prepare a personal hygiene plan and have it } approved by the safety office and suggest that her doctor have a look at } it to see if he/she has any concerns. } I had two pregnant workers in the lab at the same time, there were no } problems. } :
How long is it going to take to go through each step of the procedure mentioned above? By the end of that procedure it might be too late for the fetus.
I believe that as soon as a lady learned or suspect she is pregnant she should stop working with radiation stuff. There is no safe dose of radiation. Risk might be small but it is always positive.
Kind regards, Ayten.
} Just one personal aside, it is much easier to manage an employee if you } know that she is pregnant, some have little to no problems, but others } can have a very hard time. If you do not know what is going on, the } possibility is there to misjudge the performance of that individual, but } the law allows for the worker to privacy. Just one more thing to deal } with in the modern world. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } -----Original Message----- } } From: Cochran [mailto:fisher-at-meganet.net] } Sent: Thursday, January 02, 2003 6:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Pregnancy & SEM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } } I followed the discussions concerning pregnancy and sem operation with } interest in the past. Unfortunately, I did not save the replies as it } seemed unlikely to occur in our lab. We currently have a pregnant } occassional operator. At her request, the decision has been made to } transfer sem work to another individual for the duration of the } pregnancy. } } When I mentioned that the list server has covered this topic in the } past, I was asked if I the information was still available. I welcome } any new infomation and hope that some one can pass along archived info. } } Thanx, } Ray Cochran } } } }
Chris Jeffree wrote: ========================================================== Dental silicone is excellent for replicating skin. The product I am familiar with is Kerr Reflect, though there are others no doubt. The setting time at body temperature is rapid, the material is comparatively non-toxic, releases very cleanly and the replica contains information with resolution approaching 100nm. Positive replicas can be made with epoxy, but I have had better results using hot-melt plastics such as polycarbonate. My method was fairly unrefined - the silicone negative replica was placed in an oven at just above the melting point of the plastic, a piece of 3mm plastic sheet placed on top and weighted with a brass weight. PTFE or silicone sheet was used to prevent adhesion between plastic and weight. The specimens were allowed to cool to room temperature with the weights in place before separating positive from negative. With care more than one copy can be made if necessary, though I daresay fine detail is gradually degraded by this. I think the main reason this has worked better for me than replicating with epoxy is that I use it for replicating leaves, which are covered with wax crystals. The silicone picks up these waxes, so epoxy only sees their undersides, especially when cold- polymerized. At high temperature, polycarbonate displaces the molten wax and can replicate the negative replica of the outer surface. This may not be an issue for you in replicating skin, but the combination of high temperature and pressure also helps displace air from features like hairs, which epoxy can be reluctant to enter. Vacuum infiltration of epoxy at high temperature may improve this though. ================================================================ The polymerization time of the Kerr "Reflect" system is longer than the system I described a few days ago as the SPI Wet Replica Kit. I agree that the faster polymerization time for the replication of leaves might not be important, and a longer time might even be at times an advantage. But once the silicone is applied to skin, it literally stops all transepidermal water loss. And the trapped water has no where to go but to build up in the stratum corneum (the outer most layer of skin, or the dead skin layer), creating an experimentally induced artifact effect. This effect is often times more profound than the subtle difference one is trying to discern in terms of product efficacy or whether one product is better than another. That is one reason why so many times projects of this type, where one is looking for differences fail because the transepidermal moisture buildup makes everything look the same!
We have not looked lately at this particular Kerr product but we have looked at other dental silicones and in order to impart to them the kind of dimensional stability needed for dental impressions, they are loaded with an inorganic additive system to give them that desired dimensional stability. When one starts going up in magnification, about about 500X, the additives from the negative replicating material can be resolved (as an artifact effect). The SPI Wet Replica Kits does not have to have such good dimensional stability, and therefore has a different type and loading of the inorganic additives, and one can generally achieve magnifications (before artifacts set in) about 1.5X higher, to about 750X. While this might not sound very high in magnification in SEM terms, it tends to be more than high enough for visualizing the effects of cosmetic and topically applied pharmaceutical products to human skin. It has been our experience that most meaningful work is done not higher than about 300X although for some kinds of work up to about 600X.
The advantage of a much faster polymerizing system is that the replica essentially "cures" before the body generated moisture buildup starts to create effects all of its own. And that is what makes possible the visualization of very subtle effects from the application of a product, either short term of long term.
The negatives made this way are understandably fragile but the non-wetting characteristics of the silicone on skin, even on dry flaky skin, leads to an easy lift-off without much in the way of disruption. We have found that on most types of skin, and in particular, dry skin, a first positive is made and thrown out; its function is to serve as a "cleaning replica", to clean off any remains of skin flakes. The second replica is what is then used for the microscopy. The low melting polyolefinic very fine powder used for the positive replicas will flow pretty easily into the cylindrical "holes" that would be representing a shaft of hair. The entrapment of air bubbles is not a serious problem.
We have found that for skin work, epoxies and just about anything else much more brittle than the polyolefin used for the positive does not work as well ; that way even curved channels and irregularly shaped volumes can be successfully transmitted to the positive replica where as if done in other materials, for example, an epoxy, such micro-volumes would be snapped off then the two replicas were separated.
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 ######################## ============================================
Dick I understand that the Kerr Reflect product range has been replaced by a new range known as Extrude. The closest equivalent to the Kerr Reflect Wash (low viscosity formulation) that I used is Kerr Extrude Wash. Check out the other impression materials in their range. There is, for example, a product based on sodium alginate that is aqueous, hydrophilic, and has faster setting times than the Kerr silicone. For details and suppliers contact Kerr Dental:
www.KERRDENTAL.com Mailing Address : 1717 West Collins Orange, CA 92867 Phone : (800) KERR-123 (800-537-7123), (714) 516-7400
Silicone impression materials are supplied by most dental laboratory supply dealers throughout UK, and probably in US too.
Chris
----- Original Message ----- } From: "Richard Gordon" {gordonr-at-Ms.UManitoba.CA} To: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} Cc: "Richard Gordon" {gordonr-at-Ms.UManitoba.CA} Sent: Saturday, January 04, 2003 11:51 PM
Here is my reply to this, with some additions:
One way to do this would be to freeze the tissue in the cryo-prep chamber of cryoSEM, then observe frozen at low kV. Or plunge freeze in LN2, cryoplane, transfer to cryostage, etch, observe. If the wood is dry, freezing is probably unnecessary. And if you just want to look at cell size on a surface, you may get away without coating at very low kV. Depends on how much you are able to process this wood. Roger Heady - Roger.Heady-at-anu.edu.au, http://sres.anu.edu.au/people/headyr.html - is an expert in examining wood in SEM and could give much better advice - I don't think he's on this email list. cheers, Rosemary } } } Hi, Lou } } I'm fairly interested in this, too, would you please copy } responses to me? } } I don't know how others feel, but I'm always a bit disappointed } when people ask for off-list replies, as it shuts me out of the } loop, having given a tantalising initial glimpse of a thread. It } seems to me that one of the wonderful things about this list is } the discussions, and I would hate for it to be merely a bulletin } board on which people posted their requests for information. } } Commercial and personal privacy issues occasionally preclude } open discussion, I understand that, but that's pretty rare. } } Happy New Year } } rtch } } } } } } Hi, } } } } We have a researcher on campus who would like to examine the cell size } } of 3000 year old wood. Any ideas on the sample preparation protocol to } } preserve the cell structure without shrinking? } } } } Please direct your responss to Cheryl Jensen (jensenc-at-missouri.edu). } } } } Thanks in advance. } } Lou Ross } } -- } } Senior Electron Microscope Specialist } } Electron Microscopy Core Facility } } W136 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211-5120 } } (573) 882-4777, fax 884=5414 } } email: rosslm-at-missouri.edu } } web: www.biotech.missouri.edu/emc } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Allen Sampson {ars-at-sem.com} To: "'Sergey Ryazantsev'" {sryazant-at-ucla.edu} , "microscopy-at-sparc5.microscopy.com" {microscopy-at-sparc5.microscopy.com} 12/22/02 05:26 AM cc: Please respond to Subject: RE: Evaporator cold trap "ars-at-sem.com"
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Sergey,
That is a strong pump for this system - the normally configured system uses
a 250 L/sec pump. A diffusion pump, as any pump, is a differential system - The ultimate pressure achieved is a balance between the leak rate in the input, the back pressure in the output and the bypass rate in the pump (how
much gas can pass from the output to the input for a given pressure differential, giving rise to the mechanical pump oil contamination in a system). Although these pumps are rated up to 10-9 pressures, this is never seen in operational systems. If you were to cap the diffusion pump with a metal sealed, solid metal cap and had an enormous backing pump, you may come close to the rated ultimate rating. In practical systems, these levels are but a dream.
I'd be interested in knowing what roughing pump you are using.
No vacuum system has no leaks, particularly when elastomeric seals are used. The standard Denton, with its manual valves and bell jar seal, has a
lot of elastomeric seals. And a cold cathode vacuum gauge accuracy is debatable at 10-6 or less.
I've only recommended and used Santovac 5 for over twenty years. It is a great diffusion pump oil. My main reason for recommending it is not the ultimate vacuum attainable (debatable considering the greater temperature apparently needed by Santovac, although I've never had a problem using it in any diffusion pump), but rather it's relative insensitivity to sudden air inrushes when hot. That means that it will 'crack' and polymerize less
than other oils. In other words - it will last longer and cause less hard deposits on the pump.
As far as o-rings, I've found Buna to be quite acceptable, lightly coated with Brayco, for static seals. Where dynamic seals are used (rotational or
translational forces are common), I use Buna with Apiezon (the waxier Apiezon has more staying power, although it will also hold particulate contaminants more). The reason for cracked o-rings is neglect, not the suitability of vacuum greases. The reason for greasing o-rings is to provide a light coating that preserves the qualities of the elastomeric material, not to make up for insufficiencies in the vacuum sealing surfaces. In this respect, the better vacuum greases do a good job and do not compromise either Buna or Viton. I have many 25+ year old systems that
have original o-rings that are indistinguishable from new, in appearance, shape or conformance.
The deformability of Buna is actually a plus, at least in systems that are mostly held at vacuum. You may have noticed that a system that was just rebuilt with new or rebuilt o-rings takes some time to come to an ultimate vacuum equilibrium. That, of course, involves the outgassing of the system
components after being exposed to atmosphere for some time during the rebuild. But it also includes the time required for the elastomeric vacuum
seals to be 'sucked' into place. A well designed o-ring seal will depend on the mechanical pressures on the o-ring, but will ultimately depend on the o-ring's conformance under the gas pressures it's subjected to.
In my experience, the Buna o-rings will deform to provide a good seal faster and, properly maintained, will continue to conform to a shape that best seals. I generally use Viton for it's improved resistance to high temperatures. In either case, I use an acetone wipe for cleaning o-rings every time I recondition them. It tends to swell the o-rings with two effects - it restores them to their original shape and helps to provide a quick seal when the system is pumped down again.
BTW, I've cleaned more DPs and refurbished more vacuum systems than I'd like to elucidate. In my business, I tend to get the instruments that the manufacturers don't service anymore, didn't service properly or have been neglected for some time.
Just a few ruminations from a long career of servicing many vacuum instruments.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Sunday, December 22, 2002 3:12 AM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } Allen } } DV502A has 800 l/sec DP!!! It enough to pump down a huge volume. If you
} don't have any leaks in your system, the vacuum would be directly dependent } from the ultimate vacuum for the pump (which is somehow dependent from } pump's actual construction/quality and DP Oil). In most cases leaks are the } reason of vacuum degradation. Any normally serviced vacuum evaporator } should deliver at least very good 10-6 torr. 10-5 is very bad and actually } you may not use it for biological sample preparation. In my particular } case, I've replaced ALL O-rings on viton with touch of Apiezon L, cleaned
} and polished DP and used Santavac-5. Santavac-5 is great DP oil. You have } to try. On my DV502A I have 2*10-6 after about 40 min pumping (no LN2) and } better than 5*10-7 overnight. I am using MKS cold cathode gauge with } sensitivity up to 10-9 torr (which I don't need). I think, the vacuum } quality is mostly a function of how clean your system and how many 10-year } old cracked "buna" O-rings is inside your system. The problem with "buna" } - it does not hold the shape and easily deformed even if it's not old. It's } also destroyed by vacuum grease (does not matter what manufacturers told } you). Viton is much better. Since I spent $500 on Santavac-5 8 years ago, } I never touch my DP again. By the way, I have another vacuum system, which } I build by myself. It has exact the same volume and similar amount of } O-rings but 400 l/s TP from SEIKO. That system is oil-free. I mean, it } has scroll pump as a backing device and TP itself does not have any oil } (it's magnetically levitated beauty). So, when I build the system, I was
} expecting similar productivity for this system as for DV502A but } oil-free. I was completely wrong! This "baby" easily delivered to me } 5*10-7 torr in 20 (yes, twenty) min! After a few hours, it's going into } 10-8. So, my "theory" is that in the standard setup, oil from mechanical
} pump contaminated the whole system (and your sample!) and adsorbs a lot of } air, which slowly released during the high vacuum pumping cycle. Because
} my new system is oil-free, it's much faster. It has 12x12" Bell Jar with
} 6' collar. Another example: my old Polaron with 100 l/s DP (Santavac-5 } again) and 12x12" Bell Jar. This system is very comfortable with 2*10-6 } (no LN2) after I repaired manufacturers defect in DP. All tricks how to get } good vacuum perfectly described in the Wil Bigelow book. Have a great } Holidays (don't start cleaning DP- it's messy)! Sergey } } } At 08:13 PM 12/20/02 -0600, you wrote: } } Sergey, } } } } Have you verified that vacuum level with an independent, calibrated vacuum } } gauge? Vacuum sounds way to high for that system. I agree with all of } } your recommendations, but case in point, the vacuum you claim is on an } } order of what we normally find in a valve isolated electron gun (much } } smaller vacuum and seal volumes) pumped by a 20 L/sec ion pump. The vacuum } } level you stated for the start of your work is far closer to the best I } } have seen out of this particular evaporator after a complete rebuild. } } } } Allen R. Sampson } } Advanced Research Systems } } 317 North 4th. Street } } St. Charles, Illinois 60174 } } } } phone (630) 513-7093 fax (630) 513-7092 http://www.sem.com } } } } } } On Friday, December 20, 2002 3:01 PM, Sergey Ryazantsev } } [SMTP:sryazant-at-ucla.edu] wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FA Q.html } } } -----------------------------------------------------------------------. } } } } } } } } } Randy } } } I am sorry for late comments. I agree with Wil Bigelow that LN2 trap will } } } reduce the pump performance. If you need just to increase the system's } } } overall performance, I would suggest you have to do very good service
for } } } it first (replace all suspicious O-rings, clean everything up). I highly } } } recommend to use Santovac-5 DP (I assume, it's DP based system, because } } TP } } } don't need LN2). When I come in UCLA, I had DV-502A vacuum system with } } } 5*10-5 torr. I cleaned up DP (messy work and a lot of solvents), } } replaced } } } all O-rings (the system was 10 y.o. at the moment) and used } } } Santovac-5. Since that I do have 5*10-7 with no LN2 (and no further } } } service for more than 5 years). As an alternative, you may install some } } } "cold finger" with protective screens near your sample in the Bell } } } Jar. You really need LN2 trap over DP if you pump a lot of water in your } } } experiments. Sergey } } } } } } At 06:09 PM 12/16/02 -0600, 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 Hitachi HUS-4 vacuum evaporator. I'm hoping to put a LN2 } } cold } } } } trap } } } } on it, in an attempt to clean up the vacuum a tad more. There is a 3.5 } } inch } } } } by 4.5 inch plate right above the diffusion stack which looks to be a } } great } } } } place to attach a cold trap. Rumor has it that this was an option. Are } } there } } } } any old traps kicking around that one can acquire? I could have our } } machine } } } } shop manufacture one, but was hoping for a cheaper route. } } } } Thanks in advance, } } } } Randy Nessler } } } } 319-335-8142 } } } } } } ------------------------------------------------------ } } } } } } Sergey Ryazantsev, Ph.D. } } } Electron Microscopy } } } Department of Biological Chemistry, School of Medicine } } } University of California, Los Angeles } } } Box 951737 } } } Los Angeles, CA 90095-1737 } } } } } } (310) 825-1144 (office) } } } Pager: (310) 845-0248 } } } FAX: (310) 206-5272 (departmental) } } } mailto:sryazant-at-ucla.edu } } } } } } } } ------------------------------------------------------ } } Sergey Ryazantsev, Ph.D. } Electron Microscopy } Department of Biological Chemistry, School of Medicine } University of California, Los Angeles } Box 951737 } Los Angeles, CA 90095-1737 } } (310) 825-1144 (office) } Pager: (310) 845-0248 } FAX: (310) 206-5272 (departmental) } mailto:sryazant-at-ucla.edu } } }
Depending on the motivation of the pregnant worker, and the quality of your safety personnel, about a week. In the interim, the person can be given other duties, or in our case just carefully follow the procedures already in place, which have been designed to protect. The biggest question is: Will the lab administration give you the time to get set up before the fact. In a real sense, it is always too late to begin preparations after definitive knowledge of pregnancy is know. In Kathy's case se would have been better off to wait to tell everyone of her condition, this is a shame, however, and we have not come as far as we would like to think with women's rights. In my case it was very beneficial to know that the worker was pregnant, her work dropped off substantially and she napped at the micortome. If I was unaware of the situation. I might have reacted in a negative sense, but instead, I was able to protect her from those who would not understand.
Bill
-----Original Message----- } From: celik ayten [mailto:celik-at-ews.uiuc.edu] Sent: Friday, January 03, 2003 1:20 PM To: William McManus Cc: fisher-at-meganet.net; microscopy-at-sparc5.microscopy.com
On Fri, 3 Jan 2003, William McManus wrote:
[skip]
} 6. Once you know that you have a pregnant worker, contact your safety } people and have the worker prepare a personal hygiene plan and have it } approved by the safety office and suggest that her doctor have a look at } it to see if he/she has any concerns. } I had two pregnant workers in the lab at the same time, there were no } problems. } :
How long is it going to take to go through each step of the procedure mentioned above? By the end of that procedure it might be too late for the fetus.
I believe that as soon as a lady learned or suspect she is pregnant she should stop working with radiation stuff. There is no safe dose of radiation. Risk might be small but it is always positive.
Kind regards, Ayten.
} Just one personal aside, it is much easier to manage an employee if you } know that she is pregnant, some have little to no problems, but others } can have a very hard time. If you do not know what is going on, the } possibility is there to misjudge the performance of that individual, but } the law allows for the worker to privacy. Just one more thing to deal } with in the modern world. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } -----Original Message----- } } From: Cochran [mailto:fisher-at-meganet.net] } Sent: Thursday, January 02, 2003 6:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Pregnancy & SEM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } } I followed the discussions concerning pregnancy and sem operation with } interest in the past. Unfortunately, I did not save the replies as it } seemed unlikely to occur in our lab. We currently have a pregnant } occassional operator. At her request, the decision has been made to } transfer sem work to another individual for the duration of the } pregnancy. } } When I mentioned that the list server has covered this topic in the } past, I was asked if I the information was still available. I welcome } any new infomation and hope that some one can pass along archived info. } } Thanx, } Ray Cochran } } } }
I've recently developed a problem with a newly purchased Kodak EM film. The EM film box says "new formulation" on it.
Well, this new formulation on film is lowering my contrast and I get some funky fog on it.
I was wondering if anyone else have experienced this problem and if so what can we do?
I've used old film ( old formulation ) and experience no fogging and get nice contrast. Its also not a scope specific problem or light leaks or developer. Its the film.
Rajesh Patel Robert Wood Johnson Medical School Department of Pathology 675 Hoes Lane Piscataway, NJ 08854
Say, can anyone recommend a super-reliable and quiet brand (and size) of air compressor to use with a Philips EM400T?
Unfortunately, in addition to being very, very loud, the low grade air compressor we've been using has the intermittent habit of blowing it's circuit breaker (twice in the last couple of weeks), which has set us way back on getting our "new" microscope fully evacuated for the first time.
I'm waiting for a catalog from Jun-Air, but would like to know what model/size has been used reliably by others over the years. The Philips installation manual doesn't give much more than the air pressure requirement, so I'm uncertain what is ideal in terms of pump and tank capacities. I've asked the folks at Jun-Air, but they are reluctant to make any recommendation based solely on the pressure spec.
Thanks for any tips!
Best regards, Eric Anderson Southern CT State University Physics Dept. 501 Crescent Street, JE108B New Haven, CT 06515 203-392-6468
Those of you planning to attend the 2003 Microscopy and Microanalysis meeting August 3-7, 2003 in San Antonio meeting
I am soliciting presentations for the following session at this upcoming meeting. Related topics described below, as well as outreach programs for grades 6 and above, would be considered. Please contact me should you wish to participate.
See you in Texas!
41. Teaching Microscopy and Imaging in the Digital Age
Organizer: Steve Barlow
Computer technology is changing the ways we access equipment, view samples, and record or manage images. Computer-controlled microscopes and digital imaging have created many new training methods. In addition, these changes make it possible for classroom students to analyze microscopy data and images without requiring access to expensive microscopes. This session will look at new training approaches, such as virtual microscopes and telemicroscopy, and new teaching tools, such as CDs, DVDs, and computer software. The session will discuss using digital technology to create laboratory training protocols and to teach classroom analysis of data contained in digital images. -- 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/
Rajesh If it's film, nothing to do with it! May be for "new formulation" they suggested "new" developing procedure? Check Instruction and definitely contact Kodak. If this product going under "old" trade name, like "4489 film" it should be exact the same every time. I would suggest, you may return the whole batch of the "new formulation" film back to manufacturer. Kodak sold film through distributors, so it's possible that film was stored improperly before shipped to you (another reason to return it back). Sergey
At 10:53 AM 1/6/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Dear colleague From time to time the issue of EM safety appeared on this ListServer. Somehow, the biggest concern was the safety of the (already) pregnant women. In my point of view, when women IS pregnant and fetus is already developed, IT'S TOO LATE TO TALK (or do something) about safety in the EM Lab! Most our "dangerous" stuff has tendency to be accumulated in the body over time (UA, Pb and so on). Some substances are carcinogens (epoxy resins for instance) and therefore has "long-term" effect to us as well. Breathing the oil mist from rotary pumps (my biggest concern) may cause lung cancer many years later... So, radiation in this '"row" of "dead substances" is nearly negligible - it's easy to detect (how you could detect epoxy on your skin or U in the bones?) and most institutions already has taught regulations on radiation issue. Safety in EM Lab should complain with all local and federal regulations ANY time, not only if pregnant women decided to do EM.
If exposure to the dangerous substances (not only in EM Lab) is a concern, the couple (both male&female) should avoid danger well before they decided to have baby. I would suggest at LEAST 6 month before fertilization they should avoid driving car on freeways (carbon dioxide, poly-carbons, nitric oxide), do not stay close to the buildings with granite decorative panels (some banks has luxury to use granite for ex/in-teriors) - radioactivity!; do not fly (radioactivity again), do not eat fast-food ( carcinogens in the overheated oil). This "poor" couple should also seriously inspect their living place. If place is 20-30 years old, it's very possible that painting contains Pb, some flooring - asbestos (lung cancer) and the wood impregnated with poly-phenols (carcinogen). Who knows, may be 10 years ago somebody broken body-thermometer with mercury and mercury is still here (may cause mental disorders). Synthetic carpet delivers static electricity which may interfere with fertilization (sperm may be shocked by high-voltage discharge!!!). If this hypothetical couple will manage to avoid all those dangerous things, THEN we should start worrying about safety at the working place: computer CRT monitors (radiation), dust, clearness of the drinking water in the fountains/bottled (very questionable), artificial illumination (baby needs natural light to be correctly developed), cubicles (claustrophobia, bad for developed fetus), equipment's noise (may scary the fetus, I am serious), THEN - exposure to the chemicals from neighbor's Lab (somebody spoil acetic acid on the floor) and so on. Each chemical/instrument coming in our Labs has detailed Instruction how to use it. You just has to follow instructions and comply with local and federal regulations.
So, the bottom line is: work in the EM Lab should be SAFE any time and to EVERY person at the level of the local and federal regulations. Each professions has some 'professional risk', which we could not avoid. Signing working contract (or accepting monthly salary payment) we automatically agree with some risk, connected to the duties you suppose to perform. Another issue here, that institution offering the job should clearly state what the professional risk is for this particular job and if for some reasons you don't want to take this risk, you have to quick.
People who seriously concern about pollution should go to Russia: because of economic problems, most industrial manufacture is shout down or working at 5-10% of full power. Russia also has much less cars and widely uses hydro-powered electricity generation (not good for eco-systems, but pollution-free). Russia was only the country at Kioto Conference who DECREASED air pollution over last 10 years. Another advantage being in Russia (I really miss it), the ticket in Bolshoi is $5 and most museums available for free (or 5-10 cents ticket).
Sergey
At 07:22 AM 1/6/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I have followed your submissions to this thread with great interest. It's a new terrain for me. I have a related question.
What advice do you have for a teacher whose pregnant grad student has on-going electron microscopy (SEM/TEM) research project? I would lik to read you views on this.
Many thanks in advance.
Ike Oguocha
--- William McManus {billemac-at-biology.usu.edu} wrote: } } Depending on the motivation of the pregnant worker, and the quality } of } your safety personnel, about a week. In the interim, the person can } be } given other duties, or in our case just carefully follow the } procedures } already in place, which have been designed to protect. The biggest } question is: Will the lab administration give you the time to get set } up } before the fact. In a real sense, it is always too late to begin } preparations after definitive knowledge of pregnancy is know. In } Kathy's case se would have been better off to wait to tell everyone } of } her condition, this is a shame, however, and we have not come as far } as } we would like to think with women's rights. In my case it was very } beneficial to know that the worker was pregnant, her work dropped off } substantially and she napped at the micortome. If I was unaware of } the } situation. I might have reacted in a negative sense, but instead, I } was } able to protect her from those who would not understand. } } Bill
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I have followed your submissions to this thread with great interest. It's a new terrain for me. I have a related question.
What advice do you have for a teacher whose pregnant grad student has on-going electron microscopy (SEM/TEM) research project? I would lik to read you views on this.
Many thanks in advance.
Ike Oguocha
--- William McManus {billemac-at-biology.usu.edu} wrote: } } Depending on the motivation of the pregnant worker, and the quality } of } your safety personnel, about a week. In the interim, the person can } be } given other duties, or in our case just carefully follow the } procedures } already in place, which have been designed to protect. The biggest } question is: Will the lab administration give you the time to get set } up } before the fact. In a real sense, it is always too late to begin } preparations after definitive knowledge of pregnancy is know. In } Kathy's case se would have been better off to wait to tell everyone } of } her condition, this is a shame, however, and we have not come as far } as } we would like to think with women's rights. In my case it was very } beneficial to know that the worker was pregnant, her work dropped off } substantially and she napped at the micortome. If I was unaware of } the } situation. I might have reacted in a negative sense, but instead, I } was } able to protect her from those who would not understand. } } Bill
__________________________________________________ Do You Yahoo!? Everything you'll ever need on one web page from News and Sport to Email and Music Charts http://uk.my.yahoo.com
Invitacion para integrar el Grupo de Trabajo de Telemedicina del Plan Organizacional y Estratégico para la Sociedad de la Información en Argentina
Durante el mes de Diciembre, comienzó la actividad de los Grupos de Trabajo (GT) para la elaboración colectiva de un plan estratégico para el desarrollo de la Sociedad de la Información en Argentina (POESIA). El objetivo fundamental del plan es establecer bases que permitan contribuir a la generación y puesta en marcha de propuestas concretas que contribuyan a la transformación estructural que la Argentina requiere para salir de la crisis.
Se crearon cuatro grupos de trabajos en las áreas elegidas; e-gobierno, e-educación, e-salud (telemedicina) y e-economía. Como parte de las actividades a realizarse en forma on line y en forma presencial, todos los participantes podrán presentar los temas principales que segun su visión, constituirán los ejes tematicos de cada grupo de trabajo. Posterior debate y puesta en comun, los primeros resultados se darán a conocer en un evento presencial a realizarse en abril de 2003 con la presencia de todos los miembros participantes de los GTŽs.
Quienes deseen ampliar la información sobre los requerimientos y mecanismos de participación en el POESIA pueden consultar el sitio web de la nueva asociación (http://www.links.org.ar), en donde podrán solicitar a cada coordinador del GT su inclusión en el grupo de su interés e interiorizarse sobre la metodología de trabajo.
Sobre Links LINKS, la primera Asociación para el Estudio y Promoción de la Sociedad de la Informacion en el pais, fue creada en octubre de 2002. Esta entidad cuenta como socios fundadores a distinguidos investigadores, intelectuales, técnicos y organizadores sociales, especializados en estudios y acciones sobre la Sociedad Informacional, que unen sus esfuerzos con el fin de contribuir a la construcción de una sociedad innovadora, usando las herramientas de las tecnologías de información y comunicación (TIC). Entre los objetivos de la entidad se destacan el estudio y la promoción de la Sociedad de la Información, las acciones para integrar a la población a las nuevas oportunidades, y la transferencia de los conocimientos de la Sociedad Informacional a la comunidad. LINKS pretende constituirse en referente en temas concernientes a la aplicación de una adecuada política de Estado en materia de la Sociedad de la Información tanto en Argentina como en América Latina
Contacto Dra. Lucia E. Muñiz lmuniz-at-links.org.ar
I can remember a couple of messages Robert Grassucci has mailed to the 3D-EM newsgroup in October 2001 and March 2002 concerning a new emulsion for Kodak SO 136 EM-film. He reported a decrease in sensitivity of app. 10% for the new emulsion. He also mentioned reports about an increase in the fog level. To access the original messages you can visit the 3dem messages archive: http://3dem.sdsc.edu/list/maillist.html
Matthias Floetenmeyer
Centre for Microscopy & Microanalysis University of Queensland, Queensland 4072, Brisbane, Australia
I bought a Jun-Air unit (oil type, not oil-less) for use as a dry air source for an FTIR (consumption rate may be higher than for an EM) and found it very tricky. The unit has worked well but the specs are very misleading and they charge extra for every little nut and bolt beyond the basic unit.
If you are using the air only to operate valves, consumption rate is probably not a problem. However if you need a steady stream of air, be aware that the rated flow volume is cited while running full-out.... but the compressor cannot run at more than a 50% duty cycle - so you have to halve the delivery volume right off the bat. Next, the compressor has very small cooling fins and gets excessively hot even on a 50% duty cycle in an ambient of 60 degrees Fahrenheit. When the intake air is preheated (expanded) on the way in, the flow volume drops off very steeply (and the duty cycle becomes excessive and ... there you go again). Therefore you have to have forced cooling. Although it has an overtemp cutout, that apparently applies only to the electrical motor because the compressor head is not supposed to exceed 55 Celsius. Before the fan, I discovered it was running too hot to touch, I would guess about 80 Celsius. They sell an add-on fan but by then my wallet was tapped out and I added my own. The oil darkens a lot in use but so far seems to be functioning properly. I needed two oil mist separators (.5 micron last). Those do seem to function well. The unit is quiet, as described, but draws down the line voltage sharply when it kicks in, even though it is well below the rating for the circuit that it's on. For some reason the starting capacitors are not properly selected for the motor, I guess.
In summary, I read all the specs and went to the local distributor expecting to spend $1,400. I ended up spending $3,000; timing the cycle with a stop watch, and rigging a fan.
John Twilley
Eric Anderson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } New Year's Greetings! } } Say, can anyone recommend a super-reliable and quiet brand (and size) of } air compressor to use with a Philips EM400T? } } Unfortunately, in addition to being very, very loud, the low grade air } compressor we've been using has the intermittent habit of blowing it's } circuit breaker (twice in the last couple of weeks), which has set us } way back on getting our "new" microscope fully evacuated for the first } time. } } I'm waiting for a catalog from Jun-Air, but would like to know what } model/size has been used reliably by others over the years. The Philips } installation manual doesn't give much more than the air pressure } requirement, so I'm uncertain what is ideal in terms of pump and tank } capacities. I've asked the folks at Jun-Air, but they are reluctant to } make any recommendation based solely on the pressure spec. } } Thanks for any tips! } } Best regards, } Eric Anderson } Southern CT State University Physics Dept. } 501 Crescent Street, JE108B } New Haven, CT 06515 } 203-392-6468
if you use a safelight have you done a check on this with the new film? I usually slightly pre-fog test film (with a light or better still about 1/4 normal e.m. exposure) and then place something on the film a bit nearer to the safelight for several minutes. If you get any shadow of the object, there is a problem. This may require a change of distance, filter or bulb.
The only other obvious problems I can think of is if the developer has gone off or is the wrong one for the new film.
The drop in contrast may just be related to the fogging.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: Rajesh Patel {rpatel-at-umdnj.edu}
Rajesh, I did have a box of film (SO163) just before Christmas which gave very poor results. I suspect that the notch was in the wrong place, which meant that it was loaded emulsion side down in the microscope. What is the accelerating voltage of your microscope? If it's 200 kV or more I guess this would be less noticeable than the blank plates I had. I've never seen this happen before & I guess I've processed 20,000 TEM negatives or so in my career. I imagine film quality control is becoming poorer, since the volume is decreasing due to the rise of digital image capture systems - just as the quality of records went off in the later days of vinyl as CDs became dominant.
Richard _______________________________ Richard Beanland Analytical Services Bookham Technology plc Caswell, Towcester, Northamptonshire NN12 8EQ UK Tel: +44 (0) 1327 356362 Fax: +44 (0) 1327 356775 http://www.bookham.com
-----Original Message----- } From: Rajesh Patel [mailto:rpatel-at-umdnj.edu] Sent: 06 January 2003 18:53 To: Microscopy-at-sparc5.microscopy.com
I've recently developed a problem with a newly purchased Kodak EM film. The EM film box says "new formulation" on it.
Well, this new formulation on film is lowering my contrast and I get some funky fog on it.
I was wondering if anyone else have experienced this problem and if so what can we do?
I've used old film ( old formulation ) and experience no fogging and get nice contrast. Its also not a scope specific problem or light leaks or developer. Its the film.
Rajesh Patel Robert Wood Johnson Medical School Department of Pathology 675 Hoes Lane Piscataway, NJ 08854
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The next major EPSRC FIB meeting is on 4th April 2003 in Cambridge, UK. This will take place directly after Microscopy of Semiconductors MSM XIII.
NanoFIB 2003 will provide an exciting overview of active FIB research both in the UK and abroad. Registration and abstract submission will be via the RMS (www.rms.org.uk).
The website for the new EPSRC NanoFIB network is now running at : www.nanofib.org. Please look for info on FIB meetings, Network members, links to FIB websites. The website is to link together people active in FIB. If you would like something added to the website, such as links to your own websites, send us your suggestions!
Dr. B.J. Inkson Dept. of Engineering Materials The University of Sheffield Sir Robert Hadfield Building, Mappin Street, Sheffield, S1 3JD Tel: 0114 222 5925 FAX: 0114 222 5943 www.nanofib.org
------------------------------------------------- This mail sent through IMP: http://horde.org/imp/
} } I've recently developed a problem with a newly purchased Kodak EM film. The } EM film box says "new formulation" on it. } } Well, this new formulation on film is lowering my contrast and I get some } funky fog on it. } } I was wondering if anyone else have experienced this problem and if so what } can we do? } } I've used old film ( old formulation ) and experience no fogging and get } nice contrast. Its also not a scope specific problem or light leaks or } developer. Its the film. *************************** Rajesh, I have not personally experienced thisa, since I'm still working off an old supply of film, but another EM lab here has been going nuts for the past few months with this same issue. I can't offer any advice, but I will print out & pass along your posting to them. There is comfort in not being alone. Hopefully, we as a large group of interested users can find a solution to this problem, or push Kodak to solve it. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Hi, I would just like to put in my 2 cents worth here. I have been "doing" electron microscopy for over 25 years. I am the mother of a healthy, normal 10 year old boy. He was the end result of a few years worth of false starts. Of course I can never know if the 2 miscarriages were genetic or the result of environmental influences, they were early (6 & 12 weeks) and my vote is with something genetic....I was no spring chicken at the time! During the 3+ years that my quest for motherhood took, I made extra sure that I followed good lab safety procedures. I did double glove when working with fixatives, etc., had the fume hood in my lab inspected and avoided acrylic resins (personal bias?). My 'scope is a 100CX-II that was installed in 1982. It has always been maintained by JEOL and I had no concerns about radiation leaks. I agree with the idea that good lab safety is good lab safety, and if rules are set to protect the most delicate among us, the unborn, and everyone follows them at all times, there should not be a concern. Napping at the microtome, or putting one's feet up when seated at a desk may raise some eyebrows, but it does help you get through the day! 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 Analytical Imaging Facility of the Albert Einstein College of Medicine has an opening for an experienced electron microscopy technician. For details please see the posting on the MSA Placement Office Job Listings page.
http://www.msa.microscopy.com/PlacementOffice/JobListings.html **************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
Listers; We have been microtoming samples of high impact polystyrene (HIPS) that contains butadiene rubber moieties. These samples have been treated as follows prior to sectioning: 1) Trim block face to prescribed shape and size 2) Harden & stain block by immersing in 2% OsO4 overnight.
Most of the blocks will section fine. However, there is a fair amount of compression of the resulting sections. The sections expand slightly when exposed to chloroform vapors and a lot when exposed to xylene vapors. Sometimes it is necessary to use both to get the sections to expand. However, we are concerned about over expansion and possible swelling of components in the sample. Is there another solvent that could be used to relax the sections without concern for over expansion or swelling of components?
Also, is there someone who does cryo-sectioning of HIPS or similar materials that would be able to do this as a service project for an outside group.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
On Monday, January 6, 2003, at 12:41 PM, Eric Anderson wrote:
Dear Eric,
} Say, can anyone recommend a super-reliable and quiet brand (and size) } of } air compressor to use with a Philips EM400T?
Since the FEI people are here installing our microscope, I was able to ask about this. The Jun-Air model 6-25 is the one used both on the new scopes and the older 400 series. } } Thanks for any tips! } You're welcome. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
On Monday, January 6, 2003, at 10:53 AM, Rajesh Patel wrote:
Dear Rajesh,
} I've recently developed a problem with a newly purchased Kodak EM } film. The } EM film box says "new formulation" on it.
Is this SO163? Our newly-purchased SO163 does not say this, but it would be good to know that the new formulation is coming. } } Well, this new formulation on film is lowering my contrast and I get } some } funky fog on it. } } I was wondering if anyone else have experienced this problem and if so } what } can we do? } } I've used old film ( old formulation ) and experience no fogging and } get } nice contrast. Its also not a scope specific problem or light leaks or } developer. Its the film. } Have you tried developing the film in complete darkness? Very sensitive EM films can be fogged by safelights, and sometimes the filter on the safelight can crack and let shorter wavelengths out. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Dear Richard, I had a run of about 6 to 8 boxes of Kodak 4489 film with the notch on the wrong side, three years ago. I noticed because the emulsion side of the film is lighter under the safelight than the back side, so I loaded them right side up. I sent in the customer reply card, but never heard anything.
} Rajesh, } I did have a box of film (SO163) just before Christmas which gave } very poor results. I suspect that the notch was in the wrong place, which } meant that it was loaded emulsion side down in the microscope. What is the } accelerating voltage of your microscope? If it's 200 kV or more I guess } this would be less noticeable than the blank plates I had. } I've never seen this happen before & I guess I've processed 20,000 TEM } negatives or so in my career. I imagine film quality control is becoming } poorer, since the volume is decreasing due to the rise of digital image } capture systems - just as the quality of records went off in the later days } of vinyl as CDs became dominant. } } Richard } _______________________________ } Richard Beanland
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
Debby Sherman wrote: ============================================================== Listers; We have been microtoming samples of high impact polystyrene (HIPS) that contains butadiene rubber moieties. These samples have been treated as follows prior to sectioning: 1) Trim block face to prescribed shape and size 2) Harden & stain block by immersing in 2% OsO4 overnight.
Most of the blocks will section fine. However, there is a fair amount of compression of the resulting sections. The sections expand slightly when exposed to chloroform vapors and a lot when exposed to xylene vapors. Sometimes it is necessary to use both to get the sections to expand. However , we are concerned about over expansion and possible swelling of components in the sample. Is there another solvent that could be used to relax the sections without concern for over expansion or swelling of components?
Also, is there someone who does cryo-sectioning of HIPS or similar materials that would be able to do this as a service project for an outside group. ================================================================== The laboratory of Structure Probe, Inc. which is the "parent" of SPI Supplies, has been doing cryo sectioning of HIPS for industry for more than thirty years. You are quite correct to be concerned about compression effects in these kinds of samples, because they can create the illusion of orientation when in fact such apparent "orientation" could be an artifact.
There could be many reasons for the compression effects, which could include such factors as the diamond knife itself, the conditions of the ultramicrotomy, temperature of the sectioning ,etc.
Please contact me off-line and describe what the client is trying to accomplish, because that becomes important in terms of determing what orientation the sections should have relative to the injection direction of the molded piece. We can quote you a fixed price to do the entire job. I can not recall any HIPS sample in recent years that could not be treated as a "routine" sample, even those that have been modified with other polymers.
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 ######################## ============================================
IOWA STATE UNIVERSITY An Equal Opportunity/Affirmative Action Employer Vacancy Announcement RESEARCH ASSOCIATE II Vacancy Number: 023649 Department: BIOTECHNOLOGY Proposed Start Date: MARCH, 2003 Appointment Conditions: Continuous, 12 Months, Full Time
Special Conditions: Description: This individual will be responsible for carrying out a variety of laboratory techniques and procedures for the preparation of biological specimens leading to their microscopic observations and analyses for scientists at Iowa State University, and for off-campus scientists/clients. This individual will be responsible for assisting both the Assistant Scientist III and Director of Bessey Microscopy Facility (BMF) in maintaining the laboratory and work environment for all of the researchers who use it. Individual will need to effectively work with researchers and develop a work schedule that fits needs of BMF. Required Qualifications: Bachelor's degree in a biological sciences discipline and one year of laboratory experience that demonstrates knowledge in general and quantitative chemistry, physics, operation and maintenance of light and electron microscopes, ancillary equipment, and all phases of handling chemicals for preparing specimens. Preferred Qualifications: Master's degree in biological sciences discipline, practical experiences in use of light and electron microscopes, rotary and ultramicrotomes, digital imaging and PC computers and BEMT Certification (MSA). Salary/Wage: $30,128 minimum Application Deadline: To guarantee consideration, application must be received by January 30, 2003. Application Instructions: Send cover letter explaining interest, email address, resume, and the names, professional titles and phone numbers of three references to: Dr. Harry T. Horner, Bessey Microscopy Facility, Room 3A Bessey Hall, Iowa State University, Ames, IA 50011-1020, fax: 515-294-1337, or email: hth-at-iastate.edu.
Tracey M. Pepper Supervisor Bessey Microscopy Facility Iowa State University Ames, IA 50011-1020 p. 515.294.3872 f. 515.294.1337
Rajesh: We use Kodak 4489 EM film. The "new formulation" is definitely different in our hands. Our negatives are too thin when we use our standard microscope exposure and chemical processing methods. I have made an acceptable correction by increasing the development time of the film by about 10%, decreasing the exposure of the paper by about 1/3 stop and shortening the development time of the paper by about 10%. I have made no change in the microscope's exposure setting. We use Kodak Kodabrome II RC F4 paper and process both the 4489 film and the Kodabrome paper in a Colex automatic processor using Kodak Polymax RT chemistry. The same correction scheme should work if you are processing in D19 but the percentages will need adjusting.
I wonder what Kodak has done?
Bob -- C. Robert Bagnell, Jr., Ph.D. Associate Professor Director, Microscopy Services Laboratory Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill Chapel Hill, NC 27599 phone 919-966-2413 fax 919-966-6718 e-mail bagnell-at-med.unc.edu
I have not used TEM film before, but have used lots of photo film. First off, I would never load film under a safe light--it is not safe. Litho film could be handled in yellow light without problem. If the TEM film is pan film, it seems to me that a safe light is not advised.
The flip side of the film is always an issue. If the TEM film is like traditional film, there is a blueish anti-halation coating. That side is the back side. When the film is fixed, the coating is removed. Check an un-developed piece of film and see which is which. Another clue is which side is grainy looking and which is shiny. Grainy is the emulsion side. Probably no surprise to anyone.
When a new box of 4x5 or 220 roll film was purchased, I always developed/processed an unexposed sheet or roll. This was then measured on a densitometer for baseline fog D value. If the D value was lower than normal, the media could be pre-exposed (fogged) to bring the background up to that of other media. The idea is to get the media at a point where the lazy S D/exposure curve was at a point above the toe. Then, there is a large linear range of exposure before the knee, where saturation occurs.
One might try this on old media and the "new" stuff. This would tell whether the new formula is a little different or radically different.
gary g.
At 11:48 AM 1/7/2003, you wrote:
} Dear Richard, } I had a run of about 6 to 8 boxes of Kodak 4489 film with the notch on the } wrong side, three years ago. I noticed because the emulsion side of the film } is lighter under the safelight than the back side, so I loaded them right } side up. I sent in the customer reply card, but never heard anything. } } } } Rajesh, } } I did have a box of film (SO163) just before Christmas which gave } } very poor results. I suspect that the notch was in the wrong place, which } } meant that it was loaded emulsion side down in the microscope. What is } the } } accelerating voltage of your microscope? If it's 200 kV or more I guess } } this would be less noticeable than the blank plates I had. } } I've never seen this happen before & I guess I've processed 20,000 TEM } } negatives or so in my career. I imagine film quality control is becoming } } poorer, since the volume is decreasing due to the rise of digital image } } capture systems - just as the quality of records went off in the later } days } } of vinyl as CDs became dominant. } } } } Richard } } _______________________________ } } Richard Beanland } } Mary Mager } Electron Microscopist } Metals and Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchange.ubc.ca
David, Thanks for such nice comments. My major point was that the Lab should be safe in all possible ways anytime and for EVERYONE. Another point was, that people should think about baby well before they actually get it. The exposure to radiation/chemicals/etc of male and female BEFORE fertilization may cause serious defects in the fetus later. Actually, male's reproductive organs tends to be more sensitive to radiation and some particular chemicals than female's.
From another hand, employees should clearly understand the health risk factor associated with job. In your particular case, those individuals, who is oversensitive to formaldehyde should quit, because this hypersensitivity is a very bad sign: they could develop unusual allergic reactions with very serious result to health in future. I wish all of us to have nice safe conditions in the Lab. Sergey
At 10:46 AM 1/7/2003, you wrote: } I agree in substance, Just a few quick notes. Your concerns } prior to conception are most valid as well as the points that often the } labs are safer than home. There is only so much we can do, but we should } do what we can. There is/should be no legal way to discriminate based on } risks to fetus. So the lab must be made safe for all. } I would disagree on several points: } Federal, State and local safety laws are made for the "Average, } healthy, normal individuals. The TLV's, for example, are created for } someone in this class. Few if any body falls into this class, especially } someone who has just conceived. The presence of formaldehyde is totally } undetectable in the lab, approximately 2 times less than the TLV, yet } without special precautions, I have individuals sensitive to formaldehyde } who react to the small amounts present with skin reactions, or breathing } problems. We attempt to lower formaldehyde vapors to below the limits of } those using the lab, but they are the most sensitive indicators of the } presence of aldehyde vapors, there aren't tests sensitive enough. Kind } of a catch 22. } Oil from vacuum pumps, compared with other risks is relatively } mild. New oil (I know, I know, the breakdown products) has repeatedly } been checked free from carcinogens. (and we still exhaust to the hood system) } At least from my information, Uranyl Acetate is not a bone seeker } like Strontium is. I was told that the biological half life of soluble } uranium was in the order of 6 weeks. The acute danger to kidneys and } liver from the heavy metal toxicity may be as great as the radioactive } hazard to the body. Yes, don't drink it (and especially don't inhale the } dust, there the damage is greater and the biological half life much } longer) but I agree there are many more hazardous chemicals in a EM } Lab. (Cacodylate scares the living hell out of me every time I need to } use it, and I typically don't let any others use it but do the work for them.) } Although the persons accept the risks (making the assumption that } all managers are as careful as you and I in outlining the risks to the } employees and users and training them to avoid them) the bottom line is: } regardless what we have them sign or tell them, the legal and moral } responsibility to protect the employees and users of the laboratory rests } firmly on the manager and it is the managers responsibility to make the } lab safe for all who would wish to use it, regardless of preexisting } conditions, gender etc. } } At 05:40 PM 1/6/2003 -0800, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Why not do an exposure series on the scope with the new film to get more signal (more illumination) so that you don't have to monkey with both film development and print exposure? What you need is a properly exposed negative so that you can get the most out of your prints. If the problem is truly film formulation (slower film) and not some fogging problem, I¹d go for the increased beam intensity.
This is not the first time Kodak has changed formulations and or film format. It doesn't surprise me. We haven't ordered film lately, but it's useful to have this problem discussed here so the rest of us can beware when we do get a new batch.
Thanks, Sara Miller
On Tue, 7 Jan 2003, Robert Bagnell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Rajesh: } We use Kodak 4489 EM film. The "new formulation" is definitely } different in our hands. Our negatives are too thin when we use our } standard microscope exposure and chemical processing methods. I have } made an acceptable correction by increasing the development time of } the film by about 10%, decreasing the exposure of the paper by about } 1/3 stop and shortening the development time of the paper by about } 10%. I have made no change in the microscope's exposure setting. We } use Kodak Kodabrome II RC F4 paper and process both the 4489 film and } the Kodabrome paper in a Colex automatic processor using Kodak } Polymax RT chemistry. The same correction scheme should work if you } are processing in D19 but the percentages will need adjusting. } } I wonder what Kodak has done? } } Bob } -- } C. Robert Bagnell, Jr., Ph.D. } Associate Professor } Director, Microscopy Services Laboratory } Department of Pathology and Laboratory Medicine } University of North Carolina at Chapel Hill } Chapel Hill, NC 27599 } phone 919-966-2413 } fax 919-966-6718 } e-mail bagnell-at-med.unc.edu } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Though my issue was not regarding pregnancy, I had my concerns over spending a lot of time being bathed in a strong magnetic field. Some studies suggested correlations to increased health problems, whereas other studies suggest some magnetic fields were actually beneficial.?????
Anyway, we went through the lab with a trifield meter to ensure that no one was being unduly exposed to strong emf. While EMs have high voltage and you sit right next to those power supplies, you could reasonably assume that microscope is more sensitive to emf than your body and the manufacturers provide good shielding for image quality.
Alan Stone ASTON Metallurgical Services
At 09:50 AM 1/7/2003 -0500, you wrote: } microscopy-at-sparc5.microscopy.com
Gary TEM film is blue-sensitive, orthochromatic not panchromatic, and red or brown safelights are generally safe if chosen and used according to the manufacturer's instructions. Whether safelight is safe or not depends on film type, safelight wavelength, intensity, time, development, EM image exposure, etc. If you have any anxieties about safe exposure times better to experiment with these and determine what actually happens. Chris
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 08, 2003 12:30 AM
Hi,
I am in the middle of writing a joint grant proposal for an optical disk jukebox / image archiving solution which should hopefully hold up to ten years microscopy data. This system hopefully will aid in 'traceability' of raw data and the image processed result. I was wondering what other research institutes have in place for image archiving regarding traceability of images.
Techniques of image manipulation that, in the past, could only be done by few, and with great care, are now routine, consequently readily available software has also created ample opportunities to falsify images. Obviously the raw data and the final data to be published should be stored long term, but what about the steps in-between? As most research institutes now are multi-user facilities, the onus of scientific proof should be on the owner of the data, but since students are replaced every three - four years there must also be a burden of proof on the research institute especially if the images are to be reused at a later date.
Should a multi-user microscopy facility demonstrate traceability of data or should we rely on the users?
Steve
Steve Bagley Associate Scientist Applied Imaging Facility Paterson Institute For Cancer Research Cancer Research UK Christie Hospital, Wilmslow Rd, Manchester. M20 9BX, UK
This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute For Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.
Sarah's suggestion is a good one and in fact is what we did first. However, changing the exposure of the film in this case did not improve the result. We got more dense negatives but the contrast remained poor. I've used the old photographer's trick of decreasing exposure (in this case keeping it the same) and increasing development (of the film) to improve contrast. This definitely works for the new 4489 formulation, what ever it is.
Bob -- C. Robert Bagnell, Jr., Ph.D. Associate Professor Director, Microscopy Services Laboratory Department of Pathology and Laboratory Medicine University of North Carolina at Chapel Hill Chapel Hill, NC 27599 phone 919-966-2413 fax 919-966-6718 e-mail bagnell-at-med.unc.edu
Just a quick note: if you can wait a bit to buy, I would. Microtek (I'm 98% sure it's Microtek) has just come out with a flatbed scanner that uses ICE technology. This is used in the Nikon and other 35mm slide scanners to remove scratches and dust on the film from the scanned image. It's a hardware+software system, not just software. I don't know anything about this scanner yet, other than that it exists, but it would be worth checking out.
Phil
Hope 2003 is good for you all!!
We are looking at purchasing a photo scanner (Probably an EPSON Perfection 2450 Photo). We will be using it to scan old 35mm slides for new Powerpoint presentations.
Since it will also scan negatives and produce positive immages, I was wondering whether it could scan Electron Microscope negatives as well. This would save a good deal of effort and money in enlarging and printing onto photographic paper.
Has anyone tried this? Any recommendations?
Thanks for your consideration,
Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: (02) 9845 3306 Fax: (02) 9845 3318
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-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Rajesh: We are having the same problems with the "new" film. We tested the scope, camera, safe light, enlarger, desecator, grid holder, and developing solution. We even tested the film in a new scope at Montclair University and also concluded it is the film. We contacted kodak directly, I think you should do the same.
Omayra Velez Electron Microscopy Specialist Pathology Department Weill Cornell Medical College New York, New York 212-746-6437
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I have some problems to prepare the surface of AlMg3 for surface investigations on the one hand and TEM measurements on the other hand.
TEM is less worse because I currently have an electrolyte based on perchloric acid which lets me get satisfying specimen. Most specimen are ion milled afterwards anyway. If there is a better way (without ion milling to save time ;^)) I nevertheless would be very interested.
The other purpose is some more interesting for me: On the one hand I need to investigate the grain structure of my specimen after cold rolling and donŽt get proper grain boundaries to do that. On the other hand I am very interested in proper surfaces for surface observation in DF. I also tried to get a smooth surface with an electrolyte composed of perchloric acid and methanol, but its quite explosive and therefore has to be cooled by solid carbon dioxide...
Does anybody know any other electrolyte or acids to get the results mentioned above?
Thanks in advance,
Hanno Dierke
-- Hanno Dierke, Inst. for Metal Physics an Nuclear Condensed Matter Physics TU Braunschweig, Braunschweig, Germany h.dierke-at-tu-braunschweig.de
I have been working on phase imaging of HIPS with AFM TappingMode for a while since we do not have a TEM here. I tried different microtoming conditions, with a 35-degree diamond knife on a microtone for light microscope samples (minimum 0.5 um sectioning), for the best cutting faces and therefore the imaging from them, but with limited success. I found ca. -30 to -50 C is probably the "right" temperature, but the resulted facing quality varies from sample to sample. Defects are primarily chatters and local compressions on polystyrene matrix. I also found difficulties in parameter settings of the DI AFM for different HIPS samples. However, the same operation is very successful for ICPs (impact copolymer of polypropylene and polyethylene).
Does anybody have any experiences, suggestions or comments? Many thanks in advance.
Debby, did you try AFM on HIPS?
Jiang Liu, PhD Research and Technology Center Atofina Petrochemicals, Inc. Tel: 281-884-0529 email: jiang.liu-at-atofina.com
} From: Debby Sherman {dsherman-at-purdue.edu} } To: "message to: MSA list" {microscopy-at-sparc5.microscopy.com} } Subject: Sectioning HIPS } Date: Tue, 07 Jan 2003 11:35:41 -0500 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi Steve, As the digital revolution has progressed, I have thought image tracability will become a major issue. I have wondered if anyone has developed a method of tagging the original data file in a way that you could always identify the original? So far people have relied on image analysis techniques to determine originals from manipulations and/or actual theft of images or parts of images.
Robert Underwood U of Washington Seattle
On Wed, 8 Jan 2003, Steve Bagley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi, } } I am in the middle of writing a joint grant proposal for an optical disk } jukebox / image archiving solution which should hopefully hold up to ten } years microscopy data. This system hopefully will aid in 'traceability' of } raw data and the image processed result. I was wondering what other research } institutes have in place for image archiving regarding traceability of } images. } } Techniques of image manipulation that, in the past, could only be done by } few, and with great care, are now routine, consequently readily available } software has also created ample opportunities to falsify images. Obviously } the raw data and the final data to be published should be stored long term, } but what about the steps in-between? As most research institutes now are } multi-user facilities, the onus of scientific proof should be on the owner } of the data, but since students are replaced every three - four years there } must also be a burden of proof on the research institute especially if the } images are to be reused at a later date. } } Should a multi-user microscopy facility demonstrate traceability of data or } should we rely on the users? } } } } Steve } } } Steve Bagley } Associate Scientist } Applied Imaging Facility } Paterson Institute For Cancer Research } Cancer Research UK } Christie Hospital, Wilmslow Rd, } Manchester. M20 9BX, UK } } } } } } } } } } } } } } } } } } } } } This email is confidential and intended solely for the use of the Person(s) } ('the intended recipient') to whom it was addressed. Any views or opinions } presented are solely those of the author and do not necessarily represent } those of the Paterson Institute For Cancer Research or the Christie Hospital } NHS Trust. It may contain information that is privileged & confidential } within the meaning of applicable law. Accordingly any dissemination, } distribution, copying, or other use of this message, or any of its contents, } by any person other than the intended recipient may constitute a breach of } civil or criminal law and is strictly prohibited. If you are NOT the } intended recipient please contact the sender and dispose of this e-mail as } soon as possible. } }
University of Connecticut Institute of Materials Science
Postdoctoral Position Displacive Phase Transformations
A post-doctoral position is available immediately within the Institute of Materials Science at the University of Connecticut to work on displacive phase transformations in crystalline solids. The systems to be studied include metallic pseudoelastic alloys and ferroelectric thin films. The ideal candidate will have experience in both microstructural characterization (particularly TEM) and phenomenological/crystallographic modeling of phase transformations. This position is a one-year appointment, with possible renewal for a second year.
To apply, please send a complete resume, together with a list of publications and contact details for 3 references to either Prof. S. Pamir Alpay (p.alpay-at-ims.uconn.edu) or Prof. Mark Aindow (m.aindow-at-uconn.edu). Screening of applications will begin immediately, and will continue until the position is filled. We encourage applications from under-represented groups, including minorities, women and people with disabilities.
Mark Aindow, Associate Professor, Department of Metallurgy and Materials Engineering Institute of Materials Science, 97 North Eagleville Road, Unit 3136 University of Connecticut, Storrs, CT 06269-3136, USA
} Tony, } } Just a quick note: if you can wait a bit to buy, I would. Microtek } (I'm 98% sure it's Microtek) has just come out with a flatbed scanner } that uses ICE technology. This is used in the Nikon and other 35mm } slide scanners to remove scratches and dust on the film from the } scanned image. It's a hardware+software system, not just software. } I don't know anything about this scanner yet, other than that it } exists, but it would be worth checking out.
But be aware that any camera or scanner hardware/software that "removes dust and scratches" usually does so by blurring and eroding and then unblurring, or similar processing, to despeckle or hide items identified as dust or scratches, thereby changing image pixels and altering your data. These programs may identify "real" objects in electron micrographs as dust and delete them! The technology is great for restoring that old photograph of your great-grandma, but has the potential to change your scientific data. Try to find out how the hardware/software works before deciding which to buy.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Ann: The new film has the letters "New Formulation" written in white over a red slip, on the side or top of the box. Kodak already contacted us today and we are in process to send them our test material(negatives, prints, and info). If you do not have this film then you are lucky, is been a nut house here, with these unprintable negatives.
Omayra Velez Electron Microscopy Specialist Weill Cornell Medical College New York, New York. 010021 212-746-6437
--- "R. Ann Bliss" {bliss5-at-llnl.gov} wrote: } Omayra: } } After following the thread about the "new" film for } a few days now, I } finally went to check the new shipment of film I } received last week. } Where does one find the words "New Formulation" or } whatever it was? i } looked all over the outside of the box and did not } see those words. } Am I lucky and don't have the problem (yet) or is it } on the paper } work I never read on the inside? } } -- } } +++++++++++++++++++++++++++++ } } R. Ann Bliss } Technician, Chemistry and Materials Science } Materials Science and Technology Division } Lawrence Livermore National Laboratory } } _____________________________
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you are touching on an issue that is probably of interest to a much larger audience. There are at least two areas, where this has been an issue for a while: Forensics and Pharmaceuticals. The Pharmaceutical industry in the US is required by the FDA to keep track of data because lives may depend on it. It used to be lab books that needed to be kept in certain ways, but now that much of the data is electronic, the FDA has published requirements for the safekeeping of data (21 CFR rule 11). It may be interesting for you to see if this answers some of your questions (caution: 21 CFR rule 11 is kind of messy). But even rule 11 relies on SOPs to ensure data safety and security. Traceability and audit features are part of 21 CFR rule 11.
21 CFR rule 11 may be overkill for many settings, and a relatively simple image archive or data base might suffice. Set it up so that certain information must be entered (for example sample number, patient ID, etc.), keep the images on a central server and have all imaging instruments store their data there, and it might do what you want.
I don't think that you can have a 100% watertight system without some decisions by the users, because it would require, that ALL images are stored, regardless of content. And even then, the choice of what images to take still lies with the individuals. You probably have to set up some SOPs that tell the users that they need to save the images within a system that allows some measure of control, be it in an archive or save them to CD-R which then have to be turned in for safekeeping. The question then turns to finding a system that supports the control you are looking for.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Steve Bagley [mailto:SBagley-at-picr.man.ac.uk] Sent: Wednesday, January 08, 2003 5:49 AM To: 'Micro Discussion Group'
Hi,
I am in the middle of writing a joint grant proposal for an optical disk jukebox / image archiving solution which should hopefully hold up to ten years microscopy data. This system hopefully will aid in 'traceability' of raw data and the image processed result. I was wondering what other research institutes have in place for image archiving regarding traceability of images.
Techniques of image manipulation that, in the past, could only be done by few, and with great care, are now routine, consequently readily available software has also created ample opportunities to falsify images. Obviously the raw data and the final data to be published should be stored long term, but what about the steps in-between? As most research institutes now are multi-user facilities, the onus of scientific proof should be on the owner of the data, but since students are replaced every three - four years there must also be a burden of proof on the research institute especially if the images are to be reused at a later date.
Should a multi-user microscopy facility demonstrate traceability of data or should we rely on the users?
Steve
Steve Bagley Associate Scientist Applied Imaging Facility Paterson Institute For Cancer Research Cancer Research UK Christie Hospital, Wilmslow Rd, Manchester. M20 9BX, UK
This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute For Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.
The videotapes from the 2002 MSA tutorials in Quebec City are now available. We are now in a positions to offer the entire library on DVD as well as VHS. We expect to phase out the VHS versions as the current supply that is on hand is exhausted. The most up to date catalog can be viewed at the MSA website or go directly to this URL: http://www.biotech.ufl.edu/sems/newtape00.htm As time permits, I am trying to make abstracts of each of the tapes available online as well so that one has a better idea of what was covered in the tutorial. If you are unable to access this site I can provide a printed copy or I can email a copy of the catalog. We are now also able to accept VISA or Mastercard for payment. We will also accept institutional purchase orders for which we will invoice. As you will notice many of the tapes are quite old and the topics covered certainly deserve being updated. The Education Committee invites interested parties to present tutorials or short courses on these topics, or topics never covered that might be appropriate for tutorials. The committee may be contact at this address MSAEdCommittee-at-MSA.Microscopy.Com
Please feel free to contact me concerning the video library.
Greg Erdos Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295
It is the ScanMaker 6800. This scanner will scan reflective media up to 8.5x11" and transmissive media up to 4x5".
http://www.microtekusa.com/indexICEflash.html
It uses Digital ICE, as do the high end Nikon CoolScan scanners. The principle of image correction is based on a separate IR channel. It scans once to pick up greyscale or RGB info, then a second time for IR. Then the image is processed and "fixed" if bad. One web link says two passes while another says only one pass. ?? Anyway, about D-ICE... Sometimes it works really good. Other times not so good. So, case by case basis. I use the Nikon Super CoolScan 8000 for slides and medium format media. It is optical 4K x 4K dpi with D-ICE and GEM.
It (D-ICE) does not work on old Kodachrome transparencies since they block the IR. Other media seem OK. It has a bit of difficulty with b/w slides. The 6800 is 4800x2400 dpi optical, which ought to be OK for most applications. With Firewire and USB for MSRP $399, that is pretty good.
gary g.
At 09:49 AM 1/8/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings, Tina's comment is right on. Note also that the original poster was seeking in part to move their 'good old' projection slides over to powerpoint. Well, if you are like me, over the years you have been showing your favorite slides they have accumulated plenty of dust. If its ok in the slide projector, why worry about it in powerpoint? It will give your presentation a comfy well worn look like an elbow patch on a tweed jacket. You can of course also do nice dodging and burning in photoshop or equivalent to remove egregious dust specks before dropping them into powerpoint.
Just a thought, Tobias
} } } } On Wed, 8 Jan 2003, Philip Oshel wrote: } } } Tony, } } } } Just a quick note: if you can wait a bit to buy, I would. Microtek } } (I'm 98% sure it's Microtek) has just come out with a flatbed scanner } } that uses ICE technology. This is used in the Nikon and other 35mm } } slide scanners to remove scratches and dust on the film from the } } scanned image. It's a hardware+software system, not just software. } } I don't know anything about this scanner yet, other than that it } } exists, but it would be worth checking out. } } But be aware that any camera or scanner hardware/software that "removes } dust and scratches" usually does so by blurring and eroding and then } unblurring, or similar processing, to despeckle or hide items identified } as dust or scratches, thereby changing image pixels and altering your } data. These programs may identify "real" objects in electron micrographs } as dust and delete them! The technology is great for restoring that old } photograph of your great-grandma, but has the potential to change your } scientific data. Try to find out how the hardware/software works before } deciding which to buy. } } Aloha, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
Many, many thanks for all your thoughts and generous replies!
Since the budget allows it, (at the moment), I'm going to follow the crowd (and FEI technicians) who have suggested the Jun-Air 6-25. (Many wrote just to me and not to the whole group. Jim, unfortunately the folks at Jun-Air would not recommend a model suited to my application when I asked them. They only offered to send a catalog.)
Hopefully, my particular application and environment won't generate the problems John experienced. We really need the quiet operation (specs say 45 dB), especially because the unexpected cycling of a louder compressor is scaring folks out of their wits. Not at all a good situation for handling delicate specimens/procedures.
I'm very tempted to just use nitrogen tanks, but the old 400 has a couple of slow leaks which I might not get to fixing right away, plus we'll probably have it running continuously for long periods, so I'd rather not worry about tank deliveries and running out at a bad time. (One of the same reasons I switched to natural gas from oil for home heating..., plus cleanliness of course.)
Vitaly, thanks for the reasonably priced alternative that employs a modified refrigeration compressor. I love being creative, but just don't have the time right now. Will keep it in mind though.
Thanks & best regards to all! -Eric
} } } Eric Anderson wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } New Year's Greetings! } } } } Say, can anyone recommend a super-reliable and quiet brand (and size) of } } air compressor to use with a Philips EM400T? } } } } Unfortunately, in addition to being very, very loud, the low grade air } } compressor we've been using has the intermittent habit of blowing it's } } circuit breaker (twice in the last couple of weeks), which has set us } } way back on getting our "new" microscope fully evacuated for the first } } time. } } } } I'm waiting for a catalog from Jun-Air, but would like to know what } } model/size has been used reliably by others over the years. The Philips } } installation manual doesn't give much more than the air pressure } } requirement, so I'm uncertain what is ideal in terms of pump and tank } } capacities. I've asked the folks at Jun-Air, but they are reluctant to } } make any recommendation based solely on the pressure spec. } } } } Thanks for any tips! } } } } Best regards, } } Eric Anderson } } Southern CT State University Physics Dept. } } 501 Crescent Street, JE108B } } New Haven, CT 06515 } } 203-392-6468
I have a GATAN Cryo-Transfer cold stage for a Philips EM420 TEM. The crystal needs to be replaced and I was wondering if anyone has knowledge of a third party who does this type of repair?
Thanks in advance.
Tom -- Thomas M. Murray Phone: (206)543-2836 Manager, Electron Microscopy Center Fax: (206)543-3100 Materials Science & Engineering email: murraytm-at-u.washington.edu 302 Roberts Hall, Box 352120 University of Washington Seattle, WA 98195-2120
before you rush out and buy the Scanmaker 6800 for its Digital ICE scratch correction you should be aware that this is only available for prints and not negatives. Although the scanner takes negatives, there is no infra red light source which is required for scratch correction on film - see PC magazine review at: http://www.microtekusa.com/images/pcmag6800.pdf This is pointed to on the Microtek page: http://www.microtekusa.com/indexICEflash.html
I also assume that negatives are scanned on a glass plate rather than a glassless carrier of the Microtek Scanmaker 8700 or 1800f types so you will get more dust marks etc.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: Gary Gaugler {gary-at-gaugler.com}
Morning Folks,
I am in the process of restarting a lab which has a Zeiss EM900 TEM. When I got here, I found the manual waterlogged! Another flood in an EM Lab! I am in need of an operators and any other electrical and mechanical manuals that pertain to this scope. I am willing to pay copying and shipping costs.
Thanks,
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 calomeni-1-at-medctr.osu.edu
I've run into compressor issues with every Philips scope I've worked with, what has always gotten me though is the fact that reliable quite air compressors are reality not fantasy since JEOL provides air compressors that are just that, and the HVAC systems for most buildings also using 24/7 air compressor systems for the thermostats (the little hiss when you change the thermostat on the wall?) as seem to work well.
Our current system is on a Philips 505. But after having tried several compressors and repairs, I have wound up building my own compressor system out of various components.
(1) Using 20 gallon storage tank from previous compressor
(2) Added a Thomas Compact Oilless Reciprocating Air Compressor ($400). Go well over pressure and volumne needed so as to not stress the compressor. Remeber your really don't need much volumne.
(3) Built a mounting plate out of aluminium and large rubber bushings
(4) Using high quality air hose as vibration isolation and copper tubing.
(5) Using an Ashcroft indusdrial pressure switch for turning compressor on/off ($130). Mounted on the wall off the compressor (vibration its a killer) - NOTE: **Do not** go cheap on this component, trust me! I run the compressor pressure just high enough to minimize cycling and provide the needed pressure at the scope without relying heavily on pressure regulators. Pushing the compressors to their "rated" pressure mark and using a regulator will reduce cycling, but it adds wear to the compressor (Yes, you can run your car just below red line on the tach, but for how long, eh?)
(6) a few oil filled air pressure gauges
(7) Reusing an exisiting air drying system.
(8) Installed the compressor in the service chaseway behind the scope for more noise isolation. (I do recommend getting it out of the scope room if possible, along with the water chillers. Soldered copper piping workes well.)
---} This system has been running (knock-on-wood) very well for over four years now without a problem. Cycle duty is less than 20% even with heavy microscope use. The noise is very low, next to the compressor, and barely noticable in surrounding rooms (don't have a dB meter).
---} Get the air leaks fixed on your 400!
Good luck.
} } Many, many thanks for all your thoughts and generous replies! } } Since the budget allows it, (at the moment), I'm going to follow the crowd (and } FEI technicians) who have suggested the Jun-Air 6-25. (Many wrote just to me and } not to the whole group. Jim, unfortunately the folks at Jun-Air would not } recommend a model suited to my application when I asked them. They only offered } to send a catalog.) } } Hopefully, my particular application and environment won't generate the } problems John experienced. We really need the quiet operation (specs say 45 } dB), especially because the unexpected cycling of a louder compressor is scaring } folks out of their wits. Not at all a good situation for handling delicate } specimens/procedures. } } I'm very tempted to just use nitrogen tanks, but the old 400 has a couple of } slow leaks which I might not get to fixing right away, plus we'll probably have } it running continuously for long periods, so I'd rather not worry about tank } deliveries and running out at a bad time. (One of the same reasons I switched } to natural gas from oil for home heating..., plus cleanliness of course.) } } Vitaly, thanks for the reasonably priced alternative that employs a modified } refrigeration compressor. I love being creative, but just don't have the time } right now. Will keep it in mind though. } } Thanks & best regards to all! } -Eric } } } } } } } Eric Anderson wrote: } } } } } ------------------------------------------------------------------------ The } } } Microscopy ListServer -- Sponsor: The Microscopy Society of America To } } } Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line } } } Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } New Year's Greetings! } } } } } } Say, can anyone recommend a super-reliable and quiet brand (and size) of air } } } compressor to use with a Philips EM400T? } } } } } } Unfortunately, in addition to being very, very loud, the low grade air } } } compressor we've been using has the intermittent habit of blowing it's } } } circuit breaker (twice in the last couple of weeks), which has set us } } } way back on getting our "new" microscope fully evacuated for the first } } } time. } } } } } } I'm waiting for a catalog from Jun-Air, but would like to know what } } } model/size has been used reliably by others over the years. The Philips } } } installation manual doesn't give much more than the air pressure } } } requirement, so I'm uncertain what is ideal in terms of pump and tank } } } capacities. I've asked the folks at Jun-Air, but they are reluctant to make } } } any recommendation based solely on the pressure spec. } } } } } } Thanks for any tips! } } } } } } Best regards, } } } Eric Anderson } } } Southern CT State University Physics Dept. } } } 501 Crescent Street, JE108B } } } New Haven, CT 06515 } } } 203-392-6468 } } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
Hello. I am a graduate student at Hofstra University, currently working on a retinal analysis of a deep water shark. I am headed on my first specimen collection trip this week and have a question about my fixation protocol. I will be using a mix of paraformaldehyde/gluteraldehyde in cacodylate buffer at pH 7.2. Should I be using sucrose, CaCl2, etc..to adjust the tonicity?
Any advice would be appreciated. Thank you.
Jill Olin Graduate Student Department of Biology Hofstra University Hempstead, NY 11549
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{color} {param} 0100,0100,0100 {/param} {bigger} Hi everybody,
We are looking into buying a new vacuum evaporator. We would it like to be equipped in two electrodes (for carbon and metal evaporation), rotary/tilt stage and thickness monitor. Our laboratory deals mainly with biological/veterinary samples. I would like to hear your recommendations regarding the purchase: equipment servicing and make (which one is good which is not). Is Denton the best available on the market? Do they still supply DV 502A?
Contact Tom Schmelzer at TGS Technologies. And/or see his ad in the November Microscopy Today on page 44.
He can be reached at 724 453 3865
-----Original Message----- } From: Tom Murray [mailto:murraytm-at-u.washington.edu] Sent: Wednesday, January 08, 2003 7:59 PM To: Microscopy
Hi All,
I have a GATAN Cryo-Transfer cold stage for a Philips EM420 TEM. The crystal needs to be replaced and I was wondering if anyone has knowledge of a third party who does this type of repair?
Thanks in advance.
Tom -- Thomas M. Murray Phone: (206)543-2836 Manager, Electron Microscopy Center Fax: (206)543-3100 Materials Science & Engineering email: murraytm-at-u.washington.edu 302 Roberts Hall, Box 352120 University of Washington Seattle, WA 98195-2120
The consensus seems to be that Tom Schmelzer at TGS Technologies is a good choice.
Thanks for the replies.
Tom -- Thomas M. Murray Phone: (206)543-2836 Manager, Electron Microscopy Center Fax: (206)543-3100 Materials Science & Engineering email: murraytm-at-u.washington.edu 302 Roberts Hall, Box 352120 University of Washington Seattle, WA 98195-2120
The EM Facility at Utah State University is being closed, March 1st 2003 due to budget cuts to the University. I am looking for other facilities which will do service work, and pricing rates for that work. Some of the work will be for University personnel, but most will be for off-site industrial users. Please let me know if you can do work for one group or the other or both. This includes both biological and materials; TEM and SEM (high res, and environmental) and edx.
Thank you for any help in this matter.
Bill
William McManus Supervisor Microscopy Facility Utah State University Logan UT 84322-5305
Tina's comments are quite on the mark. I have a Nikon Coolscan 4000 ED with the scratch removal software (ice cube). I do not use it at all after some tests. The effect is usually too strong and there is no preview of the effect as it is possible in photoshop. So I scan in regular 35 mm slides with no scratch removal, and then take them over to photoshop, use "Dust and Scratches" diameter usually 2 pixel, threshold 8 to 12, and check on some areas with particular detail. Occasionally even that is too much, and then it is back to the cloning stamp and manual removal of dust and scratches. Use a feathered cloning stamp: I like diameter 9 - 13 pixels and 75% hardness, 5% spacing. The feathered tool avoids hard edges around the cloned areas. Takes a while on a 55-60 MB file (RGB TIFF). Note, that with the fine removal of dust, you may not be able to get the larger pieces, which still have to be removed manually. Also, take some time in prepping your hard copy images: use a dust removal spray, and possibly a fine optical lens brush. The 10 seconds spent on that are well worth the time not spent messing around in photoshop.
I would think that the scratch removal effects will be much more disastrous on TEM images, in which the images is directly lens formed (i.e., not pixelated), than with SEM images. In the latter, on good old 4 x 5" negatives you get glorified pixels on a fine grain emulsion, so scratch removal should be quite effective on SEM negatives, but may pose problems with TEM images.
Best wishes Daniel
At 7:49 AM -1000 1/8/03, Tina Carvalho wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} But be aware that any camera or scanner hardware/software that "removes } dust and scratches" usually does so by blurring and eroding and then } unblurring, or similar processing, ...
Not exactly true ... as mentioned by another post, if this scanner uses the ICE procedure, then it uses an infrared scan to detect dust and thereby corrects only the areas within the image which needs it.
What needs to be mentioned in addition is the fact IR cannot see through a silver emulsion, and will therefore see all exposed areas as needing correction. IR aided correction will only work with color transparencies and chromes.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
We do all manner of biological TEM and would be happy to help out if the need arises.
Sara Miller Contact info in footer
On Thu, 9 Jan 2003, William McManus wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear group: } } The EM Facility at Utah State University is being closed, March 1st 2003 } due to budget cuts to the University. I am looking for other facilities } which will do service work, and pricing rates for that work. Some of } the work will be for University personnel, but most will be for off-site } industrial users. Please let me know if you can do work for one group } or the other or both. This includes both biological and materials; TEM } and SEM (high res, and environmental) and edx. } } Thank you for any help in this matter. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Our industry is currently implementing systems to deal with regulatory agency requirements as stated by Mike Bode in his recent reply. There are a few companies working on database solutions to solve some of the traceability issues related to electronic raw data and data manipulation. I am aware of 2 companies that are marketing database applications to address these issues (Cyberlab / Nugenesis(SP?)), and there are probably many more companies. Keep in mind that these applications are not specifically targeted at imaging data, but rather will address most file types. In short, the raw data is sent to a server, the database then archives the file to the database (at some user specified interval). To view the information the files are "restored" to a specified location where they can be manipulated. These manipulated files are then returned to the database using version control methodologies. An audit trail is maintained which allows for complete tracking of raw data access and manipulations. In our installation the archived files are maintained on servers making raw data retrieval a simple affair i.e. no CD's or tapes need be changed / loaded.
In my opinion this type of archival system would be very cumbersome and prohibitively expensive for most EM facilities to implement AND maintain.
One final note: It's a sad state of affairs when the assumption is made that these systems are required to prevent fraud. Dishonest people will always find a way to "beat" the system. It was true for wet chemistry methods and it is true for digital imaging as well. In my mind these systems and procedures are required to prevent the inadvertent, unknowing alteration of raw data. Accidentally deleting or overwriting a file, "misplacing" a file, or possibly confusing a raw data file with one that you knowingly manipulated during analysis. I am speculating but I would assume that most of us have experienced some or possibly all of these situations at one point in our digital careers. Honest mistakes, yes, but troubling errors that must be avoided. They waste time, effort, money, and can be very costly in terms of your reputation and the credibility of your work.
My two cents worth.
John Robson Boehringer Ingelheim Pharmaceuticals, Inc. PO Box 368 900 Ridgebury Rd Ridgefield, CT 06877
Phone (203)798-5640 Fax (203)798-5698
e-mail jrobson-at-RDG.boehringer-ingelheim.com
-----Original Message----- } From: Steve Bagley [mailto:SBagley-at-picr.man.ac.uk] Sent: Wednesday, January 08, 2003 7:49 AM To: 'Micro Discussion Group'
Hi,
I am in the middle of writing a joint grant proposal for an optical disk jukebox / image archiving solution which should hopefully hold up to ten years microscopy data. This system hopefully will aid in 'traceability' of raw data and the image processed result. I was wondering what other research institutes have in place for image archiving regarding traceability of images.
Techniques of image manipulation that, in the past, could only be done by few, and with great care, are now routine, consequently readily available software has also created ample opportunities to falsify images. Obviously the raw data and the final data to be published should be stored long term, but what about the steps in-between? As most research institutes now are multi-user facilities, the onus of scientific proof should be on the owner of the data, but since students are replaced every three - four years there must also be a burden of proof on the research institute especially if the images are to be reused at a later date.
Should a multi-user microscopy facility demonstrate traceability of data or should we rely on the users?
Steve
Steve Bagley Associate Scientist Applied Imaging Facility Paterson Institute For Cancer Research Cancer Research UK Christie Hospital, Wilmslow Rd, Manchester. M20 9BX, UK
This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute For Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.
We are going to process a biofilm with benign strain of E. coli on surface of catheters for TEM. We are reluctant to scrape the biofilm off, so we are going to process the biofilm while attached to the catheter. We do not know if the biofilm, catheter and resin (Glauert Med Embed) will embed well and the sections will come out good. We appreciate any suggestions before we try this out.
Thank you,
Claire Haueter
Claire Haueter EM Technician Integrated Microscopy Core Baylor College of Medicine 713-798-4952
I am a graduate student with the University of Auckland requiring advice on (1) the post-fixation stage for TEM prep., (2) any advice concerning the treatment of fragile sample material for TEM (standard viewing or shadowing).
I am using TEM to look at freshly grown microstromatolitic films from a geothermal brine where the primary mineral component is colloidal silica, with a minor component of sulphur. The objective is to study biomineralisation. We have scrape one set of my samples from their glass substrate (a terribly destructive stage as far a texture goes as the samples are very brittle and porous), dehydrated them through an ethanol series and added osmium tetroxide as a post-fixative (after initial fixation with 2.5% glutaraldehyde/buffer), however in similar studies the osmium tetroxide stage has been omitted - any ideas why?
I would be greatful for any advice.
Kind Regards, Kim -- Kim M. Handley Geology Department University of Auckland k.handley-at-auckland.ac.nz
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} Steve:
There was symposium on this very topic at the MSA meeting in Quebec City this past summer. I taped most of it and when I find the time (ha!) the plan is to publish a synopsis in Microscopy Today and on the web.
The bottom line if I recall correctly, is that current Copyright Law currently provides most - but not all - protection....at least in the US. Probably there will be variations country-to-country though.
Peter Ingram Public Policy Chair and Legal Liaison Officer, MSA
} } } } } } } } } } } } } } } } } } This email is confidential and intended solely for the use of the Person(s) } ('the intended recipient') to whom it was addressed. Any views or opinions } presented are solely those of the author and do not necessarily represent } those of the Paterson Institute For Cancer Research or the Christie Hospital } NHS Trust. It may contain information that is privileged & confidential } within the meaning of applicable law. Accordingly any dissemination, } distribution, copying, or other use of this message, or any of its contents, } by any person other than the intended recipient may constitute a breach of } civil or criminal law and is strictly prohibited. If you are NOT the } intended recipient please contact the sender and dispose of this e-mail as } soon as possible.
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
We are going to process a biofilm with benign strain of E. coli on surface of catheters for TEM. We are reluctant to scrape the biofilm off, so we are going to process the biofilm while attached to the catheter. We do not know if the biofilm, catheter and resin (Glauert Med Embed) will embed well and the sections will come out good. We appreciate any suggestions before we try this out.
Thank you,
Claire Haueter
Claire Haueter EM Technician Integrated Microscopy Core Baylor College of Medicine 713-798-4952
Briefly, you can set the resin in a section of catheter. Then peel off the catheter and reembed your core in a suitable mold. A lot of the biofilm should remain in the resin.
Dave
On Thu, 9 Jan 2003 20:22:49 -0600 chaueter {chaueter-at-bcm.tmc.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Listers, } } We are going to process a biofilm with benign strain of E. coli on surface of } catheters for TEM. We are reluctant to scrape the biofilm off, so we are } going to process the biofilm while attached to the catheter. We do not know } if the biofilm, catheter and resin (Glauert Med Embed) will embed well and the } sections will come out good. We appreciate any suggestions before we try this } out. } } Thank you, } } Claire Haueter } } Claire Haueter } EM Technician } Integrated Microscopy Core } Baylor College of Medicine } 713-798-4952 } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
As for your evaporator enquiry on the best evaporator, we have reason to believe Ladd Research provides the best vacuum evaporator available and has for many years.
Naturally I must confess to a little bias on the subject, but not too much. You can check our website for more information or contact us.
Disclaimer: Ladd sells all microscopy supplies and accessories, including the Ladd Vacuum Evaporator.
Best regards, John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- From: Dorota Wadowska To: microscopy-at-sparc5.microscopy.com Sent: Thursday, January 09, 2003 8:10 AM Subject: vacuum evaporator
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society 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 everybody, We are looking into buying a new vacuum evaporator. We would it like to be equipped in two electrodes (for carbon and metal evaporation), rotary/tilt stage and thickness monitor. Our laboratory deals mainly with biological/veterinary samples. I would like to hear your recommendations regarding the purchase: equipment servicing and make (which one is good which is not). Is Denton the best available on the market? Do they still supply DV 502A? Thanks Dorota
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
Try to avoid disturbing the biofilm if yu substatum is cheap by adding resin etc to it. People have done this with cell cultures on coverslips. The resin and mold can be separated from the glass using liquid nitrogen. The biofilm should stay with the resin.
Dave
On Fri, 10 Jan 2003 16:18:33 +1300 Kim Handley {khan030-at-ec.auckland.ac.nz} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } } I am a graduate student with the University of Auckland requiring advice on (1) } the post-fixation stage for TEM prep., (2) any advice concerning the treatment } of fragile sample material for TEM (standard viewing or shadowing). } } I am using TEM to look at freshly grown microstromatolitic films from a } geothermal brine where the primary mineral component is colloidal silica, with } a minor component of sulphur. The objective is to study biomineralisation. } We have scrape one set of my samples from their glass substrate (a terribly } destructive stage as far a texture goes as the samples are very brittle and } porous), dehydrated them through an ethanol series and added osmium tetroxide } as a post-fixative (after initial fixation with 2.5% glutaraldehyde/buffer), } however in similar studies the osmium tetroxide stage has been omitted - any } ideas why? } } I would be greatful for any advice. } } Kind Regards, } Kim } -- } Kim M. Handley } Geology Department } University of Auckland } k.handley-at-auckland.ac.nz } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Hi all A happy new year to all. May this year be the year your lottery numbers come up!
Anaspec is a EM Technical support company operating from South Africa but supporting clients around the world. We are about to open an office in Sydney, Australia and so need a few new support engineers. We realise that engineers in this field are as scarce as hens teeth or the correct lottery numbers these days, but we thought we would at least post a notice on this list server so that none of our friends and clients around the world can say, "But we wanted that job!" afterwards.
As with any technical support job the pay is never enough, the hours are long and the abuse is standard, but you get to work with some real characters and see laboratories all around the world.
If you do still have a slight interest in joining Anaspec, you can visit our web site for more details on our company and then send us an email.
Thanks for your time and hope to see some of you at a conference or laboratory this year. Cheers
Luc Harmsen ANASPEC South Africa www.anaspec.co.za Tel: +27 11 794 8340 Fax: +27 11 794 8349 P.O. Box 2561 Honeydew 2040 Gauteng, South Africa
We have kinda side stepped the issue of traceability by ensuring the original data is available. All images are collected into the Analysis database by Soft Imaging System. There are others out there, this one happened to be floating around the lab at the time I arrived and seems to work well. The database is automatically split into CD size chunks which serve as our archived originals. Users are given read access via a web browser interface. They can snatch their images when they want, and yes, they can do what they want with them, but they must always know that an original exists under my control if there is ever a question.
As an aside for your storage solution, after CD archiving I move the data to a Snap server (or NAS) and lock the database. Essentially it is a series of Raid 5 hard drives and an ethernet card. I can keep about 5 years on at all times (240G) for under $2k and it is completely scalable. Took 5 min to set up and another 5 to set proper access permissions. It was far cheaper than the jukeboxes we looked into and definitely faster serving the images. It works withing most environments PC Mac, unix.... Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Peter Ingram (by way ofMicroscopyListServer) {p.ingram-at-cellbio.duke.edu} 01/10/03 12:51AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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} Steve:
There was symposium on this very topic at the MSA meeting in Quebec City this past summer. I taped most of it and when I find the time (ha!) the plan is to publish a synopsis in Microscopy Today and on the web.
The bottom line if I recall correctly, is that current Copyright Law currently provides most - but not all - protection....at least in the US. Probably there will be variations country-to-country though.
Peter Ingram Public Policy Chair and Legal Liaison Officer, MSA
} } } } } } } } } } } } } } } } } } This email is confidential and intended solely for the use of the Person(s) } ('the intended recipient') to whom it was addressed. Any views or opinions } presented are solely those of the author and do not necessarily represent } those of the Paterson Institute For Cancer Research or the Christie Hospital } NHS Trust. It may contain information that is privileged & confidential } within the meaning of applicable law. Accordingly any dissemination, } distribution, copying, or other use of this message, or any of its contents, } by any person other than the intended recipient may constitute a breach of } civil or criminal law and is strictly prohibited. If you are NOT the } intended recipient please contact the sender and dispose of this e-mail as } soon as possible.
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
I might be missing something here and wondered if some of you could answer a question for me. If we were to add EDS to our ESEM, we could minimize scattering effects by using helium as the charge suppression gas. It small size and low z won't produce spurious x-rays, has minimal beam spreading, and is ionizable for charge suppression. More importantly, many of you swear by it. Now for the question. How do you keep it out of the detector?? Aren't they sealed, and won't helium pass through the window?? Is it going to trash the detector vacuum?? or is my understanding of defector design flawed?? Maybe the pumps recover the vacuum after while as it diffused back out?? Manufacturers encouraged to chime in here. Thanks
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
Dear all, We have two PhD positions available at the moment. My (rough) translation of the original advert is below, followed by the original in German, for those who prefer that.
are available in the Structural Research Group of the Department of Materials and Geological Sciences, as part of the Special Research Area 595 Fatigue of Functional Ceramics. Employment is limited to 4 years.
Applicants should have a Bachelors or Masters Degree in Materials Science, Physics, or Chemistry, and a desire to work towards a PhD in the interdisciplinary field of piezoelectric ceramics. The primary methods used in the work shall be X-ray diffraction, Transmission electron microscopy and synchrotron radiation (at HASYLAB, Hamburg). There will be opportunities to spend time overseas at partner Institutions.
The Technical University of Darmstadt seeks to increase the proportion of women in the staff, particularly in technical subjects and suitably qualified female candidates are therefore especially encouraged to apply.
Suitably qualified disabled persons will be given preferential treatment.
Payment will be calculated using the BAT scale (pay scale for public sector workers in Germany).
Applications should be sent together with Curriculum Vitae to: Das Präsidium der TU Darmstadt, Karolinenplatz 5, 64289 Darmstadt
All applications should be received before 10th February 2003
Stellenausschreibung
Im Fachbereich Material- und Geowissenschaften, Fachgebiet Strukturforschung sind im Rahmen des Sonderforschungsbereichs 595 Ermüdung von Funktionskeramiken zwei Stellen für
Wissenschaftliche Mitarbeiter(innen) - halbtags
(Kenn-Nr. 1) in einem befristeten Arbeitsverhältnis (4 Jahre) zu besetzen.
Gesucht werden Materialwissenschaftler, Physiker oder Chemiker mit abgeschlossenem Diplom, die eine Dissertation in dem praxisnahen, interdisziplinären Arbeitsgebiet piezoelektrischer Keramiken mitarbeiten möchten. Eingesetzte Methoden sind Röntgenbeugung, Transmissionselektronenmikroskopie und Synchrotronstrahlung (am HASYLAB, Hamburg). Gelegenheit zu längeren Auslandsaufenthalten wird gegeben.
Die Technische Universität Darmstadt strebt eine Erhöhung des Anteils der Frauen am Personal, insbesondere in den technischen Bereichen an und fordert deshalb qualifizierte Frauen nachdrücklich auf, sich zu bewerben.
Schwerbehinderte werden bei gleicher Eignung bevorzugt.
Die Vergütung erfolgt nach dem BAT.
Bewerbungen sind mit den üblichen Unterlagen an das Präsidium der TU Darmstadt, Karolinenplatz 5, 64289 Darmstadt, zu senden.
Bewerbungsfrist: 10. Februar 2002
-- Ian MacLaren Strukturforschung, Materialwissenschaft, TU-Darmstadt, Germany ian.maclaren-at-physics.org / http://homepages.tu-darmstadt.de/~maclaren/
} On Thursday, January 9, 2003, at 05:10 AM, Dorota Wadowska wrote: } } } We are looking into buying a new vacuum evaporator. We would it like } } to be equipped in two electrodes (for carbon and metal evaporation), } } rotary/tilt stage and thickness monitor. Our laboratory deals mainly } } with biological/veterinary samples. I would like to hear your } } recommendations regarding the purchase: equipment servicing and make } } (which one is good which is not). Is Denton the best available on the } } market? Do they still supply DV 502A? } } Dear Dorota, } I have also been investigating this, and I've gotten info about } Cressington, Denton, and Ladd evaporators. The final decision of what } to buy depends on what you want to do. For routine carbon and metal } evaporation, Cressington makes a relatively inexpensive (~$4k) benchtop } carbon coater, which uses only a rotary pump (sold separately) and } works at a vacuum of ~10^-2 torr. Their high-end turbopumped system } goes for ~$30k. Denton makes a benchtop turbo system that will handle } both carbon and metals for about $20k, and their high-end system for } high-resolution coating--evaporation or sputtering--goes for ~$70k. } Ladd makes a free-standing system with a diffusion pump that will also } handle both carbon and metals for about $20k. Other manufacturers: } Please feel free to contact me off-list. Your requirements for } uniformity of coating and preference for benchtop vs free-standing } models--as well as your budget--will determine your choice.
Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Our service lab does TEM processing and examination of nearly all kinds of biological samples. We have particular expertise with human muscle, nerve, and kidney biopsies, and are a CAP certified facility. We also do a lot of work with Immunobed and JB-4 resins. We are happy to work with other universities and private industry. For specific information about pricing, turn-around time, etc., please contact our lab administrator, Donna Craft.
Ralph Common Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824
ralph.common-at-ht.msu.edu
Donna Craft Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824
craftd-at-msu.edu
517-355-7558 -------------
On Thu, 9 Jan 2003, William McManus wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear group: } } The EM Facility at Utah State University is being closed, March 1st 2003 } due to budget cuts to the University. I am looking for other facilities } which will do service work, and pricing rates for that work. Some of } the work will be for University personnel, but most will be for off-site } industrial users. Please let me know if you can do work for one group } or the other or both. This includes both biological and materials; TEM } and SEM (high res, and environmental) and edx. } } Thank you for any help in this matter. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Scott's got the right idea as far as I'm concerned. Whenever I finish a report I provide a CDROM with the report and [digital copies of] all the original images obtained during the project. I also include in the names of the edited images the serial numbers of the originals from which they were prepared. And I make only globally applied changes to contrast, brightness, tone, etc. in addition to rotation and cropping or enlargement. That means the originals (including the Mavica's thumbnails and HTML index) are available for all to compare with my versions. And I place another copy of each CDROM with the file in case the computer should go belly up.
Best regards, George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
Okay folks - you know the drill. A client walks into a materials lab with a biological problem or vice versa. I work in a materials lab with SEM and LM and had a client walk in today with a biological question. So I am out of my element. Can you help?
It seems this man's daughter is suffering from some kind of skin ailment and she has dug stuff out of MANY of the sores on her skin. The doctors in her HMO have not offered any relief. Her father (from another department on this campus) is quite determined to find out what the problem might be. He would take no for an answer and would accept whatever help we could give him.
I examined the samples he brought in have posted images on our web server at ftp://www.marl.iastate.edu/What_is_it/. Many of them seem to be either bundles or wads of fibers or else little nodules of "stuff" with the fibers running throughout them. I suppose the stuff was secreted by the body, but can't say for sure.
LM pictures show the fibers to be both clear and darkly colored. BSE images and EDS from the SEM show the fibers to be organic in nature. (The nodules contain notable amounts of Na, Cl, and K along with some Si. But the fibers are primarily C and O.) The images don't appear to be like any natural or manmade fibers that I have ever encountered before. I can't find a match in our copy of the Particle Atlas.
Have any of you in all your experience come across such things? I would appreciate whatever expertise you could lend.
Thanking you in advance, Warren
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
A client has approached us asking for assistance in examining some "mercury-wetted" relay contacts. They have had problems with the contacts sticking and would like us to examine a few sets of contacts to see if we can identify the source of the problem.
My question is whether I should be concerned with inserting such samples in the microscope?
I don't intend to parts to be obviously wetted with mercury. However, I presume whatever Hg goes in will probably get volatilized and run out through our diffusion and rotary pump. We have mist traps on the outlet of our rotary pump. Do I need to be concerned about it reacting in bad ways with either the pump oil or components in the scope? (This would be in our conventional SEM rather than our VP-SEM. Its EDS system is down at the moment.)
Thanks in advance for your advice.
Warren
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
The Helium will diffuse through the EDS window and slowly ruin the vacuum in the EDS detector. Fortunately you are running in a vacuum and not applying 1 atm He to the window.
When I was in ED XRF back in Be window days, we would say you could use He instead of vacuum to avoid the absorption of low energy x-rays ( {3 KeV) by air. But those customers who did so eventually had to send their detectors back for repumping. Today's UTWs also leak Helium. You will have slow EDS dewar vacuum degradation if you do this. Another negative is encountered if there is a Ion Pump on the electron gun. Ion pumps do not pump the noble gases well and get flooded.
Ron Vane XEI Scientific
----- Original Message ----- } From: "Scott Whittaker" {Whittaker.scott-at-nmnh.si.edu} To: {microscopy-at-microscopy.com} Sent: Friday, January 10, 2003 6:32 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (medvitz-at-pitt.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, January 10, 2003 at 13:13:07 ---------------------------------------------------------------------------
Email: medvitz-at-pitt.edu Name: Neil Medvitz
Organization: University of Pittsburgh
Education: Graduate College
Location: Pittsburgh, PA USA
Question: What is the advantage if any to using gold grids instead of Nickel for post-embedding immunochem.
I'd like to jump in here with a comment or two on Cyberlab that we are developing for ERES compliance (Electronic Record/Electron Signature) as part of validation efforts to be in compliance with FDA regulations.
Cyberlab will store any file type as far as I know, please correct me if you know otherwise. The server ("archive") system can be set up to poll the camera workstation computer whenever and however often the user or "management" decide. This allows for deletion of images that are of no use, such as poor exposure, poor focus, vibration without having to go through all the documentation for a totally useless image.
Once the image is taken off the camera workstation computer it is "archived" by the Cyberlab server "immediately" (users cannot set that on our system) and can be retrieved at a later time; the original can be retained or automatically deleted from the workstation after some user defined period of time. Once retrieved from the server to perform analysis, etc., if it is stored by the same file name, it is given a version number. If it has a different name or file extension it is stored by that new name but linkage to the original is set up. How we will ultimately store terabytes of data over the years remains to be seen as the issue of long term retrieval capability as hardware changes have to be addresssed.
I agree with your statement about assumptions of guilt but we have only to look around us in the business world to see that the need exists. This will only make it more difficult to "beat" the system. Since such systems are designed so that users don't have access to the servers or storage, well that is only a small part of the security of the data that we have to set up. We also have to have workstation computer security so that only authorized users can get access and only to their folders etc. so that the servers see the data as coming from that user. Much more has to be done to reach compliance or whatever rules are ultimately decided on.
Damian Neuberger Senior Research Scientist Baxter Healthcare Corp. PO Box 490 Round Lake, IL 6003 damian_neuberger-at-baxter.com
} Steve, } } Our industry is currently implementing systems to deal with regulatory } agency requirements as stated by Mike Bode in his recent reply. There are a } few companies working on database solutions to solve some of the } traceability issues related to electronic raw data and data manipulation. } I am aware of 2 companies that are marketing database applications to } address these issues (Cyberlab / Nugenesis(SP?)), and there are probably } many more companies. Keep in mind that these applications are not } specifically targeted at imaging data, but rather will address most file } types. In short, the raw data is sent to a server, the database then } archives the file to the database (at some user specified interval). To } view the information the files are "restored" to a specified location where } they can be manipulated. These manipulated files are then returned to the } database using version control methodologies. An audit trail is maintained } which allows for complete tracking of raw data access and manipulations. In } our installation the archived files are maintained on servers making raw } data retrieval a simple affair i.e. no CD's or tapes need be changed / } loaded. } } In my opinion this type of archival system would be very cumbersome and } prohibitively expensive for most EM facilities to implement AND maintain. } } One final note: It's a sad state of affairs when the assumption is made } that these systems are required to prevent fraud. Dishonest people will } always find a way to "beat" the system. It was true for wet chemistry } methods and it is true for digital imaging as well. In my mind these } systems and procedures are required to prevent the inadvertent, unknowing } alteration of raw data. Accidentally deleting or overwriting a file, } "misplacing" a file, or possibly confusing a raw data file with one that you } knowingly manipulated during analysis. I am speculating but I would assume } that most of us have experienced some or possibly all of these situations at } one point in our digital careers. Honest mistakes, yes, but troubling } errors that must be avoided. They waste time, effort, money, and can be } very costly in terms of your reputation and the credibility of your work. } } My two cents worth. } } } John Robson } Boehringer Ingelheim Pharmaceuticals, Inc. } PO Box 368 } 900 Ridgebury Rd } Ridgefield, CT 06877 } } Phone (203)798-5640 } Fax (203)798-5698 } } e-mail jrobson-at-RDG.boehringer-ingelheim.com
If they do, they could plug in a flash memory puck and dump up to a gig of data and walk out with it. Does your system prevent this?
Just a thought.
gary g.
At 10:40 PM 1/10/2003, you wrote:
} John, } } I'd like to jump in here with a comment or two on Cyberlab that we are } developing for ERES compliance (Electronic Record/Electron Signature) as } part of validation efforts to be in compliance with FDA regulations. } } Cyberlab will store any file type as far as I know, please correct me if you } know otherwise. The server ("archive") system can be set up to poll the } camera workstation computer whenever and however often the user or } "management" decide. This allows for deletion of images that are of no use, } such as poor exposure, poor focus, vibration without having to go through } all the documentation for a totally useless image. } } Once the image is taken off the camera workstation computer it is "archived" } by the Cyberlab server "immediately" (users cannot set that on our system) } and can be retrieved at a later time; the original can be retained or } automatically deleted from the workstation after some user defined period of } time. Once retrieved from the server to perform analysis, etc., if it is } stored by the same file name, it is given a version number. If it has a } different name or file extension it is stored by that new name but linkage } to the original is set up. How we will ultimately store terabytes of data } over the years remains to be seen as the issue of long term retrieval } capability as hardware changes have to be addresssed. } } I agree with your statement about assumptions of guilt but we have only to } look around us in the business world to see that the need exists. This will } only make it more difficult to "beat" the system. Since such systems are } designed so that users don't have access to the servers or storage, well } that is only a small part of the security of the data that we have to set } up. We also have to have workstation computer security so that only } authorized users can get access and only to their folders etc. so that the } servers see the data as coming from that user. Much more has to be done to } reach compliance or whatever rules are ultimately decided on. } } Damian Neuberger } Senior Research Scientist } Baxter Healthcare Corp. } PO Box 490 } Round Lake, IL 6003 } damian_neuberger-at-baxter.com } } } } } Steve, } } } } Our industry is currently implementing systems to deal with regulatory } } agency requirements as stated by Mike Bode in his recent reply. There are } a } } few companies working on database solutions to solve some of the } } traceability issues related to electronic raw data and data manipulation. } } I am aware of 2 companies that are marketing database applications to } } address these issues (Cyberlab / Nugenesis(SP?)), and there are probably } } many more companies. Keep in mind that these applications are not } } specifically targeted at imaging data, but rather will address most file } } types. In short, the raw data is sent to a server, the database then } } archives the file to the database (at some user specified interval). To } } view the information the files are "restored" to a specified location } where } } they can be manipulated. These manipulated files are then returned to the } } database using version control methodologies. An audit trail is } maintained } } which allows for complete tracking of raw data access and manipulations. } In } } our installation the archived files are maintained on servers making raw } } data retrieval a simple affair i.e. no CD's or tapes need be changed / } } loaded. } } } } In my opinion this type of archival system would be very cumbersome and } } prohibitively expensive for most EM facilities to implement AND maintain. } } } } One final note: It's a sad state of affairs when the assumption is made } } that these systems are required to prevent fraud. Dishonest people will } } always find a way to "beat" the system. It was true for wet chemistry } } methods and it is true for digital imaging as well. In my mind these } } systems and procedures are required to prevent the inadvertent, unknowing } } alteration of raw data. Accidentally deleting or overwriting a file, } } "misplacing" a file, or possibly confusing a raw data file with one that } you } } knowingly manipulated during analysis. I am speculating but I would } assume } } that most of us have experienced some or possibly all of these situations } at } } one point in our digital careers. Honest mistakes, yes, but troubling } } errors that must be avoided. They waste time, effort, money, and can be } } very costly in terms of your reputation and the credibility of your work. } } } } My two cents worth. } } } } } } John Robson } } Boehringer Ingelheim Pharmaceuticals, Inc. } } PO Box 368 } } 900 Ridgebury Rd } } Ridgefield, CT 06877 } } } } Phone (203)798-5640 } } Fax (203)798-5698 } } } } e-mail jrobson-at-RDG.boehringer-ingelheim.com
It should not be necessary to scrape the sample off the glass to process it. There was a discussion on this point recently, but it may not have been on this list - I subscribe to several. You can fix in a primary aldehyde fixative, followed by a buffer rinse and then secondary fixation with osmium tetroxide in an aqueous buffer . After rinsing in water, dehydrate through graded alcohols finishing with several changes of dry absolute alcohol or with propylene oxide, then in a disposable dish pour a layer of Epon (or other epoxy resin) onto the slide. Change the Epon at least once, then use Epon plus accelerator. Cut the bottom off a Beem capsule and place the capsule tube over the area of interest, then polymerise the whole thing. Next day, fill the Beem capsule with accelerated Epon and polymerise. Pulling the Beem capsule off the glass after cooling should bring the sample with it, or it is possible to chip the glass away before sectioning. I am assuming the glass is a slide; if it's not then it should be possible to modify this technique. Use a fume hood and glove;, all the reagents except alcohol are dangerous, especially OsO4. Hope this helps.
Lesley Weston.
on 09/01/2003 7:18 PM, Kim Handley at khan030-at-ec.auckland.ac.nz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } } I am a graduate student with the University of Auckland requiring advice on } (1) } the post-fixation stage for TEM prep., (2) any advice concerning the treatment } of fragile sample material for TEM (standard viewing or shadowing). } } I am using TEM to look at freshly grown microstromatolitic films from a } geothermal brine where the primary mineral component is colloidal silica, with } a minor component of sulphur. The objective is to study biomineralisation. } We have scrape one set of my samples from their glass substrate (a terribly } destructive stage as far a texture goes as the samples are very brittle and } porous), dehydrated them through an ethanol series and added osmium tetroxide } as a post-fixative (after initial fixation with 2.5% glutaraldehyde/buffer), } however in similar studies the osmium tetroxide stage has been omitted - any } ideas why? } } I would be greatful for any advice. } } Kind Regards, } Kim } -- } Kim M. Handley } Geology Department } University of Auckland } k.handley-at-auckland.ac.nz } } } }
Neil Medvitz asked: ========================================================================= Question: What is the advantage if any to using gold grids instead of Nickel for post-embedding immunochem. =========================================================================== There are several advantages which could be of greater or lesser importance depending on individual situations:
a) When a pair of normal antimagnetic stainless steel tweezers are used to hold a typical nickel grid during the typical immunoreaction, you have two dissimilar metals in contact in a low pH environment and you get a current flow (this is the basis of electrochemistry). And this current flow is thought to stunt the strength of the immuno reaction. I don't know to what degree that is published but this is what customers have told me over the years as to why they have concerns about nickel grids.
b) Gold grids don't suffer from this problems and they are far more inert under these same conditions than nickel. Consequently, there is for some a preference for gold. But gold is softer, and the grids tend to be a bit less self -supporting and therefore more difficult to work with, whereas nickel is more stiff, and does not bend as readily, that being the reason why some workers, at the end of the day, prefer nickel.
One can get around the electrochemistry issues when using Ni grids, however, if instead of using the normal antimagnetic stainless steel tweezer, gold plated tweezers are used. See URL http://www.2spi.com/catalog/tweezers/precision.html The gold plating acts as a passivation layer on the antimagnetic stainless steel tweezers, killing off the chances for an electro chemical reaction, also leading sometimes to corrosion product in one's samples.
Disclaimer: SPI Supplies offers both gold and nickel grids as well a range of tweezers including gold plated tweezers.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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Warren, I've not experienced it myself, but I've heard of the gun being shorted by the Hg vapor generated when the beam hits the Hg and heats it. Your VP system might be a better bet with the increased pressure and some sort of through-put of gas.
Ken Converse owner Quality Images third party SEM service Delta, PA
Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A client has approached us asking for assistance in examining some } "mercury-wetted" relay contacts. They have had problems with the } contacts sticking and would like us to examine a few sets of contacts } to see if we can identify the source of the problem. } } My question is whether I should be concerned with inserting such } samples in the microscope? } } I don't intend to parts to be obviously wetted with mercury. However, } I presume whatever Hg goes in will probably get volatilized and run } out through our diffusion and rotary pump. We have mist traps on the } outlet of our rotary pump. Do I need to be concerned about it reacting } in bad ways with either the pump oil or components in the scope? (This } would be in our conventional SEM rather than our VP-SEM. Its EDS } system is down at the moment.) } } Thanks in advance for your advice. } } Warren } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of } materials } Computer applications and networking } } } }
We put an Oxford GEM detector on our VP-SEM back in 1994. We routinely run Helium as the gas when we are running the scope. We switch over to air for idle periods. I suppose the column is filled with helium about 20 hours per week on average. (Sometimes it is operated in high vacuum mode and sometimes it is plain idle.) I don't think we had any changes in LN2 consumption over the years. We refilled the dewar twice a week and could probably count on it going 5 days between fills before going dry. The low LN2 alarm would go off before that happened.
The helium atmosphere certainly does much to improve the quality of the image and it also reduces the scattering effect on the x-ray signal. I can pass on details and images to prove the point. In fact I had to prove the point to one of our service engineers. They hadn't heard that the type of atmosphere made that big a difference.
During the same time we had a Kevex Quantum detector on our conventional SEM. We have 170 liter LN2 dewars in each scope room for filling the EDS dewars and we can count on both of the big dewars to run out within days of each other. I don't know how dependent that is on evaporation from the big dewars and how much of it is due to consumption in the EDS systems.
So I too now wonder what the manufacturers would say. Are some detector systems immune from He diffusion while components in other system render them more susceptible?
Warren
At 07:48 PM 1/10/03 -0800, you wrote:
} Scott, } } The Helium will diffuse through the EDS window and slowly ruin the } vacuum in the EDS detector. Fortunately you are running in a vacuum } and not applying 1 atm He to the window. } } When I was in ED XRF back in Be window days, we would say you could } use He instead of vacuum to avoid the absorption of low energy x-rays } ( {3 KeV) by air. But those customers who did so eventually had to send } their detectors back for repumping. Today's UTWs also leak Helium. } You will have slow EDS dewar vacuum degradation if you do this. } Another negative is encountered if there is a Ion Pump on the electron } gun. Ion pumps do not pump the noble gases well and get flooded. } } Ron Vane } XEI Scientific } } } ----- Original Message ----- } } From: "Scott Whittaker" {Whittaker.scott-at-nmnh.si.edu} } } To: {microscopy-at-microscopy.com} } } Sent: Friday, January 10, 2003 6:32 AM } } } } I might be missing something here and wondered if some of you could } } answer a question for me. If we were to add EDS to our ESEM, we could } } minimize scattering effects by using helium as the charge suppression } } gas. It small size and low z won't produce spurious x-rays, has minimal } } beam spreading, and is ionizable for charge suppression. More } } importantly, many of you swear by it. Now for the question. How do you } } keep it out of the detector?? Aren't they sealed, and won't helium pass } } through the window? Is it going to trash the detector vacuum? or is my } } understanding of detector design flawed? Maybe the pumps recover the } } vacuum after while as it diffused back out? Manufacturers encouraged to } } chime in here. } } Thanks } } } } Scott Whittaker } } Laboratories of Analytical Biology } } Smithsonian Institution } } National Museum of Natural History } } PO Box 37012 MRC104 } } Washington DC 20013-7012 } } 202-357-1651
This is very odd. I looked at the SEM images wondering if the fibres would look fungal. They do not look fungal to me. They look more like paper or cotton and some synthetic fibres (ribbed longitudinally). Do you think we are looking mostly at the material used to collect the samples?
Dave
On Fri, 10 Jan 2003 17:32:00 -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 } -----------------------------------------------------------------------. } } } Okay folks - you know the drill. A client walks into a materials lab with a } biological problem or vice versa. I work in a materials lab with SEM and LM } and had a client walk in today with a biological question. So I am out of } my element. Can you help? } } It seems this man's daughter is suffering from some kind of skin ailment } and she has dug stuff out of MANY of the sores on her skin. The doctors in } her HMO have not offered any relief. Her father (from another department on } this campus) is quite determined to find out what the problem might be. He } would take no for an answer and would accept whatever help we could give him. } } I examined the samples he brought in have posted images on our web server } at ftp://www.marl.iastate.edu/What_is_it/. Many of them seem to be either } bundles or wads of fibers or else little nodules of "stuff" with the fibers } running throughout them. I suppose the stuff was secreted by the body, but } can't say for sure. } } LM pictures show the fibers to be both clear and darkly colored. BSE images } and EDS from the SEM show the fibers to be organic in nature. (The nodules } contain notable amounts of Na, Cl, and K along with some Si. But the fibers } are primarily C and O.) The images don't appear to be like any natural or } manmade fibers that I have ever encountered before. I can't find a match in } our copy of the Particle Atlas. } } Have any of you in all your experience come across such things? I would } appreciate whatever expertise you could lend. } } Thanking you in advance, } Warren } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Gold grids are a good choice for on-grid immunodetection when using conventional immunogold reagents (with a size suited for visualization without enhancement, for instance 10 nm gold particles). Gold grids are virtually chemically inert and thus won't easily interfere with components in incubation solutions. There is one exception: silver enhancement. Since gold particles act as catalysts in the deposition of metallic silver, gold grids may become covered with metallic silver as well, locally exhausting the enhancement reagents and leading to non-reproducible results. Nickel, as far as we have tested, does not seem to act as a catalyst for silver enhancement and in a practical sense is also chemically inert towards incubation solutions. In our experience handling such grids with non-magnetic tweezers (or better, with platinum loops) has never been a reason for doubting the immuno results. The main downside to using nickel grids is that they influence the electron beam and cause astigmatism making more frequent adjustments necessary while observing specimens in the TEM.
Electrochemical phenomena may already occur whenever a metal surface is brought into contact with a solution. It does not necessarily involve a second (different) metal. Whether this actually results in a chemical reaction depends a.o. on the redox potential of the components involved and whether the overall conditions are favorable for a reaction to occur.
I would be interested to learn if there are any listers who would be willing to comment on the issue of electrochemical reactions influencing immunodetection. Any documented experience may teach us how to obtain better results.
Jan Leunissen Aurion - ImmunoGold Reagents and Accessories http://www.aurion.nl
Dear Sirs, I would like to know if somebody can help me to find the procedures for the baking of this field emission (JSM6400F). We have the connections to the heaters, we need to know if we have to dismount something in the gun before baking and if there is a difference between baking time on the gun and the column. All other information are well accepted.
I have no experience with the JSM6400F, however that said, I would strongly suspect that the gun bake time should be the longest in the system. The reason is that residual gas will tend to be driven from the hottest to the coolest region of the system, and for any microscope the best vacuum is desired in the gun. For comparison, the bakeout procedure on our SEM (CamScan 44FE) is: entire system, pumped by diffusion pump /w cold trap, all heaters on - 1 day cool lower ion pump; isolate from chamber; pump column+gun+upper ion pump with lower ion pump - 1 day cool upper ion pump; seal upper/lower pumping line; pump with both ion pumps - 1 day finally, cool gun assembly.
hope this helps, Ben Simkin (simkin-at-egr.msu.edu)
I am looking for a manual (operation instructions and technical specs.) for an Edwards E306 thermal evaporator. Of course I will pay for copy-costs and postage. If you can help me with that, please contact me directly: stefan.geimer-at-uni-koeln.de
Morning Warren, While on one of my searches to attempt to insure that our new FEI ESEM remains as uncontaminated as possible, I came across the following related to mercury in amalgams and SEM/vacuum:
http://www.gbg.bonet.se/bwf/art/instability.html
I can't speak to the validity of the experimentation, but I have bookmarked the site.
Hope this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Friday, January 10, 2003 6:39 PM To: Microscopy-at-sparc5.microscopy.com
A client has approached us asking for assistance in examining some "mercury-wetted" relay contacts. They have had problems with the contacts sticking and would like us to examine a few sets of contacts to see if we can identify the source of the problem.
My question is whether I should be concerned with inserting such samples in the microscope?
I don't intend to parts to be obviously wetted with mercury. However, I presume whatever Hg goes in will probably get volatilized and run out through our diffusion and rotary pump. We have mist traps on the outlet of our rotary pump. Do I need to be concerned about it reacting in bad ways with either the pump oil or components in the scope? (This would be in our conventional SEM rather than our VP-SEM. Its EDS system is down at the moment.)
Thanks in advance for your advice.
Warren
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Why not just call JEOL and talk to a service engineer? They are very helpful. You do not have to be on a service contract to get their help either.
Ron L
-----Original Message----- } From: Benjamin - Simkin [mailto:simkin-at-egr.msu.edu] Sent: Monday, January 13, 2003 7:02 AM To: Microscopy-at-sparc5.microscopy.com; b.rapillo-at-mclink.it
I have no experience with the JSM6400F, however that said, I would strongly suspect that the gun bake time should be the longest in the system. The reason is that residual gas will tend to be driven from the hottest to the coolest region of the system, and for any microscope the best vacuum is desired in the gun. For comparison, the bakeout procedure on our SEM (CamScan 44FE) is: entire system, pumped by diffusion pump /w cold trap, all heaters on - 1 day cool lower ion pump; isolate from chamber; pump column+gun+upper ion pump with lower ion pump - 1 day cool upper ion pump; seal upper/lower pumping line; pump with both ion pumps - 1 day finally, cool gun assembly.
hope this helps, Ben Simkin (simkin-at-egr.msu.edu)
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (burtonjoyner-at-insightbb.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, January 13, 2003 at 09:22:02 ---------------------------------------------------------------------------
Question: I am new to electron microscopy and have the opportunity to use the TEM at UK. Are there any comprehensive TEM sample preparation references in print? I am not having much luck finding good resources.
We've never had any trouble using Ni for immuno. Cu sometimes can react with reagents, particularly if you incubate overnignt, leaving blobs of copper sulfate (they turn greenish). As for Au, use them for growing cells on support films because the Au is not toxic to the cells. However, I think Au is overkill for immuno; Au grids are EXPENSIVE. If you have some sort of really corrosive treatment (why would you???), you might consider Au.
You should use several grids (at least 2; we use 3) for every different condition (different antibody--positive & negative, different antibody dilution, different tissue/cell type, etc). This means you'll be using lots of grids, which would be expensive if you're using Au ones.
Good luck, Sara Miller
On Fri, 10 Jan 2003, by way of MicroscopyListServer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (medvitz-at-pitt.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, } January 10, 2003 at 13:13:07 } --------------------------------------------------------------------------- } } Email: medvitz-at-pitt.edu } Name: Neil Medvitz } } Organization: University of Pittsburgh } } Education: Graduate College } } Location: Pittsburgh, PA USA } } Question: What is the advantage if any to using gold grids instead of } Nickel for post-embedding immunochem. } } --------------------------------------------------------------------------- } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
As always, Chuck has some good ideas. One additional one is that we don't use forceps at all. We use loops made with either Ni or Au wire (like the wire you put into the vacuum evaporator) glued onto sticks (we use plastic disposable loops normally used for spreading bacteria onto plates--with the plastic loop cut off). At the very end, we pick up the grid with forceps and dry it.
The loops are made with 1-2 inches of wire, wrapped around a drill bit just larger than the grid and twisted tightly to make a loop at one end. The other end is wrapped around the end of a plastic handle and glued with epoxy.
The advantages are that loops are reusable, and there's less likelyhood of bending the grids. Just make sure you wash grids between solutions sufficiently. We use 5-6 washes (drops of buffer--which is also probably overkill).
Good luck, Sara Miller
On Sat, 11 Jan 2003, Garber, Charles A. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Neil Medvitz asked: } ========================================================================= } Question: What is the advantage if any to using gold grids instead of } Nickel for post-embedding immunochem. } =========================================================================== } There are several advantages which could be of greater or lesser importance } depending on individual situations: } } a) When a pair of normal antimagnetic stainless steel tweezers are used to } hold a typical nickel grid during the typical immunoreaction, you have two } dissimilar metals in contact in a low pH environment and you get a current } flow (this is the basis of electrochemistry). And this current flow is } thought to stunt the strength of the immuno reaction. I don't know to what } degree that is published but this is what customers have told me over the } years as to why they have concerns about nickel grids. } } b) Gold grids don't suffer from this problems and they are far more inert } under these same conditions than nickel. Consequently, there is for some a } preference for gold. But gold is softer, and the grids tend to be a bit } less self -supporting and therefore more difficult to work with, whereas } nickel is more stiff, and does not bend as readily, that being the reason } why some workers, at the end of the day, prefer nickel. } } One can get around the electrochemistry issues when using Ni grids, however, } if instead of using the normal antimagnetic stainless steel tweezer, gold } plated tweezers are used. See URL } http://www.2spi.com/catalog/tweezers/precision.html } The gold plating acts as a passivation layer on the antimagnetic stainless } steel tweezers, killing off the chances for an electro chemical reaction, } also leading sometimes to corrosion product in one's samples. } } Disclaimer: SPI Supplies offers both gold and nickel grids as well a range } of tweezers including gold plated tweezers. } } Chuck } } PS: Remember that we are striving to be 100% paperless, therefore there } are no paper copies kept of this correspondence. Please be sure to always } reply by way of "reply" on your software so that the entire string of } correspondence can be kept in one place. } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.2spi.com } ######################## } ============================================ } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Have you considered putting the device into a vacuum system first to get any residual Hg off the surfaces prior to EM? That is, a vacuum system that is not the SEM column.
Peter Tomic Anadigics, Inc.
-----Original Message----- } From: qualityimages [mailto:qualityimages-at-netrax.net] Sent: Saturday, January 11, 2003 5:31 PM To: Warren E Straszheim; Microscopy
Warren, I've not experienced it myself, but I've heard of the gun being shorted by the Hg vapor generated when the beam hits the Hg and heats it. Your VP system might be a better bet with the increased pressure and some sort of through-put of gas.
Ken Converse owner Quality Images third party SEM service Delta, PA
Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A client has approached us asking for assistance in examining some } "mercury-wetted" relay contacts. They have had problems with the } contacts sticking and would like us to examine a few sets of contacts } to see if we can identify the source of the problem. } } My question is whether I should be concerned with inserting such } samples in the microscope? } } I don't intend to parts to be obviously wetted with mercury. However, } I presume whatever Hg goes in will probably get volatilized and run } out through our diffusion and rotary pump. We have mist traps on the } outlet of our rotary pump. Do I need to be concerned about it reacting } in bad ways with either the pump oil or components in the scope? (This } would be in our conventional SEM rather than our VP-SEM. Its EDS } system is down at the moment.) } } Thanks in advance for your advice. } } Warren } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of } materials } Computer applications and networking } } } }
I am interested in your opinions on the various brands of automatic black and white processors that produce a dry print. For the present, our Center must rely in part on silver emulsion processing as well as digital images. The processors I am looking at are the Colex, the Fujimoto CP31 and CP51. I would consider other brands.
Your comments, if you wish, can be sent to my e-mail address. In advance, I than you for your time.
Dr. Lewis B. Coons, Director Integrated Microscopy Center The University of Memphis Memphis, TN e-mail lcoons-at-memphis.edu
I have read with some interest the recent discussions of methods for cleaning cold cathode discharge gauges. In this connection, it might be of interest to note that there are two different types of CCD gauges. These are described in Sect 3.2.2 of my book Vacuum Methods in Electron Microscopy (for a description see http://www.2spi.com/catalog/books/book48.html). One type, perhaps the first type developed, is the classical 'Penning Gauge'. In modern gauges of this type the anode is usually a loop of tungsten wire that is located in the center of a surrounding cylindrical cathod, as shown in Fig. 3.12. The other type is commonly known as the 'Magneton Gauge". In this gauge the cylindrical case of the gauge is the anode, and the cathode is a rod with circular end plates that is located along the axis of the case, as shown in Fig. 3.13. Magnetron gauges are usually considered to be capable of recording considerably lower pressures than Penning gauges, and to be more stable and more accurate.
It sounds to me as though most of those who commented on the problem were describing procedures for refurbishing a Magnetron gauge, and indeed, my experience also suggests that it is best to replace the central element of this type gauge (the cathode) if it shows significang erosion, and that a local machine shop can produce a new one for you much cheaper than you can purchase one from the gauge manufacturer.
Happy New Year!! -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
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----- Original Message ----- } From: Nanovision S.R.L. {b.rapillo-at-mclink.it} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, January 13, 2003 5:36 AM
Hi Warren
The first care could be to limit evaporation, by use of a cold stage. The Pelltier stage from an ESEM schould be enought, to be just a bit above the melting point of Hg.
The second care could be the use of a cold trap in front of the sample, one like these used in HR-SEM, mounted on the objectif lens, or better, inserting a cold finger in the scope chamber and tilting the sample in it direction. This last configuration would limite the risk to evaporate something in the electron tube. Of coarse, this trap have to be cleaned after the observation, and the mounting of such a cold finger depends of what free port you have on the scope in tilting direction.
Third, use the lowest beam current possible (imaging or EDS ?).
An other care could be to pack the sample in Al foil, with a hole at the place you want to observe, to limit direct relation between the rest of the sample and the SEM vaccum. The vapor flux is proportionnal to the surface of metal exposed to the vaccum.
I don't think that you will have a strong evaporation rate in the vacuum level of a SEM, at room temperature or at -20C. The 10 to 200 pA beam current from a SEM in current observation conditions are far from the that of a MBE gun. Have a look at the vapor pressure curves, or at Holland's book about thin film technologie. Have a look at the vapor pressure curves, or at Holland's books about thin film technology. He gives the way to calculate the vapor flux.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} } } -----Original Message----- } } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] } Sent: Friday, January 10, 2003 6:39 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM: Examining mercury-wetted contacts } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A client has approached us asking for assistance in examining some } "mercury-wetted" relay contacts. They have had problems with the contacts } sticking and would like us to examine a few sets of contacts to see if we } can identify the source of the problem. } } My question is whether I should be concerned with inserting such samples in } the microscope? } } I don't intend to parts to be obviously wetted with mercury. However, I } presume whatever Hg goes in will probably get volatilized and run out } through our diffusion and rotary pump. We have mist traps on the outlet of } our rotary pump. Do I need to be concerned about it reacting in bad ways } with either the pump oil or components in the scope? (This would be in our } conventional SEM rather than our VP-SEM. Its EDS system is down at the } moment.) } } Thanks in advance for your advice. } } Warren } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of } materials } Computer applications and networking } } }
You'd better restrict yourself to techniques that are specific to the materials you need to prepare for TEM. Talk to someone knowledgeable at UK. Tell the Microscopy list the kind of materials you need to work on.
At 9:26 AM -0600 1/13/03, by way of MicroscopyListServer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ----------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center Argonne National Laboratory Materials Science Division, Building 212 9700 South Cass Avenue Argonne, IL 60439-4838 USA Telephone: 630-252-7194 FAX: 630-252-4289 recook-at-anl.gov
I have always just used scissors to cut mica to the desired shape and size.
Roger Moretz, PhD Dept of Toxicology BI Pharmaceuticals -- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 familiar with the techniques of cleaving mica, but what is the } recommended method for cutting mica to a desired shape or size? } } Linda Rinko } Design Engineer } Harvard Medical School } 060 Seeley G. Mudd } Boston, MA 02115 } 617-432-2052 } } }
there is one more possibility - gold-gilded copper grids by Gilder. I buy mine from Electron Microscopy Sciences and like them for most of post-embedding labeling, except silver-enhancement of course. They have proved sturdy and easy to handle and do not seem to react with PBS. They are not much more expensive than Ni grids. I dislike Ni grids for the reasons Jan and others listed, but pure gold ones were too flimsy.
Vlad
} Email: medvitz-at-pitt.edu } Name: Neil Medvitz } } Organization: University of Pittsburgh } } Education: Graduate College } } Location: Pittsburgh, PA USA } } Question: What is the advantage if any to using gold grids instead } of Nickel for post-embedding immunochem. } } ---------------------------------------------------------------------------
-- -------------------------------------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
We recently obtained an older metallograph, namely an Aus Jena Neophot 21, originally sold by Leco Corp. One of the lamps uses a halogen bulb with an extended base. We are having trouble finding a supplier for this type of bulb. If there are any Neophot 21 users out there that have found a bulb supplier, a recommendation would be greatly appreciated. Thanks in advance.
Joseph
Joseph M. Oparowski Center for Materials Science - Consulting and Failure Analysis Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone:(508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
Jibao He wrote: ============================================================================ ==== A my friend wants to know which company can produce High Oriented Pyrolyzed Graphite slides used for AFM? or where he can order it? ============================================================================ ==== SPI Supplies has offered highly ordered pyrolytic graphite (HOPG) for many years for AFM applications, the details for which can be found on URL http://www.2spi.com/catalog/new/hopgsub.shtml
A number of custom sizes are shown, but we can make just about any specific size one might want. It is not easy to cut down to size without the special equipment and experience, so it is better to order to the size you want instead of a larger size you might try to cut down.
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 ######################## ============================================
Linda Rinko wrote: ============================================================================ == I am familiar with the techniques of cleaving mica, but what is the recommended method for cutting mica to a desired shape or size? ============================================================================ == One must first understand that mica is very very hard and cutting a thin sheet of mica is nearly like cutting a strip of equal thickness of steel! So no matter what the edge is that you would be using for the cutting, it is going to lose its sharpness very quickly, but just how quickly will depend on the thickness of the mica being cut.
All of the mica pieces described on the SPI Supplies website See URL http://www.2spi.com/catalog/submat/mic_shet.shtml are prepared by die cutting, other wise there will be delamination at the edges and also, a large generation of particulates. Carbide tools are always used for the die cutting, and that is why there is sometimes a not-so -small tooling charge when special sizes or shapes are needed. This is also why the mica sheets and discs sold by SPI Supplies have edges far more free of fracturing and splitting vs. other methods of cutting.
We are often times asked how mica purchased from SPI Supplies can be cut down to smaller pieces. And the answer depends on the thickness of the piece being cut.
Thickness is less than 15 mils (0.015" or 0.381 mm): For sheets of this thickness or less, one can use either a very sharp scissors or an Exacto type knife or even a single edge razor blade. Be sure to wear good eye protection (which should also be worn by any one nearby watching). One can expect some edge fracturing and splitting , the amount depending on the sharpness of the scissors or blade. The splitting can be minimized by holding down the mica, as it is being cut with a ruler or other "straight edge" instrument, the higher the pressure applied , the better the result (e.g. less fracturing). The thinnest mica cleavings can be scissors cut almost as if they were a piece of paper.
Thickness is greater than 15 mils (0.015" or 0.381 mm) but less than 30 mils (0.030" or 0.762 mm): Scissors and razor blades will not work. The only possibility is to use a guillotine type cutter perhaps even a paper cutter. There will be a strong tendency for the mica to edge fracture, but that can be minimized by applying higher downward pressures to the side of the mica on the flat cutting surface. It can be further minimized by being sure that the blade is the sharpest possible.
Thickness is greater than 30 mils (0.030" or 0.762 mm): We do not believe that anyone can do a credible job cutting such mica pieces with out resorting to the use of good tooling and die cutting.
Just remember that any cutting edge that you might use to cut the mica is going to be become dull quickly. While razor blades and even scissors can be discarded and new ones then used, one normally does not think of a paper cutter as being a disposable instrument. At the same time, we are not aware of any service that resharpens the guillotine blades from worn out paper cutters.
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 ######################## ============================================
Try Kreonite and Ilford processors. Used ones can be found at:
http://dunningphoto.com/used.html
http://www.kreonite.com/procpm2.htm
Kreonite has been around a long time.
gary g.
At 09:32 AM 1/23/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } } We recently obtained an older metallograph, namely an Aus Jena Neophot 21, } originally sold by Leco Corp. One of the lamps uses a halogen bulb with an } extended base. We are having trouble finding a supplier for this type of } bulb. If there are any Neophot 21 users out there that have found a bulb } supplier, a recommendation would be greatly appreciated. Thanks in advance. } } } Joseph } } Joseph M. Oparowski } Center for Materials Science - Consulting and Failure Analysis } Bose Corporation Joseph_Oparowski-at-bose.com } The Mountain, M/S 415 Phone:(508) 766-1371 } Framingham, MA 01701-9168 Fax: (508) 766-1313
Also check out Mohrpro processors. They are available in various sizes. Reliable and easy to use. http://www.mohrpro.com/
George
George Laing National Graphic Supply ph 800.223.7130 x3109 f 800.832.2205 email scisales-at-ngscorp.com
Dear Listers:
I am interested in your opinions on the various brands of automatic black and white processors that produce a dry print. For the present, our Center must rely in part on silver emulsion processing as well as digital images. The processors I am looking at are the Colex, the Fujimoto CP31 and CP51. I would consider other brands.
Your comments, if you wish, can be sent to my e-mail address. In advance, I than you for your time.
Dr. Lewis B. Coons, Director Integrated Microscopy Center The University of Memphis Memphis, TN e-mail lcoons-at-memphis.edu
I don't know if they supply the bulb you are looking for, but we buy most of our bulbs from Gray Supply. You can find them at 1-800-TOP-BULB (1-800-867-2852) or on the web at www.topbulb.com . They have a large supply of specialty bulbs and one of them may fit your application.
Mike
=========================================== Michael D. Standing Electron Microscopy Technologist Brigham Young University 401 WIDB Provo, UT 84602
-----Original Message----- } From: Oparowski, Joseph [mailto:Joseph_Oparowski-at-bose.com] Sent: Tuesday, January 14, 2003 11:44 AM To: 'microscopy-at-sparc5.microscopy.com' Cc: Shaw, Douglas
Greetings,
We recently obtained an older metallograph, namely an Aus Jena Neophot 21, originally sold by Leco Corp. One of the lamps uses a halogen bulb with an extended base. We are having trouble finding a supplier for this type of bulb. If there are any Neophot 21 users out there that have found a bulb supplier, a recommendation would be greatly appreciated. Thanks in advance.
Joseph
Joseph M. Oparowski Center for Materials Science - Consulting and Failure Analysis Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone:(508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
On Tuesday, January 14, 2003, at 06:58 AM, Linda Rinko wrote:
} I am familiar with the techniques of cleaving mica, but what is the } recommended method for cutting mica to a desired shape or size? } Dear Linda, I buy mica in ~1 cm x 3 cm x 1 mm pieces, then just use a scissors to cut a piece lengthwise. I also cut off a corner so I can tell which side has been carbon coated. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
For more than ten years we've been using a Hammamatsu SIT camera for imaging at video rates at low light fluorescence. However,the camera has serious vignetting and noise. Also, the tube has gotten dirty (dust) and we can't clean much of it.
We'd like to replace this camera with one with a flatter field, similar sensitivity, and at least 15 FPS at 640X480 or greater.
Yes, the Roper/Cooke/Hammamatsu $16k cooled CCDs could do this in many of the situation we image, but what about for much less money?
Thanks!
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
I am doing work on fluorescent staining of blood vessel tumor cross-sections. I was told that blood vessels will autofluoresce without the aid of staining. Is this true and if so at what wavelength will it fluoresce? Currently we are double staining to get our results (staining vessel cells and proteins inside of the cells), but if we only have to stain once, it would be beneficial. If the autofluorescing does not work well enough, does anyone have any ideas on what colors/stains would work for the double staining of cells that makeup the blood vessels (endothelial) and then tagging something within those cells? Right now we are using texas red and I am getting a ton of background noise (red) that makes it to difficult to see. Any information can be mailed to me directly, please, at bened008-at-umn.edu
Thanks,
Ryan Benedict Oral Sciences Department University of Minnesota 612-626-3562
I had to face the same problem a while ago (got an old Neophot 21). What I figured out is that the prices up here in Canada is much lower, compared to the States. The company I bought the bulbs from is Microlites Scientific. You can call them at: 1-800-263-8902 or 416-2995301. Speaking about the scope, does anybody have one for parts or has some spare parts for the machine?
Regards,
Vlad Igoshev, Ph.D.
Toronto, Canada
----- Original Message ----- } From: "Oparowski, Joseph" {Joseph_Oparowski-at-bose.com} To: {microscopy-at-sparc5.microscopy.com} Cc: "Shaw, Douglas" {Douglas_Shaw-at-bose.com} Sent: Tuesday, January 14, 2003 1:44 PM
Wil-
May be you could explain to me what mean "Inverted Magnetron" (or point some page in your book)? I got AIM-x (Active Inverted Magnetron) Gauge from Edwards a few years ago. Edwards claimed that this gauge is good up to 10-9 mbar. In recent discussion, somebody (I am sorry, I don't remember the name) claimed that "cold cathode gauge" could not perform well over 10-5 torr. How it may happening that my "inverted magnetron" gauge performed perfectly up to 10-8 torr? Thanks, Sergey.
At 12:45 PM 1/13/2003, 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
Michael, you are unlikely to find a camera sensitive enough to decently record fluorescence image at 15 frames per second, with software, for much less than $16K. However if, you agree to wait for 12 long seconds while image downloads from the camera, then check out http://www.starlight-xpress.co.uk/ . They have Peltier cooled 1300 x 1030 CCD camera with SONY ICX085AL sensor, at $2,500. Ask for industrial version, it is designed for long duty cycle. That's the least expensive one of such resolution on the market. And if you use it in 650 x 515 mode, (which you are looking for), the download takes only 3 seconds. Another company, SBIG, offers the least expensive 640 x 480 camera with Texas Instrument TC37 sensor, also Peltier cooled, and also very sensitive, with integrated color filter wheel, at $1,700. You can choose from a huge variety of filters. Check www.sbig.com . Both cameras come as a complete package with the power supply, camera/filters control and image processing software, and all cables. SBIG software is somewhat better for processing. Control capabilities are similar.
Disclaimer: I have no commercial interest in neither Starlight Express nor SBIG companies, other than being a satisfied customer.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: Michael Cammer {cammer-at-aecom.yu.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 15, 2003 4:31 PM
Dear All,
We are disposing of a trusty old friend, a Cambridge S250 MkIII. I am keeping several vacuum components and the BSE detector but the remainder could form useful spares for someone. Removal/shipping costs only.
Stu ---------------------- SL Kearns, Earth Sciences University of Bristol Bristol, UK +44 (0)117 954 5421 Stuart.Kearns-at-bristol.ac.uk
We are shortly saying goodbye to two Philips 301 transmission electron microscopes. One has a double tilt facility - the other one is not so high-powered, but might be considered as a source of parts for the first.
Would anyone be interested? If so, please let me know and I will take the matter higher up as regards arrangements.
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
We, too, have the same metallograph (AusJena Neophot 21 from LECO) in our lab. If the other leads listed on the Microscopy Listserver don't pan out, you may want to contact the contractors that service our equipment:
Hi-Point Optical Calibration 567 North Park Road Bellefontaine, Ohio 43311 (937) 592-2641
Bob & Mark Pickering (specialists) Julia Pickering (office clerk)
They purport to be very familiar and experienced with this particular microscope. You may want to keep them in mind for any future spare parts needed for your AusJena metallograph.
Regards,
Stu Smalinskas Senior Metallurgist SKF USA Plymouth, Michigan
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Hi Ryan, Indeed, blood vessels will be autofluorescent and the problem is that they will autofluoresce in all the channels we are usually using e.g. red, green and blue. I am facing a similar problem right now and to solve it you would have to go with a fluorophore that will emit at a wavelenght close to the infra red end of the spectrum. That would mean around 670-700 nm or higher. You would have to avoid the 450-615 part of the spectrum were a lot of biological material will naturally fluoresce. The problem now is to find the (secondary or tertiary) antibody coupled with the probe you have selected (ex: Cy5, Cy 5.5 or others ...coupled Ab) and the other problem is that you can't see the labeling with your eyes because we can't see the infra-red or close wavelenght. So you need a digital camera linked to a computer to see anything. Obviously you also need the right filter on your microscope... Hope this helps. Emmanuelle
I have inherited a EMITECH freeze drier K775X which has never been setup. The instructions are not very good for someone who has never used any kind of freeze drier. It would be helpful to have step by step instructions for this instrument. Actually it looks pretty simple, but that is relative, isn't it?
Can someone help me? You may respond back to me directly.
Thanks.
Winnie
Edwina W. Westbrook Electron Microscopy Laboratories Agriculture Research Station P.O.Box 9061 Virginia State University Petersburg, VA 23806 (804)-524-5659
I created a Power point presentation last November 2002 consisting of several fluorescence micrographs. The file which was rather large ( 90 Mb) was left in my laptop all this time. When I opened it again this week I find that the images are now too dark and I needed to increase brightness by 3- 4 clicks on the brightness icon of Power point.I increased brightness of all the micrographs and copied the file to a CD hoping that the image will not deteriorate there and then compare it with the one in my laptop several weeks from now. Has this happened to anyone else? Is there something I should have done to prevent this?
Any suggestions or comments will be greatly appreciated.
Let me assure you that the images on your laptop have not degraded. If they had, you would have probably encountered an error reading the file, or you might have found a bunch of junk pixels in your images.
The culprit here is probably the brightness setting on your laptop display. Others who do this more than me will quickly tell you that it can be a real bugaboo trying to get monitors set right so that the image displays correctly on them and also prints out correctly on the various printers. I understand that some folks command a pretty good fee for calibrating monitors and printers for print houses.
I would suggest getting a test image to use for setting your monitor. I have a grid of gray swatches ranging from black to white. I set up the controls so black is black, white is white, and I have a pretty good range of mid-tones.
There is also a control panel from Adobe to help generate an ICM file to account for monitor characteristics. I have used it some, but am not real confident about its use.
I have made a test file (testgrad.gif) and the Adobe control panel available on our server. You may reach them by pointing your browser at ftp://www.marl.iastate.edu/ and then navigating to the "Test images" folder. You will need to drop the control applet into your Windows\System folder and then open up the Control Panel. I use it under Windows 98. It should work under other versions, but I cannot vouch for it. You will be on your own.
Hopefully, these will help you consistently setup you laptop so that the appearance stops changing from month to month.
Warren
At 11:11 AM 1/16/03 -0600, you wrote:
} I created a Power point presentation last November 2002 consisting of } several fluorescence micrographs. The file which was rather large ( 90 } Mb) was left in my laptop all this time. When I opened it again this week } I find that the images are now too dark and I needed to increase } brightness by 3- 4 clicks on the brightness icon of Power point.I } increased brightness of all the micrographs and copied the file to a CD } hoping that the image will not deteriorate there and then compare it with } the one in my laptop several weeks from now. Has this happened to anyone } else? Is there something I should have done to prevent this? } } Any suggestions or comments will be greatly appreciated. } } Cora Bucana
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Hello Everyone, I came across a new problem with our Electroscan E3 ESEM, that I've not been able to track down. At times, the image will become very noisy and streaky with really bright streaks across the image making it near impossible to focus. The problem goes away with time by venting and repumping the chamber, or fiddling with the detector ESD hook wire - but I'm not sure exactly what is causing it. Anyone ever seen this before? 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
Following the comments I made a few days ago about magnetron cold cathod discharge gauges, several people have asked about 'inverted magnetron gauges'.
As it turns out, in addition to the two most common types of cold cathode discharge gauges that I described in my book, there are at least three others: (1) inverted magnerton gauges; (2) Redhead magnetron gauges; and (3) alphatron CCD gauges. For those interested, these are described in reasonable detail in Sect. 6.7.3 of the book 'Vacuum Technology' (2nd. Ed.) by Alexander Roth (ISBN 0-444-86027-4 (Elsevier))
Briefly, the inverted magnetron has a central anode rod that is surrounded by a cylindrical cathode, as in the ordinary magnetron gauge, but in addition there is an auxiliary cylindrical cathode that surrounds theis 'inner cathode', which modifieds the field distribution at the ends of the anode. These gauges are said to be useful over the range from 10-4 to 10-13 Torr.
The Redhead magnetron gauge is a modification of the inverted magnetron which has perforations in the cathode cylinder to improve conductance. It reportedly is useful down to 10-10 Torr.
The alphatron gauge contains a radioactive source of alpha particles that produce ionazition of gas molecules at high pressures, allowing measurements in the range from 10-3 up to 40 Torr.
May all your vacuum leaks be small ones,
Wil Bigelow -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
The only explanation for what you are observing is that the screen of your laptop has deteriorated or changed. It is statistically impossible that an image "darkens" while stored on the computer. An image on the computer consists basically of millions of numbers that describe the intensity at each point. It is of course possible, that a number gets corrupted and you get a different brightness at a pixel, but that all pixels deteriorate in the same fashion is statistically impossible. Another possibility could be, that your settings for the brightness in Powerpoint changed.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Corazon D. Bucana [mailto:bucana-at-audumla.mdacc.tmc.edu] Sent: Thursday, January 16, 2003 10:11 AM To: Microscopy-at-sparc5.microscopy.com
I created a Power point presentation last November 2002 consisting of several fluorescence micrographs. The file which was rather large ( 90 Mb) was left in my laptop all this time. When I opened it again this week I find that the images are now too dark and I needed to increase brightness by 3- 4 clicks on the brightness icon of Power point.I increased brightness of all the micrographs and copied the file to a CD hoping that the image will not deteriorate there and then compare it with the one in my laptop several weeks from now. Has this happened to anyone else? Is there something I should have done to prevent this?
Any suggestions or comments will be greatly appreciated.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gtg187h-at-mail.gatech.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, January 16, 2003 at 10:28:03 ---------------------------------------------------------------------------
Email: gtg187h-at-mail.gatech.edu Name: Leslie Smith
Organization: Georgia Instituite of Technology
Education: Undergraduate College
Location: Atlanta, GA, USA
Question: My BioMedical Engineeering class is working ona problem dealing with Electonic Muscle Stimulation. we were wonderng what types of microscopy would be best for viewing muscles. Thanks so much Leslie Smith
Did your gamma change? i.e., brightness and/or contrast? That will make a huge difference. It is totally unlikely that the native files are changed or perturbed. your viewing situation is more likely the problem. Try using the Adobe gamma adjust app to set your display's gamma. Also note that LCDs are not all that great for pictures. Active matrix LCDs are better. CRTs are best...IMO.
gary g.
At 09:11 AM 1/16/2003, you wrote:
} I created a Power point presentation last November 2002 consisting of } several fluorescence micrographs. The file which was rather large ( 90 } Mb) was left in my laptop all this time. When I opened it again this week } I find that the images are now too dark and I needed to increase } brightness by 3- 4 clicks on the brightness icon of Power point.I } increased brightness of all the micrographs and copied the file to a CD } hoping that the image will not deteriorate there and then compare it with } the one in my laptop several weeks from now. Has this happened to anyone } else? Is there something I should have done to prevent this? } } Any suggestions or comments will be greatly appreciated. } } Cora Bucana }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 created a Power point presentation last November 2002 consisting of } several fluorescence micrographs. The file which was rather large ( 90 Mb) } was left in my laptop all this time. When I opened it again this week I } find that the images are now too dark and I needed to increase brightness } by 3- 4 clicks on the brightness icon of Power point.I increased brightness } of all the micrographs and copied the file to a CD hoping that the image } will not deteriorate there and then compare it with the one in my laptop } several weeks from now. Has this happened to anyone else? Is there } something I should have done to prevent this? } } Any suggestions or comments will be greatly appreciated. } } Cora Bucana } }
Compliments for the season to all. And for all TEM fans " May all darkness be filled with stable clear reproducible publishable results!" We are if full swing here in Gaborone hiding from the scorching 42 degree Celsius heat in the cold EM lab battling with all the weird and wonderful samples frown at us a "quick and easy to do"
Please give help in getting a cross section of Poly polysulfone membrane microdialysis filter to reveal the structure for SEM investigation. We would not like to embed the filter. All sectioning with blades, scalpels and under LN did not result in the expected crisp clean filter surface. Fracture mess and badly smeared surfaces is all we get!
Thanks in advance.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
No experience with Electros can models. , but on an XL-30 it sounds like electrical breakdown in the gas field. The "noise" is a good indication you are very close to full breakdown and arcing is imminent. Bright pulses indicate arcing. Is detector voltage too high or sample too close to the detector. From your message it sounds like gas regulation might be flaky if imaging conditions and sample are similar to what you used in the past. . Sample charging +or- may also play a role. Again no experience with an E3 though and I almost never use the hook and cone detector system you describe. Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Gordon Vrololjak {gvrdolja-at-nature.Berkeley.EDU} 01/16/03 02:42PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Everyone, I came across a new problem with our Electroscan E3 ESEM, that I've not been able to track down. At times, the image will become very noisy and streaky with really bright streaks across the image making it near impossible to focus. The problem goes away with time by venting and repumping the chamber, or fiddling with the detector ESD hook wire - but I'm not sure exactly what is causing it. Anyone ever seen this before? 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
Gordon, Sounds like a dirty hi-vac gauge - Cold Cathode, Penning, etc.. Follow the tips on the List Server lately about cleaning Discharge gauges.
Gary M. Easton, Pres. Scanners Corporation SEM Service/EDS/Digital Imaging Products 410-857-7633
----- Original Message ----- } From: "Gordon Vrololjak" {gvrdolja-at-nature.Berkeley.EDU} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 16, 2003 2:42 PM
This same phenomenon happens with my reports in Microsoft Word. In addition to images darkening, they sometimes shift to other pages and/or change size and dimension. Adding tables and text boxes to the report adds to the fun.
The software that we use is networked. Our IS department has told us that the networked software has a bug that causes these things to happen when file sizes increase, and that there is not a patch for it. So we have just have to deal with it.
Jeff Oakley
-----Original Message----- } From: Corazon D. Bucana [mailto:bucana-at-audumla.mdacc.tmc.edu] Sent: Thursday, January 16, 2003 11:11 AM To: Microscopy-at-sparc5.microscopy.com
I created a Power point presentation last November 2002 consisting of several fluorescence micrographs. The file which was rather large ( 90 Mb) was left in my laptop all this time. When I opened it again this week I find that the images are now too dark and I needed to increase brightness by 3- 4 clicks on the brightness icon of Power point.I increased brightness of all the micrographs and copied the file to a CD hoping that the image will not deteriorate there and then compare it with the one in my laptop several weeks from now. Has this happened to anyone else? Is there something I should have done to prevent this?
Any suggestions or comments will be greatly appreciated.
In principle, the beam current measured on the CUP should be the same as the current measured using a faraday cup on the sample holder (the "ABS" or absorbed current). Also, as one increases the beam current, the ratio between the two (ABS/CUP), which should be close to one, should not change. At least that's what we thought.
When we did a very simple and quick test, we found that at low currents ( {1 na) the ABS current was slightly (a few percent) lower than the CUP current. Then, as we increased current and compared the two, the ratio ABS/CUP increased to a value of one at a few na, then went above one (to about 1.02) with increasing current. In other words, the ratio wasn't always one, and wasn't constant - it varied systematically, increasing several percent over a change in current from {1 to a few hundred na.
This only becomes a problem if the analyst calibrates at one current and analyzes at another (which we sometimes do for reasons I won't go into) and if it is the CUP current that is causing this variation (which we are convinced it is). So we're trying to sort it out.
I'm looking for others on microprobes to run the same test: simply put a good faraday cup on your sample holder (you can make one with an old objective aperture mounted on top of a larger hole drilled part way into brass), then at different currents, measure the ratio of ABS/CUP. Then plot them versus CUP.
Please send results as an Excel file (if possible) directly to me (so as not to bore the rest of the list population), and I'll compile results to share with everyone later.
If you run this test, please let me know what sort of machine you are using (we have a 15-yr old JEOL 733).
In advance, thanks,
Dave -- _________________________________________________ David A. Wark Dept of Earth & Environmental Sciences 1C16 Science Center; Rensselaer Polytechnic Institute Troy, New York 12180 office: 518-276-2674 fax: 518-276-6680 http://www.rpi.edu/~warkd/wark.html
Have you tried Focussed Ion Beam (FIB)? I didn't take the time to look up the Tg of polysulfone but would expect it to be rather high. FIB has been shown useful in such samples for preparing sections or flat faces for TEM and SEM.
Good luck,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Coetzee, Mr S. H Physics Science" To: "Listserver Microscopy (E-mail)" {COETZEES-at-mopipi.u {Microscopy-at-sparc5.microscopy.com} b.bw} cc: Subject: Cross section of Poly polysulfone membrane microdialysis filter - SEM 01/17/03 03:57 AM
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Dear All
Compliments for the season to all. And for all TEM fans " May all darkness be filled with stable clear reproducible publishable results!" We are if full swing here in Gaborone hiding from the scorching 42 degree Celsius heat in the cold EM lab battling with all the weird and wonderful samples frown at us a "quick and easy to do"
Please give help in getting a cross section of Poly polysulfone membrane microdialysis filter to reveal the structure for SEM investigation. We would not like to embed the filter. All sectioning with blades, scalpels and under LN did not result in the expected crisp clean filter surface. Fracture mess and badly smeared surfaces is all we get!
Thanks in advance.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
I'm sort of conducting a survey - how do you freeze blocks prior to cryostat sectioning? A PI in our department is writing a grant and offered to include a freezing set-up (we just use a dewar of LN2 now) - the only equipment I've come across on the web is the giant NesLab bath. Is a styrofoam cooler of LN2 with a beaker of some solvent still a reasonable way to go, or is there some spiffy gizmo we can buy?
Thanks! (Sorry about the cross-post for those of you on both lists)
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
A Microwave Specimen Preparation Workshop sponsored by the Life Science Microscopy Facility in conjunction with Ted Pella, Inc. will be held at Purdue University (West Lafayette, IN) on March 20-22, 2003. The workshop will be conducted by Mr. Rick Giberson and will be structured as follows:
Thursday, March 20: Introduction to microwave preparation theory. Preparation of samples Friday, March 21: Use of the microwave for Immunocytochemical localization for LM & EM - theory.... practical for TEM Saturday, March 22: Evaluating samples, discussion of decalcification, and other potential uses for scientific microwave ovens.
The course will be limited to 10 hands-on participants. Course emphasis may change depending on participant's requests.
The cost for the workshop will be $350. This fee includes registration and meals (3 breakfasts, 3 lunches, 2 dinners). Participants will be responsible for their transportation and lodging.
Please contact us if you would like additional information or a registration form.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
Hi Leslie, I you were going to use the taenia coli from a mouse, then you should consider confocal laser scanning microscopy with an inverted LM. Perhaps the same would be true if you stripped the muscularis from the mouse esophagus proximal to the oral cavity. In the former you would get only smooth muscle and in the latter a transition from skeletal to smooth(I think that's true for the mouse, though I have no sections by which to check.) The simplest, multi-cellular 'smooth muscle-like' multicellular contractile system in the mammal that I have found is the so-called myoid (or peritubular) layer on the surface of the seminiferous tubule. It is aneural, thus its contractions are myogenic with potential contributions from neighbors since they are coordinated and produce a traveling wave that can be easily seen with a LM. Finally, it responds strongly to oxytocin and related octapeptides (how many amino acids?) which often leads, in a Petrie dish, to contractual extrusion of tubular contents leaving the empty peritubular cylinder. We used Hank's MEM as a medium as well as PBS, and we did simple things like varying the temp to demonstrate the effect of T on contraction rates, etc. With an isolated seminiferous tubule from a rat (they're easier to extract than those in a mouse testis), you could run some patch-clamp type electrophysiology and couple that with caged-fluorescence. Simple but sophisticated, non-neural, monolayer myo-cellular system from which you can visualize intracellular fluorescence while you are treating the cylinder as a wire!
If you use {seminiferous tubule contractility} as a search string on PubMed you should find plenty of information to demonstrate how far behind I am in the literature of the subject.
Hope this helps and good luck,
Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: gtg187h-at-mail.gatech.edu [mailto:gtg187h-at-mail.gatech.edu] Sent: Thursday, January 16, 2003 8:00 PM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gtg187h-at-mail.gatech.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, January 16, 2003 at 10:28:03 ---------------------------------------------------------------------------
Email: gtg187h-at-mail.gatech.edu Name: Leslie Smith
Organization: Georgia Instituite of Technology
Education: Undergraduate College
Location: Atlanta, GA, USA
Question: My BioMedical Engineeering class is working ona problem dealing with Electonic Muscle Stimulation. we were wonderng what types of microscopy would be best for viewing muscles. Thanks so much Leslie Smith
I've been digging through the archives looking at printer comments, and find no mention of the new (?) Epson 2200 inkjet. Has anyone used or tested the print quality of this printer? Especially as regards print resolution?
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Where can I get the Photoshop plug-in for our Sony DKC 5000 "Catseye" digital camera? It got lost while the computer was upgraded. Thanks.
Chris A. Edwards, Mgr. Microscopy and Image-analysis Laboratory Dept. Cell & Developmental Biology The University of Michigan, School of Medicine 4747 Medical Sciences II Bldg. Ann Arbor, Michigan 48109-0616 Office: 734-936-4912 Lab: 734-763-1170 FAX: 734-763-1166
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (keithi-at-svusd.k12.ca.us) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, January 18, 2003 at 01:25:37 ---------------------------------------------------------------------------
Email: keithi-at-svusd.k12.ca.us Name: Ian Keith
Organization: La Paz Intermediate School
Education: 6-8th Grade Middle School
Location: Mission Viejo CA US
Question: I am trying to find black and white drawings of some of the most common protozoans found in pond water. I have identified several single and multicellular organisms but I don't know what they are. I would like to make a guide that shows these protozoans so my students can properly identify them. Do you know where I can find such an item??
} In principle, the beam current measured on the CUP should be the same } as the current measured using a faraday cup on the sample holder (the } "ABS" or absorbed current). Also, as one increases the beam current, } the ratio between the two (ABS/CUP), which should be close to one, } should not change. At least that's what we thought. } } When we did a very simple and quick test, we found that at low } currents ( {1 na) the ABS current was slightly (a few percent) lower } than the CUP current. Then, as we increased current and compared the } two, the ratio ABS/CUP increased to a value of one at a few na, then } went above one (to about 1.02) with increasing current. In other } words, the ratio wasn't always one, and wasn't constant - it varied } systematically, increasing several percent over a change in current } from {1 to a few hundred na.
Are both of these methods using the identical circuitry and picoAmp meter. I daresay not, and small differences could be due to some impedance or resistance differences(?) In any case, conclusions from such a study would be dependent these factors and the variety between manufacturers.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
} I've been digging through the archives looking at printer comments, } and find no mention of the new (?) Epson 2200 inkjet. Has anyone used } or tested the print quality of this printer? Especially as regards } print resolution?
Relative to my experience with an Epson 1280, I believe you'll find the printer's resolution everything you require. However, there will probably be gray neutrality issues because it will print grayscale with the colored inks (if you choose "black ink only", the resolution will be worse by an approximate factor of 4). The only other issue will probably be speed (altho this printer is probably faster than my 1280).
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
I'm lookiing to get a couple of complete EDX / EDS systems suitable for an old Cambridge Instruments S-250 Mk III, and an Electroscan 2020 ESEM. Does anyone have such , or know of someone who does, and would be willing to donate to me??? Would of course pay for package and shipping, or if in the UK can collect. Anything considered,
Cheers,
Jim
Jim Buckman Pet Eng Heriot-Watt University Edinburgh Scotland
Can you also share your comments on the highest-resolution dye-sublimation printers? In particular competition to Codonics (so far unmatched). Vendors welcome !
At 04:56 PM 1/17/03 -0600 Friday, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
And here in Ottawa, Canada, we have a non-scorching -30C at the moment, so maybe if we could average things out, we'd both be better off!
A key question, is the filter large enough to cut off a piece a few mm in cross-section and a cm or so in length? If so, then microtomy (diamond knife sectioning, found perhaps in Biology or the medical school or a major hospital) could do you just fine. If large enough, but smearing when sectioned at room temperature (hopefully less than 42C!), you would require what is called cryoultramicrotomy, ie a microtome outfitted with a liquid nitrogen attachment to cool the specimen and render it more brittle, hence easier to section smoothly. As to Gary Gaugler's suggestion of using FIB, I agree that is probably the best solution, but I suspect that, at several hundred $k US, there are not too many FIBs in Botswana.
Tom
Dr. Tom Malis Scientist Advisor / Conseiller Scientifique CANMET-Mineral Technology Branch / Direction de la technologie minerale Natural Resources Canada / Ressources naturelles Canada 555 Booth St. / 555 rue Booth, Ottawa, Ontario K1A 0G1 Tel.: (613) 995-7358 FAX: (613) 947-6606 malis-at-nrcan.gc.ca
-----Original Message----- } From: Coetzee, Mr S. H Physics Science To: Listserver Microscopy (E-mail) Sent: 1/17/2003 4:57 AM
Dear All
Compliments for the season to all. And for all TEM fans " May all darkness be filled with stable clear reproducible publishable results!" We are if full swing here in Gaborone hiding from the scorching 42 degree Celsius heat in the cold EM lab battling with all the weird and wonderful samples frown at us a "quick and easy to do"
Please give help in getting a cross section of Poly polysulfone membrane microdialysis filter to reveal the structure for SEM investigation. We would not like to embed the filter. All sectioning with blades, scalpels and under LN did not result in the expected crisp clean filter surface. Fracture mess and badly smeared surfaces is all we get!
Thanks in advance.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
I don't know, Wil. Sounds like a curse to me (you know, "May you get what you wish for."). I think I'd prefer one big leak to 50 small leaks. Same net effect vacuum-wise, but usually easier to find and fix ;-)
Ken Converse owner Quality Images Third party SEM service Delta, PA
I have always made my Reynold's lead citrate using what I believe is the standard method (see below) and never tested the pH. Recently I checked it and it was just over 13. Some sources say it should be 12 +/- 0.1. What's the consensus - do people check? How do you adjust it if you find it is high - restart and use less NaOH? Thanks, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
True about the cost of a FIB. But there are probably places that would be suitable for outsourcing at a few hundred dollars. They are all over the place here in California.
gary g.
At 11:05 AM 1/18/2003, you wrote:
} And here in Ottawa, Canada, we have a non-scorching -30C at the moment, so } maybe if we could average things out, we'd both be better off! } } A key question, is the filter large enough to cut off a piece a few mm in } cross-section and a cm or so in length? If so, then microtomy (diamond } knife sectioning, found perhaps in Biology or the medical school or a major } hospital) could do you just fine. If large enough, but smearing when } sectioned at room temperature (hopefully less than 42C!), you would require } what is called cryoultramicrotomy, ie a microtome outfitted with a liquid } nitrogen attachment to cool the specimen and render it more brittle, hence } easier to section smoothly. As to Gary Gaugler's suggestion of using FIB, I } agree that is probably the best solution, but I suspect that, at several } hundred $k US, there are not too many FIBs in Botswana. } } Tom } } Dr. Tom Malis } Scientist Advisor / Conseiller Scientifique } CANMET-Mineral Technology Branch / Direction de la technologie minerale } Natural Resources Canada / Ressources naturelles Canada } 555 Booth St. / 555 rue Booth, Ottawa, Ontario K1A 0G1 } Tel.: (613) 995-7358 FAX: (613) 947-6606 } malis-at-nrcan.gc.ca } } } } -----Original Message----- } } From: Coetzee, Mr S. H Physics Science } To: Listserver Microscopy (E-mail) } Sent: 1/17/2003 4:57 AM } Subject: Cross section of Poly polysulfone membrane microdialysis filter - } SEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
keithi-at-svusd.k12.ca.us (by way of Ask-A-Microscopist) wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good luck, Nelson Conti
164 Ferne Court Palo Alto, CA 94306 Email: [ncontiSFSU-at-netscape.net]
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I am going to make cross section sample of the interface between carbon nanotube film (a few hundred micrometer thick) and Si substrate for TEM. I tried the standard method to make cross section (gluing the samples with M-bond, mechanical polishing and ion milling), but the film turned to be peeled off from substrate during polishing or ion milling, due to the large thickness of film and weak bonding between the film and substrate. Any ideas and suggestions from your experience?
Best regards,
Yiming ------------------------------------ Dr. Yiming Yao
Microscopy and Microanalysis Department of Experimental Physics Chalmers University of Technology SE-41296, Göteborg Sweden
} Email: keithi-at-svusd.k12.ca.us } Name: Ian Keith } } Organization: La Paz Intermediate School } } Education: 6-8th Grade Middle School } } Location: Mission Viejo CA US } } Question: I am trying to find black and white drawings of some of the } most common protozoans found in pond water. I have identified several } single and multicellular organisms but I don't know what they are. I } would like to make a guide that shows these protozoans so my students } can properly identify them. Do you know where I can find such an } item?? } } Thanks } } Ian -
"How to know the protozoa" is a classic; you should be able to locate a cheap copy on the web. It won't, of course, help with "multicellular organisms". But I'd really like you to get two books that are listed in the Project MICRO bibliography (URL below): ---------------------------- Loewer, P. 1996 Pond Water Zoo 90pp, 7x9", hardback, $16.00. ISBN 0-689-31736-0 Athenium Books for Young Readers, Simon & Schuster. Out of print; try a used book dealer. This is a well-written natural history of commonly encountered pond life, from monera to micro-arthropods. The illustrations are excellent black and white drawings, and it's written for young readers without being simplistic. Middle school.
Rainis, K.G. and Russell, B.J. 1996 Guide to Microlife 287pp, 5.5x8.5", paperback, $40.00. ISBN 0-531-11266-7 Franklin Watts, Danbury, CT. The price of this book is both unfortunate and understandable. Unfortunate, because it should be in the library of every class that studies the microlife of our environment; understandable, because almost every page has one or more excellent color light micrographs. It's a comprehensive field guide to the microworld. The authors say that the 115 microorganisms described comprise 75-90% of those that may be encountered in the "wild". The habitats described are diverse: the home, soils, plants and debris, and four aquatic environments, with detailed advice on collecting methods for each. Described organisms are equally diverse, ranging from monerans to millimeter-sized arthropods. Species descriptions include ecological information, advice on collection and culture, and frequent suggestions for further investigation. Middle school - adult. RECOMMENDED ---------------------- If you'd like to share your completed guide with MICRO (a pdf would be nice), contact me any time.
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
personally i prefer the venable's modification. as for pH, the alkalinity is what makes it go into solution and keeps CO3 precipitates down. but i never check it.
I seem to recall that there is a company which refurbishes ARL EPMAs and sells them.
Am I remembering right?
Who is it?
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
The 2200, and its overseas equivalent, the 2100, is a newish (2002 some time) Epson printer with 5 coloured inks and 2 shades of black. I imagine it will be at least equivalent to the 1290/1280 in resolution, etc. Have just been looking into getting a new printer and think this is the one we'll go for. cheers, Rosemary
} } } } Listers, } } } } I've been digging through the archives looking at printer comments, } } and find no mention of the new (?) Epson 2200 inkjet. Has anyone } } used or tested the print quality of this printer? Especially as } } regards print resolution? } } } } Phil } } -- } } Philip Oshel } } Supervisor, BBPIC microscopy facility } } Department of Animal Sciences } } University of Wisconsin } } 1675 Observatory Drive } } Madison, WI 53706 - 1284 } } voice: (608) 263-4162 } } fax: (608) 262-5157 (dept. fax)
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
Can You help with some prooven instruments/vendors offering TEM preparation technique as Jet electropolishing (similar like Fischione). Also appreciate Yr opinion about use of it and reliability of different models.
regards
Krzysztof Herman ================================ LABSOFT / FEI Service ul.Ba¿ancia 45A, 02-892 Warszawa tel/fax: (0 - 22) 6449753, 6449750 kom: (0- 601) 307456 E-mail: kherman-at-labsoft.com.pl ================================
Here at Millipore, we take cross sectional images of Polysulfone and Polyethersulfone just about every day. The key to getting a good freeze fracture in LN2 is to make sure the membrane is wet first. Here is the procedure we use:
Setup: Piece of membrane 1 or 2 pair(s) of forceps 1 weighing tin with Isopropyl Alcohol (just enough to cover the bottom of the tin) 1 weighing tin with DI or RO water (just enough to cover the bottom of the tin) 1 Styrofoam coffee cup trimmed down to make it about 2-3 inches high (5-8cm) LN2 Paper towel single edge razor blade
1. Cut a 5mm by 2cm portion of membrane from the sample. 2. Submerge membrane in Isopropyl Alcohol for approximately 1 minute to make sure the membrane is fully wet with IPA. 3. Submerge the sample in water. During this step, the sample will want to "jump around" in the water, so it may be necessary to hold it under the water with the forceps. I also find that dipping it repeatedly back in the IPA then back in the water helps with the exchange. Remember, there is TONS of surface area we're dealing with! 4. Once the piece of membrane appears to wet out with the water, pat the surfaces dry with a paper towel. Make sure to remove water from the forceps as well. 5. Grasp the sample around one third of the way up with the forceps and submerge into LN2 until the membrane and forceps are chilled. (the major bubbling around the forceps will stop. This takes about 10-20 seconds) 6. On this step, timing is key. With the forceps in your right hand, bring the index finger of your left hand close to the top of the cup. As soon as you remove the membrane from the LN2, apply pressure to the top of the strip of membrane with your left index finger until the sample fractures. This will yield the cleanest fracture. 7. You can now trim the fractured surface from the rest of the strip to about 1mm and mount it on the stub. Two-sided carbon tape works best for this.
Alternate fracture method: If you find the "finger" method to not work well for you, you can use 2 pairs of forceps to hold the strip in the LN2. When it is all frozen and still in the LN2, pull one pair of forceps towards you, and one away from you with a bit of a snap, and the membrane should break. The caveat with this method is that there is less control of where the break will occur, and sometimes it will break at the edge of the forceps, resulting in some smearing of the exposed surface.
I hope this is helpful to you, and provides an inexpensive way for you to achieve your desired results. If you would like to contact me further regarding this procedure, please feel free to do so!
Sincerely,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
"Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} 01/17/2003 04:57 AM
To: "Listserver Microscopy (E-mail)" {Microscopy-at-sparc5.microscopy.com} cc: Subject: Cross section of Poly polysulfone membrane microdialysis filter - SEM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear All
Compliments for the season to all. And for all TEM fans " May all darkness be filled with stable clear reproducible publishable results!" We are if full swing here in Gaborone hiding from the scorching 42 degree Celsius heat in the cold EM lab battling with all the weird and wonderful samples frown at us a "quick and easy to do"
Please give help in getting a cross section of Poly polysulfone membrane microdialysis filter to reveal the structure for SEM investigation. We would not like to embed the filter. All sectioning with blades, scalpels and under LN did not result in the expected crisp clean filter surface. Fracture mess and badly smeared surfaces is all we get!
Thanks in advance.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
I would like to mount a standard for SEM calibration, and was wondering what the best method was to do so? It appears my predesessor simply 'glued' the standard to a metal plate that was inserted into the sample holder. But it seems to me that I should use some sort of conductive adhesive? Thanks,
Jeff Roth
-- ================================================== Jeffrey Roth Science Technician James Center 205 phone: 717.245.1109 Department of Geology cell: 717.579.0644 Dickinson College fax: 717.245.1971 Carlisle, PA 17013 email: rothj-at-dickinson.edu ==================================================
I, too, have had significant problems working with images imported into Microsoft Word. However, my software is located on the PC hard drive. The biggest problem that I have encountered with images in Microsoft Word occurs when annotating images. After the image is imported into Word, annotation may be done in two ways (to my knowledge): (1) Text, arrows, etc. may be simply superimposed over the images. The problem with this approach is that the annotations are not linked to the image and may not remain superimposed on the image if the image moves. (2) Annotations can also be linked (probably not the best choice of words) to the image by double-clicking on the image to open the image field, adding the annotation, then closing the image field. These annotations are permanent unless intentionally moved or deleted.
The problems occur when one implements the second option. Comparing images before and after annotation, I found that the annotated images often sustained substantial changes in gray or color levels. Case in point, EDS maps were so badly affected that the color key was no longer correct.
My solutions follow: (1) Annotate images in Adobe Photoshop before importing into Word. Note that the effects of lossy compression on annotations (blurred edges) may be pronounced. (2) Use Microsoft PowerPoint for image presentation. I have encountered no problems with image files in PowerPoint.
Good luck to you in your endeavors.
Cheers,
"The opinions expressed are those of Gary M. Brown and do not represent the opinions of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
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This same phenomenon happens with my reports in Microsoft Word. In addition to images darkening, they sometimes shift to other pages and/or change size and dimension. Adding tables and text boxes to the report adds to the fun.
The software that we use is networked. Our IS department has told us that the networked software has a bug that causes these things to happen when file sizes increase, and that there is not a patch for it. So we have just have to deal with it.
Jeff Oakley
-----Original Message----- } From: Corazon D. Bucana [mailto:bucana-at-audumla.mdacc.tmc.edu] Sent: Thursday, January 16, 2003 11:11 AM To: Microscopy-at-sparc5.microscopy.com
I created a Power point presentation last November 2002 consisting of several fluorescence micrographs. The file which was rather large ( 90 Mb)
was left in my laptop all this time. When I opened it again this week I find that the images are now too dark and I needed to increase brightness by 3- 4 clicks on the brightness icon of Power point.I increased brightness
of all the micrographs and copied the file to a CD hoping that the image will not deteriorate there and then compare it with the one in my laptop several weeks from now. Has this happened to anyone else? Is there something I should have done to prevent this?
Any suggestions or comments will be greatly appreciated.
Hi all! The Limnology people here gave me bottles of riverine water samples + flocs. They would like SEM images of their natural riverine flocs. Do you have suggestions on how to process these floc samples? We're interested in looking at the overall floc morphology and microbial cells as well. We're also interested in looking at their inorganic matter composition.
Thanks.
Lizette Tuason Univ. of Northern British Columbia 3333 University Way Prince George, BC V2N 4Z9 Canada Phone (250) 960-5677
I would like to mount a standard for SEM calibration, and was wondering what the best method was to do so? It appears my predesessor simply 'glued' the standard to a specialized metal plate that was inserted into the sample holder. But it seems to me that I should use some sort of conductive adhesive? Thanks,
Jeff Roth
======================== Jeffrey Roth Science Technician Department of Geology Dickinson College Carlisle, PA 17013 rothj-at-dickinson.edu ========================
If you purchase a Geller MRS standard, they can be pre-mounted according to your stage's needs. These standards are for magnification and resolution cal. Mostly mag. These standards can be purchased with ISO/NIST certification. Other standards from Pella, SPI, et. al. are usually not mounted. But I suspect that you could get them mounted. There are spheres, Gold on Tin and Tin balls. These are good for checking stig and rez. Baseline at purchase and re-check periodically to see if rez or stig has shifted appreciably. Then check apertures and/or column liner.
If you buy standards, I'd recommend storing them under vacuum when not being used.
gary g.
At 05:56 AM 1/20/2003, you wrote:
} I would like to mount a standard for SEM calibration, and was wondering } what the best method was to do so? It appears my predesessor simply } 'glued' the standard to a metal plate that was inserted into the sample } holder. But it seems to me that I should use some sort of conductive } adhesive? Thanks, } } Jeff Roth } } -- } ================================================== } Jeffrey Roth Science Technician } James Center 205 phone: 717.245.1109 } Department of Geology cell: 717.579.0644 } Dickinson College fax: 717.245.1971 } Carlisle, PA 17013 email: rothj-at-dickinson.edu } ================================================== } } }
The Fuji Pictrograph 3500 blows away the Codonics and costs less, although we have had two blown circuit boards in ours.
John Mardinly Intel
-----Original Message----- } From: Marek Malecki, M.D., Ph.D., Professor [mailto:MMalecki-at-wisc.edu] Sent: Saturday, January 18, 2003 11:05 AM To: Microscopy-at-sparc5.microscopy.com
Can you also share your comments on the highest-resolution dye-sublimation printers? In particular competition to Codonics (so far unmatched). Vendors welcome !
At 04:56 PM 1/17/03 -0600 Friday, you wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I haven't been able to find any laser printer sheets of appropriately sized light microscope slide labels.
ie: 7/8" wide X 6/8" high.
As a result, we have always been forced to use rolls, and type out each label tediously on a typewriter.
I was just wondering if anyone knows of any slicker system for labelling microscope slide labels. Even upstairs with the thousands of slide labels that they make, they have to tediously type them all out 1 by 1 on the typewriter.
I have found for micrographs and negative envelopes, that a combination of Microsoft Word mail merged with an appropriate Excel data base does a nice job using Avery 5160 address labels, but unfortunately, I haven't as yet been able to find a properly sized label for microscope slides, or even one that I could cut nicely. :-(
I was just wondering how other people tackled this problem. Do you all do it the tedious way too? Here is a marketing opportunity for some ambitious person who wants to make quick money, by solving a problem in the workplace.
John, Did you mean the "Fuji Pictrography 3500"? http://www.electroimage.com/fp3k.htm -Mike
"Mardinly, John" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Fuji Pictrograph 3500 blows away the Codonics and costs less, } although we have had two blown circuit boards in ours. } } John Mardinly } Intel } } -----Original Message----- } } From: Marek Malecki, M.D., Ph.D., Professor [mailto:MMalecki-at-wisc.edu] } Sent: Saturday, January 18, 2003 11:05 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: the highest-resolution dye-sublimation printers WAS: Re: } printer hunt } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 you also share your comments on the highest-resolution } dye-sublimation printers? } In particular competition to Codonics (so far unmatched). } Vendors welcome ! } } At 04:56 PM 1/17/03 -0600 Friday, 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 } } -----------------------------------------------------------------------
} Nelson Conti wrote: } } } Dear Ian: } } One guide that I am aware of is titled "The Illustrated Guide to the Protozoa" published by the Society of Protozoologists (URL:http://www.uga.protozoa.edu) (pretty sure). I am a member of that Society which is how I know about such a guide. I have no idea how much such a guide would cost, but perhaps if you tried their site, there should be information about the guide I've suggested. } } The correct URL is www.uga.edu/~protozoa . } } } ______________________________________________________________________ } } 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-onid.orst.edu
-- _______________________________________________________________________ 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-onid.orst.edu
Yes. John Mardinly Phone: 408-765-2346 Pager: 877-277-1182
-----Original Message----- } From: Michael O'Keefe [mailto:MAOKeefe-at-lbl.gov] Sent: Monday, January 20, 2003 12:40 PM To: Mardinly, John Cc: Marek Malecki, M.D., Ph.D., Professor; Microscopy-at-sparc5.microscopy.com
Help Listservers,
I need to explain, justify, etc., to colleagues (who have no microscopy/microtomy experience or background) reasons why you should isloate TEM rooms from SEM rooms from microtome rooms? In other words, why you should avoid putting a TEM, an SEM and a cryoultramicrotome all in the same room. Thanks
Fred A. Hayes Analyst Polymer Microscopy Collins and Aikman IntelliMold Systems 4651 Platt Lane Ann Arbor, MI 48108 734-477-7029 direct 734-477-9214 fax 734-477-9212 office www.IntelliMold.net www.collinsaikman.com fred.hayes-at-colaik.com
In my last posting about the pH of Reynold's lead citrate, I didn't give the exact method we use. Here it is:
1.33 gm Pb(NO3)2 1.76 gm Na3 citrate -2H20 30 ml dH2O (all water used for this prep is freshly boiled and cooled to get rid of CO2) shake vigorously let stand 30 min Add 8 ml of freshly made 1 N NaOH dilute to 50 ml total volume.
Reynold's says the pH was "routinely found to be 12.0 +/- 0.1". The only real difference between my method and Reynold's is that he used 1 N NaOH prepared by diluting a commercial 10 N carbonate free stock. I make my NaOH from pellets immediately before use. Assuming I accurately weigh it, any error would presumably result from some water absorption by the pellets so the error would be to decrease the NaOH concentration. But the pH we get implies too much NaOH.
We got a pH of ~13.5 (with good quality pH paper - i don't want to use my pH meter for this). We cut the volume of NaOH to 7 ml and got a pH of ~12.5 or slightly less (pH paper has 0.5 unit steps). Reynold's notes that the staining intensity decreases with higher pH
My question remains - do people measure the pH of their Reynolds? do they get 12? I assume you can't adjust the pH with HCl but I am open to correction. I fully understand the concept of emphirecally testing it and seeing the results. I am not sure if my results are the best and furthermore, I would like to have a method that is consistent and that I understand so I don't need to re-invent this wheel each time. I am aware of the other lead formulations but am interested in people's experience with Reynolds. I may switch but I think switching everytime a problem comes up without attempting to understand the problem is poor science. Thanks for any comments. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I was wondering if anyone could help me with information about the proceedings for the 1993 and 1994 (51st and 52nd, respectively) Annual MSA Meetings. I am looking for the publishers for each of these years. Was it San Francisco Press or Jones & Begell? Also, I am looking for the editors for the 1993 year.
If the standard was coated after it was glued on, the coating may provide sufficient conductive pathway. If it works, it was done OK. I often mount things with Duco cement but use silver paint on edges to make sure I have a good electrical connection after coating. A conductive adhesive is another way to go, for sure.
Ron L
-----Original Message----- } From: Jeff Roth [mailto:rothj-at-dickinson.edu] Sent: Monday, January 20, 2003 1:31 PM To: Microscopy-at-sparc5.microscopy.com
I would like to mount a standard for SEM calibration, and was wondering what the best method was to do so? It appears my predesessor simply 'glued' the standard to a specialized metal plate that was inserted into
the sample holder. But it seems to me that I should use some sort of conductive adhesive? Thanks,
Jeff Roth
======================== Jeffrey Roth Science Technician Department of Geology Dickinson College Carlisle, PA 17013 rothj-at-dickinson.edu ========================
Ian, In addition to numerous sites accessible via google, I've used earlier versions of the following for many years and the current version is available at amazon.com.
Pond Life: A Guide to Common Plants and Animals of North American Ponds and Lakes (Golden Guide) by George Kell Reid, Herbert S. Zim (Editor), George S. Fichter (Editor), Jonathan P. Latimer, Karen Stray Nolting, John L. Brooks, Sally D. Kaicher (Illustrator), Tom Dolan (Illustrator) Paperback: 160 pages ; Dimensions (in inches): 0.34 x 5.94 x 3.93 Publisher: St. Martin's Press; ; Revised and Updated edition (April 2001) ISBN: 1582381305 $6.95 Rosemary
I was wondering if anyone could help me with information about the proceedings for the 1993 and 1994 (51st and 52nd, respectively) Annual MSA Meetings. I am looking for the publishers for each of these years. Also, I am looking for the editors for the 1993 year.
Check out the Struers Tenupol. Expensive, but the Hasselblad of Jet electropolishing. John Mardinly
-----Original Message----- } From: Krzysztof Herman [mailto:kherman-at-labsoft.com.pl] Sent: Monday, January 20, 2003 4:57 AM To: MSA
Hi list
Can You help with some prooven instruments/vendors offering TEM preparation technique as Jet electropolishing (similar like Fischione). Also appreciate Yr opinion about use of it and reliability of different models.
regards
Krzysztof Herman ================================ LABSOFT / FEI Service ul.Ba¿ancia 45A, 02-892 Warszawa tel/fax: (0 - 22) 6449753, 6449750 kom: (0- 601) 307456 E-mail: kherman-at-labsoft.com.pl ================================
A TEM Specimen Preparation Course Will be offered March 12-14, 2003 at the Materials Characterization Facility at the University of Central Florida, Orlando, FL USA
A review of all techniques will be given, with primary emphasis on tripod polishing, ion milling, and all latest FIB techniques
Instructors: Ron Anderson, Microscopy Today Fred Stevie, NC State Lucille Giannuzzi, UCF Brian Kempshall, UCF
For more information contact Lucille Giannuzzi, lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor Mechanical, Materials, and Aerospace Engineering University of Central Florida 4000 Central Florida Blvd. PO Box 162450 Orlando, FL 32816-2450 email lag-at-mail.ucf.edu phone 407-823-5770 fax 407-823-0208 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Rosemary, The two shades of black are for use with different papers. One black for ordinary papers, the other for glossy photo papers. You can get details on the Epson web site. Damian
} The 2200, and its overseas equivalent, the 2100, is a newish (2002 some } time) Epson printer with 5 coloured inks and 2 shades of black. I imagine } it will be at least equivalent to the 1290/1280 in resolution, etc. Have } just been looking into getting a new printer and think this is the one } we'll go for. } cheers, } Rosemary
Hi all We have a client with an old JEOL 1200 TEM that was modified by the factory to have two diffusion pumps as the vacuum system. We now have a problem that the EPROMs that control the vacuum system have gone faulty and are have a real challenge trying to get some replacements. Also the manuals are all for systems with either an Ion pump or a Turbo pump as a second pump for the gun and column. This means we are really having fun trying to work out the logics.
Can anyone help us out with a set of EPROMS, the software for the EPROMS and or manuals specific for the double diffusion pump version of the JEOL 1200?
Thanks
Luc Harmsen ANASPEC South Africa www.anaspec.co.za Tel: +27 11 794 8340 Fax: +27 11 794 8349 P.O. Box 2561 Honeydew 2040 Gauteng, South Africa
This is to inform you that the Call for Abstracts for the forthcoming International Meeting on Applied Physics (APHYS-2003), to be held in Badajoz (Spain), during October 14-18th 2003, is now opened.
Website www.formatex.org/aphys2003/aphys2003.htm
Abstracts submission deadline: March 24th 2003
As you can see, Imaging Techniques and Applied Microscopy will have very important weight in the scientific Program of APHYS-2003. Some of topics to be covered will be:
- Surfaces, Interfaces and Colloids - Imaging Techniques, Microscopy - Nano-sciences and Technologies - Materials Science - Biomedical Engineering and Biomaterials Science&Engineering - Computational Physics
In addition to the regular Scientific Program, several International Workshops will be held as pre-conference events. The following two will be of high interest for the Microscopy community:
1. Workshop on Modern Applied Microscopy in Molecular and Cell Biophysics Research(please visit the website for details)
2. Pre-conference Workshop: International Interdisciplinar Workshop on Bioengineered Non-crystalline Solids
For all the information regarding the conference, please visit www.formatex.org/aphys2003/aphys2003.htm
Potencial reviewers are kindly asked to write us with personal data, field(s) of expertise and a list of publication. The official list of reviewers will be also included in the conference publications.
Proceedings will be published within several international journals, depending on each paper topic:
- Journal of Microscopy - Journal of Non-crystalline Solids - Applied Surface Science - Physica Scripta
For issues regarding Commercial Exhibition and Sponsorship, please refer to the Conference website or contact Ines Solo de Zaldivar (secretariat-at-formatex.org )
Thank you in advance and please contact us for any suggestion or question.
Antonio Mendez-Vilas APHYS-2003 Co-ordinator secretariat-at-formatex.org www.formatex.org/aphys2003/aphys2003.htm
} Can anyone suggest a UK supplier of Polyvinyl Alcohol } at 24-32 centipoise viscosity?
Another possibility is mixing PV-acetate with ethanol. I believe my recipe is 20% by weight, which will lay down, dry and become a heat-sensitive "glue" for relatively large samples. I'd make the mixture 10% for very small particles.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
Yes, pH the stain to around 12 +/- 0.1 You do get erratic staining with a higher pH.
Best of Luck,
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-8806 calomeni-1-at-medctr.osu.edu
} } } Tom Phillips {phillipst-at-missouri.edu} 01/18/03 05:47PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have always made my Reynold's lead citrate using what I believe is the standard method (see below) and never tested the pH. Recently I checked it and it was just over 13. Some sources say it should be 12 +/- 0.1. What's the consensus - do people check? How do you adjust it if you find it is high - restart and use less NaOH? Thanks, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I am a happy user of commercial CO2-free, titrated NaOH. I buy mine from Electron Microscopy Sciences [in which I have no commercial interest, wish I had :)]. It is inexpensive, something like $10 a bottle. This *is* the consistent method, as long as you keep using the same water of course. When I was making Reynolds from pellets, sometimes I would get stain that peppers easily and sometimes the stain would come out too weak. I think NaOH stock is the key, once you have good water.
Paper shows pH of my stain about 12. I never measured with pH meter but I know there are electrodes which would allow you to do that, I once asked an Orion tech. I do remember seeing in publications that it is important that the pH be 12+/-0.1, and if I was running a teaching lab, I would probably get a proper electrode and measure and maybe experiment adjusting if the pH turned out different from 12+/-0.1. Here, however, I am content with the fact that my Reynolds stains reproducibly well.
As for the other lead stain recipes, I often use the Venable-Cogeshall [sp?] instead of Reynolds. It does not make as "sweet" a picture 20k and above as Reynolds but does seem to give more contrast. I use it for no more than 2 days, then make fresh the next time (have pre-weighed LC aliquotes sitting in 15 ml Falcons and just add water and NaOH when the time comes). Again, keeping the same water and NaOH stock makes it highly consistent.
All the best, Vlad
} In my last posting about the pH of Reynold's lead citrate, I didn't } give the exact method we use. Here it is: } } 1.33 gm Pb(NO3)2 } 1.76 gm Na3 citrate -2H20 } 30 ml dH2O (all water used for this prep is freshly boiled and } cooled to get rid of CO2) } shake vigorously let stand 30 min } Add 8 ml of freshly made 1 N NaOH } dilute to 50 ml total volume. } } Reynold's says the pH was "routinely found to be 12.0 +/- 0.1". The } only real difference between my method and Reynold's is that he used } 1 N NaOH prepared by diluting a commercial 10 N carbonate free } stock. I make my NaOH from pellets immediately before use. } Assuming I accurately weigh it, any error would presumably result } from some water absorption by the pellets so the error would be to } decrease the NaOH concentration. But the pH we get implies too much } NaOH. } } We got a pH of ~13.5 (with good quality pH paper - i don't want to } use my pH meter for this). We cut the volume of NaOH to 7 ml and } got a pH of ~12.5 or slightly less (pH paper has 0.5 unit steps). } Reynold's notes that the staining intensity decreases with higher pH } } } } My question remains - do people measure the pH of their Reynolds? } do they get 12? I assume you can't adjust the pH with HCl but I am } open to correction. I fully understand the concept of emphirecally } testing it and seeing the results. I am not sure if my results are } the best and furthermore, I would like to have a method that is } consistent and that I understand so I don't need to re-invent this } wheel each time. I am aware of the other lead formulations but am } interested in people's experience with Reynolds. I may switch but I } think switching everytime a problem comes up without attempting to } understand the problem is poor science. Thanks for any comments. Tom } } } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- -------------------------------------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
Fred, Get you hands on a copy of the book in the Glauert series :Design of the Electron Microscope Laboratory (or something very close to that name, I don't have it in from of me). I have found it of enormous value, although it remains to be seen if I too will succeed in a similar argument. 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
Post-Doctoral Research Associate Stevens Institute of Technology
There is a post-doctoral opportunity within the Department of Chemical, Biochemical, and Materials Engineering at the Stevens Institute of Technology to develop and apply advanced techniques of transmission electron microscopy to quantitatively study the morphology of both dry and hydrated synthetic and natural polymers.
The ideal candidate will have experience in electron optics, electron energy-loss spectroscopy, and cryo-TEM. Acceptable candidates will at least have: (1) experience with transmission electron microscopy, either
from a physical or biological sciences perspective; (2) a willingness to
develop a leadership role in problems associated with the nanoscale morphology of hydrated polymeric biomaterials; and (3) good communication skills.
This is an NIH-funded position associated with a new inter-institutional
NCRR on Polymeric Biomaterials. This NCRR is associated with the New Jersey Center for Biomaterials and involves core laboratories at Rutgers, NJIT, and Stevens. A multi-year appointment for this position is anticipated.
The Stevens Institute of Technology is a small private university concentrated on engineering, science, and technology management. Stevens is located in very close proximity to New York City. The Stevens electron-optics laboratory contains a Philips CM20 FEG TEM/STEM,
a Philips CM30 SuperTwin TEM, and a LEO 982 FEG SEM. The CM20 FEG TEM/STEM is equipped with a Gatan Enfina ccd PEELS system and a Gatan Multiscan digital camera. Both are interfaced to an Emispec Vision acquisition and control system. The facility is fully equipped with cryomicrotomy and cryo-transfer capabilities to deal with frozen hydrated materials.
For further information please contact:
Professor Matthew R. Libera Dept. Chemical, Biochemical, and Materials Engineering Stevens Institute of Technology Hoboken, New Jersey 07030 ph: 201-216-5259 fax: 201-216-8306 mlibera-at-stevens-tech.edu
MS Word is such a pain because of the way it works with images. A couple of tricks I've discovered over the years.
1) Place the image within a text box, rather than directly on the page, it seems to give you much greater control over the location on the page. It also makes it much easier to add text annotations that stay with the image.
If you don't want the text box to have a border you can remove it by selecting the box outline and look for the "paint brush" icon on the Draw toolbar, then use the down arrow and select "none". If you'd like the text box to be transparent, select the text box outline and look for the "paint bucket" icon on the Draw toolbar, then use the down arrow and select "none". If you discover its hard to find the text box border once you made the edge "invisible", first select the image with a single left click (it should have the solid black resizing "handles") and then use the keyboard left or right arrow keys and the selection with move out to the textbox outline (with black bordered resizing "handles").
2) Always use the INSERT | PICTURE | FROM FILE option as opposed to copying and pasting an image into Word. I find that the images are harder to work with if I paste them in. Also, you can insert TIFF or BMP images into Word, you don't have to use JPEG.
FYI: Last known, there was inexpensive software on the web that compressed bloated Powerpoint files. I also have a freeware utility on my PC called "packword" that compresses Word 97 and 2000 files. I'm sorry I don't have the URLs, I'm writing this from home.
Best regards, Doug
} -- Original Message -- } Date: Mon, 20 Jan 2003 09:30:47 -0600 } From: "gary.m.brown-at-exxonmobil.com"-at-sparc5.microscopy.com } Subject: RE: presentation images in Microsoft Word } To: oakleyj-at-rayovac.com } Cc: Microscopy-at-sparc5.microscopy.com } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } "Oakley, Jeff"
} } {oakleyj-at-rayovac.c To: {Microscopy-at-sparc5.microscopy.com} } } om} cc:
} } Subject: RE: presentation } images }
} } 01/17/03 07:50 AM
} }
} }
} } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
.......................................................................... Douglas W. Cromey, M.S., Principal Research Specialist Dept. of Cell Biology & Anatomy, University of Arizona Tucson, AZ 85724-5044 USA 520-626-2824 {cromey-at-arizona.edu} .......................................................................... http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
Usually all microscope columns are sensitive to electromagnetic fields and should be placed on distances at least 10 feet from each other and from power lines. Additionally microscopes are sources of vibrations and acoustic noise which could degrade images of high resolution instruments.
And, sure, it is absolutely inconvenient for operators to have SEM and TEM in the same room: working TEM requires full darkness and this will give SEM operator only unnecessary eye fatigue and problems with specimen replacement. I could agree that it is possible to install couple of older (and less sensitive) SEMs in the same room, but each TEM should have separate light-tight room.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} } Help Listservers, } } I need to explain, justify, etc., to colleagues (who have no } microscopy/microtomy experience or background) reasons why you should } isloate TEM rooms from SEM rooms from microtome rooms? In } other words, why } you should avoid putting a TEM, an SEM and a } cryoultramicrotome all in the } same room. Thanks } } Fred A. Hayes } Analyst } Polymer Microscopy } Collins and Aikman } IntelliMold Systems } 4651 Platt Lane } Ann Arbor, MI 48108 } 734-477-7029 direct } 734-477-9214 fax } 734-477-9212 office } www.IntelliMold.net } www.collinsaikman.com } fred.hayes-at-colaik.com } } }
I posed this question in late December when many were on Christmas vacation so will try again. Does anyone know the original citation for double stick carbon tape used on SEM specimen holders? I think it was first used in the mid 1990's but I have been unable to locate when and who introduced it.
Dear Fred, TEM rooms must be able to be completely darkened and are the ones that must be isolated from noise, vibration, air currents, etc. Microtomes would have to be vibration isolated from the pumps on SEMs and TEMs. ----- Original Message ----- } From: "Hayes, Fred" {Fred.Hayes-at-colaik.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, January 20, 2003 3:48 PM
We have a CoStar label maker that we purchased some time ago for regular mailing labels. It takes stock of various widths and factors the appropriate pitch into its software. We just run off a ribbon of labels one wide by as many long as we need.
Warren
At 02:20 PM 1/20/03 -0600, you wrote:
} I haven't been able to find any laser printer sheets of appropriately sized } light microscope slide labels. } } ie: 7/8" wide X 6/8" high. } } As a result, we have always been forced to use rolls, and type out each } label tediously on a typewriter. } } I was just wondering if anyone knows of any slicker system for labelling } microscope slide labels. Even upstairs with the thousands of slide labels } that they make, they have to tediously type them all out 1 by 1 on the } typewriter. } } I have found for micrographs and negative envelopes, that a combination of } Microsoft Word mail merged with an appropriate Excel data base does a nice } job using Avery 5160 address labels, but unfortunately, I haven't as yet } been able to find a properly sized label for microscope slides, or even one } that I could cut nicely. :-( } } I was just wondering how other people tackled this problem. Do you all do } it the tedious way too? Here is a marketing opportunity for some ambitious } person who wants to make quick money, by solving a problem in the workplace.
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Listers, We are going to try to plunge freeze and freeze substitute mouse pancreas. Now I know that the most desirable way to do this would be with high pressure freezing but that is not an option at the moment. So I am hoping to get a bit of help with what we do have available.
We have done quite a bit of plunge freezing microbes and fungi into Liquid nitrogen cooled liquid propane and then substitution in ethanol, acetone, etc. with good results. Embedding resin depends on tissue...Epoxy (LX112) or HM20 lowacryl as needed.
I would like advice on using a cryo-protectant for the mouse pancreas ..thinking of using 18% glycerol or 18% propylene glycol. Is this necessary and do you recommend one over the other?
Also for substitution, I was going to use acetone + 2% osmium for standard ultrastructure and just acetone for ICC (switched to ETOH for infiltration with HM20.
I would appreciate comments on above general approach and any other information you would like to share that may enhance contrast on final sections.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
} It seems this man's daughter is suffering from some kind of skin ailment } and she has dug stuff out of MANY of the sores on her skin. The } doctors in her HMO have not offered any relief. Her father (from another } department on this campus) is quite determined to find out what the problem } might be.
OK, I'm not a doctor and, compared to most on this list, I know very little about microscopy, but...
when I was young I had a skin ailment that sounds somewhat like what you are describing. You provided very little information on the skin ailment itself, so I may be way off base here. What I had was a rash of boils (at one point I had 42 of them at one time on various parts of my body). I would expect the doctors at your HMO to be able to recognize boils, but perhaps they're not all that common anymore. Anyway, the eruptions had a fibrous core, sometimes more than one. This core was so cohesive that when finally removed it left a cylindrical hole in the sore. Sometimes the core would come out on its own along with gobs of pus, sometimes I would grab it with a pair of tweezers and pull it out. After removal of the core the skin eruption would subside and disappear.
Hope that helps.
Bruce Girrell
BTW - The doctor prescribed moist heat to help the boils drain and would have us wash them regularly with PhisoHex (no longer available) soap. Sometimes the doctor would lance the boils to drain them. None of this did anything to prevent recurrances. My grandfather said "You need a blood cleanser. Drink burdock root tea." So I drank that nasty stuff (no sweetening or anything else) for about two weeks. It may just be a coincidence, but I have never had another boil.
I'd suggest much earlier than the mid 1990s; I first learned SEM in 1993, and my impression then was it was old hat already. Perhaps the mid 1980s? Ben Simkin (simkin-at-egr.msu.edu) dpt. Chemical Engineering and Materials Science Michigan State University
We are experiencing artifacts in thick epon-araldite resin sections and are about at our wit's end. You can view the artifact by linking to http://biotech.missouri.edu/emc/stain_artifact.jpg and noting the dark magenta blobs in the nuclear layer of the retinal tissue in the image. We have determined that the "blobs" are associated with "dimples" in the sections, but they have never shown up in hundreds of other samples done previous to this particular project. We are virtually certain this is artifact because serial sections show the blobs in different places and in different concentrations.
What we have done so far is to try varying the section thicknesses from from 0.4 microns to 3.0 microns, mixing the aqueous toluidine blue 1:1 with varying concentrations of ethanol from 20% to absolute, diluting the stain with water, mixing fresh stain, cutting on both glass and diamond histo knives, trying different temperatures on the hot plates, trying untreated slides and slides treated with BSA, staining in the microwave at varying wattages and times, and dipping slides and sections in methanol before staining, then staining normally.
So far, the only things that have made any difference are using ethanol in the stain and dipping the slides in methanol prior to staining, but neither are consistent. They might work one time, but not the next and we still get the blobs showing up in different places (but ALWAYS in the nuclear layer only) and in different numbers.
HEELLLPPP!!!
Most grateful for ANY assistance with this one!
Randy
Randy Tindall EM Specialist Electron Microscopy Core----We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Hi Debby: We just got our new HPF so that will teach you for leaving us! For freeze-sub without aldehydes or osmium, add 0.1% uranyl to the acetone since it will increase membrane contrast. a good paper on this is Moreira et al. J Histochem Cytochem 46(7):847-854. Leica's web site has a large compilation of freeze sub protocols. I have used cryoprotectants with fixed material (e.g., the Tokyasu method) but I haven't frozen unfixed material with cryo-protectants. I would worry about the osmotic stress and diffusion of the material. I am a little doubtful about the quality of freezing you will achieve with this approach. Tom
At 01:53 PM 1/21/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Major Focus has sheets of Laser printer compatible self adhesive paper labels (15/16" X 15/16") which work well on 3X1 slides, however, they are not chemical resistant. They can be reached at (828) 313-0923. I have no financial interest in this company.
Craig M. Klotz B.S., CT (ASCP) Neuromuscular Pathology cklotz-at-mcw.edu EM/Lab Tech. "Chance favors the prepared mind" - L. Pasteur
This may not help with the slide labeling problem, but if there are people out there that are still typing or writing block labels manually, I can send them a Word document which acts as a template. Using search and replace, labels can be made and printed out in a few seconds. A similar approach formatted to fit on label pages might work with the slides too. I also have a program I wrote in VBA (in Word) that lets you write negative labels with almost no typing (magnifications are entered by clicking on screen buttons). We just print them on regular paper and tape them to the negative sleeves -- not elegant, but cheap and easy. I would be happy to send this out too, but the code would have to be modified for the magnifications of other microscopes.
Since the documents will have to be sent as attachments, anybody interested will have to contact me directly.
Ralph Common Michigan State University Division of Human Pathology
-----Original Message----- } From: Garry Burgess [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Monday, January 20, 2003 3:20 PM To: microscopy-at-sparc5.microscopy.com
I haven't been able to find any laser printer sheets of appropriately sized light microscope slide labels.
ie: 7/8" wide X 6/8" high.
As a result, we have always been forced to use rolls, and type out each label tediously on a typewriter.
I was just wondering if anyone knows of any slicker system for labelling microscope slide labels. Even upstairs with the thousands of slide labels that they make, they have to tediously type them all out 1 by 1 on the typewriter.
I have found for micrographs and negative envelopes, that a combination of Microsoft Word mail merged with an appropriate Excel data base does a nice job using Avery 5160 address labels, but unfortunately, I haven't as yet been able to find a properly sized label for microscope slides, or even one that I could cut nicely. :-(
I was just wondering how other people tackled this problem. Do you all do it the tedious way too? Here is a marketing opportunity for some ambitious person who wants to make quick money, by solving a problem in the workplace.
If you want, I can get the name and address etc. for a computer run printer that prints directly to slides. It is used at Baxter Healthcare in the histology lab. They stack slides in the machine, the operator goes to the computer and types in the necessary information and it automatically begins printing on the slide; they use slides with a white end to print on.
} I haven't been able to find any laser printer sheets of appropriately sized } light microscope slide labels. } } ie: 7/8" wide X 6/8" high. } } As a result, we have always been forced to use rolls, and type out each } label tediously on a typewriter. } } I was just wondering if anyone knows of any slicker system for labelling } microscope slide labels. Even upstairs with the thousands of slide labels } that they make, they have to tediously type them all out 1 by 1 on the } typewriter. } } I have found for micrographs and negative envelopes, that a combination of } Microsoft Word mail merged with an appropriate Excel data base does a nice } job using Avery 5160 address labels, but unfortunately, I haven't as yet } been able to find a properly sized label for microscope slides, or even one } that I could cut nicely. :-( } } I was just wondering how other people tackled this problem. Do you all do } it the tedious way too? Here is a marketing opportunity for some ambitious } person who wants to make quick money, by solving a problem in the workplace. }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Fred, } Get you hands on a copy of the book in the Glauert series :Design of } the Electron Microscope Laboratory (or something very close to that } name, I don't have it in from of me). I have found it of enormous } value, although it remains to be seen if I too will succeed in a } similar argument. } good luck, } Lee
The book is :
Practical Methods in Electron Microscopy, volume 4 : Design of the Electron Microscope Laboratory Audrey M. Glauert ISBN 0 7204 4250 8, 0 7204 4259 1 or 0 444 10807 6 depending on the publisher.
My book is printed in 1975, I took a quick glance in it but I couldn't see any specific discussion about sites with multiple microscopes.
I got the first emission current from my salvaged TEM tonight. :-)
The presence of dimples suggests that tiny bubbles may have formed when the specimen is stained. Bubbles probably originate from degassing of the water when it is heated. The bubbles distort the heated plastic and could concentrate the staining by acting as "micro-dishes" to hold the stain in place even during the rinsing steps. Ethanol probably would break the tension of the water and allow a more efficient removal of the stain during the rinsing steps.
The way to check this would be to use thoroughly degassed reagents (heated/vacuumed) until after the staining step.
Good luck and let us know how you solve the problem.
Do you have any blocks (maybe different tissues) prepped in the same way that do not show this artifact?
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 ##############################################################
} } } Can anyone suggest a UK supplier of Polyvinyl Alcohol } } at 24-32 centipoise viscosity? } } Another possibility is mixing PV-acetate with ethanol. I believe my } recipe is 20% by weight, which will lay down, dry and become a } heat-sensitive "glue" for relatively large samples. I'd make the } mixture 10% for very small particles. }
I don't know the application, so it may not matter, but don't forget that PV-acetate does tend to hydrolyse in the long term and release acetic acid. PV alcohol doesn't do this, so is to be preferred for archival/longterm applications.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
In a message dated 01/20/2003 1:41:50 PM US Mountain Standard Time, GBurgess-at-exchange.hsc.mb.ca writes:
{ { I was just wondering if anyone knows of any slicker system for labelling microscope slide labels. Even upstairs with the thousands of slide labels that they make, they have to tediously type them all out 1 by 1 on the typewriter. } }
Garry,
I can't vouch for them, never having used them, but the following labels are supposed to be temperature-, solvent- and caustic- resistant, as well as waterproof, and they come in sheets that are laser-printable. They also seem to be about the same size that you are looking for (7/8" x 7/8").
If you don't want to invest in a slide labeling device, you could probably use these sheets along with Microsoft Word and do a decent job. Also, Avery used to have a plug-in for MS Word (kind of like a Wizard). It's probably still available for download from Avery's website...haven't checked lately, though.
(that's {http://www.divbio.com/slide.html} , for Diversified BioTech).
There are devices that will print on frosted slides, and some of them come w/ software that will automatically keep track of your accessioning numbers, outside labs and Docs, etc. Contact me off-list if you need details.
Cautionary note: how resistant is the ink after it's "fused" to the label? I realize that laser printers use heat to set the ink on the paper (or label), but you might want to check for chemical resistance after printing some test labels.
The new Leica IPS Slide Labeler and IPC Cassette Labeler use an inkjet system, but it's a special ink that gets passed under a Xenon strobe to polymerize the ink and set it on the frosted slide or the cassette. Other labelers use heated scribes and metallic foils to do the labeling.So you have several options!
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Hello- I have an old JEOL 100CX, Which had been relocated twice but was working fine. Recently there was a water leak and the water fell on the HT cable going down to the HT tank. After the leak repair and clearing, the system was switched on after a week's time, Upon puttng on HT there were lots of discharges with high Beam current. Does any one suspect that water may have seeped in the Tank. or there is some circuiting problem in the cable. Any advices.
Shashi Singh Scientist CCMB-Hydearbad INDIA
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I've never had a similar need, but this is what I would suggest. Avery 3383 Sticker Project Paper and one of those paper guillotines, or a better yet, a rotary paper trimmer with the perforating blade. OpenOffice (which is on this computer I'm using) has a built-in label designer where you can specify pitch, rows, columns, etc., then database fields can be used to fill in the labels. I would guess MS Office or other office suites also have this ability.
Regards, Dave Harrison
On 20 Jan 2003 at 14:20, Garry Burgess wrote:
} } } I haven't been able to find any laser printer sheets of appropriately sized } light microscope slide labels. } } ie: 7/8" wide X 6/8" high. } } As a result, we have always been forced to use rolls, and type out each } label tediously on a typewriter. }
Is it possible that you have a dirty batch of slides, so your sections are not adhering as usual, and/or the stain is picking up the dirt? Perhaps if the stain binds to the nuclear layer and there is dirt/grease underneath, then it won't rinse out properly - hence the blobs. My only suggestion is to clean the slides to death and/or try with some of those expensive pre-cleaned slides (people here use them for in situs).
Ethanol in the stain might loosen up this dirt, and methanol dipping also, but perhaps a ferocious clean of the slides might do the trick (if this is the problem, of course). From experience, a cheap brand of slides we used to get suddenly became very dirty and we had to change brands. Not sure what the problem was but we never went back to the original brand to see they recovered their quality.
my 2c worth, good luck, Rosemary
} } } Dear Listers, } } We are experiencing artifacts in thick epon-araldite resin sections and } are about at our wit's end. You can view the artifact by linking to } http://biotech.missouri.edu/emc/stain_artifact.jpg and noting the dark } magenta blobs in the nuclear layer of the retinal tissue in the image. } We have determined that the "blobs" are associated with "dimples" in the } sections, but they have never shown up in hundreds of other samples done } previous to this particular project. We are virtually certain this is } artifact because serial sections show the blobs in different places and } in different concentrations. } } What we have done so far is to try varying the section thicknesses from } from 0.4 microns to 3.0 microns, mixing the aqueous toluidine blue 1:1 } with varying concentrations of ethanol from 20% to absolute, diluting } the stain with water, mixing fresh stain, cutting on both glass and } diamond histo knives, trying different temperatures on the hot plates, } trying untreated slides and slides treated with BSA, staining in the } microwave at varying wattages and times, and dipping slides and sections } in methanol before staining, then staining normally. } } So far, the only things that have made any difference are using ethanol } in the stain and dipping the slides in methanol prior to staining, but } neither are consistent. They might work one time, but not the next and } we still get the blobs showing up in different places (but ALWAYS in the } nuclear layer only) and in different numbers. } } HEELLLPPP!!! } } Most grateful for ANY assistance with this one! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core----We're the Fun Core! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
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-- Jan Coetzee Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
Apologies for multiple postings and potentially horrible layout!
Dear All, We have a vacancy for a probe operator in our department. We are particularly keen on finding someone interested in research and particularly with an interest in studying for a higher degree. For more information on the post and what is entailed/expected please contact me directly. As you'll see, application forms are available on-line. Cheers, Malc.
RHODES UNIVERSITY
Grahamstown
DEPARTMENT OF GEOLOGY
Applications are invited from suitably qualified candidates for the following post from as early a date as possible:
PRINCIPAL TECHNICAL OFFICER : ELECTRON MICROPROBE LABORATORY
Candidates should have a minimum of a BSc(Honours) degree with a strong background in Earth Science or related disciplines and be computer literate. Primary responsibilities include the operation and basic maintenance of the Department' s JEOL 733 electron microprobe regional research facility. Duties will also include assisting staff, students, and visiting researchers in the use of the facility. Applicants should be committed to working in a productive research environment and will be strongly encouraged to further their own research interests. Candidates should have experience in a chemical analytical environment and experience with x-ray analytical techniques would be an advantage.
Applications forms, further particulars and salary details are available from http://www.ru.ac.za/jobs or by phoning 046-6038004/6038115. Completed applications should be returned to the Recruitment & Selection Section by 17 February 2003.
Curricula vitae which are not accompanied by an official Rhodes University application form will, regretfully, be returned to candidates.
SALARY RANGE: From R69612 to R99276 per annum (pension fund)
From R65676 to R93624 per annum (provident fund)
- - - - o 0 o - - - -
PRINCIPAL TECHNICAL OFFICER
ELECTRON MICROPROBE LABORATORY
Main job objectives
This is a technical position which relates to the routine maintenance and operation of the Geology Department' s electron microprobe regional analytical facility. The job exists to provide support for internal and external users, ensuring smooth and trouble-free operation of the instrument, thereby access to as many users as possible.
Key responsibility areas
maintenance - routine maintenance of the electron microprobe and allied equipment/ materials to ensure operational efficiency; planning - to organise routine services through liaison with technical service companies, and to manage the booking of time slots for instrument utilisation; supervision and training - to provide supervision and technical support for facility users. Also to provide practical training of new users of the instrument, to ensure maximum utilisation of the instrument; research - to conduct own research and to collaborate in research projects of internal and external users of the instrument as required. Own research should lead towards higher academic degrees where appropriate; consultancy - to carry out analytical work for industry and similar organisations as and when appropriate for the purpose of generating income for the regional analytical facility.
January/February 2003
-- Dr MP Roberts Phone: [+27](0)46 603 8313 (work) Dept of Geology [+27] (0)46 6361197 (home) Rhodes University Fax: [+27](0)46 622 9715 6140 Grahamstown Cell: 083 4060 262 SOUTH AFRICA e-mail: m.roberts-at-ru.ac.za
If your conference, seminar, workshop, .. is missing, use the New Submission choice (on the page mentioned) to fill in the easy form for including your meeting to the list.
Best regards, Petr Schauer +--------------------------------------------------------------------+ | Dr. Petr Schauer tel.: +420 541 514 313 | | Head of Electron Microscopy Laboratory fax : +420 541 514 404 | | INSTITUTE OF SCIENTIFIC INSTRUMENTS +420 541 514 402 | | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno petr-at-dp.fce.vutbr.cz | | Czech Republic www: http://www.petr.isibrno.cz/ | +--------------------------------------------------------------------+
These dimples are plastic's areas, which did not stick well to the glass forming sort of "pockets" which holds the stain. In most cases there was air bubble here before water is dried out on hot plate. I do have such artefacts from time to time on my semithings. It looks like drying the slides in the vacuum oven may help. I did not play so much with vacuum because those dimples don't bother me. What I figured out, that vacuum should be just a few mm Hg, otherwise it generates even more bubbles. I was using +60oC. As for special dimple's localization on your sections, I think this is because the different tissue area has different properties and impregnated with plastic differently. Plastic tends to change volume being in water. Those changes may be different from area to area (different tissue properties). In your particular case that area extended in the water (plus heat) more than others. It generates sort of folding, which made those "pockets" when dry. If you did not have it before, it, perhaps, mean, you slightly changed the protocol/chemicals/solvents. You may try to extend all dehydratation/embedding steps for more uniform plastic penetration/embedding/polymerization. I hope it helps. Good luck. Sergey
At 07:09 PM 1/21/03 -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
The stain blobs are only or mainly in the nuclear layer, not elsewhere in the section or on the background, so it is probably not dirt on the glass that is staining. Yes, its quite possible as Rosemary suggests that the sections are not uniformly adhering, and stain is being trapped. The fact that the stain is mostly in the nuclear layer and not elsewhere in the section suggests that this region is adhering to the glass least well. Why would this happen? My guess is that the section is not flat, and that the distortion is greatest in the nuclear layer. If that is so, the ripple should be detectable under the microscope - the section will be go in and out of focus. Three possible ways of curing this. 1) make sure the section gets really hot on the drying hot plate, and has enough time to relax. High plate temperature, large blob of distilled water. Thick sections may need longer to flatten fully than they get before the drop dries. 2) Use chloroform vapour to flatten section 3) Use heat pen to flatten section Cleaning the slide will help with adhesion, but if the section is rippled there will probably still be voids beneath. A simple quick way of cleaning slides is to immerse in 1:3 conc. HCl : ethanol for a few minutes, then Rinse with abs. ethanol and air dry. This can be done in a glass or plastic slide rack. hth Chris
----- Original Message ----- } From: "Rosemary White" {rosemary.white-at-csiro.au} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 22, 2003 5:10 AM
Hi Randy:
I agree with the others who responded to your plea for help, blobs of stain trapped under the section. I suspect that the problem is confined to the nuclear layer becasue infiltration is marginal here due to a high nucleus/cytoplasm ratio. So for your next processing run try more infiltration time in pure resin and another change of pure resin on a rotator or under vacuum. Also, might be time for a fresh batch of resin and accelerator? For the material you already have the other responders have good suggestions. Basically you need to give the sections more time to stretch out and flatten in warm water (or dilute alcohol or acetone) on the slide. If you look at unstained sections with phase contrast or Nomarski optics I think you will see out of focus areas that correspond to 'bubbles' in the section where the stain is accumulating due to lack of adhesion. The other thing you could try is to stain the sections free-floating, then rinse and mount on a slide. Good luck and let us know how things work out
Geoff
"Tindall, Randy D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listers, } } We are experiencing artifacts in thick epon-araldite resin sections and } are about at our wit's end. You can view the artifact by linking to } http://biotech.missouri.edu/emc/stain_artifact.jpg and noting the dark } magenta blobs in the nuclear layer of the retinal tissue in the image. } We have determined that the "blobs" are associated with "dimples" in the } sections, but they have never shown up in hundreds of other samples done } previous to this particular project. We are virtually certain this is } artifact because serial sections show the blobs in different places and } in different concentrations. } } What we have done so far is to try varying the section thicknesses from } from 0.4 microns to 3.0 microns, mixing the aqueous toluidine blue 1:1 } with varying concentrations of ethanol from 20% to absolute, diluting } the stain with water, mixing fresh stain, cutting on both glass and } diamond histo knives, trying different temperatures on the hot plates, } trying untreated slides and slides treated with BSA, staining in the } microwave at varying wattages and times, and dipping slides and sections } in methanol before staining, then staining normally. } } So far, the only things that have made any difference are using ethanol } in the stain and dipping the slides in methanol prior to staining, but } neither are consistent. They might work one time, but not the next and } we still get the blobs showing up in different places (but ALWAYS in the } nuclear layer only) and in different numbers. } } HEELLLPPP!!! } } Most grateful for ANY assistance with this one! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core----We're the Fun Core! } 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/
-- ********************************************** 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 **********************************************
Have you checked your vacuum behavior during discharge to make sure that the problem is not IN THE column ?
If the vacuum is getting worse when discharge occurs - then the problem is likely from the region around the cathode & in the column.
If there is no vacuum change then it is probably from the tank or circuitry...
Ephram Shizgal
On Tue, 21 Jan 2003, shashi singh wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } {a href="http://mail.delongamerica.com/jump/http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html"} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {/a} } -----------------------------------------------------------------------. } } } Hello- } I have an old JEOL 100CX, Which had been relocated } twice but was working fine. Recently there was a water } leak and the water fell on the HT cable going down to } the HT tank. After the leak repair and clearing, the } system was switched on after a week's time, Upon } puttng on HT there were lots of discharges with high } Beam current. Does any one suspect that water may have } seeped in the Tank. or there is some circuiting } problem in the cable. Any advices. } } Shashi Singh } Scientist } CCMB-Hydearbad } INDIA } } } __________________________________________________ } Do you Yahoo!? } Yahoo! Mail Plus - Powerful. Affordable. Sign up now. } {a href="http://mail.delongamerica.com/jump/http://mailplus.yahoo.com"} http://mailplus.yahoo.com {/a}
In response to your email regarding printers, I would like to suggest the following Sony printers as possible solutions for your applications: UPD70A, UPD50, UPDR100
Should you have any questions or need PDF's of the specifications, please feel free to contact me at your earliest convenience.
Best Regards,
Philip Slakmon Director of Sales & Marketing Arctec Technologies Inc. Innovative Solutions For Science & Technology Tel: 514-482-2856 x: 228 Fax: 514-482-2556
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Try this method on your normal slides....I feel the problem is the epoxy/araldite plastic section is not completely spread to it's maximum smoothness, thereby trapping some of your stain. The method below works because of the differences in surface tension between water and xylene.
METHOD:
Cut at 1-2microns, place the section on a drop of water on slide, and then place ONE drop of 100% xylene DIRECTLY on top of the floating section.
You will see the section get kinda of "wrinkly/crinkly" and then start to smooth out. Wait until it reaches its maximum smoothness (around 15-45 seconds).
Very carefully (sometimes I use a dissecting needle to hold a corner of the section while I drain the slide) drain the slide and let it completely dry in an upright position.
Put it on the hot plate and let heat around 1 minute then stain as you would have done before you had all these problems.
Good Luck!
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
-----Original Message----- } From: Rosemary White [mailto:rosemary.white-at-csiro.au] Sent: Wednesday, January 22, 2003 12:10 AM To: microscopy-at-sparc5.microscopy.com
Hi Randy,
Is it possible that you have a dirty batch of slides, so your sections are not adhering as usual, and/or the stain is picking up the dirt? Perhaps if the stain binds to the nuclear layer and there is dirt/grease underneath, then it won't rinse out properly - hence the blobs. My only suggestion is to clean the slides to death and/or try with some of those expensive pre-cleaned slides (people here use them for in situs).
Ethanol in the stain might loosen up this dirt, and methanol dipping also, but perhaps a ferocious clean of the slides might do the trick (if this is the problem, of course). From experience, a cheap brand of slides we used to get suddenly became very dirty and we had to change brands. Not sure what the problem was but we never went back to the original brand to see they recovered their quality.
my 2c worth, good luck, Rosemary
} } } Dear Listers, } } We are experiencing artifacts in thick epon-araldite resin sections and } are about at our wit's end. You can view the artifact by linking to } http://biotech.missouri.edu/emc/stain_artifact.jpg and noting the dark } magenta blobs in the nuclear layer of the retinal tissue in the image. } We have determined that the "blobs" are associated with "dimples" in the } sections, but they have never shown up in hundreds of other samples done } previous to this particular project. We are virtually certain this is } artifact because serial sections show the blobs in different places and } in different concentrations. } } What we have done so far is to try varying the section thicknesses from } from 0.4 microns to 3.0 microns, mixing the aqueous toluidine blue 1:1 } with varying concentrations of ethanol from 20% to absolute, diluting } the stain with water, mixing fresh stain, cutting on both glass and } diamond histo knives, trying different temperatures on the hot plates, } trying untreated slides and slides treated with BSA, staining in the } microwave at varying wattages and times, and dipping slides and sections } in methanol before staining, then staining normally. } } So far, the only things that have made any difference are using ethanol } in the stain and dipping the slides in methanol prior to staining, but } neither are consistent. They might work one time, but not the next and } we still get the blobs showing up in different places (but ALWAYS in the } nuclear layer only) and in different numbers. } } HEELLLPPP!!! } } Most grateful for ANY assistance with this one! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core----We're the Fun Core! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
we have done frozen fixation/freeze-substitution (with HPF machine) for many years. We had tried many cryo-protectants. It seems that 8% methanol in water was the best for us (plant and fungal material). We did not often see cryo damage mm deep in a leaf sample and did not have contamination either. By the freeze-substitution we uses 0.5% Osmium in acetone. We had excellent cell preservation (see Xu and Mendgen 1994, Planta 195:282). Plastic tubes(ependorf) or stainless steel containers were used. It did not give us any trouble. For morphological study we embedded the samples in epon/Araldit and for immmunohistochemical ones we embedded them in LR white (without Osmium). Good luck!
Haixin Xu University of Maryland Baltimore County Biological Sciences
On Tue, 21 Jan 2003, 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 } -----------------------------------------------------------------------. } } } Listers, } We are going to try to plunge freeze and freeze substitute mouse } pancreas. Now I know that the most desirable way to do this would be with } high pressure freezing but that is not an option at the moment. So I am } hoping to get a bit of help with what we do have available. } } We have done quite a bit of plunge freezing microbes and fungi into } Liquid nitrogen cooled liquid propane and then substitution in ethanol, } acetone, etc. with good results. Embedding resin depends on tissue...Epoxy } (LX112) or HM20 lowacryl as needed. } } I would like advice on using a cryo-protectant for the mouse pancreas } ..thinking of using 18% glycerol or 18% propylene glycol. Is this } necessary and do you recommend one over the other? } } Also for substitution, I was going to use acetone + 2% osmium for standard } ultrastructure and just acetone for ICC (switched to ETOH for infiltration } with HM20. } } I would appreciate comments on above general approach and any other } information you would like to share that may enhance contrast on final } sections. } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } West Lafayette, IN 47907 } }
We believe it originated in Japan around 1990, give or take a few years. We added it to our product line on customer demand. All we can tell you is the demand seemed to develop around the early to mid '90's.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "John Skvarla" {jskvarla-at-ou.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, January 21, 2003 11:06 AM
Hi
A quick test of your tank would be to switch off the HT and remove the HT cable from the tank. Then switch on the HT. If you still have discharge sounds the problem is the tank. If not, then the problem is your cable or the cable connection.
Steve Chapman Senior Consultant Protrain EM Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "shashi singh" {shashis_99-at-yahoo.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 22, 2003 3:55 AM
Hi Debby and others: According to an old Balzers pub., "Artefacts and Specimen Preparation Faults in Freeze Etch Technology", it was stated that using glycerol before fixation causes mitochondrial swelling (L. Bachmann and W.W. Schmitt, Proc. Nat. Acad. Sci. USA 68, 2149-2152, 1971) and transformation of the laminar system of the ER into a vesicular system. (H. Moor, Phil. Trans. Roy. Soc. Lond. 261, 121-131, 1971) . Both pubs state that fixation before freezing will prevent this occurrence. Charlie Murphy
Charles Murphy USDA, ARS, Electron Microscopy Unit Bldg. 177B, BARC East Beltsville, MD, 20705 (301) 504-8046 (301) 504-8923 fax murphyc-at-ba.ars.usda.gov
Thank you for your replies regarding a replacement for our SIT camera. We're going to try the Retiga EX.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
A colleague of mine was wondering if the Nikon 8000 ED is able to scan intact, 3.25 x 4 inch TEM negatives at high resolution (4000 ppi). Any information would be appreciated.
Thank you.
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
We use a DataMax E-4203 (w/ Thermal Transfer, 203 dpi, Firmware Ver 02.02) from a local supplier (Mintek Barcode Technologies). Ribbon and labels are from a supplier found on the internet (Electronic Imaging Materials). The Techs print directly from Meditech (our LIS). Surgical #, Pt. Name, Date, Institution, and bar code is printed on the approx. square slide label. This printer can also be accessed from word processors if necessary (it would not be difficult to make a template for Word). There is a wide range of labels available depending your budget including some which are xylene resistant.
Everyone seems very happy with it - in fact we are buying more of them.
No commercial interest, & just my opinion
Regards,
Peter O. Steele, PhD, PMIAC, CLDir Pathology and Laboratory Medicine All Children's Hospital St. Petersburg, FL, USA 727 892-4465
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No...it cannot. The largest it will do is 6x9cm. The width limit is a killer. If you center your subject, you can trim the neg and use the glass holder to scan a 6x9 field.
gary g.
At 11:58 AM 1/22/2003, you wrote:
} A colleague of mine was wondering if the Nikon 8000 ED is able to scan } intact, 3.25 x 4 inch TEM negatives at high resolution (4000 ppi). Any } information would be appreciated. } } Thank you. } } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/~image/ } ##############################################################
The machines connect to your computer for the label text/numbering. The slides are those with a white end. The Cassette labeler is for labeling embedding cassettes for tissue sample. So both the tissue and slide have the same label, if you want. You mentioned "Even upstairs with the thousands of slide labels that they make, they have to tediously type them all out 1 by 1 on the typewriter." This would be a real help for them. Disclaimer: no financial interest in TBS, etc. Damian Neuberger, PhD Senior Research Scientist Baxter Healthcare Corp Tel: 847.270.5888 Fax: 847.270.5897
} } I haven't been able to find any laser printer sheets of appropriately } sized } } light microscope slide labels. } } } } ie: 7/8" wide X 6/8" high. } } } } As a result, we have always been forced to use rolls, and type out each } } label tediously on a typewriter. } } } } I was just wondering if anyone knows of any slicker system for labelling } } microscope slide labels. Even upstairs with the thousands of slide labels } } that they make, they have to tediously type them all out 1 by 1 on the } } typewriter. } } } } I have found for micrographs and negative envelopes, that a combination of } } Microsoft Word mail merged with an appropriate Excel data base does a nice } } job using Avery 5160 address labels, but unfortunately, I haven't as yet } } been able to find a properly sized label for microscope slides, or even } one } } that I could cut nicely. :-( } } } } I was just wondering how other people tackled this problem. Do you all do } } it the tedious way too? Here is a marketing opportunity for some } ambitious } } person who wants to make quick money, by solving a problem in the } workplace. } } }
Hi Debby, There are some HPF's not far from you (Miami, OH and St Paul, MN)...it is worth the drive if you want to freeze chunks of tissue. Sounds like you know that the success of different freezing methods is sample size dependent - especially plunge freezing ( {20 um). So are you slicing and dicing the pancreas before you freeze it? If so, what solution will it be in? Ringers? I don't work on animal tissue so don't laugh too hard at that if that is way off but it would help to know your prep method. I had an animal tissue cryoprotectant recipe from Hong Yi at Emory University {hyi-at-emory.edu} but my filing system is failing me on locating that at the moment. I use a 15% dextran solution (MW 40,000 -Sigma) for HPF of plant material and fungi and I when I worked with a student on freezing frog retina slices and he used 35% dextran (178,000 MW). The reference is: M T Wilson et al. (1998) Ultrastructure of the frog retina after high pressure freezing and freeze substitution. J of Microscopy 189, 219-235. I agree with the others about adding some uranyl acetate to the substitution fluid. It really jazzes up the contrast. good luck, and I wanna know if you have success plunge freezing that pancreas. Beth
} Listers, } We are going to try to plunge freeze and freeze substitute mouse } pancreas. Now I know that the most desirable way to do this would be with } high pressure freezing but that is not an option at the moment. So I am } hoping to get a bit of help with what we do have available. } } We have done quite a bit of plunge freezing microbes and fungi into } Liquid nitrogen cooled liquid propane and then substitution in ethanol, } acetone, etc. with good results. Embedding resin depends on tissue...Epoxy } (LX112) or HM20 lowacryl as needed. } } I would like advice on using a cryo-protectant for the mouse pancreas } ..thinking of using 18% glycerol or 18% propylene glycol. Is this } necessary and do you recommend one over the other? } } Also for substitution, I was going to use acetone + 2% osmium for standard } ultrastructure and just acetone for ICC (switched to ETOH for infiltration } with HM20. } } I would appreciate comments on above general approach and any other } information you would like to share that may enhance contrast on final } sections. } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } West Lafayette, IN 47907
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) **********************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kathleen.smith-at-sjvls.org) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, January 22, 2003 at 16:06:43 ---------------------------------------------------------------------------
Email: kathleen.smith-at-sjvls.org Name: Kathleen Smith
Organization: San Joaquin Valley Information Service
Education: Graduate College
Location: Fresno, California USA
Question: Dear Microscopist:
I am a public librarian with the San Joaquin Valley Information Service in Fresno California. I have a patron who is looking for a video called "The Human Body Through the Electron Microscope." It was originally produced by Time Life Educational Films in the last 70's, possible 1979. I have not been able to find a source for this video in any library or for purchase, possible because of its age. I thought I would avail myself of this organization's specialty in this area. I did not see it listed on any of the video tape bibliographies posted on the MSA's site. Have you ever heard of this video? Do you happen to know of a source? Thanks for your help.
Kathleen Smith, Librarian San Joaquin Valley Information Service Fresno, CA 93704 Ph: 559-488-3229 Fax: 559-488-2965 Email: kathleen.smith-at-sjvls.org URL: http://www.sjvls.org/sjvis
Also, make sure that tank connector is clean, and has ~~ 1 inch (2+ cm) of transformer oil in it. Especially since water presence is suspected. Start at 20 kV, then step voltage up. Do not exceed 80 kV with that little oil in the empty connector well.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
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----- Original Message ----- } From: Steve Chapman {protrain-at-emcourses.com} To: shashi singh {shashis_99-at-yahoo.com} Cc: MSA {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 22, 2003 1:55 PM
Dear Colleagues
The organizers of the forthcoming International Meeting on Applied Physics (APHYS-2003), www.formatex.org/aphys2003/aphys2003.htm are asking for qualified Scientific Reviewers for the Microscopy, Imaging Techniques and Surfaces topics of the Conference. If you would like to participate as reviewers, please contact us at secretariat-at-formatex.org with your contact data, field(s) of expertise and a list of publications. Accepted papers after review will be published in several special issues (Journal of Microscopy, Applied Surface Science and a book edition). The official list of reviewers will be included in the publications.
Hello all, I would like to measure the particle size distribution from a batch of alumina powder with a nominal size of 1µm. I am presently measuring particle sizes from SEM images as we don't have an automated system but it is very labour intensive, slow and somewhat subjective. I have sought help from outside but it seems that our powder is of an awkward size at the lower limit of laser measurement's capabilities. Can anyone suggest a UK company or institution who could measure powder of this size range. Failing that, if anyone could give me the name of a technique it would be of great help as I would then know what to look and ask for. Thanks in advance,
Justin
---------------------------------- Justin Ritherdon, Materials Science and Engineering, Department of Engineering, University of Liverpool, LIVERPOOL, L69 3GH, United Kingdom
My name is Hans van Hirtum and I work for Hogeschool Brabant Nova Knowledge in Etten-Leur, the Netherlands. We organize International post-graduate short-courses on a variety of biomedical laboratory techniques. All international courses are organized in close collaboration with experts in the field of interest. This spring we offer the following courses on histological techniques:
- FISH Techniques in Moleculair Pathology - Confocal Light Microscopy: fundamentals and biological applications - Tissue micro-array - Multiple Staining in Immunohistochemistry
Please visit our website http://www.novaknowledge.nl/english.htm for detailed information about these courses and other courses that we offer (e.g. quantitative PCR and strategic protein purification).
Best regards, Hans van Hirtum
Ing. J.P. van Hirtum Hogeschool Brabant Nova Knowledge P.O.box 5690 4801 EB Breda, the Netherlands T: +31 (0) 76 572 2644 F: +31 (0) 76 572 2640 E: hvanhirtum-at-hsbb.nl W: http://www.novaknowledge.nl
We do have a problem with vacuum because of an inefficient Gun/Column Valve But once it attains high vacuum and ready state we do not encounter any problems. These discharges are post leak and the beam emission current increases if the HT is switched on for even 5 min, and even at 20kV. Shashi Singh
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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} If your conference, seminar, workshop, .. is missing, use the New } Submission choice (on the page mentioned) to fill in the easy form for } including your meeting to the list.
Dear Colleagues,
Thank you for the yesterday's submission of interesting meetings. All submissions have been displayed. Note that also your Laboratories can be submitted and listed in the extensive list of Microscopy Laboratories (http://www.petr.isibrno.cz/microscopy/laboratories.php). Keep in your mind that your Laboratory can be much easily searchable if listed as much as possible.
I agree with John Mardinly, the Fuji Pictography printer provides the best photographic quality print. This printer can print up to 400 DPI, and offers built-in color calibration. It only has a SCSI II connection, so must be connected to a computer, but can be networked as a shared resource. The list price for this printer is $3995, and prints are approximately $3.00 each. This printer actually uses a photographic process to print images on Fuji photo paper. The paper is fed from a roll, and the printer has an automatic sheet cutter. (http://www.fujifilm.com/JSP/fuji/epartners/Products.jsp?nav=1&parent=PRODUCT_CATEGORY_474433&product=6213061)
A less expensive alternative, although certainly not as full featured or robust, is the Olympus P400. This is a dye-sub printer, offering 314 dpi, and will print up to 7.64 in. x 10 in. images on A4 paper. This printer has USB and parallel port interfaces, and can be networked by a print server. It also accepts SmartMedia(tm) 3V (3.3V): 4, 8, 16, 32, 64, or 128MB PCMCIA PC Card Type II (ATA format or PC Card Adapter for CompactFlash or Memory Stick), to print directly from your digital camera media. The list price for this printer is $499 and prints are approximately $2.00 each. The printer has a 50 sheet paper tray. (http://www.olympusamerica.com/cpg_section/cpg_product_lobbypage.asp?l=1&bc=23&p=19&product=632)
Depending on the quality you require, an excellent alternative is the Xerox Phaser 8200. This printer uses a solid ink, and offers a 1200 dpi photo mode. This is a production printer, delivering 16 pages per minute, with the first print as fast as 9 seconds. You can print on a variety of materials ranging from 16 lb paper to 110 lb business cards or envelopes. It can be purchased with a duplex option, and two additional 500 sheet feeders, providing up to 1200 sheet paper capacity. In its least expensive form, this printer offers USB and parallel connections. The other three models have built in networking. This printer is offered in 4 models with prices ranging from $1500 to $3500. Prints are approximately $0.20 each. (http://www.officeprinting.xerox.com/perl-bin/product.pl?product=8200)
Please contact me directly if you have additional interest in any of these printers.
Thank you, Bob Voigt (bobvoigt-at-restechimage.com) Resolution Technology, Inc. (www.restechimage.com) Phone (614) 921-0045 FAX (614) 921-0046
Is there anyone out there using the Aspex PSM 75 SEM? any comments about this equipment would be helpful good or bad. I cannot find any info at the Aspex website due to under construction. Thanks as always
Robert Fowler Quality Assurance Technician (Failure Analysis) TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com
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One of our undergraduates has a project to set up a microscope with laser for an optical tweezer experiment. The highest power lens on the microscope has the following data on it:
50x / 0.80 "infinity" / 0.17-A
The first two figures I understand to be magnification and numerical aperture, but I would like to ask a few things:
(1) what does the second set of figures refer to?
(2) how would one estimate the focal length of this lens?
(3) if one is shining a parallel laser beam down into this lens from above, how could one determine the widest diameter of beam which would actually go through this lens without being stopped?
Any help would be much appreciated.
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +----------------------------------------+ Phone:+44 (0) 118 9318572 Fax: +44 (0) 118 9750203 University internal extension 7867 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +----------------------------------------+
Justin Ritherdon wrote: =============================================================== I would like to measure the particle size distribution from a batch of alumina powder with a nominal size of 1µm. I am presently measuring particle sizes from SEM images as we don't have an automated system but it is very labour intensive, slow and somewhat subjective. I have sought help from outside but it seems that our powder is of an awkward size at the lower limit of laser measurement's capabilities. Can anyone suggest a UK company or institution who could measure powder of this size range. Failing that, if anyone could give me the name of a technique it would be of great help as I would then know what to look and ask for. ================================================================ It might not be the complete solution you are asking for but it will help, "it" being our "Tacky Dot Slide" products as shown on URL http://www.2spi.com/catalog/new/tacky.shtml
If you use the 15 um dots (smallest dot size available), at the most, several particles will appear per dot, and while you will still have the "counting" issue, at least it will be far less confusing and certainly much less subjective. And if the powder is arranged on orthogonal centers, then it might even be possible to have them analyzed automatically using appropriate software.
Chuck
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Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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You may be using or have access to an older laser unit. We have a laser particle counter with both forward and right-angle scattering. The lower limit for our laser sizer is 0.05 µm. Given the very low cost of particle sizing by laser, I would find a local materials lab with a modern laser unit. If you want more details on particles sizing or morphology by other methods you can contact me off-line. Roy Nelson, PhD mtl-at-njcc.com Material Testing Lab. Pennington, NJ 08534
Justin Ritherdon wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello all, } I would like to measure the particle size distribution from a batch of } alumina powder with a nominal size of 1µm. } I am presently measuring particle sizes from SEM images as we don't have an } automated system but it is very labour intensive, slow and somewhat } subjective. } I have sought help from outside but it seems that our powder is of an } awkward size at the lower limit of laser measurement's capabilities. } Can anyone suggest a UK company or institution who could measure powder of } this size range. Failing that, if anyone could give me the name of a } technique it would be of great help as I would then know what to look and } ask for. } Thanks in advance, } } Justin } } ---------------------------------- } Justin Ritherdon, } Materials Science and Engineering, } Department of Engineering, } University of Liverpool, } LIVERPOOL, } L69 3GH, } United Kingdom } } Tel. 0151 794 5396 } Fax. 0151 794 4675 } International. +44 151 794 ....
This is just a thought but in the semiconductor industry partricle size/ distribution is critical. A device known as a "Surfscan" can do two things, measure particle size and provide a geographical distrubution with statistics on particulates residing on a flat surface. Try this website; http://ceaspub.eas.asu.edu/csser/surfscan.htm
The one caveat to this technique is that if you have particles that are co-joined, the system may not recognize them as separate and distinct from each other. This system does us a laser and claims a 90% detection probability of 0.22 uM diameter latex spheres on a flat silicon surface.
These tools are primarily found in semiconductor fabrication facilities and in some universities that have same.
Regards,
Peter Tomic
-----Original Message----- } From: Justin Ritherdon [mailto:J.Ritherdon-at-liverpool.ac.uk] Sent: Thursday, January 23, 2003 4:17 AM To: Microscopy Listserver
Hello all, I would like to measure the particle size distribution from a batch of alumina powder with a nominal size of 1µm. I am presently measuring particle sizes from SEM images as we don't have an automated system but it is very labour intensive, slow and somewhat subjective. I have sought help from outside but it seems that our powder is of an awkward size at the lower limit of laser measurement's capabilities. Can anyone suggest a UK company or institution who could measure powder of this size range. Failing that, if anyone could give me the name of a technique it would be of great help as I would then know what to look and ask for. Thanks in advance,
Justin
---------------------------------- Justin Ritherdon, Materials Science and Engineering, Department of Engineering, University of Liverpool, LIVERPOOL, L69 3GH, United Kingdom
Eighth Annual INTERNATIONAL 12-Day Short Course on
3D Microscopy of Living Cells June 15 - 26, 2003 (Pre-course: afternoon, June 14)
Seventh, Post-course Workshop on
3D Image Processing, June 28 - July 30, 2003
Organized by Prof. James Pawley, (University of Wisconsin-Madison)
in association with
The Departments of Pharmacology and Physiology and the Brain Research Centre, University of British Columbia Vancouver, BC, Canada
DATES
Applications must be received by March 15, 2003 Deposit due April 15, 2003 Registration 5:00 - 7:00 PM Saturday, June 14, 2003 First Lecture 7:30 PM Saturday, June 14, 2003 Live-cell Course ends, noon Thursday, June 26, 2003 3D Image Processing Course, June 28 - 30, 2003
APPLICATIONS
Applicants must complete a questionnaire on the web to assess knowledge level, field of interest and proposed personal, live-cell, project. www.3dcourse.ubc.ca/application.htm
Enrollment will be limited to about 24-32 participants (exact number depends on number of 3D Systems available). Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with access to basic texts to read before the course begins. Application forms can be obtained from:
Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU
Additional information is available from: www.3dcourse.ubc.ca/brochure.htm
We expect to have at least 11, 3D microscope workstations for student use during 14 3D-Lab session and there will be an international faculty of 20.
Application deadlines:
Application forms must be received for screening by March 15, 2003. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2003. In general, refunds of the deposit will only be possible if someone on the waiting list can take the place. The remainder is due before Registration.
Pre-Course Tuition (1/2 day Basic Optical principles) $120 (US) 3D Live-cell Course Tuition (includes lunches and snacks): $2450 (US) Workshop Tuition (includes lunches and snacks): $900 (US)
Room/board about $40/day (US) depending on room type.
I hope that this includes all of the information that you need, but if not, please get back to me.
Jim Pawley -- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
} Email: kathleen.smith-at-sjvls.org } Name: Kathleen Smith } } Organization: San Joaquin Valley Information Service } } Education: Graduate College } } Location: Fresno, California USA } } Question: Dear Microscopist: } } I am a public librarian with the San Joaquin Valley Information } Service in Fresno California. I have a patron who is looking for a } video called "The Human Body Through the Electron Microscope." It was } originally produced by Time Life Educational Films in the last 70's, } possible 1979. I have not been able to find a source for this video } in any library or for purchase, possible because of its age. I } thought I would avail myself of this organization's specialty in this } area. I did not see it listed on any of the video tape bibliographies } posted on the MSA's site. Have you ever heard of this video? Do you } happen to know of a source? Thanks for your help. } } Kathleen -
Congratulations on your thoroughness! I wrote one of the MSA video tape bibliographies, for Project MICRO. It's the product of a lot of searching, and I've never heard of that tape. Your patron may have an alternate title for one of Lennert Nilssen's tapes; they seem to have been issued more than once, and they definitely predate the 90s. Here's the MICRO description:
Nilssen, L. 1996 "The Photographer's Secrets" 1 hour, $19.95 from WGBH Boston Video, P.O.Box 2284, South Burlington, VT, 05407, 800-255-9424. Lennart Nilssen is famous for his beautiful images of human development. This is part three of the Nova series "Odyssey of Life" (or part 4 of the series "The Wonder of Life") It explains his use of SEM and other imaging tools that blur the line between microscopy and macrophotography; it will be particularly interesting for budding microscopists. All ages.
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 looking for help finding a source for periodic table wall charts. I'm using them for microanalysis so I need K, L & M lines in keV. I've been using the small ones, 2' x 3', that the vendors give away but I'd like to get bigger ones. I need 6 of them.
Anyone run across something like this?
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 Mailto: opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
Have you thought of using image analysis software to get size information and statistical values? Without seeing any images I can't say if it is possible in your case, but if you wish, I can provide you with the name and phone numbers of a colleague of mine in the UK. He might be able to help you directly.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Justin Ritherdon [mailto:J.Ritherdon-at-liverpool.ac.uk] Sent: Thursday, January 23, 2003 10:17 AM To: Microscopy Listserver
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
The first in the world to market a product one might call (generically) a double sided adhesive conductive carbon filled tape was a company in Japan called Oken Shoji. This would have been in the early 1980's. The owner of that firm was and still is Mr. Hisashi Sato. He also applied for and received a Japanese patent for the tape at that time. Apparently it covers only tape cut 8 mm wide and tapes either larger or smaller than that width somehow don't infringe. I know that sounds a bit irrational but that is the way it has been explained to me.
I have also been told that there is another name on the patent, that of a professor in Japan. I believe that the original user (inventor) of the tape for this application was that professor, who happened to know Mr. Sato and then it was Mr. Sato who actually commercialized the product. It is of course all in Japanese so this information comes to me second-hand.
In any case, the tape was widely used in Japanese SEM laboratories before it was even known here in the USA or in Europe. It was the Japanese service engineers who brought samples of the tape to the USA from Japan and when the US employees of the Japanese microscope firms went to Japan for training, they brought samples back with them as well. It was my further recollection that the first SEM manufacturer to be encouraging their customers to use the carbon tape as a means of promoting column cleanliness was ISI/Topcon.
SPI Supplies was introduced to the tape by the ISI people during that time frame and almost simultaneously, the tape was introduced in the USA and Canadian markets by both SPI Supplies and E. F. Fullam, Inc. Some time later after that, the tape was then offered by Ted Pella, Inc., Electron Microscopy Sciences, Inc. and later on by Energy Beam Sciences, Inc. In Europe at that time (early 1980's) the tape was offered by Polaron Equipment Ltd. and Agar Scientific Ltd.
Today, the tape is not all the same. Some is on a white plastic core and some is on a cardboard core. The tape on the cardboard core to the best of my knowledge has direct traceability to the original tape that has been produced since the early 1980's. Anyone who has used this tape knows very well how the cardboard core starts to disintegrate and particulate, hardly a good thing for EM lab cleanliness. The newest version of this tape has a white plastic core that does not shed particulates. For more information see URL http://www.2spi.com/catalog/spec_prep/cond_adhes-tapes.shtml
Today, one can use this kind of product not only in tape form but also in die cut discs and sheets. It is also available as a double sided adhesive silver filled sheet. All of these products trace their progeny back to the original carbon tape concept and patent.
Disclaimer: SPI Supplies offers all of the mentioned products so we have a vested interest in promoting their use.
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 ######################## ============================================
Good morning all, I am looking for the "one size fits all" type of embedding resin that can be used for polymers. For a number of years we have been using a two part epoxy resin which for most general purpose applications has been very satisfactory. Because the availability of these components has become uncertain I wanted to ask other users and suppliers for their suggestions or recommendations. Most of the materials that we wish to embed and section have glass transition temperatures (Tgs) between 50 and 70 degrees C. For some materials: blends, copolymers and the like cryo-sectioning may be necessary. Embedding resins that require curing at elevated temperatures are out of the question. Obviously we must be certain that any curing protocol leaves the sample unchanged. Are (is) there a single embedding resin that meets all these requirements? Thanks.
Trying to stay warm in the Great White North ... it is a cool -20C today. Thankfully there is little wind chill to worry about.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: Owen P. Mills [mailto:opmills-at-mtu.edu] Sent: Thursday, January 23, 2003 4:51 PM To: Microscopy
Dear All,
I'm looking for help finding a source for periodic table wall charts. I'm using them for microanalysis so I need K, L & M lines in keV. I've been using the small ones, 2' x 3', that the vendors give away but I'd like to get bigger ones. I need 6 of them.
Anyone run across something like this?
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 Mailto: opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
The 1st Meeting of the MSA FIB FIG will held in conjunction with the 4th Annual FIB Users Workshop at the FLAVS/Florida Society for Microscpy meeting at the University of Central Florida on Tuesday, March 18, 2003.
A List of Confirmed Speakers Include:
Joe Michael, Sandia National Lab Richard Langford, Oxford University Jeff McDowell, Sela Richard Young, FEI Company Peter Gnauk, LEO Elektronenmikroskopie Kevin McIlwraith, Hitachi Janice Lomness, UCF David Fries, USF Tom Kelly, Imago Scientific Fred Stevie, NCSU Brian Kempshall,UCF Lucille Giannuzzi,UCF
There will also be a FIB Users Workshop consisting of 10 minute presentations of theory, techniques, and/or applications of FIB/dual beam/cross beam instrumentation. For more information, or if anyone would like to participate as a presenter in the Workshop please contact:
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor Mechanical, Materials, and Aerospace Engineering University of Central Florida 4000 Central Florida Blvd. PO Box 162450 Orlando, FL 32816-2450 email lag-at-mail.ucf.edu phone 407-823-5770 fax 407-823-0208 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: Owen P. Mills [mailto:opmills-at-mtu.edu] Sent: Thursday, January 23, 2003 4:51 PM To: Microscopy
Dear All,
I'm looking for help finding a source for periodic table wall charts. I'm using them for microanalysis so I need K, L & M lines in keV. I've been using the small ones, 2' x 3', that the vendors give away but I'd like to get bigger ones. I need 6 of them.
Anyone run across something like this?
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 Mailto: opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Deadlines are getting nearer for the FOCUS ON MICROSCOPY 2003 conference 13-16 April 2003, University of Genoa, Italy. Conference location: Palazzo Ducale, Genoa. See http://www.palazzoducale.genova.it
The important deadlines are: Abstract due date: February 1st, 2003 Early registration until: February 15th, 2003
Abstract submission and registration preferably on-line through our website http://www.focusonmicroscopy.org where all information about the conference can be found.
OPERA! We have been able to reserve for the conference a number of seats for the the premiere of the opera La Boheme by Giacomo Puccini in the Genova Opera house on the evening of 15 April. http://www.carlofelice.it For more information on the opera and reservation see our web-site.
Hotels in Genoa may be a bit tight around the conference period, so timely booking is suggested.
Greetings and looking forward to see you in Genoa!
Alberto Diaspro, University of Genoa, Italy Cesare Usai, National Research Council, Italy Fred Brakenhoff, University of Amsterdam, the Netherlands
FOCUS ON MICROSCOPY 2003 16th International Conference on 3D Image Processing in Microscopy 15th International Conference on Confocal Microscopy
April 13th-16th, 2003 University of Genoa, Italy Conference location: Palazzo Ducale, Genoa
Confocal, multiphoton excitation and deconvolution imaging techniques have become indispensable tools in microscopy for the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. We see time-resolved micro-spectroscopy, FRET and related advanced type of fluorescence imaging modes combined with a trend towards faster -3D- image collection at lower specimen dosage and ever increasing resolutions. New non-linear excitation modes like harmonic and coherent anti-Stokes Raman imaging are rapidly evolving. Sophisticated data processing like Image Correlation Spectroscopy (ICS) can provide access to the statistics of the underlying specimen structures down to the ten nm range. Recent developments in these areas will be covered with an eye on their practical applications. Special sessions will be devoted to the application of 3D techniques for the microscopy of living cells and tissues and the use of GFP and other labeling techniques in these studies.
These conferences offer an efficient meeting point for developers and users working in these rapidly developing fields and play an important role in the dissemination of information about new developments in microscopy.
Further information:
http://www.focusonmicroscopy.org ==================================================================== -- +-----------------------------------------+ Prof. Dr. G.J. Brakenhoff University of Amsterdam Institute for Molecular Cell Biology Section Molecular Cytology Kruislaan 316 1098 SM Amsterdam, The Netherlands
Well we need to look at your problem step by step.
1) Switch off the HT. Then if you remove the gun cable from the tank and switch on do you hear discharges?
2) If you do not hear discharge it tells us that the problem is in the cable or the gun vacuum.
3) Do you have a penning gauge that you could fit in one of the gauge positions in the gun area?
4) If we can monitor the vacuum we will be able to find out if it is a vacuum problem you have, or a gun cable problem.
5) If we decide it is a gun cable problem you will need to contact an organisation in your country that will be able to replace the cable, using your current cable ends.
6) This type of company may make or repair x-ray sets in your hospitals, these also use high voltage cables. Another alternative is a company that makes cables for other high voltage applications. I have used both when I have had service problems around the world.
I have just returned from running courses in India it is unfortunate I did not see your posting until recently.
Please keep the information flowing so that I may guide you to a solution.
Steve Chapman Senior Consultant Protrain EM Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "shashi singh" {shashis_99-at-yahoo.com} To: {microscopy-at-sparc5.microscopy.com} Cc: {shizgal-at-delongamerica.com} Sent: Thursday, January 23, 2003 11:29 AM
Dear all,
The Department of Physics at Boston College just finished installation of a state-of-the-art JEOL 2010F FEG TEM. It is a multi-purpose ultrahigh resolution analytical electron microscope with a wide range of capabilities such as high resolution image observation, nano area X-ray analysis, versatile analysis by convergent-beam electron diffraction, and analysis of the atomic structure and/or bonding state of atoms.
The Physics Department is also equipped with a JEOL 6340F Scanning Electron Microscope (SEM), a JEOL 200CX TEM, and an X-ray diffractometer.
All the above equipment is open to external users. Universities, Institutions, corporations as well as other scientific collaborators are welcome to use our facilities. Interested party please contact Dr. Jianyu Huang at the following address:
----------------------------------------------- Department of Physics Boston College 140 Commonwealth Avenue, Higgins Hall Chestnut Hill, MA 02467
All Ladd resins require heat or generate heat upon curing, including Mercox Corrosion casting resin.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- 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: Friday, January 24, 2003 8:45 AM
} January 24, 2003 } } Good morning all, } I am looking for the "one size fits all" type of embedding resin that can be } used for polymers. For a number of years we have been using a two part epoxy } resin which for most general purpose applications has been very } satisfactory. Because the availability of these components has become } uncertain I wanted to ask other users and suppliers for their suggestions or } recommendations. Most of the materials that we wish to embed and section } have glass transition temperatures (Tgs) between 50 and 70 degrees C. For } some materials: blends, copolymers and the like cryo-sectioning may be } necessary. Embedding resins that require curing at elevated temperatures are } out of the question. Obviously we must be certain that any curing protocol } leaves the sample unchanged. Are (is) there a single embedding resin that } meets all these requirements? Thanks. } } Trying to stay warm in the Great White North ... it is a cool -20C today. } Thankfully there is little wind chill to worry about. } } Paul -
Have you ever tried Epofix? It cures at room temperature, and has good cutting qualities. All such RT polymerizations are exothermic, tho, and I haven't checked its temperature rise.
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
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nasen.baer-at-planet-interkom.de) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, January 25, 2003 at 17:03:35 ---------------------------------------------------------------------------
Email: nasen.baer-at-planet-interkom.de Name: Klaus Fischer
Education: 9-12th Grade High School
Location: Stuttgart, Germany
Question: Hello,
I'm looking for a set or single slides concerning nerve-system, sens-organs (human and animal), nerve-cells and nerve-endings in skin, sens-organs and motor nerve endings. Do you have an idea, where I can get such things?
We recently demo'ed a cryo-ultramicrotome and saw the amazing improvement that using a Diatome Static Line II ionizing gun so we bought one when we got our new cryo-ultramicrotome. I just tried out the ionizer as I sectioned some Epon sections at room temp on water. I was really surprised by how much better the block sectioned and the quality of the ribbons. I was getting great ribbons and as soon as I turned off the ionizer, the next section would crumble. This happened at least 6 times. Is this type of improvement typical? Are other users using ionizers with resin sections? Since my cryo-ultramicrotome is in a different location than my two regular ultramicrotomes, i may be forced to buy another one of these. Standard disclaimer: I have no financial interest in Diatome or much else. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Cor 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Consider the Polaroid Sprintscan 45. It will do 2000x4000 (2500 equivalent) up to 4x5".
Does one really need 4000 dpi for a TEM neg?
gary g.
At 02:36 PM 1/24/2003, you wrote: } I do not know about the Nikon but Polaroid has a scanner, the SprintScan } 4000 Plus that scans 35mm to 6x7 at 4000 x 4000 dpi. It is a great scanner! } } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Wednesday, January 22, 2003 5:57 PM } To: John J. Bozzola } Cc: MSA listserver } Subject: Re: Nikon 8000 ED Scanner } } No...it cannot. The largest it will do } is 6x9cm. The width limit is a killer. } If you center your subject, you can trim } the neg and use the glass holder to scan } a 6x9 field. } } gary g. } } } At 11:58 AM 1/22/2003, you wrote: } } } A colleague of mine was wondering if the Nikon 8000 ED is able to scan } } intact, 3.25 x 4 inch TEM negatives at high resolution (4000 ppi). Any } } information would be appreciated. } } } } Thank you. } } } } ############################################################## } } John J. Bozzola, Ph.D., Director } } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } } 750 Communications Drive - MC 4402 } } Southern Illinois University } } Carbondale, IL 62901 U.S.A. } } Phone: 618-453-3730 } } Fax: 618-453-2665 } } Email: bozzola-at-siu.edu } } Web: http://www.siu.edu/~image/ } } ##############################################################
The 2003 Spring Meeting of THE TEXAS SOCIETY FOR MICROSCOPY Embracing All Forms of Microscopy
April 3, 4, 5, 2003
Meeting will be held in Hubbard Hall on the campus of Texas Womans University, Denton, TX.
THURSDAY WORKSHOP Microwave Techniques Presented by Rick Giberson, Research and Development Manager, Ted Pella, Inc. Workshop sponsored by Ted Pella, Inc. and TSM
MSA LAS-SPONSORED SPEAKER FRIDAY, APRIL 4, 2003 HUBBARD HALL TWU - DENTON CAMPUS Dr. Lucille A. Giannuzzi, Ph.D., Associate Professor, Mechanical, Materials & Aerospace Engineering and Director, UCF/Cirent Materials Characterization Facility, University of Central Florida, Orlando, FL. Topic: "Focused Ion Beam Specimen Preparation for Everything". Focused ion beam techniques have been developed to prepare site-specific specimens in a reproducible and rapid manner for SEM and TEM analysis. The conventional and the lift-out methods of specimen preparation will be discussed and FIB applications on a wide range of materials will be presented. The usefulness for using the FIB for solving scientific research problems will be emphasized.
Registration forms, hotel information, and author's instructions can be found on our website, http://www.texasmicroscopy.org/ . Questions about the program can be directed to Jo Taylor, Program Chair, at jtaylor-at-sfasu.edu. Author's and abstract questions should be directed to our Editor, Camelia Maier, at cmaier-at-twu.edu.
There you will find a table of most intense line energies in keV, visible with EDX-spectrometer. In addition, critical excitation energies and most probable line overlaps (K/L, K/M and L/M) are presented (atomic numbers). The element names are in German (left column) and in English (right column).
Klick at the animated table image. Your browser will load the entire image in JPG-format (956 KByte). Then store the image on your computer and print it out. Use the maximum size and best resolution, available with your printer.
Sorry, until now the homepage is only in German. in a couple of weeks you will find there a English version and the program to download, from this the poster is a screen shot.
Good luck
Frank
"Owen P. Mills" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All, } } I'm looking for help finding a source for periodic table wall charts. I'm } using them for microanalysis so I need K, L & M lines in keV. I've been } using the small ones, 2' x 3', that the vendors give away but I'd like to } get bigger ones. I need 6 of them. } } Anyone run across something like this? } } Owen } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } Mailto: opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills
We have water in the HV tank, please suggest us how to remove the water and what are the parts likely to be replaced. Since we are not trained in this system, any diagram assisting to above could be sent to me personally.
since Agfa has stopped to produce the x-ray film developer G-230 I would like to ask if someone still has got the original Agfa recipe for this developer i-or the G-150 developer- in his/her hands. I am asking because I am not very satisfied with the results we obtained with Kodak's D-19 developer for our last 2000 sheets of Agfa Scientia film.
We too have had some concerns expressed to us by our customers concerning the new formulation of 4489 film. The concern has been limited in volume, but for those customers who are concerned we maintain a stock of the old formulation of the film. If you require any just be sure to ask for the old formulation.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
Do you embed all your samples? If so, why? I understand that the samples some encounter can only be addressed by embeddment. However, I routinely work with polymeric materials (films, molded materials, granules, fibers, fabrics, etc) using optical microscopy, SEM, LVSEM, TEM and AFM. I embed only when absolutely necessary. Obviously some samples must be embedded. Many, however, can be sectioned or faced after gluing onto mounts or grasped with handmade polyethylene chucks.
Consider working without epoxy wherever possible. The benefits of unembedded samples include: shortened prep; improved microtomy conditions; and elimination of the complications of epoxy that is infiltrated into or surrounding the part during the analysis (contaminates, charging, etc).
Feel free to reach me off-line for more information.
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Gerroir, Paul J" {Paul.Gerroir-at-crt. To: "Microscopy-at-MSA. Microscopy. com (E-mail)" xerox.com} {Microscopy-at-sparc5.microscopy.com} cc: Subject: TEM: Embedding Resins for Polymeric Materials 01/24/03 07:45 AM
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January 24, 2003
Good morning all, I am looking for the "one size fits all" type of embedding resin that can be used for polymers. For a number of years we have been using a two part epoxy resin which for most general purpose applications has been very satisfactory. Because the availability of these components has become uncertain I wanted to ask other users and suppliers for their suggestions or recommendations. Most of the materials that we wish to embed and section have glass transition temperatures (Tgs) between 50 and 70 degrees C. For some materials: blends, copolymers and the like cryo-sectioning may be necessary. Embedding resins that require curing at elevated temperatures are out of the question. Obviously we must be certain that any curing protocol leaves the sample unchanged. Are (is) there a single embedding resin that meets all these requirements? Thanks.
Trying to stay warm in the Great White North ... it is a cool -20C today. Thankfully there is little wind chill to worry about.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Food Structure and Functionality Symposium 2003 Held in conjunction with the 94th AOCS Annual Meeting and Expo May 4-7, 2003 Kansas City Convention Center, Kansas City, Missouri, USA
Schedule as of January 27th, 2003
Sunday, May 4, 2003 8:30 a.m. to 4:30 p.m. Short Course: Roadmap guide to Image Analysis for the Food Industry given by Dr. John Russ, Materials Science and Engineering Department, North Carolina State University, USA
Monday, May 5, 2003 Opening of Symposium
Microencapsulation of Food Ingredients and Nutrients: Opportunities and Challenges. Chairs: Moshe Rosenberg, University of California, USA and Beatrice Conde-Petit, Swiss Federal Institute of Technology, Zurich, Switzerland 8:00 a.m. - 12:00 p.m. Starch as Encapsulation Agent: Structural Properties of Starch-Flavor Inclusion Complexes. B. Conde-Petit
Water-Insoluble Microcapsules and Microspheres Consisting of Whey Proteins. M. Rosenberg
Use of Microencapsulation for the Stabilization of Oxidatively Sensitive Lipids and Nutrients, Such as Omega 3 Oils, to Significantly Improve Shelf Life and Handling Characteristics. P. Lee
Improvement of Flavor Performance by Microencapsulation.. K.B. de Roos
Agricultural Applications of Microscopy and Imaging. Chairs R. Gary Fulcher, University of Minnesota, USA and S. Shea Miller, Agriculture and Agri-Food Canada, Ottawa, Canada 2:00-4:00 p.m.
3 speakers confirmed, no further information available at this time.
Food Structure and Functionality Forum Dedicated Poster Session 4:00p.m. to 6:00p.m.
Food Structure and Functionality Forum Division Board Meeting 6:00p.m. to 7:00 p.m.
Tuesday, May 6, 2003 Colloidal and Interfacial Properties for Understanding Tailoring Food Product Behavior I - Dairy Applications 9:00 a.m. - 12:00 p.m. Chairs: Mark Auty, Dairy Products Research Centre, Ireland and John A. Lucey, University of Wisconsin, USA
The Influence of Vegetable Fat on Sensory Characteristics of a Cheese. S. Karlsson
Recent Progress on Understanding the Melting Process in Cheese. J.A. Lucey
Colloidal and Interfacial Properties of the Caseins. D.S. Horne
Role of C-terminal Region of Bovine. P. Qi
Influence of the Oil Characteristics on the Interfacial Properties on Oil-In-Water Emulsions. C. Granger
Stability of Pickering Emulsions in Tablespreads. D. Rousseau
Creaminess: Its Origin in Microstructure. M. Paques
Food Structure and Functionality Forum Division Luncheon 12:00p.m. to 2:00 p.m. Speaker: Moshe Rosenberg, University of California, USA. Topic: Science and Technology in manufacturing High Quality Cheese: Challenges and Opportunities
Colloidal and Interfacial Properties for Understanding and Tailoring Food Product Behavior II 2:00-5:00 p.m. Chairs: Marcel Paques, Friesland Coberco Dairy Foods, The Netherlands and David G. Pechak, Kraft Foods, Inc., USA
Pickering Stabilization of Water-in-Oil Emulsions. S.M. Hodge
Structural Studies of Interfaces in Frozen Dairy Desserts. H.D. Goff
Creaminess Versus Fattiness in Oil-in-Water Semi-Solid Emulsions. R. Janssen
New Information on the Internal Structure of Starch from Atomic Force Microscopy Studies. V.J. Morris
Accuracy Evaluation of Low-Resolution NMR of Oil Drop Size Distribution in Triglyceride Emulsions. N. Denkov
Interfaces and Food Functionality. M. Martin
Food Structure and Functionality Forum Division Members Meeting 5:00p.m. to 6:00p.m.
Wednesday, May 7, 2003 New Methods & Techniques for Food Structure and Functionality Analysis 8:00 a.m. - 12:00 p.m. Chairs: Kathy Groves, Leatherhead Food International, UK and Maud Langton, Swedish Institute for Food and Biotechnology, Sweden.
3-Dimensional Imaging of Lipid Crystallization by Widefield Deconvolution Microscopy. J. Litwinenko
High Pressure Homogenization of Raw Bovine Milk: Effects on Fat Globule Size Distribution, Protein Functionality and Microbial Inactivation.. J.C. Cheftel
Electron Microscopy of Wet Samples at Atmospheric Pressure: Unique Analytical Capabilities for Food, Emulsions, and Biological Specimens. O. Gileadi
Light Scattering Intensity Fluctuations and Gel Formation in Opaque Systems. D. Horne
Processed Foods: Structural, Functional and Chemical Properties 2:00-5:00 p.m. Chairs: Diana Kittleson, General Mills Technology East, USA and Bernhard Tauscher, Federal Research Centre for Nutrition, Germany.
Rheometric Measurement of Cocoa Butter Tempering Properties. J. Alander
Complex Formation of B-Cyclodextrin in Liquid Foods. K.A. Schmidt
Alteration of Food Peptides and Proteins under High Hydrostatic Pressure. A. Fernandez- Garcia
Paula M. Allan-Wojtas Research Scientist - Food Microstructure / Chercheur scientique - microstructure des aliments Food Safety and Quality Team / Salubrité et qualité des aliments Agriculture and Agri-Food Canada /Agriculture et Agroalimentaire Canada Telephone / Téléphone: 902-679-5566 Facsimile / Télécopieur: 902-679-2311 32 Main Street / 32 rue Main Kentville, Nova Scotia / Kentville (Nouvelle-Écosse) B4N 1J5 allanwojtasp-at-agr.gc.ca
Roadmap Guide to Image Analysis for the Food Industry May 4, 2003 Held in conjunction with the 94th AOCS Annual Meeting Kansas City Convention Center Kansas City, MO, USA
Course Faculty Dr. John Russ, visiting professor, Materials Science and Engineering Department, North Carolina State University, USA.
Sponsored by the Food Structure and Functionality Forum Division of the AOCS
Course Description Image analysis is an extremely valuable technique for generating data from images. The tools presented in this class can be applied to a broad range of food applications. Statistical analysis of data generated from images can be very useful in aiding product development, processing, packaging, and quality control for raw ingredients and processed foods.
This one-day short course is intended to provide a condensed guide to the process of image analysis. The class will cover the steps used to acquire, process, extract features, and make measurements from images. It will also demonstrate how image analysis can be applied to problem solving for food research, production, or quality control. Specific topics include: image acquisition, correction of image defects, detail enhancement, thresholding of image features, binary image processing, feature automation, and batch processing. Participants are invited to submit images or image analysis problems prior to the course. The images may be discussed or used in the course demonstrations. However, it may not be possible to offer a full solution due to the time constraints of a one-day short course. The class is limited to 20 participants.
For more information visit the website:
http://www.aocs.org/meetings/fsffcourse/
or contact coordinator
Diana Kittleson, General Mills Technology East
(Diana.Kittleson-at-genmills.com)
Paula M. Allan-Wojtas Research Scientist - Food Microstructure / Chercheur scientique - microstructure des aliments Food Safety and Quality Team / Salubrité et qualité des aliments Agriculture and Agri-Food Canada /Agriculture et Agroalimentaire Canada Telephone / Téléphone: 902-679-5566 Facsimile / Télécopieur: 902-679-2311 32 Main Street / 32 rue Main Kentville, Nova Scotia / Kentville (Nouvelle-Écosse) B4N 1J5 allanwojtasp-at-agr.gc.ca
Within about the next two hours (11:30 Hawaiian time, 1:30 West Coast time) I need to come up with a replacement for our digital acquisition on our FESEM. Don't you just love grant administrators?
We have a Hitachi S-800 FESEM, one of the last Hitachi SEMs without digital image acquisition. We have happily been using Printerface from GW Electronics for many years. However, it's life is limited, and I've been handed a bucket of money I can use - by noon today. I'd like to know what some of you have used on your analog SEMs (especially if you're happy).
Other places to put my money might be a new sputter coater (and I'm finally getting a new CPD).
The money can only be used for equipment over $5,000, whereas most of what I want is just under that!
Mahalo, Tina
78F degrees, sunny with a few clouds, surf coming down from 20-25 feet
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} } Dear Owen, } } please try } } http://www.mikroanalytik.de/software.phtml } } There you will find a table of most intense line energies in keV, } visible with EDX-spectrometer. In addition, critical excitation } energies and most probable line overlaps (K/L, K/M and L/M) are } presented (atomic numbers). The element names are in German (left } column) and in English (right column). } } Klick at the animated table image. Your browser will load the entire } image in JPG-format (956 KByte). Then store the image on your computer } and print it out. Use the maximum size and best resolution, available } with your printer. } } Sorry, until now the homepage is only in German. in a couple of weeks } you will find there a English version and the program to download, } from this the poster is a screen shot. } } Good luck
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hello all - have any of you any experience embedding materials (especially rocks) in sulfur for subsequent thin sectioning? Or does anyone know of good references for this technique?
Thanks!
-- {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} Rick Hugo, Ph.D. Geomicrobiology and Electron Microscopy Lab Department of Geology, Portland State University {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
Micscape Magazine online (the online magazine of Microscopy UK) has an on-line Pond Life ID kit. Here's the link - http://www.microscopy-uk.org.uk/pond/index.html
Shannan Little Electron Microscopy Technician Agriculture & Agri-Food Canada Lethbridge Research Centre
The American Chemical Society is once again offering "Applied Optical Microscopy" in conjunction with the upcoming PITTCON in Orlando Florida. This is one of the few hands-on courses remaining in the ACS curriculum and is also one of the few courses with extensive material on polarized light. Note that, while this is an offering of the Chemical Society, the course is very multi-disciplinary. Participants from all scientific disciplines are welcome.
} Dates: March 7-9, 2003 } Location: Renaissance Orlando Resort at SeaWorld, Orlando, Florida } Registration, course materials, and coffee breaks: $1345 for ACS members, $1445 for non-members } Housing and food: on your own, but PITTCON does have a housing bureau } Syllabus, course description and registration information: www.MicroscopyEducation.com } Instructors: } Barbara Foster, Microscopy/Microscopy Education, Inc. } Dr. Barry Fookes, Chemistry Dept/Criminalistics Division, University of Central Florida } Dr. Kenneth Piel, Microscopy/Microscopy Education
} Participants are encouraged to bring samples from their own work.
A verbatim testimonial from a recent student:"Doing light microscopy with taking this ACS course is like doing the laundry without a washing machine".
Best regards, Barbara Foster (organizer and principal lecturer) Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Many thanks for the recommendations. I have enough info for the budget; now I only have to cross my fingers for a couple of months. Also asking for a new CPD, sputter coater, and perhaps a backscattered electron detector.
There were enthusuastic recommendations from people using SIS's ADDA II, 4Pi, Emispec, Quartz PCI, Orion, Advanced Database Systems, and Evex. The first two got multiple "votes", and I'm sure over the course of several days there would be more. However, I've got what I need for now.
Thanks to you all!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I've been doing some EM immuno's lately and getting funky, non-specific results. My negative controls come out clean, but the immunos are so random I'm doubting myself. What I'd like to do is run a known positive control on human cell lines. I'd like something that stains some part of the cell but not all over the cell.
Suggestions?
Anything you give me is gratefully accepted and I thank you in advance, or TIA for the instant message crowd ;-)
Drowing my sorrows in cold water fish gelatin,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We have a JEOL 6301 SEM, and we use a PC with a Flashbus card (Integral Technologies, Inc) to capture digital images. The card comes with a variety of input cable types in order to accomodate the many different types of video output ports (i.e., s-video, BNC, 4-pin din, RGB, etc.) found on laboratory equipment.
It's 78 degrees here too, today!
Best Regards, Tom Barbieri
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Monday, January 27, 2003 12:18 PM To: Microscopy Listserver
Hi, All-
Within about the next two hours (11:30 Hawaiian time, 1:30 West Coast time) I need to come up with a replacement for our digital acquisition on our FESEM. Don't you just love grant administrators?
We have a Hitachi S-800 FESEM, one of the last Hitachi SEMs without digital image acquisition. We have happily been using Printerface from GW Electronics for many years. However, it's life is limited, and I've been handed a bucket of money I can use - by noon today. I'd like to know what some of you have used on your analog SEMs (especially if you're happy).
Other places to put my money might be a new sputter coater (and I'm finally getting a new CPD).
The money can only be used for equipment over $5,000, whereas most of what I want is just under that!
Mahalo, Tina
78F degrees, sunny with a few clouds, surf coming down from 20-25 feet
**************************************************************************** * 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* ****************************************************************************
As memory serves me (and it seems to serve more poorly with each passing year), Polymer Microscopy, by Grubb & Sawyer, may address it. Good luck.
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Rick Hugo {hugo-at-pdx.edu} To: Microscopy List {Microscopy-at-sparc5.microscopy.com} cc: 01/27/03 02:18 PM Subject: Sulfur embedding?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello all - have any of you any experience embedding materials (especially rocks) in sulfur for subsequent thin sectioning? Or does anyone know of good references for this technique?
Thanks!
-- {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} Rick Hugo, Ph.D. Geomicrobiology and Electron Microscopy Lab Department of Geology, Portland State University {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
The original electron microprobe was built by Raymond Castaing in Paris around 1950. It had a fixed electron beam and crystal spectrometers. A commercial version was first marketed by Cameca. When I was a new graduate student in 1967, a Cameca microprobe was being installed in my department in Oxford. It, too, had a fixed beam, two or three (I don't remember which) crystal spectrometers, and an optical microscope to set the position of the sample under the beam. I don't know how the beam was aligned or focused (I never used the instrument myself, though other members of my group did).
The SEM didn't come along until later, in the early '60's, commercialized by Cambridge Instruments. As time has passed, the "microprobe" and the "SEM" have become more alike, until today microprobes can generate respectable SEM images of a sample, and the beam can be steered electrically to the position of interest. Almost all SEMs have a port on which can be mounted a crystal (or wavelength) spectrometer. The majority of both types of instruments also typically have an EDX spectrometer.
So the difference is now mainly semantics. A microprobe is an instrument optimized for stable, high beam current and with a chamber designed for mounting several wavelength spectrometers, together with an EDX detector, while an SEM is an instrument which is fundamentally the same, but optimized for generation of fine electron probes and capable of working well at lower voltages, and with a specimen chamber designed for flexibility in mounting samples.
WDX and EDX spectroscopy are two methods of getting at the energy (or its equivalent, wavelength) of an x-ray in order to determine the chemistry of a sample. Both methods have unique attributes, which is why both techniques are still in use.
Tony Garratt-Reed.
At 10:08 AM 1/28/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
Microprobe is essentially a specialized SEM with: a) several (3-5) WDS and, possibly, EDS. b) higher beam stability c) optical scope in a specimen chamber for bringing specimen in focal plane of WDS. d) staff better qualified for X-ray microanalysis (optional).
Vladimir
} } What is Electron Microprobe? How is it different from EDS or WDS = } analysis? } } Any information is greatly appreciated. } Pavel } } } } }
We have someone who is interested in using TEM to image sooty, 0.06 micrometer particles suspended in (gasp!) lube oil from an engine. Does anyone have experience examining such samples?
My inclination is to solvent-clean the soot until the oil is removed totally and then place the particles on a carbon substrate. However, the researcher is concerned that this will remove the dispersant additives, leaving the soot free to agglomerate. I agree that this may occur, but we would be able to suspend the particles (ultrasonically, perhaps) prior to deposition on the grid.
Suggestions?
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 ##############################################################
I have not worked on a JEOL tank but assume it is similar to other Japanese models. In which case the tank components are attached to the top of the tank. Unbolting and lifting the top clear of the oil (its very heavy, a crane would be of assistance) should allow the components to drip dry. The place in which you do this must be free from dust.
I would suggest you cover the floor with plastic sheeting as this is a messy task. Once the tank top has been remove from the tank the dirt oil may be disposed of through an appropriate agency. Then fresh transformer oil may be placed in the tank after it has been thoroughly cleaned. All the tank top components should be checked and where possible cleaned with a fluff free cloth. Keep this area covered with a plastic sheet at all times, do everything in your power to prevent dust and other contaminants from settling on the components.
When these tasks have been performed the tank top needs to be very very slowly lowered into the oil. Shake the tank top as it is lowered to try to encourage all air pockets to be removed. When the top is in place and bolted down, gently shake the tank for a little while, this again is to help remove air pockets.
When you switch the HT on, do so at the lowest kV and allow it to operate under these condition for at least an hour. Repeat this procedure for each kV. If you cannot carry out this procedure in one day please allow at least 15 minutes at each kV until you reach a step you have not reached before. I would carry out this procedure with the gun cable removed so that you can be sure there are no other areas of interference.
Good luck, cleanliness is the secret for success
Steve Chapman Senior Consultant Protrain EM Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: {"venkat-at-www.ccmb.res.in"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, January 27, 2003 12:01 PM
Listers,
Hi. We (Nestor Zaluzec and I) are in the process of scanning back copies of Microscopy Today into our web page.
The plan is to have downloadable copies available on the web six months after they appear in print. Free. The entire magazine is available online--advertisements and all.
You can see what the first six months of MT 2002 look like at http://www.microscopy-today.com follow the links to the back issue Table of Contents.
Our plan is to have at least the last five years available for you to download. The entire ten years can be done if there is interest.
************************* We are missing one issue. The April 2000 issue number 3. Neither Don Grimes nor I have a copy! *************************
If anyone has a copy of the April 2000 issue of Microscopy Today (Number 03) in good condition that we can borrow for a couple of weeks please respond to this note. Don, Nestor, and I will think of some nice reward for you!
Thank you!
Ron Anderson, Editor Microscopy Today microtoday-at-attglobal.net
} What is Electron Microprobe? } See http://www.geology.wisc.edu/~johnf/empa.html
Best
Keewook Yi Research Analyst / electron microscope Indiana University School of Dentistry 415 Lansing st. Indianapolis, IN 46202-2876 kyi-at-iupui.edu tel) 317-274-2598
This is the way it's been done for billions of years. Small moves, Ellie. Small moves. - from "Contact"
} ---------- } From: atcsem } Sent: Tuesday, January 28, 2003 10:08 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: What is Electron Microprobe? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } What is Electron Microprobe? How is it different from EDS or WDS = } analysis? } } Any information is greatly appreciated. } Pavel } } } } } }
This may appear some distance from what you want, and it is not always as obvious as one might desire, however, with a little work this piece of software can provide nice help in doing XRF which is what WDS is, as I understand it.
Hoe I'm right, Ritchie, 'cause here's the link:
http://users.skynet.be/xray_corner/
Regards,
Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging
Mail to: Geology, CASI West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383
For help and information only, The CASI houses: An FEI Quanta 400 and Technai 12T, Oxford INCA Energy 400, Tousimis AutoSamdri 815 and Olympus FV-300.
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Monday, January 27, 2003 2:40 PM To: Frank Eggert; Microscopy-at-sparc5.microscopy.com
Thanks, that's a great one.
Does anyone know of a wds-friendly version?
cheers
rtch
} } Dear Owen, } } please try } } http://www.mikroanalytik.de/software.phtml } } There you will find a table of most intense line energies in keV, } visible with EDX-spectrometer. In addition, critical excitation } energies and most probable line overlaps (K/L, K/M and L/M) are } presented (atomic numbers). The element names are in German (left } column) and in English (right column). } } Klick at the animated table image. Your browser will load the entire } image in JPG-format (956 KByte). Then store the image on your computer } and print it out. Use the maximum size and best resolution, available } with your printer. } } Sorry, until now the homepage is only in German. in a couple of weeks } you will find there a English version and the program to download, } from this the poster is a screen shot. } } Good luck
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
} When I was a new graduate } student in 1967, a Cameca microprobe was being installed in my department } in Oxford. It, too, had a fixed beam, two or three (I don't remember } which) crystal spectrometers,
Two
} and an optical microscope to set the position } of the sample under the beam. I don't know how the beam was aligned or } focused (I never used the instrument myself, though other members of my } group did). }
With great difficulty, the MS42 (or was the MS46 it is so long since I used it towards the end of its life) was difficult to focus, there were phosphor screens with aperatures in there centres at various points to align the beam. And I seem to remember using cathodluminenece of thoria to focus the beam.
} The SEM didn't come along until later, in the early '60's, commercialized } by Cambridge Instruments. As time has passed, the "microprobe" and the } "SEM" have become more alike, until today microprobes can generate } respectable SEM images of a sample, and the beam can be steered } electrically to the position of interest. Almost all SEMs have a port on } which can be mounted a crystal (or wavelength) spectrometer. The majority } of both types of instruments also typically have an EDX spectrometer. } } So the difference is now mainly semantics. A microprobe is an instrument } optimized for stable, high beam current and with a chamber designed for } mounting several wavelength spectrometers, together with an EDX detector, } while an SEM is an instrument which is fundamentally the same, but } optimized for generation of fine electron probes and capable of working } well at lower voltages, and with a specimen chamber designed for } flexibility in mounting samples. }
One other important feature of the microprobe is that the stages are designed to give fixed geometry between the surface of the sample and the spectrometers (fixed 0 tilt). Also modern EPMAs have large high precision computer controlled stages to make the automatic analysis of many hundreds of points possible in a day, or the acquistion of large area element maps. (Unlike the MS4? where every thing was driven by hand and I was lucky if I got the data for more than 5 slag analyses in a day - the data which then had to be taken over the road to run through the correction programs).
-- Chris Salter Department of Materials Characterisation Services, Begbroke Business and Science Park, Sandy Lane, Yarnton, Oxford OX5 1PF.
Tels 01865 283722, EPMA 283741, Home 463424, Mobile 07776031608
Electron Microprobe is simply another term for Wavelength Dispersive Spectroscopy - WDS - they are one and the same. Many in the industry refer to WDS as EPMA - Electron Probe Microanalysis.
Hope this helps.
Sincerely,
Carol Jean Hirt Vice President Materials Research Laboratories, Inc. 290 North Bridge Street Struthers, OH 44471 www.mrllab.com 800 424-1776
} } What is Electron Microprobe? How is it different from EDS or WDS = } analysis? } } Any information is greatly appreciated. } Pavel } } } } }
I use a good old tube-insertion method to observe the cross-sectional view of multi-wall carbon nanotubes. Get a tube of metal tube, insert your sample, then soak in low-viscosity glue, like Mbond. After curing, you can handle pretty easily for grinding, lapping, dimpling, and milling. If you need more strength between the tube wall and the sample in the middle, filling the empty space with thin wires would help. If you use single-modulation from substrate side while milling, it would give better results. I can get a full view of the 200-micron-thick layer using this technique even though it has visible area at the edge. Good luck,
Young-Woon Kim, Ph.D.
Research Professor School of Materials Science and Engineering, Seoul National University Tel) +82-2-880-7977 Fax) +82-2-883-8197 e-mail) ywkim-at-gong.snu.ac.kr
-----Original Message----- } From: yimin yao [mailto:yimin-at-fy.chalmers.se] Sent: Monday, January 20, 2003 1:26 AM To: Microscopy-at-sparc5.microscopy.com
Dear all,
I am going to make cross section sample of the interface between carbon nanotube film (a few hundred micrometer thick) and Si substrate for TEM. I tried the standard method to make cross section (gluing the samples with M-bond, mechanical polishing and ion milling), but the film turned to be peeled off from substrate during polishing or ion milling, due to the large thickness of film and weak bonding between the film and substrate. Any ideas and suggestions from your experience?
Best regards,
Yiming ------------------------------------ Dr. Yiming Yao
Microscopy and Microanalysis Department of Experimental Physics Chalmers University of Technology SE-41296, Göteborg Sweden
As I know, in the current language the "electron microprobe" is the "surgery knife" you are using for EDS/WDS analysis, i.e. the fine electron beam itself, to be positioned on the area to be analysed.
Am I wrong ?
Corneliu Sarbu, PhD Department of Metallurgy and Applied Materials Science (MTM Dept.) Catholic University of Leuven (KULeuven) Kasteelpark ARENBERG nr. 44 B-3001 Heverlee-Leuven, Belgium **************************************************************** Phone: +32-16-32.1241 - office +32-16-32.1264 - secretary of department Fax: +32-16-32.1992 or +32-16-32.1270 e-mail: Corneliu.Sarbu-at-mtm.kuleuven.ac.be web-site: www.mtm.kuleuven.ac.be ****************************************************************
At 28/01/03 10:08, you wrote: } Microscopy-at-sparc5.microscopy.com
I'm looking for a cheap, fast and reliable supplier (for Europe) of good holey carbon coated grids for Cryo-TEM.
Any suggestions?
Thanks everybody,
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290
One way is to use a gas reaction cell or a controlled environment TEM. I have used ours to look at soot particulate dispersion in engine oil for a commercial company although your 60nm particles might be tricky. Specimen prep was to wipe the oil across a carbon filmed grid.
Seeing your Illinois affiliation try Univ of Illinois (Iain Robertson in Prof. Birnbaum's group) at Champagne(?) for a start.
Good luck, Ron
On Tue, 28 Jan 2003 16:14:32 -0600 "John J. Bozzola" {bozzola-at-siu.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 } -----------------------------------------------------------------------. } } } We have someone who is interested in using TEM to image sooty, 0.06 } micrometer particles suspended in (gasp!) lube oil from an engine. } Does anyone have experience examining such samples? } } My inclination is to solvent-clean the soot until the oil is removed } totally and then place the particles on a carbon substrate. However, } the researcher is concerned that this will remove the dispersant } additives, leaving the soot free to agglomerate. I agree that this } may occur, but we would be able to suspend the particles } (ultrasonically, perhaps) prior to deposition on the grid. } } Suggestions? } } 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 } ############################################################## } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
We are pleased to host EMAG'03 in Oxford. 3rd to 5th September 2003.
Hello, I recently aquired a used Reichert-Jung Biocut 2030 microtome from a lab auction. I am a bit unfamiliar with this model and am seeking advice from those list members who might use one. On the left side of the instument is a small wheel or knob that is associated with a series of three coggs within the casing of the machine. From what I understand, this is the course advance knob. However, when I turn the knob either clockwise or counter clockwise the coggs only make about a quarter of a turn. The course advance knob is held in place by two flexible pieces of metal and can thus be turned further without advancing the coggs. I have no manual so I am not sure if there is something that I should be doing make it work properly. The microtome got a good bang during shipping and I am not sure if it is damaged. I replaced the mounting bolts for the metal piece to which the coggs are mounted and everything looks alright to me. From what I can tell the fine advance is working OKay and the sections come out alright. Thanks for any enlightenment. Sincerely, Jonathan
Jonathan Wilson (PhD) CIIMAR Rua do Campo Alegre 823 4150-180 Porto, Portugal tel: 351 22 608 0470 / 71 fax: 351 22 608 0479 e-mail: wilson_jm-at-cimar.org web: www.cimar.org
I am really happy with those from Ted Pella. You can send an e-mail directly to Christel at sales-at-tedpella.com or have a look on the online catalog www.tedpella.com Danièle
-------------------------------------------- Danièle Spehner Head of the EM Department INSERM EPI 03 45 - EFS-Alsace 10 rue Spielmann - 67065 STRASBOURG tel : 03 88 21 25 25 - fax : 03 88 21 25 44 e-mail : daniele.spehner-at-efs-alsace.fr ------------------------------------
-----Message d'origine----- De : Philip Koeck [mailto:Philip.Koeck-at-biosci.ki.se] Envoyé : mercredi 29 janvier 2003 11:05 À : Microscopy-at-sparc5.microscopy.com Objet : TEM: holey carbon grids
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I'm looking for a cheap, fast and reliable supplier (for Europe) of good holey carbon coated grids for Cryo-TEM.
Any suggestions?
Thanks everybody,
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290
A recent mention of the double-stick carbon adhesive tabs is found in K. B. Bolte's Technique for Obtaining Scanning Electron Micrographs of Minute Athropods, Proc Ent Soc Ont, 127:67-87, 1996.
Bruce F. Ingber, Biologist Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
} } } "Garber, Charles A." {cgarber-at-2spi.com} 01/24/03 07:43AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
The first in the world to market a product one might call (generically) a double sided adhesive conductive carbon filled tape was a company in Japan called Oken Shoji. This would have been in the early 1980's. The owner of that firm was and still is Mr. Hisashi Sato. He also applied for and received a Japanese patent for the tape at that time. Apparently it covers only tape cut 8 mm wide and tapes either larger or smaller than that width somehow don't infringe. I know that sounds a bit irrational but that is the way it has been explained to me.
I have also been told that there is another name on the patent, that of a professor in Japan. I believe that the original user (inventor) of the tape for this application was that professor, who happened to know Mr. Sato and then it was Mr. Sato who actually commercialized the product. It is of course all in Japanese so this information comes to me second-hand.
In any case, the tape was widely used in Japanese SEM laboratories before it was even known here in the USA or in Europe. It was the Japanese service engineers who brought samples of the tape to the USA from Japan and when the US employees of the Japanese microscope firms went to Japan for training, they brought samples back with them as well. It was my further recollection that the first SEM manufacturer to be encouraging their customers to use the carbon tape as a means of promoting column cleanliness was ISI/Topcon.
SPI Supplies was introduced to the tape by the ISI people during that time frame and almost simultaneously, the tape was introduced in the USA and Canadian markets by both SPI Supplies and E. F. Fullam, Inc. Some time later after that, the tape was then offered by Ted Pella, Inc., Electron Microscopy Sciences, Inc. and later on by Energy Beam Sciences, Inc. In Europe at that time (early 1980's) the tape was offered by Polaron Equipment Ltd. and Agar Scientific Ltd.
Today, the tape is not all the same. Some is on a white plastic core and some is on a cardboard core. The tape on the cardboard core to the best of my knowledge has direct traceability to the original tape that has been produced since the early 1980's. Anyone who has used this tape knows very well how the cardboard core starts to disintegrate and particulate, hardly a good thing for EM lab cleanliness. The newest version of this tape has a white plastic core that does not shed particulates. For more information see URL http://www.2spi.com/catalog/spec_prep/cond_adhes-tapes.shtml
Today, one can use this kind of product not only in tape form but also in die cut discs and sheets. It is also available as a double sided adhesive silver filled sheet. All of these products trace their progeny back to the original carbon tape concept and patent.
Disclaimer: SPI Supplies offers all of the mentioned products so we have a vested interest in promoting their use.
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 ######################## ============================================
would 1% sucrose be a problem in frozen hydrated specimens (single particles)? If so is it necessary to dyalize it out or is it sufficient to wash it of the grid before plunge-freezing?
Thanks in advance,
Philip
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290
Rick - We have worked on embedding particles in sulfur for some time now. I have always thought to present this technique at MSA but just haven't gotten around to it yet. To the best of my knowledge, the idea to embed in sulfur was conceived by our colleague, John Bradley. Perhaps others have done so as well. We developed our own set of sulfur-embedding techniques and methods suitable to our particles. Embedding materials in sulfur is a wonderful technique to look at microtomed sections since sulfur will sublimate in vacuum and you can be left with a section on a TEM grid that is free of embedding material. We study interplanetary dust particles (IDPs - typically 5 - 10 um in size) which are composed of 1000's of mineral grains, glass and carbonaceous phases, including organic materials. These particles are often highly porous and the normal infiltrating epoxy resins have always made it difficult for us to determine the carbon compounds vs the epoxy,thus our need for this technique (or other carbon-free embedding methods).
Sulfur is a very difficult material to work with and requires fairly extensive experience and practice due to some unusual properties. For instance, liquid sulfur is one of the rare materials (the only?) whose viscosity goes up as you heat it to higher temperatures (at least up to about 190 C); this is due to increasing polymerization of sulfur ions in the melt with increasing temperature. Our technique of sulfur embedding is to place a particle (say a 10 um interplanetary dust particle) in the middle of a clean glass slide. On a second clean glass slide we heat a tiny crystal of ultra-pure sulfur above its melting point of 119 C. The sulfur will melt and after removal of the heat will supercool forming a liquid drop on the glass slide. I like to have the drop diameter 50 - 100 um or less, otherwise the liquid sulfur may spontaneously crystallize. While observing under a good binocular microscope, the particle on the clean glass slide is lowered into the sulfur drop which will wet both glass slides. When the two glass slides are separated the particle on the first glass slide will have a hemispherical liquid drop around it. At this point you need to crystallize the supercooled sulfur which surrounds the particle by touching a small sulfur crystal to the drop which will create nucleation sites and initiate crystallization. There are other ways to do this besides what I have just described. Unfortunately, fractures may develop in the sulfur during crystallization which can lead to contamination and other problems later. Fracturing of the crystallized sulfur is a problem with this technique that we haven't found a satisfactory solution to yet.
The particle in solid sulfur is then mounted with epoxy to a pre-cut epoxy cylinder which fits the chuck of the microtome. The sulfur mount must first be dislodged from the slide as the crystallized sulfur will want to stick to the glass. It can then be trimmed and microtomed in the usual way but you have to be careful as microtoming is more difficult than with epoxy resins due to the softness and fragile nature of crystallized sulfur but it can be done with practice. For us, this works best on small samples (in the 10 - 20 um range). Another important factor is that many epoxies will dissolve the sulfur while curing thus you need to experiment to determine which ones are suitable to hold your sulfur mount. I have found one or two that are suitable for our needs. Superglue can also be used.
I haven't provided all the details of our sulfur embedding technique here and there are more pitfalls than I have described. The technique is difficult and somewhat unpredictable, but if everything goes right you can end up with nice microtomed sections free of any embedding material. This is useful for observing or measuring carbon by EDX if you mount your sample on a TEM grid with an SiO film (have to watch out for carbon contamination in the microscope, though). I can imagine that there are other, perhaps better ways to mount materials in sulfur than I have described; this is just a technique that has evolved over the last several years for us.
Dave
Dave Joswiak University of Washington Dept. of Astronomy, 351580 Seattle, WA 98195 (206)543-7702 joswiak-at-astro.washington.edu
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I have used Agar and found them excellent, but not lately. John Mardinly Phone: 408-765-2346 Pager: 877-277-1182
-----Original Message----- } From: Philip Koeck [mailto:Philip.Koeck-at-biosci.ki.se] Sent: Wednesday, January 29, 2003 2:05 AM To: Microscopy-at-sparc5.microscopy.com
I'm looking for a cheap, fast and reliable supplier (for Europe) of good holey carbon coated grids for Cryo-TEM.
Any suggestions?
Thanks everybody,
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290
Dear List, I have a problem with my Microanalysis: sometimes (often) I have counts when the Beam is off. But not two or three counts ... thousands of counts for several days. Normally I try to empty and clean the Dewar to be sure I have not frozen particles of powder inside, but the problem remains. So I call Philips technicians, they make tests on the electronic parts and send the detector to Germany because they cannot find the problem. Ok, I wait one month and when the detector returns it is ok, but nobody know what the problem was. The problem has been present for 4 years and now it occurs two - three times per year. For the ventilation of the chamber I use the air to be sure I have not high pressure in the chamber wich may damage the thin window of the detector. My detector is EDAX and my SEM is PHILIPS XL 20. Please help me! Thanks.
Thanks Doug- Great tips- I'm printing them up until I memorize them. Rgds, Mike Shaw Roselle, NJ
} Gary, } } MS Word is such a pain because of the way it works with images. A couple } of tricks I've discovered over the years. } } 1) Place the image within a text box, rather than directly on the page, it } seems to give you much greater control over the location on the page. It } also makes it much easier to add text annotations that stay with the image. } } } If you don't want the text box to have a border you can remove it by selecting } the box outline and look for the "paint brush" icon on the Draw toolbar, } then use the down arrow and select "none". If you'd like the text box to } be transparent, select the text box outline and look for the "paint bucket" } icon on the Draw toolbar, then use the down arrow and select "none". If } you discover its hard to find the text box border once you made the edge } "invisible", first select the image with a single left click (it should have } the solid black resizing "handles") and then use the keyboard left or right } arrow keys and the selection with move out to the textbox outline (with black } bordered resizing "handles"). } } 2) Always use the INSERT | PICTURE | FROM FILE option as opposed to copying } and pasting an image into Word. I find that the images are harder to work } with if I paste them in. Also, you can insert TIFF or BMP } images into Word, } you don't have to use JPEG.
Dear Jonathan, have a Reichert ultracutE , I think the knob you are referring is the same this instrument has. This is the advance wheel for semithick sectioning. You move it clockwise it will advance 0.5 micron upto first cog, the next is for 1 micron and the last is for 2 microns. Shashi Singh Scientist, CCMB Hyderabad INDIA ----------------. Hello, I recently aquired a used Reichert-Jung Biocut 2030 microtome from a lab auction. I am a bit unfamiliar with this model and am seeking advice from those list members who might use one. On the left side of the instument is a small wheel or knob that is associated with a series of three coggs within the casing of the machine. From what I understand, this is the course advance knob. However, when I turn the knob either clockwise or counter clockwise the coggs only make about a quarter of a turn. The course advance knob is held in place by two flexible pieces of metal and can thus be turned further without advancing the coggs. I have no manual so I am not sure if there is something that I should be doing make it work properly. The microtome got a good bang during shipping and I am not sure if it is damaged. I replaced the mounting bolts for the metal piece to which the coggs are mounted and everything looks alright to me. } From what I can tell the fine advance is working OKay and the sections come out alright. Thanks for any enlightenment. Sincerely, Jonathan
Jonathan Wilson (PhD) CIIMAR Rua do Campo Alegre 823 4150-180 Porto, Portugal tel: 351 22 608 0470 / 71 fax: 351 22 608 0479 e-mail: wilson_jm-at-cimar.org web: www.cimar.org
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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} } ----- Original Message ----- } From: "Dr. Brendon Price" {priceb-at-uctbc1.uct.ac.za} } To: "Paula Sicurello" {patpxs-at-gwumc.edu} } Sent: Wednesday, January 29, 2003 9:50 AM } Subject: Re: Positive control } } } } Hi Paula } } } } In order to try and offer some advice, could you provide information on } the } } following: } } } } 1. What species was the antibody raised in and what } protein/ligand/compound } } was it raised against? } } 2. Have you cleaned up the antibody preparation by immunoaffinity or PEG } 6 } } kDa precipitation? } } 3. Do you get similar non-specific binding patterns in your Western } blots. } } What blocking buffers are you using? } } 4. What type of TEM immunolabelling are you doing? Cryo, resin? If resin, } } is it LR White or the Lowicryls? What fixation procedure are you using? } } 5. What concentrations of primary antibody are you using? } } 6. are you using a gold-conjugated linker antibody or [if your primary } } antibody is from rabbits] protein A gold? } } 7. When you mention that the labelling is random, do you mean that you } get } } labelling randomly throughout the cell, or that the labelling density is } } highly variable? If the density is variable, is the protein/compound } } expressed at different stages during the cell cycle? Perhaps it is } } asymmetrically distributed within the cell and this may result in variable } } labelling densities. } } } } The most important bit of information I need is the labelling pattern of } } your Western blots on reducing SDS-PAGE gels. If these are clean [i.e. } only } } band(s) at the anticipated molecular weight], then we can proceed further } in } } the troubleshooting. } } } } Regards } } } } Brendon } } ********************* } } Dr. Brendon Price } } Chief Scientific Officer } } Electron Microscope Unit } } University of Cape Town } } Private Bag } } Rondebosch } } 7701 } } Tel: +27 21 650-2819 } } Fax: + 27 21 689-1528 } } ----- Original Message ----- } } From: "Paula Sicurello" {patpxs-at-gwumc.edu} } } To: {microscopy-at-sparc5.microscopy.com} } } Sent: Tuesday, January 28, 2003 4:20 PM } } Subject: Positive control } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hello Listers, } } } } } } I've been doing some EM immuno's lately and getting funky, non-specific } } } results. My negative controls come out clean, but the immunos are so } } } random I'm doubting myself. What I'd like to do is run a known positive } } } control on human cell lines. I'd like something that stains some part } } } of the cell but not all over the cell. } } } } } } Suggestions? } } } } } } Anything you give me is gratefully accepted and I thank you in advance, } } } or TIA for the instant message crowd ;-) } } } } } } Drowing my sorrows in cold water fish gelatin, } } } } } } Paula :-) } } } } } } Paula Sicurello } } } George Washington Univ. Medical Center } } } Dept. of Pathology, Ross Hall rm 505 } } } Electron Microscope Lab } } } 2300 Eye St. } } } Washington, DC 20037 } } } 202-994-2930 phone } } } 202-994-2518 fax } } } } } } } } }
It's 8 degrees Fahrenheit here in New Jersey. Try Quartz PCI as a capture device for slow scan SEM image capture.
Peter
-----Original Message----- } From: Barbieri Thomas-r53545 [mailto:Thomas.Barbieri-at-motorola.com] Sent: Tuesday, January 28, 2003 10:31 AM To: 'Tina Carvalho'; Microscopy Listserver
Hello Tina,
We have a JEOL 6301 SEM, and we use a PC with a Flashbus card (Integral Technologies, Inc) to capture digital images. The card comes with a variety of input cable types in order to accomodate the many different types of video output ports (i.e., s-video, BNC, 4-pin din, RGB, etc.) found on laboratory equipment.
It's 78 degrees here too, today!
Best Regards, Tom Barbieri
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Monday, January 27, 2003 12:18 PM To: Microscopy Listserver
Hi, All-
Within about the next two hours (11:30 Hawaiian time, 1:30 West Coast time) I need to come up with a replacement for our digital acquisition on our FESEM. Don't you just love grant administrators?
We have a Hitachi S-800 FESEM, one of the last Hitachi SEMs without digital image acquisition. We have happily been using Printerface from GW Electronics for many years. However, it's life is limited, and I've been handed a bucket of money I can use - by noon today. I'd like to know what some of you have used on your analog SEMs (especially if you're happy).
Other places to put my money might be a new sputter coater (and I'm finally getting a new CPD).
The money can only be used for equipment over $5,000, whereas most of what I want is just under that!
Mahalo, Tina
78F degrees, sunny with a few clouds, surf coming down from 20-25 feet
**************************************************************************** * 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 had a problem like this. Turned out to be caused by the infra-red leds of my chamber camera. When the camera is switched off, the counts (3-5k per second) return close to zero.
Chris
----- Original Message ----- } From: "by way of MicroscopyListserver" {Stefano_Maretti-at-sacmi.it} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 30, 2003 2:10 AM
Here is a mess of language. According to the rules of science language "microprobe" is a composite word, from "micro = of a micrometer dimension" and "probe", i.e. "microprobe = a probe of micrometer dimension". To call "Microprobe" an instrument (notice the uppercase) means "instrument = a probe of micrometer dimensions". So, finally, we get to the same point: microprobe is the instrument of micrometer dimensions we use in order to perform X-rays EDS/WDS analysis. That "instrument" is the electron beam itself, as the "whole" instrument is not of micrometer dimensions at all.
Am I right ?
Corneliu
At 29/01/03 09:48, you wrote: } I think, you are right. } } A Microprobe is an instrument that is very similar to an SEM, except that it } usually is not optimized for imaging (you can do that, too), but for } elemental analysis using WDX or EDX specroscopy. In many cases these } instruments are computer controlled to allow the automatic movement of the } stage to collect reference spectra under identical conditions, and more } emphasis is placed on beam current and stability. } } The "probe" in microprobe is of course the electron beam, which can be } focused on microscopical areas (hence the name). } } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 12596 West Bayaud Avenue } Suite 300 } Lakewood, CO 80228 } =================================== } 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: corneliu sarbu [mailto:corneliu.sarbu-at-mtm.kuleuven.ac.be] } Sent: Wednesday, January 29, 2003 9:27 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: What is Electron Microprobe? } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America To } Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line } Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } As I know, in the current language the "electron microprobe" is the "surgery } knife" you are using for EDS/WDS analysis, i.e. the fine electron beam } itself, to be positioned on the area to be analysed. } } Am I wrong ? } } Corneliu Sarbu, PhD } Department of Metallurgy and Applied Materials Science (MTM Dept.) Catholic } University of Leuven (KULeuven) Kasteelpark ARENBERG nr. 44 B-3001 } Heverlee-Leuven, Belgium } **************************************************************** } Phone: +32-16-32.1241 - office } +32-16-32.1264 - secretary of department } Fax: +32-16-32.1992 or +32-16-32.1270 } e-mail: Corneliu.Sarbu-at-mtm.kuleuven.ac.be } web-site: www.mtm.kuleuven.ac.be } **************************************************************** } } At 28/01/03 10:08, you wrote: } } Microscopy-at-sparc5.microscopy.com
Corneliu Sarbu, PhD Department of Metallurgy and Applied Materials Science (MTM Dept.) Catholic University of Leuven (KULeuven) Kasteelpark ARENBERG nr. 44 B-3001 Heverlee-Leuven, Belgium **************************************************************** Phone: +32-16-32.1241 - office +32-16-32.1264 - secretary of department Fax: +32-16-32.1992 or +32-16-32.1270 e-mail: Corneliu.Sarbu-at-mtm.kuleuven.ac.be web-site: www.mtm.kuleuven.ac.be ****************************************************************
We are able to supply reliable holey carbon films on copper, nickel or gold grids with reasonable delivery. Please contact our Swedish dealer Analytical Standards AB on the following:
Tel 031 880 810 Fax 031 880 886
They should be able to supply a copy of our Microscopy catalogue on CD ROM. Any problems please contact me direct by e-mail at: } Terry.Cooper-at-btinternet.com
Best regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, England Tel ++44 (0)118 981 7775 Fax ++44 (0)118 981 7881 e-mail sales-at-taab.co.uk } www.taab.co.uk
I can share a recent example of how the file size in Word is a huge function of how the images are imported into it.
A customer sent me a three page memo with 4 of my SEM images in it for my review. The original image files were all jpgs about 150 kB in size. The file he sent me was 4.5MB! By clogging up the email system, it's a pain to work with that large a file. I tried a simple test by deleting the four images from the memo and using the Insert, Picture, From File command to put the same four images into Word. I then used the Format Picture menu to resize the images down to fit side by side in the memo as before and saved the file. The new file size was only 550 kB! Not tiny, but more in line with what I would expect.
I informed the customer about this technique and he admitted to simply cut and pasting them into Word. That is a sure recipe for making huge files unnecessarily. It's kind of curious that there is such a large difference, but I have seen it over and over.
Richard Shalvoy Arch Chemicals, Inc. 350 Knotter Drive Cheshire, CT 06410 (203) 271-4394 rbshalvoy-at-archchemicals.com
-----Original Message----- } From: "Microshaw-at-aol.com"-at-sparc5.microscopy.com [mailto:"Microshaw-at-aol.com"-at-sparc5.microscopy.com] Sent: Wednesday, January 29, 2003 9:42 PM To: cromey-at-Arizona.edu; Microscopy-at-sparc5.microscopy.com
Thanks Doug- Great tips- I'm printing them up until I memorize them. Rgds, Mike Shaw Roselle, NJ
} Gary, } } MS Word is such a pain because of the way it works with images. A couple } of tricks I've discovered over the years. } } 1) Place the image within a text box, rather than directly on the page, it } seems to give you much greater control over the location on the page. It } also makes it much easier to add text annotations that stay with the image. } } } If you don't want the text box to have a border you can remove it by selecting } the box outline and look for the "paint brush" icon on the Draw toolbar, } then use the down arrow and select "none". If you'd like the text box to } be transparent, select the text box outline and look for the "paint bucket" } icon on the Draw toolbar, then use the down arrow and select "none". If } you discover its hard to find the text box border once you made the edge } "invisible", first select the image with a single left click (it should have } the solid black resizing "handles") and then use the keyboard left or right } arrow keys and the selection with move out to the textbox outline (with black } bordered resizing "handles"). } } 2) Always use the INSERT | PICTURE | FROM FILE option as opposed to copying } and pasting an image into Word. I find that the images are harder to work } with if I paste them in. Also, you can insert TIFF or BMP } images into Word, } you don't have to use JPEG.
There is an excellent paper, part of which looks at the soot with the TEM: SAE # 2002-01-1672
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. www.cst.cmich.edu/users/willi1gl/BDMF/BDMF.htm
} -----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] } Sent: Tuesday, January 28, 2003 5:15 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM imaging of sooty oil specimen } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have someone who is interested in using TEM to image sooty, 0.06 } micrometer particles suspended in (gasp!) lube oil from an engine. } Does anyone have experience examining such samples? } } My inclination is to solvent-clean the soot until the oil is removed } totally and then place the particles on a carbon substrate. However, } the researcher is concerned that this will remove the dispersant } additives, leaving the soot free to agglomerate. I agree that this } may occur, but we would be able to suspend the particles } (ultrasonically, perhaps) prior to deposition on the grid. } } Suggestions? } } 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 } ##############################################################
Philip, I came exactly to the same conclusion and since I use Ted Pella's I have no problem. .. and I do not know for your country but in France, shipment included is Ted Pella really sheaper than Agar. Another important point is that you can negotiate the prices for big orders and discuss with them whenever you have to. I have no interest in this compagny, I am a just a very satisfied customer. Danièle
-----Message d'origine----- De : Mardinly, John [mailto:john.mardinly-at-intel.com] Envoyé : mercredi 29 janvier 2003 22:57 À : Philip Koeck; Microscopy-at-sparc5.microscopy.com Objet : RE: holey carbon grids
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I have used Agar and found them excellent, but not lately. John Mardinly Phone: 408-765-2346 Pager: 877-277-1182
-----Original Message----- } From: Philip Koeck [mailto:Philip.Koeck-at-biosci.ki.se] Sent: Wednesday, January 29, 2003 2:05 AM To: Microscopy-at-sparc5.microscopy.com
I'm looking for a cheap, fast and reliable supplier (for Europe) of good holey carbon coated grids for Cryo-TEM.
Any suggestions?
Thanks everybody,
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290
Since we are planning to implement a new ZEISS 510 confocal microscope in our lab, I would like to receive some comments from confocal users having already practical experience with the LSM 510 META. Is the method of emission fingerprinting and the linear unmixing program really as useful as it sounds to discriminate overlapping signals?
Sincerely yours,
Gilbert Engler -- ================================================================== Gilbert Engler DEPARTMENT OF PLANT SYSTEMS BIOLOGY Fax:32 (0)9 2645349 GHENT UNIVERSITY, K. L. Ledeganckstraat 35, B-9000 Gent, Belgium Vlaams Instituut voor Biotechnologie VIB mailto:gieng-at-gengenp.rug.ac.be http://www.psb.rug.ac.be ==================================================================
Chris, your system must be better than ours {g} - when we turn on our infra-red chamber scope, we get tens of thousands of counts and totally swamp our detector. It is definitely necessary to make sure the chamber scope is turned off.
Stefano, I wonder if your problem might also lie with the amplifier discriminators, particularly the fast discriminator. EDS systems deal with very small pulses so that the signal threshold needs to be set just above the noise level. If that threshold has shifted, you could very well find yourself detecting noise.
What sort of spectrum do you find under these circumstances? I would expect that you would see a substantial peak at the left side of the spectrum. If it is a signal from acoustic vibrations or from the infra-red camera, I would expect the signal to tail off to the right for many channels. If it is a problem due to the discriminator setting, I would expect only the first channel or two to have counts. You would then need to adjust your amplifier.
Unfortunately, I have no idea how the EDAX amps are adjusted. I have seen amps with trim pots that control the discriminators and I have seen ones where the controls are done through software. But EDAX should be able to tell you the proper procedure, and it should be described in the setup or calibration section of your manual.
Good luck. Warren
At 10:03 AM 1/30/03 +0000, you wrote:
} I had a problem like this. } Turned out to be caused by the infra-red leds of my chamber camera. } When the camera is switched off, the counts (3-5k per second) return } close to zero. } } Chris } } ----- Original Message ----- } } From: "by way of MicroscopyListserver" {Stefano_Maretti-at-sacmi.it} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Thursday, January 30, 2003 2:10 AM } Subject: EDAX: Counts with Electron-Beam off } } } } } Dear List, I have a problem with my Microanalysis: sometimes (often) } I have } } counts when the Beam is off. But not two or three counts ... } thousands of } } counts for several days. Normally I try to empty and clean the Dewar } to be } } sure I have not frozen particles of powder inside, but the problem } remains. } } So I call Philips technicians, they make tests on the electronic } parts and } } send the detector to Germany because they cannot find the problem. } Ok, I } } wait one month and when the detector returns it is ok, but nobody } know what } } the problem was. The problem has been present for 4 years and now it } occurs } } two - three times per year. For the ventilation of the chamber I use } the } } air to be sure I have not high pressure in the chamber wich may } damage the } } thin window of the detector. My detector is EDAX and my SEM is } PHILIPS XL } } 20. } } Please help me! Thanks. } } } } Stefano Maretti } } R&D CENTRE } } SACMI Imola - Italy - } }
I believe part of the problem has to do with how MS handles the cut and paste. If the images are copied, they are likely placed on the clipboard according to your display setup. Windows has prepared an image at whatever color depth you are using. It bothers me to see a black and white image turn into a 24-bit color image just because that is what my display is set for.
I think new versions of Windows and Word or Excel may have improved their handling of such images. I know I used to have the same problem when clients would paste their XRD patterns into Word as a bitmap, but now it doesn't seem to be such a problem. It seems MS might now be doing some compression of their own.
Either way, your suggestion to insert the picture directly from the JPG file is definitely the better way to go.
Warren
At 07:22 AM 1/30/03 -0600, you wrote:
} I can share a recent example of how the file size in Word is a huge function } of how the images are imported into it. } } A customer sent me a three page memo with 4 of my SEM images in it for my } review. The original image files were all jpgs about 150 kB in size. The } file he sent me was 4.5MB! By clogging up the email system, it's a pain to } work with that large a file. I tried a simple test by deleting the four } images from the memo and using the Insert, Picture, From File command to put } the same four images into Word. I then used the Format Picture menu to } resize the images down to fit side by side in the memo as before and saved } the file. The new file size was only 550 kB! Not tiny, but more in line } with what I would expect. } } I informed the customer about this technique and he admitted to simply cut } and pasting them into Word. That is a sure recipe for making huge files } unnecessarily. It's kind of curious that there is such a large difference, } but I have seen it over and over. } } Richard Shalvoy } Arch Chemicals, Inc. } 350 Knotter Drive } Cheshire, CT 06410 } (203) 271-4394 } rbshalvoy-at-archchemicals.com
Two things immediately come to mind. First, if there is an "in chamber" I-R camera it must be turned off or you'll get exactly what you are seeing, several thousand cps with no beam. It will also intefere with some backscattered detectors. Second, be sure you have no coupled vibration into the detector from hoses, machinery, chillers etc.
Peter Tomic Anadigics, Inc.
-----Original Message----- } From: Stefano_Maretti-at-sacmi.it [mailto:Stefano_Maretti-at-sacmi.it] Sent: Wednesday, January 29, 2003 9:11 PM To: Microscopy-at-sparc5.microscopy.com
Dear List, I have a problem with my Microanalysis: sometimes (often) I have counts when the Beam is off. But not two or three counts ... thousands of counts for several days. Normally I try to empty and clean the Dewar to be sure I have not frozen particles of powder inside, but the problem remains. So I call Philips technicians, they make tests on the electronic parts and send the detector to Germany because they cannot find the problem. Ok, I wait one month and when the detector returns it is ok, but nobody know what the problem was. The problem has been present for 4 years and now it occurs two - three times per year. For the ventilation of the chamber I use the air to be sure I have not high pressure in the chamber wich may damage the thin window of the detector. My detector is EDAX and my SEM is PHILIPS XL 20. Please help me! Thanks.
Jpeg are compressed files and inserting them into MSWord would uncompress the files. So it's better to send the files separately as attachments rather than in the document.
Pavel
----- Original Message ----- } From: "Shalvoy, Richard B **CHES" {RBShalvoy-at-archchemicals.com} To: {"Microshaw-at-aol.com"-at-sparc5.microscopy.com} ; {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 30, 2003 8:22 AM
Hello,
I hope the weather is okay in beautiful Italy.
My first approach, after all you have already done, would be to get a Geiger Counter and check the area for any radioactivity. Also, if you are using xray diffraction equipment nearby, see if the mysterious counts correspond to when that equipment is energized.
Good luck.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. 1807 West Slaughter Lane, Number 200-499 Austin, Texas 78748-6200 Phone 512/282-5507 FAX 512/280-0702
Sustaining Member - MICROSCOPY SOCIETY OF AMERICA ----- Original Message ----- } From: "by way of MicroscopyListserver" {Stefano_Maretti-at-sacmi.it} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 29, 2003 8:10 PM
Dr, Sarbu;
You may be confusing an "electron microprobe" with an FIB [Focused Ion Beam] which is in fact like a surgical knife in some respects in that it removes material.
Peter Tomic Anadigics
-----Original Message----- } From: corneliu sarbu [mailto:corneliu.sarbu-at-mtm.kuleuven.ac.be] Sent: Wednesday, January 29, 2003 3:27 AM To: Microscopy-at-sparc5.microscopy.com
As I know, in the current language the "electron microprobe" is the "surgery knife" you are using for EDS/WDS analysis, i.e. the fine electron beam itself, to be positioned on the area to be analysed.
Am I wrong ?
Corneliu Sarbu, PhD Department of Metallurgy and Applied Materials Science (MTM Dept.) Catholic University of Leuven (KULeuven) Kasteelpark ARENBERG nr. 44 B-3001 Heverlee-Leuven, Belgium **************************************************************** Phone: +32-16-32.1241 - office +32-16-32.1264 - secretary of department Fax: +32-16-32.1992 or +32-16-32.1270 e-mail: Corneliu.Sarbu-at-mtm.kuleuven.ac.be web-site: www.mtm.kuleuven.ac.be ****************************************************************
At 28/01/03 10:08, you wrote: } Microscopy-at-sparc5.microscopy.com
Now I have to ask if this can all be distilled into one general statement. Would not a SEM, no matter what the detection system is [EDS, WDS] be generally called an electron microprobe? In my mind it's any instrument that uses an accelerated electron striking a sample to generate quantitative or qualitative data about the specimen.
Peter
-----Original Message----- } From: Dusevich, Vladimir [mailto:dusevichv-at-umkc.edu] Sent: Tuesday, January 28, 2003 3:56 PM To: Microscopy-at-sparc5.microscopy.com
Microprobe is essentially a specialized SEM with: a) several (3-5) WDS and, possibly, EDS. b) higher beam stability c) optical scope in a specimen chamber for bringing specimen in focal plane of WDS. d) staff better qualified for X-ray microanalysis (optional).
Vladimir
} } What is Electron Microprobe? How is it different from EDS or WDS = } analysis? } } Any information is greatly appreciated. } Pavel } } } } }
Dear Stefano, I have had this problem in the past. It turned out to be a ground loop fault on the cover of the pre-amp on the detector. The aluminum of the pre-amp box had oxidized and was no longer providing the proper electrical ground. This can also happen if the detector touches something inside the SEM chamber or if you get a ground loop between the EDX and SEM. You really should have it checked in your lab, set up on your SEM. Regards, Mary ----- Original Message ----- } From: "by way of MicroscopyListserver" {Stefano_Maretti-at-sacmi.it} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 29, 2003 6:10 PM
Thanks to all. You are very kind.
David, Alex (The weather is good: cold and sunny), Mike, Randy, Paul J., Chuck, Jim and Chris: I have not IR camera or light inside or near my chamber. I have not radioactive samples around and I have the problem also when the chamber is empty.
Russ: My EDAX system is DX-4 and I have a pre - Sapphire detector (I think).
Peter: Yes, I always have the backscattered detector installed (do you think it is the cause of my problem?) and perhaps pre-vacuum pump gives bad vibration.
Paul: I want to verify your opinion but I need a bit time. Thank you.
Dear Stefano, I have had this problem in the past. It turned out to be a ground loop fault on the cover of the pre-amp on the detector. The aluminum of the pre-amp box had oxidized and was no longer providing the proper electrical ground. This can also happen if the detector touches something inside the SEM chamber or if you get a ground loop between the EDX and SEM. You really should have it checked in your lab, set up on your SEM. Regards, Mary ----- Original Message ----- } From: "by way of MicroscopyListserver" {Stefano_Maretti-at-sacmi.it} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 29, 2003 6:10 PM
Dear Corneliu, Microprobe is a shorthand word for "Electron Probe Micro Analyser", which is the proper name of the instrument. It is the analyser which is the instrument. Mary ----- Original Message ----- } From: "corneliu sarbu" {corneliu.sarbu-at-mtm.kuleuven.ac.be} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 30, 2003 2:49 AM
Corneliu,
One big mistake that a person can make is trying to systematically make sense of the English language, even when dealing with scientific words.
Another one that will throw you for a loop is the FIB (Focused Ion Beam). The term that should be reserved for the small probe of gallium atoms has been given to the entire instrument. Of course the better term of "ion mill" had already been assigned to something else.
On the lighter side, English is the language that has us going home, going TO church, and going TO A movie, and going TO THE store. Explain that one. Also, feet smell and noses run.
Don't try to figure it out, just accept it!
Jeff Oakley
-----Original Message----- } From: corneliu sarbu [mailto:corneliu.sarbu-at-mtm.kuleuven.ac.be] Sent: Thursday, January 30, 2003 4:50 AM To: Microscopy-at-sparc5.microscopy.com
Here is a mess of language. According to the rules of science language "microprobe" is a composite word, from "micro = of a micrometer dimension" and "probe", i.e. "microprobe = a probe of micrometer dimension". To call "Microprobe" an instrument (notice the uppercase) means "instrument = a probe of micrometer dimensions". So, finally, we get to the same point: microprobe is the instrument of micrometer dimensions we use in order to perform X-rays EDS/WDS analysis. That "instrument" is the electron beam itself, as the "whole" instrument is not of micrometer dimensions at all.
Am I right ?
Corneliu
At 29/01/03 09:48, you wrote: } I think, you are right. } } A Microprobe is an instrument that is very similar to an SEM, except that it } usually is not optimized for imaging (you can do that, too), but for } elemental analysis using WDX or EDX specroscopy. In many cases these } instruments are computer controlled to allow the automatic movement of the } stage to collect reference spectra under identical conditions, and more } emphasis is placed on beam current and stability. } } The "probe" in microprobe is of course the electron beam, which can be } focused on microscopical areas (hence the name). } } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 12596 West Bayaud Avenue } Suite 300 } Lakewood, CO 80228 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } ===================================
} } I had a problem like this. } Turned out to be caused by the infra-red leds of my chamber camera. } When the camera is switched off, the counts (3-5k per second) return } close to zero. } } Chris }
What would be the mechanism of this effect, I wonder?
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Another trick is to put the images and text into a Word table, if you size the cells in the table to about the size you want, the images will insert to fit the cell size.
And if copying and pasting, make sure you do "Paste Special" and unclick the box that says "float over text". That way the images always stay in the same spot with respect to the surrounding text and don't mysteriously jump around when you insert or delete text. cheers, Rosemary } } Thanks Doug- } Great tips- I'm printing them up until I memorize them. } Rgds, } Mike Shaw } Roselle, NJ } } } Gary, } } } } MS Word is such a pain because of the way it works with images. A couple } } of tricks I've discovered over the years. } } } } 1) Place the image within a text box, rather than directly on the page, it } } seems to give you much greater control over the location on the page. It } } also makes it much easier to add text annotations that stay with the image. } } } } } } If you don't want the text box to have a border you can remove it by } } selecting } } the box outline and look for the "paint brush" icon on the Draw toolbar, } } then use the down arrow and select "none". If you'd like the text box to } } be transparent, select the text box outline and look for the "paint bucket" } } icon on the Draw toolbar, then use the down arrow and select "none". If } } you discover its hard to find the text box border once you made the edge } } "invisible", first select the image with a single left click (it should have } } the solid black resizing "handles") and then use the keyboard left or right } } arrow keys and the selection with move out to the textbox outline (with } } black } } bordered resizing "handles"). } } } } 2) Always use the INSERT | PICTURE | FROM FILE option as opposed to copying } } and pasting an image into Word. I find that the images are harder to work } } with if I paste them in. Also, you can insert TIFF or BMP } } images into Word, } } you don't have to use JPEG.
I need to frequently move two loaded 5 1/2"L x 4 1/2"W x 3"H EM film boxes through a lighted hallway. Any suggestions for a light tight carrier would be appreciated. I have seen a rather complex 'portable darkroom' bag but can use something simpler.
Thank you. Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
Don't know, but I would be interested to know. We need a physicist here! I am also intrigued to note that some of my specimens show cathodoluminescence outputs sufficiently bright for the position of the scanning beam to be visible on the chamber cam. How bright does it need to get to have an impact on counts, and what artefacts might result?
Chris
----- Original Message ----- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 30, 2003 9:38 PM Subject: IR LEDS/EDS
} } } } } I had a problem like this. } } Turned out to be caused by the infra-red leds of my chamber camera. } } When the camera is switched off, the counts (3-5k per second) return } } close to zero. } } } } Chris } } } } What would be the mechanism of this effect, I wonder? } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
You can sublimate the sulfur by placing your TEM grid into a vacuum (roughing pump range OK) for a few minutes, room temperature is fine. Another possible way to remove the sulfur is to gently heat your sample at atmosphere for an hour or so. Of course, if temperature sensitive organic compounds are present you might choose the vacuum method instead.
Many of the IDPs that we process have very high porosities. Surface tension wicks the liquid sulfur inside the voids which simply goes away during sublimation leaving a clean section, at least in theory. You need to watch for contamination that is present in the sulfur to begin with though, especially organic contaminants. I would recommend that you purchase the very cleanest sulfur you can get and then gently distill it onto a watch glass or petri dish which you can keep covered.
Dave
Dave Joswiak University of Washington Dept. of Astronomy, 351580 Seattle, WA 98195 (206)543-7702 joswiak-at-astro.washington.edu
} Hello Dave, thank you for this information - it's extremely helpful. } The prospect of magically removing the embedding medium for TEM } examination is really cool! You should definitely write this up. } Some random questions: At what temperature do you sublimate the } sulfur? Do you experience any degradation of the organics at this } temperature? Do your IDP's have any porosity? Our samples are often } quite porous, so infiltration of the sulfur is an issue. } } Thanks again, } Rick } } joswiak-at-orca.astro.washington.edu wrote: } } } Rick - We have worked on embedding particles in sulfur for some } } time now. I have always thought to present this technique at MSA } } but just haven't gotten around to it yet. To the best of my } } knowledge, the idea to embed in sulfur was conceived by our } } colleague, John Bradley. Perhaps others have done so as well. We } } developed our own set of sulfur-embedding techniques and methods } } suitable to our particles. Embedding materials in sulfur is a } } wonderful technique to look at microtomed sections since sulfur } } will sublimate in vacuum and you can be left with a section on a } } TEM grid that is free of embedding material. We study } } interplanetary dust particles (IDPs - typically 5 - 10 um in size) } } which are composed of 1000's of mineral grains, glass and } } carbonaceous phases, including organic materials. These particles } } are often highly porous and the normal infiltrating epoxy resins } } have always made it difficult for us to determine the carbon } } compounds vs the epoxy,thus our need for this technique (or other } } carbon-free embedding methods). } } } } Sulfur is a very difficult material to work with and requires } } fairly extensive experience and practice due to some unusual } } properties. For instance, liquid sulfur is one of the rare } } materials (the only?) whose viscosity goes up as you heat it to } } higher temperatures (at least up to about 190 C); this is due to } } increasing polymerization of sulfur ions in the melt with } } increasing temperature. Our technique of sulfur embedding is to } } place a particle (say a 10 um interplanetary dust particle) in the } } middle of a clean glass slide. On a second clean glass slide we } } heat a tiny crystal of ultra-pure sulfur above its melting point of } } 119 C. The sulfur will melt and after removal of the heat will } } supercool forming a liquid drop on the glass slide. I like to have } } the drop diameter 50 - 100 um or less, otherwise the liquid sulfur } } may spontaneously crystallize. While observing under a good } } binocular microscope, the particle on the clean glass slide is } } lowered into the sulfur drop which will wet both glass slides. } } When the two glass slides are separated the particle on the first } } glass slide will have a hemispherical liquid drop around it. At } } this point you need to crystallize the supercooled sulfur which } } surrounds the particle by touching a small sulfur crystal to the } } drop which will create nucleation sites and initiate } } crystallization. There are other ways to do this besides what I } } have just described. Unfortunately, fractures may develop in the } } sulfur during crystallization which can lead to contamination and } } other problems later. Fracturing of the crystallized sulfur is a } } problem with this technique that we haven't found a satisfactory } } solution to yet. } } } } The particle in solid sulfur is then mounted with epoxy to a } } pre-cut epoxy cylinder which fits the chuck of the microtome. The } } sulfur mount must first be dislodged from the slide as the } } crystallized sulfur will want to stick to the glass. It can then } } be trimmed and microtomed in the usual way but you have to be } } careful as microtoming is more difficult than with epoxy resins due } } to the softness and fragile nature of crystallized sulfur but it } } can be done with practice. For us, this works best on small } } samples (in the 10 - 20 um range). Another important factor is } } that many epoxies will dissolve the sulfur while curing thus you } } need to experiment to determine which ones are suitable to hold } } your sulfur mount. I have found one or two that are suitable for } } our needs. Superglue can also be used. } } } } I haven't provided all the details of our sulfur embedding } } technique here and there are more pitfalls than I have described. } } The technique is difficult and somewhat unpredictable, but if } } everything goes right you can end up with nice microtomed sections } } free of any embedding material. This is useful for observing or } } measuring carbon by EDX if you mount your sample on a TEM grid with } } an SiO film (have to watch out for carbon contamination in the } } microscope, though). I can imagine that there are other, perhaps } } better ways to mount materials in sulfur than I have described; } } this is just a technique that has evolved over the last several } } years for us. } } } } } } Dave } } } } Dave Joswiak } } University of Washington } } Dept. of Astronomy, 351580 } } Seattle, WA 98195 } } (206)543-7702 } } joswiak-at-astro.washington.edu } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We use a Pelican case - one of those cases used for camera equipment where you want to keep dust, light and/or water out. They come in various sizes and have foam inserts that where you can either remove cubes or cut holes to fit your equipment or film boxes. They are not cheap. They work very well.
I believe there is at least one other brand as well - inquire at your local camera shop or check the web.
Pelican Products http://www.casesbypelican.com
} I need to frequently move two loaded 5 1/2"L x 4 1/2"W x 3"H EM film boxes } through a lighted hallway. Any suggestions for a light tight carrier would } be appreciated. I have seen a rather complex 'portable darkroom' bag but } can use something simpler.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} I need to frequently move two loaded 5 1/2"L x 4 1/2"W x 3"H EM film boxes } through a lighted hallway. Any suggestions for a light tight carrier would } be appreciated. I have seen a rather complex 'portable darkroom' bag but } can use something simpler.
I use an empty cardboard box that previously contained 500 sheets of 8.5 x 11 photographic paper. When you put the EM film boxes inside of the black-lined cardboard box, it is completely light tight. You can't beat the price. ############################################################## 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/~image/ ##############################################################
} } My first approach, after all you have already done, would be to get a } Geiger Counter and check the area for any radioactivity. Also, if you } are using xray diffraction equipment nearby, see if the mysterious } counts correspond to when that equipment is energized. }
Oh come on!
If there were to be sufficient ambient X-radiation to get into the sample chamber of an SEM from outside, the staff would be all dead by now.
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
What on earth would be the point of 'distilling it into one general statement'?
About as much as trying to distill into one general statement the description of, say, a cat.
By now there have been sufficient sensible replies for us all to know what an electron microprobe is, haven't there?
It's not some metaphorical, etymological, or philosophical mystery.
Look, as someone suggested, at http://www.geology.wisc.edu/~johnf/empa.html.
Or look at the JEOL website.
Just read what Vladimir said. It's like an SEM, with better X-Ray detection, better stability, an optical microscope, and, (I like this one) better operator training.
Of course an SEM can not be generally called an electron microprobe!
That would be like saying there was no difference between a JSM-840 and a JXA-840.
rtch
} From: Peter Tomic {PTomic-at-anadigics.com} To: Microscopy-at-sparc5.microscopy.com
Another thing to watch out for is re-using old MSWord reports as templates. In some cases the file size does not shrink accordingly, and you can get a document with one 250k image and a file size of 10MB! This problem seemed to go away with one MSWord update, then come back again with another.
-----Original Message----- } From: Shalvoy, Richard B **CHES [mailto:RBShalvoy-at-archchemicals.com] Sent: 30 January 2003 13:23 To: '"Microshaw-at-aol.com"-at-sparc5.microscopy.com'; Microscopy-at-sparc5.microscopy.com
I can share a recent example of how the file size in Word is a huge function of how the images are imported into it.
A customer sent me a three page memo with 4 of my SEM images in it for my review. The original image files were all jpgs about 150 kB in size. The file he sent me was 4.5MB! By clogging up the email system, it's a pain to work with that large a file. I tried a simple test by deleting the four images from the memo and using the Insert, Picture, From File command to put the same four images into Word. I then used the Format Picture menu to resize the images down to fit side by side in the memo as before and saved the file. The new file size was only 550 kB! Not tiny, but more in line with what I would expect.
I informed the customer about this technique and he admitted to simply cut and pasting them into Word. That is a sure recipe for making huge files unnecessarily. It's kind of curious that there is such a large difference, but I have seen it over and over.
Richard Shalvoy Arch Chemicals, Inc. 350 Knotter Drive Cheshire, CT 06410 (203) 271-4394 rbshalvoy-at-archchemicals.com
-----Original Message----- } From: "Microshaw-at-aol.com"-at-sparc5.microscopy.com [mailto:"Microshaw-at-aol.com"-at-sparc5.microscopy.com] Sent: Wednesday, January 29, 2003 9:42 PM To: cromey-at-Arizona.edu; Microscopy-at-sparc5.microscopy.com
Thanks Doug- Great tips- I'm printing them up until I memorize them. Rgds, Mike Shaw Roselle, NJ
} Gary, } } MS Word is such a pain because of the way it works with images. A couple } of tricks I've discovered over the years. } } 1) Place the image within a text box, rather than directly on the page, it } seems to give you much greater control over the location on the page. It } also makes it much easier to add text annotations that stay with the image. } } } If you don't want the text box to have a border you can remove it by selecting } the box outline and look for the "paint brush" icon on the Draw toolbar, } then use the down arrow and select "none". If you'd like the text box to } be transparent, select the text box outline and look for the "paint bucket" } icon on the Draw toolbar, then use the down arrow and select "none". If } you discover its hard to find the text box border once you made the edge } "invisible", first select the image with a single left click (it should have } the solid black resizing "handles") and then use the keyboard left or right } arrow keys and the selection with move out to the textbox outline (with black } bordered resizing "handles"). } } 2) Always use the INSERT | PICTURE | FROM FILE option as opposed to copying } and pasting an image into Word. I find that the images are harder to work } with if I paste them in. Also, you can insert TIFF or BMP } images into Word, } you don't have to use JPEG.
Hi, We had just the same problem with high counts some times ago. Service engineers without any results several times took out the detector from the column. The last serviceman grounded the detector to the microscope column and the problem vanished. All calibration passed well and the system is just the same as before the trouble. However, no one has explained us why the detector has to be connected to the column (the wrong way according to manuals).
Best regards from Prague. Oldrich
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-241062399 Fax: +420-241062347 WEB: http://www.biomed.cas.cz/mbu/lem113/lem.htm
I am doing a double tilting experiment with TEM. From diffraction zone A to diffraction zone B, I tilted X axis for xx degree and Y axis for yy degree. Could anyone let me know how to calculate the angle between diffraction zones A and B.
For the situation you are talking about, the total tilt, theta, generated from your two independent rotations, xx and yy (about perpendicular axes) is:
cos(theta) = cos(xx)cos(yy)
(for xx and yy { 90 degrees)
Dick Fonda
At 9:54 AM +0000 1/31/03, Ji, Ying wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ________________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 ________________________________________________________________
The point is to understand that the term "electron microprobe" is being used to describe multiple instruments and is not just a SEM, EDX, WDS, TEM etc. I agree, enough already it's Friday.
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Thursday, January 30, 2003 9:40 PM To: Peter Tomic; Microscopy-at-sparc5.microscopy.com
Stefano Maretti writes ...
} I have not IR camera or light inside or near my chamber. } I have not radioactive samples around and I have the problem } also when the chamber is empty.
At what energy are these x-rays measured? If they're at the low end I suspect the chip may be warm because of a poor vacuum(?) The improvement after a 4-month wait could be because the detector could be equalizing with the chamber vacuum(?)
Another cheap container is a new, unused 1 gallon paint can. You can get these at Lowes or Home Depot or any paint store. It is wise to attach a church key opener to the can so you always have it with you. Greg Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295
We have an infra red chamberscope camera for checking specimen height and users keep forgetting to turn it off before taking X-ray analyses - it also affects our solid state back-scatter detector.
The x-ray detector is a thin window light element detector and I suspect that they are more prone to infra red than beryllium windows.
Malcolm Haswell University of Sunderland UK
----- Original Message ----- } From: Chris Jeffree {c.jeffree-at-ed.ac.uk}
Jim,
When we routinely used film plates for TEM, we carried loaded canisters in a box for Kodak 8x10" paper. I reinforced the corners with black electrical tape to ensure it was light-tight. This solution never failed me.
Another solution that I used for desiccating film may come in handy to some: We all know that EM film plates should be desiccated before use in the TEM. One problem that can occur is that the paper box containing the film sucks up lots of moisture that defeats the desiccant. I solved this by storing my unused film in a developing can made to hold two 35-mm reel. The can has a light-tight top to allow desiccation.
My belief is that the simplest solutions are the best. Good luck.
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Jim Romanow {bsgphy3-at-uconnvm.u To: microscopy-at-sparc5.microscopy.com conn.edu} cc: Subject: light tight container?
01/30/03 03:36 PM
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To all,
I need to frequently move two loaded 5 1/2"L x 4 1/2"W x 3"H EM film boxes through a lighted hallway. Any suggestions for a light tight carrier would be appreciated. I have seen a rather complex 'portable darkroom' bag but can use something simpler.
Thank you. Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
Chris- I have run across instances where the sample's cathodoluminescence has interfered with my EDX analysis. I first noticed it when I was analyzing a phosphor and the EDX peaks were (uniformly) shifted to higher energies. Further experiments showed that it took quite a bit of CL (visible on IR chamber camera) to produce this effect , and a little more would result in 100% dead time. I was unable to reproduce the effect on our other SEM/EDX so I am fairly confident it is a complicated function of thresholds, time constants, window materials, CL wavelength, etc. In addition, I could fine tune the effect by dimming the IR chamberscope LEDs to the proper (low) level to affect the peak shift. Of course, normal IR chamberscope LED levels blind the EDX detector completely. In any case, it is easy to avoid this CL artifact by reducing the beam energy and/or current. I found that Zinc Silicate (fluorescent light phosphor) and synthetic sapphire insulators (small red, yes red, balls) produced enough CL for this effect in my SEM/EDX system. It did, however, require that I hit them hard with approximately 1nA at 25kV (I don't remember exactly). I have received conflicting answers when I have asked whether exposing the biased EDX detector to bright light, such as the IR chamberscope, for long periods of time will cause damage so I try to avoid the situation. I know that my SUTW should be exposed to bright light, but I expect that is really intense light which could melt it due to poor thermal conductivity to a heat sink. Does anyone out there know the answer to this question?
Sincerely, Matthew Ervin, Ph.D. (301)394-0017 phone, (301)394-1559 fax MErvin-at-ARL.Army.mil
M/S: AMSRL-SE-RL US Army Research Laboratory 2800 Powder Mill Road Adelphi, MD 20783-1197
Disclaimer: The opinions and views expressed above are those of the author and do not necessarily represent those of the U.S. Army Research Laboratory or any other government agency
Chris Jeffree {c.jeffree-at-ed.ac.uk}
01/30/2003 05:51 PM Please respond to Chris Jeffree
To: microscopy-at-sparc5.microscopy.com cc: Subject: Fw: IR LEDS/EDS
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Don't know, but I would be interested to know. We need a physicist here! I am also intrigued to note that some of my specimens show cathodoluminescence outputs sufficiently bright for the position of the scanning beam to be visible on the chamber cam. How bright does it need to get to have an impact on counts, and what artefacts might result?
Chris
----- Original Message ----- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 30, 2003 9:38 PM Subject: IR LEDS/EDS
} } } } } I had a problem like this. } } Turned out to be caused by the infra-red leds of my chamber camera. } } When the camera is switched off, the counts (3-5k per second) return } } close to zero. } } } } Chris } } } } What would be the mechanism of this effect, I wonder? } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
If you need no more than to move the film between rooms then a black polythene bag will be more than adequate, and cheap. A good source would be the packaging of large size photographic paper or xray film, so contact your local photographer. Some of these bags are foil-lined - even better. Alternatively, thick black bags are widely sold as rubble sacks in hardware stores.
Chris
On 30 Jan 03, at 14:11, 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 } ---------------------------------------------------------------------- } -. } } } We use a Pelican case - one of those cases used for camera equipment } where you want to keep dust, light and/or water out. They come in } various sizes and have foam inserts that where you can either remove } cubes or cut holes to fit your equipment or film boxes. They are not } cheap. They work very well. } } I believe there is at least one other brand as well - inquire at your } local camera shop or check the web. } } Pelican Products http://www.casesbypelican.com } } } I need to frequently move two loaded 5 1/2"L x 4 1/2"W x 3"H EM film } } boxes through a lighted hallway. Any suggestions for a light tight } } carrier would be appreciated. I have seen a rather complex 'portable } } darkroom' bag but can use something simpler. } } Aloha, } Tina } } } ********************************************************************** } ****** * Tina (Weatherby) Carvalho * } tina-at-pbrc.hawaii.edu * * Biological Electron Microscope } Facility * (808) 956-6251 * * University of Hawaii at } Manoa * http://www.pbrc.hawaii.edu/bemf* } ********************************************************************** } ****** } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 /8669 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
As a committed user of Spurr's resin, I have the task now of correlating Xray diffraction values with measured structures in Spurr's embedded skeletal muscle samples. Using the 43nm periodicity of C-protein (Myosin Binding Protein C) in the A-band of the sarcomere and the 38.5nm periodicity in the thin filaments, I'm routinely seeing a shrinkage artifact of approximately 15% in the periodicities of the thick and thin filaments, as well as in the A-band width (1.37 microns in Spurr's embedded samples vs the generally accepted 1.6 microns for A-bands in LM or laser diffraction) . For those of you out there that use Spurr's resin and have done morphometry, what percentage of shrinkage have you commonly seen? Is this about right? Thanks in advance, Patrick C. Nahirney, Ph.D. Laboratory of Muscle Biology National Institute of Arthritis and Musculoskeletal and Skin Diseases National Institutes of Health
The goniometer-measured angle bteween the zones can be calculated by using standard rotation matrices for rotation around x and y-axes in a Cartesian basis. The orientation of the specimen before, g, and after tilt, g', is then related by the compound orthonormal rotation matrix R=RxRy: g' = Rg.
The measured tilt angle, Y, between the zones are then - according to my calculations -
cos y = (cos a)(cos b) / sqrt( (cos^2(a) sin^2(b)) + (sin^2(a) cos^2(b)) )
where a and b are the measured angles tilted around the two goniometer axes.
Paul
======================= Paul Baggethun Engineer Alcoa Technical Center Alcoa Center, PA 15069 USA =======================
-----Original Message----- } From: Ji, Ying [mailto:y.ji-at-imperial.ac.uk] Sent: Friday, January 31, 2003 4:55 AM To: 'microscopy-at-sparc5.microscopy.com'
Dear All
I am doing a double tilting experiment with TEM. From diffraction zone A to diffraction zone B, I tilted X axis for xx degree and Y axis for yy degree. Could anyone let me know how to calculate the angle between diffraction zones A and B.
Matt That's interesting. With my system, with beam off the chambercam leds induce an artefactual peak which starts just below Carbon ka and ramps up to the left (ie peaks at lower energies) and is worth a few k counts, depending on detector parameters selected. The other day I was looking at a cathodoluminescent rock specimen, producing a visible signal on the chamber cam (but with LEDs off) and at times detected a small, apparently artefactual peak at a similar position. Background information on the specimen indicates no elements in the "detected" range. I take it this could be an artefact arising from specimen cathodoluminescence? Best wishes Chris
On 31 Jan 03, at 10:05, Matt Ervin wrote:
} Chris- } I have run across instances where the sample's } cathodoluminescence } has interfered with my EDX analysis. I first noticed it when I was } analyzing a phosphor and the EDX peaks were (uniformly) shifted to } higher energies. Further experiments showed that it took quite a bit } of CL (visible on IR chamber camera) to produce this effect , and a } little more would result in 100% dead time. I was unable to reproduce } the effect on our other SEM/EDX so I am fairly confident it is a } complicated function of thresholds, time constants, window materials, } CL wavelength, etc. In addition, I could fine tune the effect by } dimming the IR chamberscope LEDs to the proper (low) level to affect } the peak shift. Of course, normal IR chamberscope LED levels blind } the EDX detector completely. In any case, it is easy to avoid this CL } artifact by reducing the beam energy and/or current. I found that } Zinc Silicate (fluorescent light phosphor) and synthetic sapphire } insulators (small red, yes red, balls) produced enough CL for this } effect in my SEM/EDX system. It did, however, require that I hit them } hard with approximately 1nA at 25kV (I don't remember exactly). I have } received conflicting answers when I have asked whether exposing the } biased EDX detector to bright light, such as the IR chamberscope, for } long periods of time will cause damage so I try to avoid the } situation. I know that my SUTW should be exposed to bright light, but } I expect that is really intense light which could melt it due to poor } thermal conductivity to a heat sink. Does anyone out there know the } answer to this question? } } Sincerely, } Matthew Ervin, Ph.D. } (301)394-0017 phone, (301)394-1559 fax } MErvin-at-ARL.Army.mil } } M/S: AMSRL-SE-RL } US Army Research Laboratory } 2800 Powder Mill Road } Adelphi, MD 20783-1197 } } Disclaimer: The opinions and views expressed above are those of the } author and do not necessarily represent those of the U.S. Army } Research Laboratory or any other government agency } } } } } Chris Jeffree {c.jeffree-at-ed.ac.uk} } } 01/30/2003 05:51 PM } Please respond to Chris Jeffree } } } To: microscopy-at-sparc5.microscopy.com } cc: } Subject: Fw: IR LEDS/EDS } } } ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } } Don't know, but I would be interested to know. We need a physicist } here! I am also intrigued to note that some of my specimens show } cathodoluminescence outputs sufficiently bright for the position of } the scanning beam to } be visible on the chamber cam. } How bright does it need to get to have an impact on counts, and what } artefacts might result? } } Chris } } ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; } {microscopy-at-sparc5.microscopy.com} } Sent: Thursday, January 30, 2003 9:38 PM } Subject: IR LEDS/EDS } } } } } } } } } } I had a problem like this. } } } Turned out to be caused by the infra-red leds of my chamber } camera. } } } When the camera is switched off, the counts (3-5k per second) } return } } } close to zero. } } } } } } Chris } } } } } } } What would be the mechanism of this effect, I wonder? } } } } rtch } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } } The University of Auckland email : r.sims-at-auckland.ac.nz } } Private Bag 92019 } } Auckland } } New Zealand } } } } } } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 /8669 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
This listserver is set up to encompass "basically any question/comment/observation or general information/announcement which involves microscopy and/or microanalysis." As long as a discussion thread is being actively participated in, I would say that it is a valid posting. There is a very easy method to rid oneself of postings one is not interested in. It's called the delete function.
Shannan Little
This is getting a bit tiresome.
What on earth would be the point of 'distilling it into one general statement'?
About as much as trying to distill into one general statement the description of, say, a cat.
By now there have been sufficient sensible replies for us all to know what an electron microprobe is, haven't there?
It's not some metaphorical, etymological, or philosophical mystery.
Look, as someone suggested, at http://www.geology.wisc.edu/~johnf/empa.html.
Or look at the JEOL website.
Just read what Vladimir said. It's like an SEM, with better X-Ray detection, better stability, an optical microscope, and, (I like this one) better operator training.
Of course an SEM can not be generally called an electron microprobe!
That would be like saying there was no difference between a JSM-840 and a JXA-840.
What usually happens is that the document's file becomes fragmented. In this case, more sectors on disk are used (wasted) and leads to a larger file size. If you routinely keep disk fragmentation low, then do a Save As with the same file name. This should put the new save in a contiguous area.
gary g
At 11:17 PM 1/30/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think you would have to worry about two problems - (1) shrinkage during the actual polymerization step and (2) compression during thin sectioning. I don't have the reference off the top of my head but Daniel Studer has a nice paper (J. Microscopy about 1-2 years ago, I think) in which he shows significant (up to 50% if i remember correctly) along the cutting axis. his paper showed this could be avoided using an oscillating diamond knife. this paper has important implications for high resolution measurements since the compression would be different in the different axes.
At 10:08 AM 1/31/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Please help. I'll pay any price for a decent copy of a Reichert OM-U3 ops/service manual. Someone was nice enough to send a copy of a copy, of a copy, etc., but all the illustrations are illegible. That was an ops manual, and I'll still need a service manual, or at least a schematic or wiring diagram. Anyone with some spare specimen block holders would be helpful also. The instrument that I have only came with one flat block holder. I have lots of lab stuff I would be happy to trade.
--
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Dear Jim, I use the black plastic bags that the photographic paper comes wrapped in to transport my film boxes to the dark room. I discovered the hard way that the boxes themselves were not light tight. Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Jim Romanow" {bsgphy3-at-uconnvm.uconn.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 30, 2003 1:36 PM
Dear Ritchie, I believe the Si(Li) detector is sensitive to visible light, as it is to the x-ray photons. With a Be window the visible light could not penetrate to the detector, but with the new thin membrane windows the IR gets right through. Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 30, 2003 1:38 PM
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More than 25 years ago we had our machine shop make a rectangular box and cover of sheet stainless steel, complete with handles. It holds both a loaded film box and an exposed film box from a JEOL 100 CX. It still looks and works like new and was worth the expense.
Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Avenue Argonne, Il., 60439
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id MAA00103 for dist-Microscopy; Fri, 31 Jan 2003 12:30:36 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id MAA00096 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Fri, 31 Jan 2003 12:30:05 -0600 (CST) Received: from mailhub-3.iastate.edu (mailhub-3.iastate.edu [129.186.140.13]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id MAA00080 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 31 Jan 2003 12:29:50 -0600 (CST) Received: from mailout-1.iastate.edu (mailout-1.iastate.edu [129.186.140.1]) by mailhub-3.iastate.edu (8.9.3/8.9.3) with SMTP id MAA01830 for {Microscopy-at-msa.microscopy.com} ; Fri, 31 Jan 2003 12:24:26 -0600 Received: from spare.marl.iastate.edu(129.186.227.3) by mailout-1.iastate.edu via csmap id 24931; Fri, 31 Jan 2003 12:26:17 -0600 (CST) Message-Id: {5.1.1.5.2.20030131095902.045b9170-at-pop-2.iastate.edu} X-Sender: wesaia-at-pop-2.iastate.edu X-Mailer: QUALCOMM Windows Eudora Version 5.1.1
I always understood this to be due to "photons is photons". Granted that the energy of an IR photon is much less than an x-ray photon. (What is the wavelength cutoff for generating an electron-hole pair?) However, there are just so darn many of those IR photons. So not only do we get the very low energy counts due to the photons, but also we get a lot of dead-time and pile-up of photons. I usually see our dead time max out when the light comes on. It takes several seconds for the circuitry to get back to normal after we shut the light off.
I know x-ray windows are supposed to be treated to render them opaque to infra-red. However, that treatment appears to be more effective in some cases than other. Both of our detectors are affected at least some when the cameras are turned on.
Regarding cathodoluminescence, I wonder if the signal is too weak to be of concern. A 20-kV beam at 1 nA is only dumping 20 micro-watts of power into the sample. Only a small fraction of that gets converted over to light. I don't know the power rating of our IR bulb on our chamber scope, but I am sure it is quite a few orders of magnitude more powerful.
Warren
At 10:51 PM 1/30/03 +0000, you wrote:
} Don't know, but I would be interested to know. We need a physicist } here! } I am also intrigued to note that some of my specimens show } cathodoluminescence outputs sufficiently bright for the position of } the scanning beam to } be visible on the chamber cam. } How bright does it need to get to have an impact on counts, and what } artefacts might result? } } Chris } } ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; } {microscopy-at-sparc5.microscopy.com} } Sent: Thursday, January 30, 2003 9:38 PM } Subject: IR LEDS/EDS } } } } I had a problem like this. } } } Turned out to be caused by the infra-red leds of my chamber } camera. } } } When the camera is switched off, the counts (3-5k per second) } return } } } close to zero. } } } } } } Chris } } } } } } } What would be the mechanism of this effect, I wonder? } } } } rtch } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } } The University of Auckland email : r.sims-at-auckland.ac.nz } } Private Bag 92019 } } Auckland } } New Zealand
} } Don't know, but I would be interested to know. We need a physicist } here! } ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; } } } } } What would be the mechanism of this effect, I wonder? } } } } rtch }
I expected the usual suspects to leap on this, but since it hasn't happened, I will chime in. I are an engineer, not a physicist; take with the appropriate grain of salt, but I have worked in pulse processor design.
The Al coatings on thin window detectors do a good job of keeping out visible light, but are fairly transparent to IR. The IR photons are absorbed just like X-rays, but because their energy is so low and the flux high compared to X-ray photons, the result is a nearly continuous flow of current through the detector which is indistinguishable (to the pulse processing electronics) from a "leaky detector". Essentially, high leakage current raises the noise threshold setting required to prevent false triggering of the pulse processor. Since thin-window detectors are intended to trigger on low-energy X-rays, that threshold in a properly funtioning detector is set fairly low. Therefore, the IR- induced "leakage current" causes the pulse processing electronics to start triggering continuously on noise, causing a large peak at the low end of the spectrum.
Chris: light shining on a semiconductor (which the detector is) generates electron-hole pairs, just like an x-ray does. But the magnitude of the number of these pairs is much higher because there are usually more photons in a light beam than characteristic x-rays being generated by a sample. This flood of electron-hole pairs overwhelms the amplifier and the counting electronics. Some detectors have metallized (mere atoms of metal, so they won't interfere w/ analysis) windows to keep out the 'ambient' light, such as maybe generated by CL. But if your chamberscope's light source is oriented so that it shines directly on the detector, enough light can get in to cause problems. The collimator on the detector nose can also help keep light out. I've worked on SEMs that have had the chamberscope light source directly opposite the detector and had to be vigilant about turning it off when is was not needed. I now have a scope that has the light source behind the detector and it give no trouble at all. For the curious, on the Oxford Instruments site, on the page for their EDX hardware (http://www.oxford-instruments.com/ANLPDP174.htm) near the bottom right hand side is a list of "Related PDFs" and the last one is "EDX Hardware Explained". If you've ever wondered what's in the snout of a detector and how it works its magic, this is a good read. NOTE: I have no interest, financial or otherwise, in Oxford Instruments. Just a satisfied customer.
Chris Jeffree wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Don't know, but I would be interested to know. We need a physicist } here! } I am also intrigued to note that some of my specimens show } cathodoluminescence outputs sufficiently bright for the position of } the scanning beam to } be visible on the chamber cam. } How bright does it need to get to have an impact on counts, and what } artefacts might result? } } Chris } } ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; } {microscopy-at-sparc5.microscopy.com} } Sent: Thursday, January 30, 2003 9:38 PM } Subject: IR LEDS/EDS } } } } } } } } } I had a problem like this. } } } Turned out to be caused by the infra-red leds of my chamber } camera. } } } When the camera is switched off, the counts (3-5k per second) } return } } } close to zero. } } } } } } Chris } } } } } } } What would be the mechanism of this effect, I wonder? } } } } rtch } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } } The University of Auckland email : r.sims-at-auckland.ac.nz } } Private Bag 92019 } } Auckland } } New Zealand } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
As I understood it at M&M 2002 in Quebec City, the Diatome oscillating knife was close to market, as in the 'fine-tuning' stage. You might want to inquire of Diatome US as to the state of that knife. The reduction in compression for soft materials was phenomenal.
Tom Malis
Dr. Tom Malis Scientist Advisor Natural Resources Canada Govt. of Canada 613-995-7358 malis-at-nrcan.gc.ca
-----Original Message----- } From: Tom Phillips To: Nahirney, Patrick (NIH/NIAMS) Cc: Microscopy-at-sparc5.microscopy.com Sent: 1/31/2003 11:49 AM
I think you would have to worry about two problems - (1) shrinkage during the actual polymerization step and (2) compression during thin sectioning. I don't have the reference off the top of my head but Daniel Studer has a nice paper (J. Microscopy about 1-2 years ago, I think) in which he shows significant (up to 50% if i remember correctly) along the cutting axis. his paper showed this could be avoided using an oscillating diamond knife. this paper has important implications for high resolution measurements since the compression would be different in the different axes.
At 10:08 AM 1/31/2003 -0500, you wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Peter, In that case, a TEM would also be an electron microprobe. I think it is better if the names are used specifically. Electron microprobes are designed quite differently from SEMs. Call an SEM with WDS an SEM with WDS.
At 11:12 AM 1/30/2003 -0500, Peter Tomic wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Black plastic bags win the popularity (and economy) contest!
Quick list of light tight container suggestions:
Black plastic photo paper bag with or without cardboard box Ammunition storage box Paint can Pelican camera equipment case Spring loaded paper safe box Plastic tool/tote box Custom made stainless steel box
Thank you very much for all of the feedback. I am leaning toward the ammo box because it might hold up well under student abuse; the most bullet proof solution:}
Regards, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
the resultant y" axis orientation (equivalent to the specimen normal in this case) is given as:
y" = -x sin(a)cos(b) + y cos(a)cos(b) + z sin(b)
The total tilt is found from the arccos of the dot product of the initial y axis and the final y" axis, which gives (assuming unit vectors)
cos(theta) = cos(a)cos(b)
which is what I stated this morning.
Dick Fonda
At 9:54 AM +0000 1/31/03, Ji, Ying wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We're staining COS cells with Alexa 647 phalloidin (Mol. Probes A-22282 lot 41B1-1). The f-actin is staining fine and looks great when we excite at 633 nm and detect through the Leica AOBS or normal Cy5 filter set.
However, when we excite at 488 nm, we're seeing the f-actin staining appearing in the green channel too. We checked on different microscopes.
Has anybody else seen something as weird as this? We suspect contamination, but don't know where it would have come from unless it is an isomer or something from the manufacturing process.
One of our controls is the Alexa-phalloidin staining alone without any antibodies, so we know it's not some weird antibody binding artifact.
Any help appreciated.
Thanks.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
On Friday, January 31, 2003, at 01:54 AM, Ji, Ying wrote:
} I am doing a double tilting experiment with TEM. From diffraction zone } A to } diffraction zone B, I tilted X axis for xx degree and Y axis for yy } degree. } Could anyone let me know how to calculate the angle between diffraction } zones A and B. } } Thank you very much in advance! } Dear Yun, I don't have my spherical trigonometry book here, having moved recently, but the way to approach the problem is to imagine the electron beam incident on the North pole with the Greenwich meridian facing you, then perform the tilts and locate the position of the incident beam after these have been done. If you have a double tilt holder like the one I used when I was in Albany NY, the tilts can be done in either order, so tilt in Y (keeping the Greenwich meridian facing you) so the beam is now incident on latitude 90-yy, then tilt through xx degrees along the great circle perpendicular to the Greenwich meridian and passing through it at latitude 90-yy. You will have a right spherical triangle; the hypotenuse is the angle you're looking for. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
A colleague of mine recently asked about a type of "Laser Scanning Phase Contrast Microscopy", for which I do not have much knowledge.
Please advise: What's the major difference between this and the conventional LSCM that we use? Which institution in the East Coast might have such a facility?
Any advice is highly appreciated and will be forwarded to this colleague. Have a great weekend!
QC
Qian-Chun Yu, MB, Ph.D. Director Biomedical Imaging Core Laboratory University of Pennsylvania Department of Pathology & Lab Medicine School of Medicine 110 Richards Building Philadelphia, PA 19104
We use tin cookie boxes. They come in all different sizes and before christmas, 2001 we asked regular lab users to save their cookie boxes if they received any as christmas gift. They are light tight and so far have been working great.
By the way, we just asked for used, empty boxes, no cookies.
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
Gary I think, you wrong: fragmented file occupied more physical space on HD (yes) but fragmented/non-fragmented files has the same bit-size. So fragmentation may affect the reading/writing time only, which on the modern computers is negligibly. If the disk seriously fragmented, yes, it may affect computer's performance in general. "Save as" command not necessary writes in non-fragmented space. NTFS usually decently manage this issue and keep fragmentation at relatively low level, but it's OS decision where to place your file (for instance, if file is small, it would be placed in the NTFS analog of FAT at the beginning of drive- this is special precaution against fragmentation). If the HD is seriously fragmented, even "Save as" will cause the file fragmentation. Sergey
At 08:18 AM 1/31/2003, 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
To All, } } The company I work for currently utilizes a coated glass slide that is } amine reactive for printing our microarrays. The supplier of the } substrate is reticent to divulge any information regarding the chemical } composition, thickness, and QC measures employed in their process. } Subsequently, we would like to ascertain as much info as possible } regarding the coating thickness, surface topography, etc. and were } hoping that experts in the field of microscopy might offer some } suggestions or be willing to undertake such a project. } } } Chris Michaelson }
Hi- we are in the process of purchasing a new confocal, and are looking at a Zeiss510 Meta or a Leica AOBS or SP2. I think that in the demo process we are seeing what we need to, and are impressed by the capabilities of each. I would appreciate user perspectives that we cannot get from the manufacturers though- are there instrument aspects of either microscope that are repeatedly problematic or limiting? Do multi-user facilities have particular difficulty with either? I do not expect an obvious "winner" for an answer, but am more interested in hearing what some of the ups and down are, and seeing if those problems or circumstances would be issues for us. Thanks for any experience you an offer, I am sure it will be useful.
Betsey ************************************************************************ Betsey Pitts Research Associate/Facilities Manager, Microscopy The Center for Biofilm Engineering
366 EPS Building, Montana State University Bozeman, MT 59717
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I always understood this to be due to "photons is photons". Granted that the energy of an IR photon is much less than an x-ray photon. (What is the wavelength cutoff for generating an electron-hole pair?) However, there are just so darn many of those IR photons. So not only do we get the very low energy counts due to the photons, but also we get a lot of dead-time and pile-up of photons. I usually see our dead time max out when the light comes on. It takes several seconds for the circuitry to get back to normal after we shut the light off.
I know x-ray windows are supposed to be treated to render them opaque to infra-red. However, that treatment appears to be more effective in some cases than other. Both of our detectors are affected at least some when the cameras are turned on.
Regarding cathodoluminescence, I wonder if the signal is too weak to be of concern. A 20-kV beam at 1 nA is only dumping 20 micro-watts of power into the sample. Only a small fraction of that gets converted over to light. I don't know the power rating of our IR bulb on our chamber scope, but I am sure it is quite a few orders of magnitude more powerful.
Warren
At 10:51 PM 1/30/03 +0000, you wrote:
} Don't know, but I would be interested to know. We need a physicist } here! } I am also intrigued to note that some of my specimens show } cathodoluminescence outputs sufficiently bright for the position of } the scanning beam to } be visible on the chamber cam. } How bright does it need to get to have an impact on counts, and what } artefacts might result? } } Chris } } ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; } {microscopy-at-sparc5.microscopy.com} } Sent: Thursday, January 30, 2003 9:38 PM } Subject: IR LEDS/EDS } } } } I had a problem like this. } } } Turned out to be caused by the infra-red leds of my chamber } camera. } } } When the camera is switched off, the counts (3-5k per second) } return } } } close to zero. } } } } } } Chris } } } } } } } What would be the mechanism of this effect, I wonder? } } } } rtch } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } } The University of Auckland email : r.sims-at-auckland.ac.nz } } Private Bag 92019 } } Auckland } } New Zealand
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More than 25 years ago we had our machine shop make a rectangular box and cover of sheet stainless steel, complete with handles. It holds both a loaded film box and an exposed film box from a JEOL 100 CX. It still looks and works like new and was worth the expense.
Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Avenue Argonne, Il., 60439
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id AAA13221 for dist-Microscopy; Sun, 2 Feb 2003 00:36:44 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id AAA13141 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Sun, 2 Feb 2003 00:36:11 -0600 (CST) Received: from mailgw.solutia.com (mailgw.solutia.com [205.191.166.80]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id AAA13100 for {Microscopy-at-sparc5.microscopy.com} ; Sun, 2 Feb 2003 00:35:55 -0600 (CST) Received: from mail pickup service by mailgw.solutia.com with Microsoft SMTPSVC; Sun, 2 Feb 2003 01:30:32 -0500 Received: from soidmzvw01.solutia.com ([205.191.64.37]) by mailgw.solutia.com with Microsoft SMTPSVC(5.0.2195.5329); Sat, 1 Feb 2003 06:14:11 -0500 Received: from 206.69.208.10 by soidmzvw01.solutia.com (InterScan E-Mail VirusWall NT); Sat, 01 Feb 2003 06:09:35 -0500 Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id WAA07909 for dist-Microscopy; Fri, 31 Jan 2003 22:10:01 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id WAA07902 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Fri, 31 Jan 2003 22:09:28 -0600 (CST) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id WAA07895 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 31 Jan 2003 22:09:13 -0600 (CST) Received: from dns1.nmsu.edu (dns1.NMSU.Edu [128.123.3.5]) by bubba.NMSU.Edu (8.10.2+Sun/8.9.0) with ESMTP id h1143lc08143 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 31 Jan 2003 21:03:47 -0700 (MST) Received: from hector.NMSU.Edu (hector.NMSU.Edu [128.123.34.9]) by dns1.nmsu.edu (8.9.3/8.9.0) with ESMTP id VAA01052 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 31 Jan 2003 21:03:50 -0700 (MST) Received: from localhost (sghoshro-at-localhost) by hector.NMSU.Edu (AIX4.3/8.9.3/8.9.0) with ESMTP id VAA27220 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 31 Jan 2003 21:03:49 -0700 X-Authentication-Warning: hector.NMSU.Edu: sghoshro owned process doing -bs
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We use tin cookie boxes. They come in all different sizes and before christmas, 2001 we asked regular lab users to save their cookie boxes if they received any as christmas gift. They are light tight and so far have been working great.
By the way, we just asked for used, empty boxes, no cookies.
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
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Chris: light shining on a semiconductor (which the detector is) generates electron-hole pairs, just like an x-ray does. But the magnitude of the number of these pairs is much higher because there are usually more photons in a light beam than characteristic x-rays being generated by a sample. This flood of electron-hole pairs overwhelms the amplifier and the counting electronics. Some detectors have metallized (mere atoms of metal, so they won't interfere w/ analysis) windows to keep out the 'ambient' light, such as maybe generated by CL. But if your chamberscope's light source is oriented so that it shines directly on the detector, enough light can get in to cause problems. The collimator on the detector nose can also help keep light out. I've worked on SEMs that have had the chamberscope light source directly opposite the detector and had to be vigilant about turning it off when is was not needed. I now have a scope that has the light source behind the detector and it give no trouble at all. For the curious, on the Oxford Instruments site, on the page for their EDX hardware (http://www.oxford-instruments.com/ANLPDP174.htm) near the bottom right hand side is a list of "Related PDFs" and the last one is "EDX Hardware Explained". If you've ever wondered what's in the snout of a detector and how it works its magic, this is a good read. NOTE: I have no interest, financial or otherwise, in Oxford Instruments. Just a satisfied customer.
Chris Jeffree wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Don't know, but I would be interested to know. We need a physicist } here! } I am also intrigued to note that some of my specimens show } cathodoluminescence outputs sufficiently bright for the position of } the scanning beam to } be visible on the chamber cam. } How bright does it need to get to have an impact on counts, and what } artefacts might result? } } Chris } } ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; } {microscopy-at-sparc5.microscopy.com} } Sent: Thursday, January 30, 2003 9:38 PM } Subject: IR LEDS/EDS } } } } } } } } } I had a problem like this. } } } Turned out to be caused by the infra-red leds of my chamber } camera. } } } When the camera is switched off, the counts (3-5k per second) } return } } } close to zero. } } } } } } Chris } } } } } } } What would be the mechanism of this effect, I wonder? } } } } rtch } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } } The University of Auckland email : r.sims-at-auckland.ac.nz } } Private Bag 92019 } } Auckland } } New Zealand } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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Peter, In that case, a TEM would also be an electron microprobe. I think it is better if the names are used specifically. Electron microprobes are designed quite differently from SEMs. Call an SEM with WDS an SEM with WDS.
At 11:12 AM 1/30/2003 -0500, Peter Tomic wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Dear Ritchie, I believe the Si(Li) detector is sensitive to visible light, as it is to the x-ray photons. With a Be window the visible light could not penetrate to the detector, but with the new thin membrane windows the IR gets right through. Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 30, 2003 1:38 PM
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Gary I think, you wrong: fragmented file occupied more physical space on HD (yes) but fragmented/non-fragmented files has the same bit-size. So fragmentation may affect the reading/writing time only, which on the modern computers is negligibly. If the disk seriously fragmented, yes, it may affect computer's performance in general. "Save as" command not necessary writes in non-fragmented space. NTFS usually decently manage this issue and keep fragmentation at relatively low level, but it's OS decision where to place your file (for instance, if file is small, it would be placed in the NTFS analog of FAT at the beginning of drive- this is special precaution against fragmentation). If the HD is seriously fragmented, even "Save as" will cause the file fragmentation. Sergey
At 08:18 AM 1/31/2003, 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
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Please help. I'll pay any price for a decent copy of a Reichert OM-U3 ops/service manual. Someone was nice enough to send a copy of a copy, of a copy, etc., but all the illustrations are illegible. That was an ops manual, and I'll still need a service manual, or at least a schematic or wiring diagram. Anyone with some spare specimen block holders would be helpful also. The instrument that I have only came with one flat block holder. I have lots of lab stuff I would be happy to trade.
--
My address is: Equipment Resurrection 1005 Terra Nova Boulevard, Suite 2 Pacifica, CA 94044. The phone number is 650-738-0351 and web address, http://equiprx.net/.
EQUIPMENT RESURRECTION purchases only the highest quality surplus and used laboratory equipment. We do repairs and refurbishments to factory specifications, where and when necessary. Only when the instrument meets ALL our quality control specifications, do we resell to labs and scientists interested in high quality, used equipment that is, as good as new in performance, but may be lacking the all the latest features.
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MOST ORDERS SHIPPED WITHIN 24 HOURS PERSONAL CHECKS WELCOME DISCOVER, VISA AND MASTER CARD ARE ACCEPTED FOR ORDERS OVER $25. SORRY, I CANNOT ACCEPT CREDIT CARDS THAT DO NOT HAVE AN ADDRESS WITHIN THE USA.
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I think you would have to worry about two problems - (1) shrinkage during the actual polymerization step and (2) compression during thin sectioning. I don't have the reference off the top of my head but Daniel Studer has a nice paper (J. Microscopy about 1-2 years ago, I think) in which he shows significant (up to 50% if i remember correctly) along the cutting axis. his paper showed this could be avoided using an oscillating diamond knife. this paper has important implications for high resolution measurements since the compression would be different in the different axes.
At 10:08 AM 1/31/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
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Dear Jim, I use the black plastic bags that the photographic paper comes wrapped in to transport my film boxes to the dark room. I discovered the hard way that the boxes themselves were not light tight. Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Jim Romanow" {bsgphy3-at-uconnvm.uconn.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 30, 2003 1:36 PM
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On Friday, January 31, 2003, at 01:54 AM, Ji, Ying wrote:
} I am doing a double tilting experiment with TEM. From diffraction zone } A to } diffraction zone B, I tilted X axis for xx degree and Y axis for yy } degree. } Could anyone let me know how to calculate the angle between diffraction } zones A and B. } } Thank you very much in advance! } Dear Yun, I don't have my spherical trigonometry book here, having moved recently, but the way to approach the problem is to imagine the electron beam incident on the North pole with the Greenwich meridian facing you, then perform the tilts and locate the position of the incident beam after these have been done. If you have a double tilt holder like the one I used when I was in Albany NY, the tilts can be done in either order, so tilt in Y (keeping the Greenwich meridian facing you) so the beam is now incident on latitude 90-yy, then tilt through xx degrees along the great circle perpendicular to the Greenwich meridian and passing through it at latitude 90-yy. You will have a right spherical triangle; the hypotenuse is the angle you're looking for. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
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As I understood it at M&M 2002 in Quebec City, the Diatome oscillating knife was close to market, as in the 'fine-tuning' stage. You might want to inquire of Diatome US as to the state of that knife. The reduction in compression for soft materials was phenomenal.
Tom Malis
Dr. Tom Malis Scientist Advisor Natural Resources Canada Govt. of Canada 613-995-7358 malis-at-nrcan.gc.ca
-----Original Message----- } From: Tom Phillips To: Nahirney, Patrick (NIH/NIAMS) Cc: Microscopy-at-sparc5.microscopy.com Sent: 1/31/2003 11:49 AM
I think you would have to worry about two problems - (1) shrinkage during the actual polymerization step and (2) compression during thin sectioning. I don't have the reference off the top of my head but Daniel Studer has a nice paper (J. Microscopy about 1-2 years ago, I think) in which he shows significant (up to 50% if i remember correctly) along the cutting axis. his paper showed this could be avoided using an oscillating diamond knife. this paper has important implications for high resolution measurements since the compression would be different in the different axes.
At 10:08 AM 1/31/2003 -0500, you wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Light tight container for film transport?
Black plastic bags win the popularity (and economy) contest!
Quick list of light tight container suggestions:
Black plastic photo paper bag with or without cardboard box Ammunition storage box Paint can Pelican camera equipment case Spring loaded paper safe box Plastic tool/tote box Custom made stainless steel box
Thank you very much for all of the feedback. I am leaning toward the ammo box because it might hold up well under student abuse; the most bullet proof solution:}
Regards, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
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Dear Fellow Microscopists,
A colleague of mine recently asked about a type of "Laser Scanning Phase Contrast Microscopy", for which I do not have much knowledge.
Please advise: What's the major difference between this and the conventional LSCM that we use? Which institution in the East Coast might have such a facility?
Any advice is highly appreciated and will be forwarded to this colleague. Have a great weekend!
QC
Qian-Chun Yu, MB, Ph.D. Director Biomedical Imaging Core Laboratory University of Pennsylvania Department of Pathology & Lab Medicine School of Medicine 110 Richards Building Philadelphia, PA 19104
the resultant y" axis orientation (equivalent to the specimen normal in this case) is given as:
y" = -x sin(a)cos(b) + y cos(a)cos(b) + z sin(b)
The total tilt is found from the arccos of the dot product of the initial y axis and the final y" axis, which gives (assuming unit vectors)
cos(theta) = cos(a)cos(b)
which is what I stated this morning.
Dick Fonda
At 9:54 AM +0000 1/31/03, Ji, Ying wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Chris Jeffree wrote:
} } Don't know, but I would be interested to know. We need a physicist } here! } ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; } } } } } What would be the mechanism of this effect, I wonder? } } } } rtch }
I expected the usual suspects to leap on this, but since it hasn't happened, I will chime in. I are an engineer, not a physicist; take with the appropriate grain of salt, but I have worked in pulse processor design.
The Al coatings on thin window detectors do a good job of keeping out visible light, but are fairly transparent to IR. The IR photons are absorbed just like X-rays, but because their energy is so low and the flux high compared to X-ray photons, the result is a nearly continuous flow of current through the detector which is indistinguishable (to the pulse processing electronics) from a "leaky detector". Essentially, high leakage current raises the noise threshold setting required to prevent false triggering of the pulse processor. Since thin-window detectors are intended to trigger on low-energy X-rays, that threshold in a properly funtioning detector is set fairly low. Therefore, the IR- induced "leakage current" causes the pulse processing electronics to start triggering continuously on noise, causing a large peak at the low end of the spectrum.
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We're having an odd problem.
We're staining COS cells with Alexa 647 phalloidin (Mol. Probes A-22282 lot 41B1-1). The f-actin is staining fine and looks great when we excite at 633 nm and detect through the Leica AOBS or normal Cy5 filter set.
However, when we excite at 488 nm, we're seeing the f-actin staining appearing in the green channel too. We checked on different microscopes.
Has anybody else seen something as weird as this? We suspect contamination, but don't know where it would have come from unless it is an isomer or something from the manufacturing process.
One of our controls is the Alexa-phalloidin staining alone without any antibodies, so we know it's not some weird antibody binding artifact.
Any help appreciated.
Thanks.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
Those of you in the path of the debris feild of the tragic break up of the space shuttle Colombia have a unique opportunity to try to catch microscopic debris from it over the next few days as the winds move it over the south east part of the United States. This is the largest event of its kind and first chance for this kind of research.
Pans of glycerin, water or some other liquid or possibly the sticky side of tap or some kind of gel to trap the particles on roofs and in the open at ground level should catch this debris. Some kind of baffles to protect the pans from wind should help catch small particles and protect from contamination from the surrounding area.
Reports in California by an astronomer of small flashes following the shuttle as it pass over may extend the area were debris can be found.
The current winds aloft should carry the debris along the Gulf Coast and across central Florida if they continue as they are now.
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
I'm trying to set up the column alignment of my JSM 840, and rotation of the Y stigmator gives such a lot of image shift that it's hard to use. The X stig gives almost none.
There are trim pots which balance the currents to the stig coils, but the amount of image shift is way more than can be corrected for by the trim pots.
The currents flowing in all of the 8 coils (4 'X', 4 'Y') are all very similar, and they all change similarly when the stig controls and the trim pots are turned, so it seems that the stig electronics and the coils are all OK.
Is this effect likely to be caused by a gross misalignment of the column?
Any tips on how to remedy?
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
In my last order of Kodak 4489 film, the boxes were marked "new formulation". Since it has been some time since I've needed to order film, I was wondering if any of you have noticed sufficient differences in exposure, density, etc. with this new film that required re-calibration of the photographic parameters on your microscopes. The last time Kodak re-formulated their emulsion, we needed to do extensive testing and calibration of our TEMs to obtain photos with the desired exposure levels.
Thanks.
Valerie M. Knowlton Research Assistant/Teaching Technician Center for Electron Microscopy 1219 Gardner Hall, Box 7615 North Carolina State University Raleigh, NC 27695
Richard Fonda is right. I made a foolish mistake when multiplying the two rotation matrixes together.
My sincerest apologies. Paul
======================= Paul Baggethun Engineer Alcoa Technical Center Alcoa Center, PA 15069 USA =======================
-----Original Message----- } From: Richard W. Fonda [mailto:fonda-at-anvil.nrl.navy.mil] Sent: Friday, January 31, 2003 4:20 PM To: Ji, Ying; 'microscopy-at-sparc5.microscopy.com'
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There was some debate about the answer I gave to this question this morning, so I looked into the math a bit further.
If you consider the standard rotation of cartesian coordinates about the z axis by an angle, a, as follows:
x' = x cos(a) + y sin(a) y' = -x sin(a) + y cos(a) z' = z
and follow this by a rotation about the x' axis by an angle, b:
the resultant y" axis orientation (equivalent to the specimen normal in this case) is given as:
y" = -x sin(a)cos(b) + y cos(a)cos(b) + z sin(b)
The total tilt is found from the arccos of the dot product of the initial y axis and the final y" axis, which gives (assuming unit vectors)
cos(theta) = cos(a)cos(b)
which is what I stated this morning.
Dick Fonda
At 9:54 AM +0000 1/31/03, Ji, Ying wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings Betsey, It is a wise idea to contact confocal facilities and get some input regarding experiences with established instrumentation. Another forum you may wish to query is the Confocal Listserver {confocal-at-listserv.buffalo.edu} , and you may wish to look at the archives (http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal); there was a pertinent thread discussing the two platforms last week. There is a listserver set up as a resource for administrators of Leica CFM equipment as well which you may wish to address--subscribers include 60 or so facility coordinators around the globe who manage Leica confocal microscopes. If you would like I can forward your query to this listserver (you have to be subscribed to post as an anti-spam measure). Lastly, you might try to find facilities on the web (eg. enter "Leica SP-2" into a Google search and see what comes up). Most facilities are happy to share their experiences. No high tolerance instrument will perform without incident indefinitely and intelligent purchase decisions take this into account. Sophisticated platforms such as the SP-2 and the META showcase synergy between a number of instrument sub-systems; seemingly minor problems or mis-calibrations in the imaging pipeline can wreak havoc with important measurements. It is important that problems can be addressed effectively and efficiently--for this it is necessary to have sincere support from a particular manufacturer. An instrumentation purchase choice implies a commitment to a relationship with a particular brand for the useful lifespan of the instrument. It is important for suppliers of confocal instrumentation to remain abreast of cutting edge technology, but it is also important for demonstrate a commitment to regular maintenance and dedication to support of established instruments. Four areas you may wish to get candid information on include:
1.) Availability of service personnel. Service engineers should have effective support with scheduling, and should be able to provide an accurate timeline to the facility visit.
2.) Effective communication between the domestic and global service infrastructure as well as between service management and facility managers.
3.) Parts availability. Replacement parts should be available, and these parts should be ensured to perform acceptably before installation
4.) Preventative maintenance. Heavily used, research critical LSM instrumentation should be overhauled in a disciplined manner at regular intervals to ensure maximum reliability. The rate at which certain components deteriorate should be predictable, and a thorough checklist which ensures that all the components are performing up to specification at regular intervals would do much to bolster end user confidence.
That being said, I feel that the Leica SP-2 showcases some elegant engineering solutions and I'm not infrequently re-impressed with the instrument's capabilities. When the platform is in top working condition I would put it up against any other laser scanning microscope (the newer AOBS feature seems to be a promising development as well, but we don't presently have that capability). I haven't had nearly as much experience with the META at this point, but I'm sure there are facilities which would be happy to provide you with input about instrument performance.
Good luck in your decision--feel free to contact me directly if you have specific questions or concerns.
Best Regards, Karl G.
On Saturday, February 1, 2003, at 01:27 PM, Pitts, Betsey wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } } Hi- } we are in the process of purchasing a new confocal, and are looking } at a Zeiss510 Meta or a Leica AOBS or SP2. I think that in the demo } process } we are seeing what we need to, and are impressed by the capabilities of } each. I would appreciate user perspectives that we cannot get from the } manufacturers though- are there instrument aspects of either } microscope that } are repeatedly problematic or limiting? Do multi-user facilities have } particular difficulty with either? I do not expect an obvious } "winner" for } an answer, but am more interested in hearing what some of the ups and } down } are, and seeing if those problems or circumstances would be issues for } us. } Thanks for any experience you an offer, I am sure it will be useful. } } Betsey } *********************************************************************** } * } Betsey Pitts } Research Associate/Facilities Manager, Microscopy } The Center for Biofilm Engineering } } 366 EPS Building, Montana State University } Bozeman, MT 59717 } } betsey_p-at-erc.montana.edu } Ph (406) 994-7813 } Fax (406) 994-6098 } } *********************************************************************** } * } } Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, Champaign 61801 Office: B650J Phone: 217-244-6292 Fax: 217-244-6219
Hello Everyone I may be opening a can of worms here but....
What do TEM labs do about the dust created when polymerised resin is cut with a hacksaw or rasp or razor blade. Any resin - Spurr's, Epon, HM20, LR Gold, LR White, etc etc.
Is there a vaccum system recommended? Or do you just use a wet towel system. Or brush the dust into the garbage can and create dust in the atmosphere and not worry about it. Do you have a policy of only cutting resin down to manageable size in a fume hood and then vaccum up the dust? Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility First Vice President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
Hi All: I am looking for online responses as suggestions of KNOWN suppliers of Denka M3 Lab6 filaments for a JEOL TEM. I would invite offline responses as to specific prices. (Essentially, I am comparison shopping for the supplier with the best prices). Thanks in advance, Michael Coviello Lab Manager, UT Arlington
Hello Everyone The annual meeting of the Microscopical Society of Canada is meeting in Vancouver this year June 4-6, 2003 at the University of British Columbia.
This is a call for papers for two concurrent session in the Biological and Physical Sciences
Instructions for authors, registration and accomodation can be found on the website http://www.emlab.ubc.ca
The list of workshops, exhibitors, local University tours, and invited speakers should be on the website soon. There are links to Tourism Vancouver should you wish to extend your visit to one of the most beautiful places on this planet. (You can probably tell - I am biased).
And there are links to the International Cryo EM course which will be in the following week. Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility First Vice President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
} Is there a vaccum system recommended? Or do you just use a wet towel } system. Or brush the dust into the garbage can and create dust in the } atmosphere and not worry about it. Do you have a policy of only } cutting resin down to manageable size in a fume hood and then vaccum } up the dust?
I personally use the wet paper towel system. Wrap it up well and then throw it in the trash. Of course, then I wonder about its ultimate fate. Over here a large proportion of our wastes go to an incinerator to generate electrical power, and then I worry about toxic vapors released into the atmosphere. Can't decide if that's better or worse than landfill, where stuff can leach into our water system!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Sergey is correct, disk fragmentation has little effect on file size. If you have the program preferences set to "allow fast saves" MS Word will save the recent changes made to your file, along with the original file, thereby making the total file size ever larger (until a certain size is reached after which the file is saved as an original). If you choose "Save as" then the document is saved as an original, minimizing the file size. You can force the smallest size files by de-selecting "allow fast saves"; it will take only a fraction of a second longer to save each time (unless you are writing a book and have a huge file).
Kim {} {} {} {} {} Kim Rensing PhD Department of Botany, UBC 6270 University Blvd. Vancouver BC, Canada V6T 1Z4
On Friday, January 31, 2003, at 08:43 PM, Sergey Ryazantsev wrote: } Gary } I think, you wrong: fragmented file occupied more physical space on HD } (yes) but fragmented/non-fragmented files has the same bit-size. So } fragmentation may affect the reading/writing time only, which on the } modern computers is negligibly. If the disk seriously fragmented, } yes, it may affect computer's performance in general. "Save as" } command not necessary writes in non-fragmented space. NTFS usually } decently manage this issue and keep fragmentation at relatively low } level, but it's OS decision where to place your file (for instance, if } file is small, it would be placed in the NTFS analog of FAT at the } beginning of drive- this is special precaution against fragmentation). } If the HD is seriously fragmented, even "Save as" will cause the file } fragmentation. Sergey } } At 08:18 AM 1/31/2003, you wrote: } } } } What usually happens is that the document's file } } becomes fragmented. In this case, more sectors } } on disk are used (wasted) and leads to a larger } } file size. If you routinely keep disk fragmentation } } low, then do a Save As with the same file name. } } This should put the new save in a contiguous } } area. } } } } gary g } }
Hello all, first off, I am a complete novice, last time I looked through a microscope was in high school, too many years ago to mention. I have done some research though-so I'm not completely in the dark. (Besides, I'm a geek in disguise anyway.) What I want to do is to be able to examine samples of my sourdough starter to see if, and if possible, what kinds of lactobacillis are present in the starter. I have a scope w/capability of 970 power (10x/97xOil)... I was able to stain a sample of yogurt w/methylene blue sucessfully and see some rod shaped bacteria in that slide. They were smaller than I thought they would be, but I could distinguish them. My attempts w/the sourdough are less sucessful. I take a sample of the starter and dilute it and then put a drop on the slide, dry it, pass it through the flame, and stain it. I've found instructions for preparing slide with crystal violet that are a little more involved, using counterstaining etc. Would I be better off getting some cyrstal violet and using this procedure, rather than the more simple method w/the methylene blue, or am I just trying to do something that isn't really possible w/a scope w/only 970 capability. Any advice or direction to documents that would help would be preciated. -cj-
Hello Elaine Nice to hear from you. I was on your Cryo-EM course in the summer last year and still not received yet the samples, which supposed to be processed on that course. If I do remember correctly you promised to me to sent those samples ASAP. Is it still possible to get them back? Your course was very costly to me. The samples were (it was discussed with you prior I signed for the course) from trangenic mice with BM transplantation. The cost of such mice is a few thousand $$ each... As a matter of fact I was disappointed how the Cryo-course where handled. The samples were not processed in time (was sitting in refrigerator waiting for what?), specific antibodies were not ordered in time and then where lost, we did not have necessary reagents in time and equipment was constantly busy... I understand, it's normal laboratory life when you have to make reagent at the last moment (in the cosy environment of your own Lab, not in yours) or wait for equipment available, but it was Cryo-course I paid for that nearly from my pocket a few thousand $$. I was expecting to have at least attention to my needs at such price. I am sorry I made this public statement, but it seems to me, people should know what they may expect to have from the course and I am still WANT BACK MY SAMPLES! Sergey
At 03:05 PM 2/3/2003, 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
Because of a recent posting I find it appropriate to remind you all of the Listserver Rules, which you all received copies of upon subscription confirmation. In particuliar I draw your attention to #4. If you have a problem with an organization, this is NOT the place to air it.
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Nestor just informed me that my posting (see below) is inappropriate on this ListServer, because I violate the rule. Rule #4 states:
# 4.) This forum is not to be used as a platform to accuse or defame any individual or organization in any negative manner. You may disagree with any comment posted and post a reply, but this server may not be used to spread misleading, derogatory or disparaaging comments under any conditions.
I am deeply apologize, I broke the rule and let you to be exposed to my message. I did not intend to hurt anybody, but express my sincere impression about mentioned here CryoEM course. I also deeply concerned I could not receive my very valuable samples for more than 6 month. Again, I apologize, I posted my message against the rules. I already contact to Nestor, so he could help me to understand what was wrong in my message and how I could avoid mistakes in the future. Sincerely, Sergey Ryazantsev.
At 08:38 PM 2/3/2003, 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
Hi Folks, Is there any one out there who knows how to refurbish windows for gas flow counters? These are for a JEOL 733. I've a heap of mylar and a stack of old windows and no idea what to do. I would like to know, how to clean the old stuff off, how thick the mylar should be, how to attach a new bit and whether these need C coating afterwards and if so how thick a layer. Cheers, Malc -- Dr MP Roberts Phone: [+27](0)46 603 8313 (work) Dept of Geology [+27] (0)46 6361197 (home) Rhodes University Fax: [+27](0)46 622 9715 6140 Grahamstown Cell: 083 4060 262 SOUTH AFRICA e-mail: m.roberts-at-ru.ac.za
I always try to use an appropriate size of embedding mold to minimise the need for cutting a lot of resin and make sure that the specimen is as close to the tip as possible. Trimming should then be possible by razor blade or glass knife on the microtome and I haven't used a hacksaw on a resin block for some time.
In the unlikely event that I need to saw a specimen I would do so in the fume hood using a small detachable vice to hold the block. The dust would be collected using a damp paper towel, placed in a pot and then dried. This can easily be made safe later by topping up with some waste unpolymerised resin and polymerising for disposal as normal waste.
Many years ago we used an ordinary portable vacuum cleaner but very quickly realized that this would churn out the most hazardous particle sizes of dust. There are apparently lots of new vacuum cleaners with filters in them but I don't know whether their performance would be guaranteed for potentially carcinogenic dust. I have, however, seen a couple of portable machines advertised for disposal of photocopier toner dust which may be suitable. I still don't think that resin dust should be encouraged even if you intend to clean it up later because you don't know how much is already airborne.
I have little worry about the slivers and chips of resin created by razor and glass knife cutting because they are too big to be inhaled (as far as I know), settle very quickly and can be brushed into a waste pot easily. But I would be very concerned about brushing, blowing or vacuuming ~ micron size particles. Of course 20+ years ago information was a bit sparce about the hazards of resins and their dusts and we treated them with a lot less care.
Good luck
Malcolm
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK ----- Original Message ----- } From: Elaine Humphrey {ech-at-interchange.ubc.ca}
Thanks everybody, for your responses. we finally found water in the HT tank. Trying to arrange for oil to change it.
Shashi Singh Scientist CCMB, Hyderabad INDIA
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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The program committee for M&M 2003 would like to remind everyone that presentation submissions are due in less than two weeks (February 17). Instructions for submission and details about the meeting are available on-line at:
The meeting is shaping up to be very exciting with particular focus on Nanotechnology and Optical Microscopy, as well as the broad coverage of Electron Microscopy techniques and applications that you expect at M&M. The program committee encourages you to participate by submitting a paper on your work, and we hope to see you in San Antonio!
I just want to say that the recent comment by Sergey to Elaine Humphrey re: problems with the Cryo course at last year's MSC meeting was inappropriate. He has a personal complaint that should have been directed specifically to Elaine. It was totally uncalled for to air it to the whole ListServer community.
I've noticed that this type of grudge comments do occur occasionally on the ListServer and take up valuable time and space. Please limit comments like that to the person involved.
Peggy Sherwood
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
In our lab we have an plain old household vaccum cleaner that is used to suck up the dust. The hose used to be set up on a stand so that it could suck up the dust as it is made, but the stand wasn`t functional when I got here 2 years ago, so I couldn`t tell you how it worked. These days I just vaccum the dust up as I go along and try not to let it get all over the place (we don`t work in a fume hood). The vaccum bag has never needed to be emptied in my time here, so I have no idea how the waste is dealt with.
I hope this helps, Robin __________________________ Robin Elizabeth Young Laboratoire de Jacques Paiement Université de Montréal re.young-at-umontreal.ca
} What do TEM labs do about the dust created when polymerised resin is } cut with a hacksaw or rasp or razor blade. Any resin - Spurr's, Epon, } HM20, LR Gold, LR White, etc etc. } } Is there a vaccum system recommended? Or do you just use a wet towel } system. Or brush the dust into the garbage can and create dust in the } atmosphere and not worry about it. Do you have a policy of only } cutting resin down to manageable size in a fume hood and then vaccum } up the dust?
Elaine and Robin: If you are using a plain old vacuum, I would bet you are capturing the big particles and simply exhausting the small, more dangerous ones. that's why a regular vacuum is worse for some allergic to dust. They make vacuums with HEPA filters now but I don't know if they are really effective. the water trap ones are not according to studies i have read. the best option for vacuums is one that vents to the outside (e.g., a built-in whole house vacuum) and these are widely recommended for those with bad dust allegies. The bottom line is that you may be making things worse since you are distributing them into the air.
We use a Dremel moto-tool for trimming our blocks. Vastly superior to a hacksaw or razor blade in my opinion but it generates a ton of dust. I would never do it outside the fume hood. My guess is you would be risking the equivalent of silicosis. Tom
At 10:49 AM 2/4/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Dear Malc, It has been a long time since I did this on my JEOL JXA-3A, but we used to use 4 or 6 micron mylar, stretch it over the window and glue it on with 5-minute epoxy, then trim off the excess. I believe we sanded off the old epoxy from the brass window holder. The windows are usually then lightly aluminum-coated so they won't charge. Good luck Mary Mager Electron Microscopist Metals and Materials Eng. UBC 6350 Stores Rd. Vancouver, B.C. CANADA tel: 604-822-5648 fax: 604-822-3619 ----- Original Message ----- } From: "Malc" {m.roberts-at-ru.ac.za} To: "Microscopy discussion group" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 04, 2003 1:57 AM
The answer is that Alexa 647 has a component that, when excited at 470 to 490 nm, has an emission very similar to FITC. This has been confirmed by our spectrophotometer, a colleague with the Zeiss Meta (posted on the list) and Molecular Probes themselves.
Practically, this means that while the dye works great at 647 nm, it cannot be used with GFP or other staining in the green range.
We're gonna try Alexa 633.
Thanks!! ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
If there is texture on the surface, what it the general size of the features? Also, what is the thickness specification for the coating?
If you want to do thickness measurements non-destructively, have you considered Raman confocal?
There are a number of approaches to this problem. Please feel free to call me off-line for further discussion.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
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At 05:21 PM 1/31/03 -0600, Chris Michaelson (by way of wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } The Al coatings on thin window detectors do a good job of } keeping out visible light, but are fairly transparent to IR. } The IR photons are absorbed just like X-rays, but because } their energy is so low and the flux high compared to X-ray } photons, the result is a nearly continuous flow of current } through the detector which is indistinguishable (to the } pulse processing electronics) from a "leaky detector".
Just off-the-cuff, and in ignorance, I'm surprised that IR has enough energy to produce any elextrons/holes at all, as I didn't think it would get through the Au layer.
I guess this must be the right explanation, though
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Has anyone tried the Leica EM Trim specimen trimmer?
Uses a milling tool to trim down blocks, while observing througha stereo microscope head. Has a vacuum connection to trim away the nasties.
Thinking of buying one so any feed back would be appreciated.
Allan -- ------------------------------------------------- Allan Mitchell Technical Manager Otago Centre for Electron Microscopy C/-Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
I want to thank all who responded with offers for manuals. Someone was able send me a scanned copy that is as close to an original as I could hope for. I understand that the arm bearing surfaces are probably damaged. The instrument was transported without using the shipping blocks, even though they had them in the drawers. So my question is, does anyone know of a source for replacement specimen arm bearings? I already called Leica and they said good luck. Very best regards to all who replied, Steve D'Angelo
--
Equipment Resurrection 1005 Terra Nova Boulevard, Suite 2 Pacifica, CA 94044. 650-738-0351 http://equiprx.net/
I bought a hand held domestic vacuum cleaner. Then one day I was using it over a black bag and noticed that it appeared to be depositing fine dust.
I returned to the old method. I do sawing inside a large plastic bag in a fume hood. I knot the bag and store it the cupboard (which is vented by the system) under the hood. When it is full in oh... 2024 when I retire I will polymerise it.
Dave
On Tue, 04 Feb 2003 10:42:28 -0600 Tom Phillips {phillipst-at-missouri.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Elaine and Robin: If you are using a plain old vacuum, I would bet you are } capturing the big particles and simply exhausting the small, more dangerous } ones. that's why a regular vacuum is worse for some allergic to } dust. They make vacuums with HEPA filters now but I don't know if they are } really effective. the water trap ones are not according to studies i have } read. the best option for vacuums is one that vents to the outside (e.g., a } built-in whole house vacuum) and these are widely recommended for those } with bad dust allegies. The bottom line is that you may be making things } worse since you are distributing them into the air. } } We use a Dremel moto-tool for trimming our blocks. Vastly superior to a } hacksaw or razor blade in my opinion but it generates a ton of dust. I } would never do it outside the fume hood. My guess is you would be risking } the equivalent of silicosis. Tom } } } } At 10:49 AM 2/4/2003 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi Elaine, } } } } In our lab we have an plain old household vaccum cleaner that is used to } } suck up the dust. The hose used to be set up on a stand so that it could } } suck up the dust as it is made, but the stand wasn`t functional when I got } } here 2 years ago, so I couldn`t tell you how it worked. These days I just } } vaccum the dust up as I go along and try not to let it get all over the } } place (we don`t work in a fume hood). The vaccum bag has never needed to be } } emptied in my time here, so I have no idea how the waste is dealt with. } } } } I hope this helps, } } Robin } } __________________________ } } Robin Elizabeth Young } } Laboratoire de Jacques Paiement } } Université de Montréal } } re.young-at-umontreal.ca } } } } } What do TEM labs do about the dust created when polymerised resin is } } } cut with a hacksaw or rasp or razor blade. Any resin - Spurr's, Epon, } } } HM20, LR Gold, LR White, etc etc. } } } } } } Is there a vaccum system recommended? Or do you just use a wet towel } } } system. Or brush the dust into the garbage can and create dust in the } } } atmosphere and not worry about it. Do you have a policy of only } } } cutting resin down to manageable size in a fume hood and then vaccum } } } up the dust? } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Steve: I don't know where you are located, but Helmut Patzig, of MOC, Valley Cottage, NY, keeps the OMU-3 here running with the odd bits. Phone number is 845-268- 6450. He might have the arm or bearing (I damaged one of those many, many, many years ago!!) and some other bits you will likely need. However, _no one_ seems to have any more of the drive belts!
Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals -- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 want to thank all who responded with offers for manuals. } Someone was able send me a scanned copy that is as close to an original } as I could hope for. } I understand that the arm bearing surfaces are probably damaged. } The instrument was transported without using the shipping blocks, even } though they had them in the drawers. } So my question is, does anyone know of a source for replacement specimen } arm bearings? } I already called Leica and they said good luck. } Very best regards to all who replied, } Steve D'Angelo } } -- } } } Equipment Resurrection } 1005 Terra Nova Boulevard, Suite 2 } Pacifica, CA 94044. } 650-738-0351 } http://equiprx.net/ } } }
I have just seen a note in Microscopy Today, (downloaded from http://www.microscopy-today.com follow the links to the back issue Table of Contents), (March/April 2002).
Karen Pawlowski uses a method that traps the dust in water in a flask.
Dave
On Tue, 04 Feb 2003 10:42:28 -0600 Tom Phillips {phillipst-at-missouri.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Elaine and Robin: If you are using a plain old vacuum, I would bet you are } capturing the big particles and simply exhausting the small, more dangerous } ones. that's why a regular vacuum is worse for some allergic to } dust. They make vacuums with HEPA filters now but I don't know if they are } really effective. the water trap ones are not according to studies i have } read. the best option for vacuums is one that vents to the outside (e.g., a } built-in whole house vacuum) and these are widely recommended for those } with bad dust allegies. The bottom line is that you may be making things } worse since you are distributing them into the air. } } We use a Dremel moto-tool for trimming our blocks. Vastly superior to a } hacksaw or razor blade in my opinion but it generates a ton of dust. I } would never do it outside the fume hood. My guess is you would be risking } the equivalent of silicosis. Tom } } } } At 10:49 AM 2/4/2003 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi Elaine, } } } } In our lab we have an plain old household vaccum cleaner that is used to } } suck up the dust. The hose used to be set up on a stand so that it could } } suck up the dust as it is made, but the stand wasn`t functional when I got } } here 2 years ago, so I couldn`t tell you how it worked. These days I just } } vaccum the dust up as I go along and try not to let it get all over the } } place (we don`t work in a fume hood). The vaccum bag has never needed to be } } emptied in my time here, so I have no idea how the waste is dealt with. } } } } I hope this helps, } } Robin } } __________________________ } } Robin Elizabeth Young } } Laboratoire de Jacques Paiement } } Université de Montréal } } re.young-at-umontreal.ca } } } } } What do TEM labs do about the dust created when polymerised resin is } } } cut with a hacksaw or rasp or razor blade. Any resin - Spurr's, Epon, } } } HM20, LR Gold, LR White, etc etc. } } } } } } Is there a vaccum system recommended? Or do you just use a wet towel } } } system. Or brush the dust into the garbage can and create dust in the } } } atmosphere and not worry about it. Do you have a policy of only } } } cutting resin down to manageable size in a fume hood and then vaccum } } } up the dust? } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I hate the way Word deals with images. It makes huge files too. I would recommend Adobe Pagemaker or Deneba Canvas for much more controllable combination or text and graphics. If you have Acrobat (full version) installed you can then also export the files to pdf format and share them easily on the web. Of course, these software packages are not so ideal for presentations and I have not yet found an alternative to Powerpoint for that.
(P.S. This is not just Microsoft bashing, I wish they would do a better job of Graphics handling in word. In my opinion, the competition is better at this just now. As a pure Word processor I find MS Word very good).
Best wishes
Ian
On Mon, 20 Jan 2003 09:30:47 -0600, {"gary.m.brown-at-exxonmobil.com"-at-sparc5.microscopy.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Jeff, } } I, too, have had significant problems working with images imported into } Microsoft Word. However, my software is located on the PC hard drive. The } biggest problem that I have encountered with images in Microsoft Word } occurs when annotating images. After the image is imported into Word, } annotation may be done in two ways (to my knowledge): (1) Text, arrows, } etc. may be simply superimposed over the images. The problem with this } approach is that the annotations are not linked to the image and may not } remain superimposed on the image if the image moves. (2) Annotations can } also be linked (probably not the best choice of words) to the image by } double-clicking on the image to open the image field, adding the } annotation, then closing the image field. These annotations are permanent } unless intentionally moved or deleted. } } The problems occur when one implements the second option. Comparing } images } before and after annotation, I found that the annotated images often } sustained substantial changes in gray or color levels. Case in point, EDS } maps were so badly affected that the color key was no longer correct. } } My solutions follow: (1) Annotate images in Adobe Photoshop before } importing into Word. Note that the effects of lossy compression on } annotations (blurred edges) may be pronounced. (2) Use Microsoft } PowerPoint } for image presentation. I have encountered no problems with image files } in } PowerPoint. } } Good luck to you in your endeavors. } } Cheers, } } "The opinions expressed are those of Gary M. Brown and do not represent } the } opinions of ExxonMobil Corporation nor its affiliates." } } Gary M. Brown } ExxonMobil Chemical Company } Baytown Polymers Center } 5200 Bayway Drive } Baytown, Texas 77520-2101 } phone: (281) 834-2387 } fax: (281) 834-2395 } e-mail: Gary.M.Brown-at-ExxonMobil.com } } } "Oakley, Jeff" } {oakleyj-at-rayovac.c To: {Microscopy-at-sparc5.microscopy.com} } om} cc: } Subject: RE: presentation images } 01/17/03 07:50 AM } } } } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This same phenomenon happens with my reports in Microsoft Word. In } addition to images darkening, they sometimes shift to other pages and/or } change size and dimension. Adding tables and text boxes to the report } adds } to the fun. } } The software that we use is networked. Our IS department has told us } that } the networked software has a bug that causes these things to happen when } file sizes increase, and that there is not a patch for it. So we have } just } have to deal with it. } } Jeff Oakley } } } -----Original Message----- } } From: Corazon D. Bucana [mailto:bucana-at-audumla.mdacc.tmc.edu] } Sent: Thursday, January 16, 2003 11:11 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: presentation images } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I created a Power point presentation last November 2002 consisting of } several fluorescence micrographs. The file which was rather large ( 90 } Mb) } } was left in my laptop all this time. When I opened it again this week I } find that the images are now too dark and I needed to increase brightness } by 3-4 clicks on the brightness icon of Power point.I increased } brightness } } of all the micrographs and copied the file to a CD hoping that the image } will not deteriorate there and then compare it with the one in my laptop } several weeks from now. Has this happened to anyone else? Is there } something I should have done to prevent this? } } Any suggestions or comments will be greatly appreciated. } } Cora Bucana } } } } } } } } }
-- Ian MacLaren Technische Universität Darmstadt Material-und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany
I'm curious as to how many people have had problems using molecular sieves in dehydration solvents, with respect to knife damage. We seem to be going through diamond knives at a uncomfortable rate and we're wondering if this could be a contributing factor. We are very careful with our knives and try to minimize contact cleaning of the edges. We soak the knives often in the recommended solution in a commercial cleansing unit, and the knife manufacturer has evaluated one of our knives and confirmed that the edge is chipped, not dirty.
As a multi-user facility, we do a LOT of ultramicrotomy on a large variety of samples, so this may just be normal wear and tear, but we would sure like to minimize this expense.
Thanks much!
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
I was wondering if anyone on the list is familiar with this stain. I am trying to find a procedure for either making this stain or a commercial source for the stain. It was listed in a methods section for negative staining isolated neurofibrillary tangles (Crowther, R. A. 1991. PNAS 88:2288) but the author did not reference a source for the stain or a procedure. I would appreciate any help provided.
Thanks
Lawrence X. Oakford, Ph. D. Technical Manager Microscopy Core Facility Department of Cell Biology and Genetics UNT Health Science Center 3500 Camp Bowie Blvd. Fort Worth, TX 76107
Preparing my abstract for MM'03 I have found that: - Two page document (4 images with superimposed EDS line scans) saved in Word 2000 format was 758K size. - Saved in Word 6.0 format (as required for uploading by submission instructions) it had size of 2.93M (!). - Saved in PDF format (Acrobat 5.0) file was just 286K, but line scans, still pretty visible, were looking not nice.
I believe I should upload PDF files, since Word files anyway will be converted in PDF for CD publishing. But are not we loosing quality going digital now?
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: Rosemary White [mailto:rosemary.white-at-csiro.au] } Sent: Thursday, January 30, 2003 4:00 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: presentation images in Microsoft Word } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Another trick is to put the images and text into a Word } table, if you size } the cells in the table to about the size you want, the images } will insert } to fit the cell size. } } And if copying and pasting, make sure you do "Paste Special" } and unclick } the box that says "float over text". That way the images } always stay in } the same spot with respect to the surrounding text and don't } mysteriously } jump around when you insert or delete text. } cheers, } Rosemary } } } } Thanks Doug- } } Great tips- I'm printing them up until I memorize them. } } Rgds, } } Mike Shaw } } Roselle, NJ } } } } } Gary, } } } } } } MS Word is such a pain because of the way it works with } images. A couple } } } of tricks I've discovered over the years. } } } } } } 1) Place the image within a text box, rather than directly } on the page, it } } } seems to give you much greater control over the location } on the page. It } } } also makes it much easier to add text annotations that } stay with the image. } } } } } } } } } If you don't want the text box to have a border you can } remove it by } } } selecting } } } the box outline and look for the "paint brush" icon on the } Draw toolbar, } } } then use the down arrow and select "none". If you'd like } the text box to } } } be transparent, select the text box outline and look for } the "paint bucket" } } } icon on the Draw toolbar, then use the down arrow and } select "none". If } } } you discover its hard to find the text box border once you } made the edge } } } "invisible", first select the image with a single left } click (it should have } } } the solid black resizing "handles") and then use the } keyboard left or right } } } arrow keys and the selection with move out to the textbox } outline (with } } } black } } } bordered resizing "handles"). } } } } } } 2) Always use the INSERT | PICTURE | FROM FILE option as } opposed to copying } } } and pasting an image into Word. I find that the images } are harder to work } } } with if I paste them in. Also, you can insert TIFF or BMP } } } images into Word, } } } you don't have to use JPEG. } } } } }
I run a clinical EM lab with a geriatric JEOL 100C TEMSCAN microscope. I am campaigning for new equipment and would like to get a stand alone TEM and SEM. Currently my scope is in a rather large room that should be able to accommodate two instruments with their columns at least ten feet apart. I would hate to have new instruments installed only to find that they interfered with each other, either electrically or mechanically (vibration, etc.). I would like to hear from anyone who has wrestled with this problem....or is it a problem? I am on the second floor and not too near elevator shafts or electrical traces and vibration for my one scope has not been a problem.
Joiner Joiner Cartwright, Jr., Ph.D. Department of Pathology Baylor College of Medicine Houston, Texas U.S.A.
The New England Society for Microscopy announces its Early Spring meeting, to be held at the JEOL(USA) Inc. facility in Peabody, Massachusetts.
This is something we discovered over 20 years ago: molecular sieves work well BUT it is necessary to allow the alcohols to stand untouched for one month in order for the ceramic-like "fines" to settle out. Also, be very careful when withdrawing sieve-dried alcohols. Do not pour the alcohols but use a pipette and remove it from the top of the liquid. When the level drops to less than 1 inch above the sieves, it's time to move on to the next bottle. Basically, we would prepare about 6-10 pints of ethanol at one time, allowing them to "age" for at at least 30 days.
I must admit that now I tend to use 100% ethanol right out of freshly opened containers (individually sealed pints) except in the most critical of applications (Spurr's dehydrations, for example) and have NEVER had a problem with water.
I know of at least two investigators (not me) who have damaged diamond knives by not taking precautions with molecular sieves. Basically, once the fines get onto your specimen they are impossible to remove and will damage your diamond knife.
} I'm curious as to how many people have had problems using molecular } sieves in dehydration solvents, with respect to knife damage. We seem } to be going through diamond knives at a uncomfortable rate and we're } wondering if this could be a contributing factor. We are very careful } with our knives and try to minimize contact cleaning of the edges. We } soak the knives often in the recommended solution in a commercial } cleansing unit, and the knife manufacturer has evaluated one of our } knives and confirmed that the edge is chipped, not dirty. } } As a multi-user facility, we do a LOT of ultramicrotomy on a large } variety of samples, so this may just be normal wear and tear, but we } would sure like to minimize this expense.
############################################################## 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 ##############################################################
Thank you, Valerie. That's a good point about monitor light, etc. and perhaps a curtain arrangement might help. As for the number of people in the confined space....Is this an advantage or disadvantage??
Joiner ++++++++++++++++++++++++++++++ At 03:04 PM 02/06/2003 -0500, you wrote: We have two microscopes (a JEOL 100S TEM and a Philips 505 SEM) within a space of about 15 X 20 feet, but separated into 2 rooms, each about 15 X 10 ft. The columns are about 11 feet apart and we have absolutely no problems with interference or vibration. However, unless both of the new microscopes can be operated in room light (i.e. have computer monitors & digital capture), putting both scopes into one room could be a problem when both scopes need to be used at the same time. Also, if the space is really small, do you want that many people in there at one time? Having space for the equipment is one thing, but room for the users too, is another.....
Valerie
+++++++++++++++++++++++++++++++
Hello Listrers -
I run a clinical EM lab with a geriatric JEOL 100C TEMSCAN microscope. I am campaigning for new equipment and would like to get a stand alone TEM and SEM. Currently my scope is in a rather large room that should be able to accommodate two instruments with their columns at least ten feet apart. I would hate to have new instruments installed only to find that they interfered with each other, either electrically or mechanically (vibration, etc.). I would like to hear from anyone who has wrestled with this problem....or is it a problem? I am on the second floor and not too near elevator shafts or electrical traces and vibration for my one scope has not been a problem.
Joiner Joiner Cartwright, Jr., Ph.D. Department of Pathology Baylor College of Medicine Houston, Texas U.S.A.
Our lab purchased a Leica EM Trim about a year ago. It does a pretty good job of trimming. The vacuum cleaner (purchased separately) comes on automatically when you push the trimming button and does a good job of keeping the dust down.
Gene P. Young Sr. Analytical Technologist Analytical Sciences, Polymer Characterization The Dow Chemical Company 2301 N. Brazosport Blvd., B-1470 Freeport, Texas 77541-3257 Fax: (979) 238-0095 Phone:(979) 238-1579
-----Original Message----- } From: Allan Mitchell [mailto:allan.mitchell-at-stonebow.otago.ac.nz] Sent: Tuesday, February 04, 2003 4:05 PM To: Microscopy-at-sparc5.microscopy.com
Has anyone tried the Leica EM Trim specimen trimmer?
Uses a milling tool to trim down blocks, while observing througha stereo microscope head. Has a vacuum connection to trim away the nasties.
Thinking of buying one so any feed back would be appreciated.
Allan -- ------------------------------------------------- Allan Mitchell Technical Manager Otago Centre for Electron Microscopy C/-Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
You can set the resolution of your images higher in Adobe Acrobat. I don't know which Acrobat version you have and it is a little different in each. The easiest way is to use PDFWriter and set the resolution output to 300 dpi or a little higher. If you use Distiller, then you will need to go in and change the resolution of your gray scale and line drawings to higher and also make sure that you are using a higher resolution print option. You can set everything to 300 dpi and it should come out pretty good. You can also set your line drawing settings higher than 300 if you want.
In adobe Acrobat 5.0, I save in 4.0 format and for documents that I save for myself for reference, I use a general resolution of 600 dpi. For image compression schemes, I use bi-cubic sampling to 300 dpi for images above 350 dpi for both color and grayscale, and 600 for monochrome images. These settings work fairly well for me and the document sizes don't get too large.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Dusevich, Vladimir [mailto:dusevichv-at-umkc.edu] Sent: Thursday, February 06, 2003 11:25 AM To: Microscopy-at-sparc5.microscopy.com
Hi Listers,
Preparing my abstract for MM'03 I have found that: - Two page document (4 images with superimposed EDS line scans) saved in Word 2000 format was 758K size. - Saved in Word 6.0 format (as required for uploading by submission instructions) it had size of 2.93M (!). - Saved in PDF format (Acrobat 5.0) file was just 286K, but line scans, still pretty visible, were looking not nice.
I believe I should upload PDF files, since Word files anyway will be converted in PDF for CD publishing. But are not we loosing quality going digital now?
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: Rosemary White [mailto:rosemary.white-at-csiro.au] } Sent: Thursday, January 30, 2003 4:00 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: presentation images in Microsoft Word } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Another trick is to put the images and text into a Word } table, if you size } the cells in the table to about the size you want, the images } will insert } to fit the cell size. } } And if copying and pasting, make sure you do "Paste Special" } and unclick } the box that says "float over text". That way the images } always stay in } the same spot with respect to the surrounding text and don't } mysteriously } jump around when you insert or delete text. } cheers, } Rosemary } } } } Thanks Doug- } } Great tips- I'm printing them up until I memorize them. } } Rgds, } } Mike Shaw } } Roselle, NJ } } } } } Gary, } } } } } } MS Word is such a pain because of the way it works with } images. A couple } } } of tricks I've discovered over the years. } } } } } } 1) Place the image within a text box, rather than directly } on the page, it } } } seems to give you much greater control over the location } on the page. It } } } also makes it much easier to add text annotations that } stay with the image. } } } } } } } } } If you don't want the text box to have a border you can } remove it by } } } selecting } } } the box outline and look for the "paint brush" icon on the } Draw toolbar, } } } then use the down arrow and select "none". If you'd like } the text box to } } } be transparent, select the text box outline and look for } the "paint bucket" } } } icon on the Draw toolbar, then use the down arrow and } select "none". If } } } you discover its hard to find the text box border once you } made the edge } } } "invisible", first select the image with a single left } click (it should have } } } the solid black resizing "handles") and then use the } keyboard left or right } } } arrow keys and the selection with move out to the textbox } outline (with } } } black } } } bordered resizing "handles"). } } } } } } 2) Always use the INSERT | PICTURE | FROM FILE option as } opposed to copying } } } and pasting an image into Word. I find that the images } are harder to work } } } with if I paste them in. Also, you can insert TIFF or BMP } } } images into Word, } } } you don't have to use JPEG. } } } } }
Sounds like you have lossy image compression enabled in Acrobat, or possibly low output resolution settings.
Lossless or uncompressed options are available in Acrobat, and should give you the high quality you need for publishing. There are quality "presets" as well; "prepress" is the high quality option if I remember correctly (or just manually reduce compression and increase output resolution).
I believe there was an article with tips on using Acrobat for scientific publishing in a recent issue of "Microscopy Today". Anyone remember what issue it was?
-Kevin ------------------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 10:25 AM 2/6/03 -0600, "Dusevich, Vladimir" {dusevichv-at-umkc.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Joiner, Large rooms are a rare blessing! Generally the interference is different light requirements for the 2 instruments. I've seen a number of labs where the TEM will have a light-tight curtain suspended from an overhead rail surrounding it. I believe this is a fairly readily available darkroom curtain Also be sure that each room section has its own light switches and one doesn't interfere with the other. The TEM needs the darkness, while the newer computer based SEMs can operate in full light. An older SEM's light sensitivity is more related to each operator than to the instrument itself. Some people can see the images in lighter conditions than others.
If either instrument is going to be used a lot for high mag, then sound and other sources of vibration from the other instrument's user can be a problem. Also, computers related to EDS and image capture should be kept away from either column.
Ken Converse owner Quality Images third party SEM service Delta,PA
Joiner Cartwright, Jr., Ph.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 Listrers - } } I run a clinical EM lab with a geriatric JEOL 100C TEMSCAN microscope. } I am campaigning for new equipment and would like to get a stand alone } TEM and SEM. Currently my scope is in a rather large room that should } be able to accommodate two instruments with their columns at least ten } feet apart. I would hate to have new instruments installed only to } find that they interfered with each other, either electrically or } mechanically (vibration, etc.). I would like to hear from anyone who } has wrestled with this problem....or is it a problem? I am on the } second floor and not too near elevator shafts or electrical traces and } vibration for my one scope has not been a problem. } } Joiner } Joiner Cartwright, Jr., Ph.D. } Department of Pathology } Baylor College of Medicine } Houston, Texas U.S.A. } } }
You might want to check the recent archives, as this topic came up not too long ago. It sounds like you will be getting (if successful!) 'workhorse' instruments, which are less sensitive to fields, etc than 'high-end' instruments. The best bet might be to check with potential vendors re column distances and vibrations.
On the practical side, though, the SEM is a normal-to-low light environment, whereas the TEM has to have a low-to-no light environment. I've heard of places where they've tried to get around this with floor-to-ceiling drapes, but light spillage during plate exposure on the TEM will always be an issue. Chatting during simultaneous usage will also be an irritant. I'm sure space is at a premium, however, so why not consider a partition? One advantage of close-together instruments would be sharing a common water chiller, if that is in the budget.
Tom
Dr. Tom Malis Scientist Advisor Natural Resources Canada Govt. of Canada 613-995-7358 malis-at-nrcan.gc.ca
I run a clinical EM lab with a geriatric JEOL 100C TEMSCAN microscope. I am campaigning for new equipment and would like to get a stand alone TEM and SEM. Currently my scope is in a rather large room that should be able to
accommodate two instruments with their columns at least ten feet apart. I would hate to have new instruments installed only to find that they interfered with each other, either electrically or mechanically (vibration, etc.). I would like to hear from anyone who has wrestled with this problem....or is it a problem? I am on the second floor and not too near
elevator shafts or electrical traces and vibration for my one scope has not been a problem.
Joiner Joiner Cartwright, Jr., Ph.D. Department of Pathology Baylor College of Medicine Houston, Texas U.S.A.
Randy, I've not the foggiest as to what molecular sieves are made of. Are there any 'hard bits' in them, like fine ceramic particles? If so, that could be your problem. In many hard materials edge chipping is a reality. However, when sectioning uniformly hard materials like ceramics, the only way they will section is if one has an ultrafine facet on the block, ie the order of several microns. That methodology also has the benefit of reducing the area of damage along the edge, and one can 'walk' along the edge for some time before resharpening is needed. So if there is something hard/tough enough in your sieves to chip the edge, and you are cutting sections of hundreds of microns, you will surely get chipping spread over that length of the edge. The question than becomes, can your clients live with smaller sections?
Tom
-----Original Message----- } From: Tindall, Randy D. To: microscopy-at-sparc5.microscopy.com Sent: 2/6/2003 10:07 AM
Dear Listers,
I'm curious as to how many people have had problems using molecular sieves in dehydration solvents, with respect to knife damage. We seem to be going through diamond knives at a uncomfortable rate and we're wondering if this could be a contributing factor. We are very careful with our knives and try to minimize contact cleaning of the edges. We soak the knives often in the recommended solution in a commercial cleansing unit, and the knife manufacturer has evaluated one of our knives and confirmed that the edge is chipped, not dirty.
As a multi-user facility, we do a LOT of ultramicrotomy on a large variety of samples, so this may just be normal wear and tear, but we would sure like to minimize this expense.
Thanks much!
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! 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/
Hello Randy, We use molecular sieves in our 100% ethanol used in dehydration series and in 15 years have never had a problem associated with this practice. We also use only plastic disposable pipettes to avoid any glass splinters in any of our bottles. Do users in your lab share knives? We once had a student in the lab who rapidly destroyed several knives when he got his hands on them. He continually reported that there was something wrong with the knife! We don't know what he did, but we knew who did it, and we took measures to prevent further damage. Dean Abel Biological Sciences 141 BB University of Iowa Iowa City IA 52242-1324
Were the 4 images imported by reference or included in the final document? Big difference.
gary g.
At 08:25 AM 2/6/2003, you wrote:
} Hi Listers, } } Preparing my abstract for MM'03 I have found that: } - Two page document (4 images with superimposed EDS line scans) } saved in Word 2000 format was 758K size. } - Saved in Word 6.0 format (as required for uploading by } submission instructions) it had size of 2.93M (!). } - Saved in PDF format (Acrobat 5.0) file was just 286K, } but line scans, still pretty visible, were looking not } nice. } } I believe I should upload PDF files, since Word files } anyway will be converted in PDF for CD publishing. But are not } we loosing quality going digital now? } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } } } -----Original Message----- } } From: Rosemary White [mailto:rosemary.white-at-csiro.au] } } Sent: Thursday, January 30, 2003 4:00 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Re: presentation images in Microsoft Word } } } } } } -------------------------------------------------------------- } } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } } ---------. } } } } } } Another trick is to put the images and text into a Word } } table, if you size } } the cells in the table to about the size you want, the images } } will insert } } to fit the cell size. } } } } And if copying and pasting, make sure you do "Paste Special" } } and unclick } } the box that says "float over text". That way the images } } always stay in } } the same spot with respect to the surrounding text and don't } } mysteriously } } jump around when you insert or delete text. } } cheers, } } Rosemary } } } } } } Thanks Doug- } } } Great tips- I'm printing them up until I memorize them. } } } Rgds, } } } Mike Shaw } } } Roselle, NJ } } } } } } } Gary, } } } } } } } } MS Word is such a pain because of the way it works with } } images. A couple } } } } of tricks I've discovered over the years. } } } } } } } } 1) Place the image within a text box, rather than directly } } on the page, it } } } } seems to give you much greater control over the location } } on the page. It } } } } also makes it much easier to add text annotations that } } stay with the image. } } } } } } } } } } } } If you don't want the text box to have a border you can } } remove it by } } } } selecting } } } } the box outline and look for the "paint brush" icon on the } } Draw toolbar, } } } } then use the down arrow and select "none". If you'd like } } the text box to } } } } be transparent, select the text box outline and look for } } the "paint bucket" } } } } icon on the Draw toolbar, then use the down arrow and } } select "none". If } } } } you discover its hard to find the text box border once you } } made the edge } } } } "invisible", first select the image with a single left } } click (it should have } } } } the solid black resizing "handles") and then use the } } keyboard left or right } } } } arrow keys and the selection with move out to the textbox } } outline (with } } } } black } } } } bordered resizing "handles"). } } } } } } } } 2) Always use the INSERT | PICTURE | FROM FILE option as } } opposed to copying } } } } and pasting an image into Word. I find that the images } } are harder to work } } } } with if I paste them in. Also, you can insert TIFF or BMP } } } } images into Word, } } } } you don't have to use JPEG. } } } } } } } } } }
WE ARE MEMBERS OF A SPECIAL COMMITTEE FOR BUDGET AND PLANNING OF THE NIGERIAN NATIONAL PETROLEUM CORPORATION (NNPC). THIS COMMITTEE IS PRINCIPALLY CONCERNED WITH CONTRACT AWARDS AND APPROVAL. WITH OUR POSITIONS, WE HAVE SUCCESSFULLY SECURED FOR OURSELVES THE SUM OF TWENTY-ONE MILLION, FIVE HUNDRED THOUSAND UNITED STATES DOLLARS(US$21.5M). THIS AMOUNT WAS CAREFULLY MANIPULATED BY OVER-INVOICING OF AN OLD CONTRACT.
BASED ON INFORMATION GATHERED ABOUT YOU, WE BELIEVE YOU WOULD BE IN A POSITION TO HELP US IN TRANSFERRING THIS FUND (US$21.5M) INTO A SAFE ACCOUNT. IT HAS BEEN AGREED THAT THE OWNER OF THE ACCOUNT WILL BE COMPENSATED WITH 20% OF THE REMITTEDFUNDS, WHILE WE KEEP 70%, AND 10% WILL BE SET ASIDE TO OFFSET EXPENSES AND PAYTHE NECESSARY TAXES.
ALL MODALITIES OF THIS TRANSACTION HAVE BEEN WORKED OUT AND ONCE STARTED WILL NOT TAKE MORE THAN 10 WORKING DAYS, WITH YOUR FULL SUPPORT. THIS TRANSACTIONIS 100% RISK FREE.
IF THIS PROPOSAL SATISFIES YOU, PLEASE REACH US ONLY BY FAX OR PHONE,FOR MORE INFORMATION. IT MIGHT BE DIFFICULT TO GET THROUGH TO ME, BECAUSE OF POOR TELECOMMUNICATION SYSTEM HERE. PLEASE KEEP TRYING, YOU WILL DEFINITELY GET THROUGH. PLEASE TREAT AS URGENT AND VERY CONFIDENTIAL. YOURS FAITHFULLY, DR.REGINALD AKON NB.: FOR CONFIDENTIAL REASONS AND DUE TO THE POOR COMMUNICATION SYSTEM IN MY COUNTRY, MOST OFTEN FOREIGN CALLS COULD BE DIVERTED TO THE WRONG PERSON. SO FOR YOU TO BE VERY SURE YOU ARE RIGHTLY SPEAKING WITH ME, IT IS VERY IMPORTANT THATWHEN YOU CALL AND ASK FOR ME, THE MOMENT I PICK UP THE PHONE, YOU SHOULD ASK ME FORTHE "CODE WORD" AND MY ANSWER WOULD BE "BORNGREAT" BEFORE WE PROCEED DISCUSSIONS, BUT IF I DO NOT SAY "BORNGREAT",THAT MEANS YOU ARE NOT SPEAKING WITH ME JUST DISCONNECT THE LINE AND CALL ME BACK TILL I GIVE YOU THE CODE WORD.
WE ARE MEMBERS OF A SPECIAL COMMITTEE FOR BUDGET AND PLANNING OF THE NIGERIAN NATIONAL PETROLEUM CORPORATION (NNPC). THIS COMMITTEE IS PRINCIPALLY CONCERNED WITH CONTRACT AWARDS AND APPROVAL. WITH OUR POSITIONS, WE HAVE SUCCESSFULLY SECURED FOR OURSELVES THE SUM OF TWENTY-ONE MILLION, FIVE HUNDRED THOUSAND UNITED STATES DOLLARS(US$21.5M). THIS AMOUNT WAS CAREFULLY MANIPULATED BY OVER-INVOICING OF AN OLD CONTRACT.
BASED ON INFORMATION GATHERED ABOUT YOU, WE BELIEVE YOU WOULD BE IN A POSITION TO HELP US IN TRANSFERRING THIS FUND (US$21.5M) INTO A SAFE ACCOUNT. IT HAS BEEN AGREED THAT THE OWNER OF THE ACCOUNT WILL BE COMPENSATED WITH 20% OF THE REMITTEDFUNDS, WHILE WE KEEP 70%, AND 10% WILL BE SET ASIDE TO OFFSET EXPENSES AND PAYTHE NECESSARY TAXES.
ALL MODALITIES OF THIS TRANSACTION HAVE BEEN WORKED OUT AND ONCE STARTED WILL NOT TAKE MORE THAN 10 WORKING DAYS, WITH YOUR FULL SUPPORT. THIS TRANSACTIONIS 100% RISK FREE.
IF THIS PROPOSAL SATISFIES YOU, PLEASE REACH US ONLY BY FAX OR PHONE,FOR MORE INFORMATION. IT MIGHT BE DIFFICULT TO GET THROUGH TO ME, BECAUSE OF POOR TELECOMMUNICATION SYSTEM HERE. PLEASE KEEP TRYING, YOU WILL DEFINITELY GET THROUGH. PLEASE TREAT AS URGENT AND VERY CONFIDENTIAL. YOURS FAITHFULLY, DR.REGINALD AKON NB.: FOR CONFIDENTIAL REASONS AND DUE TO THE POOR COMMUNICATION SYSTEM IN MY COUNTRY, MOST OFTEN FOREIGN CALLS COULD BE DIVERTED TO THE WRONG PERSON. SO FOR YOU TO BE VERY SURE YOU ARE RIGHTLY SPEAKING WITH ME, IT IS VERY IMPORTANT THATWHEN YOU CALL AND ASK FOR ME, THE MOMENT I PICK UP THE PHONE, YOU SHOULD ASK ME FORTHE "CODE WORD" AND MY ANSWER WOULD BE "BORNGREAT" BEFORE WE PROCEED DISCUSSIONS, BUT IF I DO NOT SAY "BORNGREAT",THAT MEANS YOU ARE NOT SPEAKING WITH ME JUST DISCONNECT THE LINE AND CALL ME BACK TILL I GIVE YOU THE CODE WORD.
On 6 Feb 2003, at 12:47, Joiner Cartwright, Jr., Ph.D. wrote:
} I run a clinical EM lab with a geriatric JEOL 100C TEMSCAN microscope. } I am campaigning for new equipment and would like to get a stand alone } TEM and SEM. .
We have three EM's (2 x TEM and 1 x SEM) in an area of about 10m x 5m, separated only by floor to ceiling curtains. This works well but remember that the air-conditioning system must be geared to handle the heat output from these instruments, peripheral PC's, etc (we have 9 monitors in that area), AND the personnel using them. We have no problem from interference (vibration or magnetic field) between the instruments.
An advantage of this arrangement is that it is very useful for exhibitions, teaching, etc.
Regards
Rob
ps. 39C here yesterday, and heading that way, or more, already today - airconditioning HAS to work well.
=====================================
Rob Cross Director : EM Unit, Rhodes University tel: (046) 603 8168/9
We have just had a look at some Poly polysulfone filters. Thanks for the list on the sample prep. We found Al, Cu, Fe, Ni, Cr particles and even a W and Bi particle as well in the unused filters! Some leftovers from the processing plant? This might be the case with your filters as well.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
} -----Original Message----- } From: Malis, Tom [mailto:malis-at-nrcan.gc.ca] } Sent: Friday, February 07, 2003 2:01 AM } To: 'Tindall, Randy D. '; 'microscopy-at-sparc5.microscopy.com ' } Subject: RE: ultramicrotomy: knife damage } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Randy, I've not the foggiest as to what molecular sieves are } made of. Are } there any 'hard bits' in them, like fine ceramic particles? } If so, that } could be your problem. In many hard materials edge chipping } is a reality. } However, when sectioning uniformly hard materials like } ceramics, the only } way they will section is if one has an ultrafine facet on the } block, ie the } order of several microns. That methodology also has the } benefit of reducing } the area of damage along the edge, and one can 'walk' along } the edge for } some time before resharpening is needed. So if there is something } hard/tough enough in your sieves to chip the edge, and you are cutting } sections of hundreds of microns, you will surely get chipping } spread over } that length of the edge. The question than becomes, can your } clients live } with smaller sections? } } Tom } } -----Original Message----- } } From: Tindall, Randy D. } To: microscopy-at-sparc5.microscopy.com } Sent: 2/6/2003 10:07 AM } Subject: TEM: ultramicrotomy: knife damage } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Dear Listers, } } I'm curious as to how many people have had problems using molecular } sieves in dehydration solvents, with respect to knife damage. We seem } to be going through diamond knives at a uncomfortable rate and we're } wondering if this could be a contributing factor. We are very careful } with our knives and try to minimize contact cleaning of the edges. We } soak the knives often in the recommended solution in a commercial } cleansing unit, and the knife manufacturer has evaluated one of our } knives and confirmed that the edge is chipped, not dirty. } } As a multi-user facility, we do a LOT of ultramicrotomy on a large } variety of samples, so this may just be normal wear and tear, but we } would sure like to minimize this expense. } } Thanks much! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core---We're the Fun Core! } 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/ } }
Works on windows, linux, solaris, etc. Has the same fonctionnality than StarOffice 5.2, is free, and saves files in an zipped XML format. It is able to read and save in MSoffice and StarOffice formats. It have a few bugs, but not more than MS Office, and works well. It have an writer, a mathematical formules editor, drawer (lie Corel draw), a calculator like Excel, and a présentation tool like PowerPoint. I did only try the presentation soft, but a texte file has a quarter size than a Word97/2000 one, and half than a rtf.
And last but not least, it is free.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I hate the way Word deals with images. It makes huge files too. I would } recommend Adobe Pagemaker or Deneba Canvas for much more controllable } combination or text and graphics. If you have Acrobat (full version) } installed you can then also export the files to pdf format and share them } easily on the web. Of course, these software packages are not so ideal for } presentations and I have not yet found an alternative to Powerpoint for } that. } } (P.S. This is not just Microsoft bashing, I wish they would do a better } job of Graphics handling in word. In my opinion, the competition is better } at this just now. As a pure Word processor I find MS Word very good). } } Best wishes } } Ian } } On Mon, 20 Jan 2003 09:30:47 -0600, } {"gary.m.brown-at-exxonmobil.com"-at-sparc5.microscopy.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Jeff, } } } } I, too, have had significant problems working with images imported into } } Microsoft Word. However, my software is located on the PC hard drive. The } } biggest problem that I have encountered with images in Microsoft Word } } occurs when annotating images. After the image is imported into Word, } } annotation may be done in two ways (to my knowledge): (1) Text, arrows, } } etc. may be simply superimposed over the images. The problem with this } } approach is that the annotations are not linked to the image and may not } } remain superimposed on the image if the image moves. (2) Annotations can } } also be linked (probably not the best choice of words) to the image by } } double-clicking on the image to open the image field, adding the } } annotation, then closing the image field. These annotations are permanent } } unless intentionally moved or deleted. } } } } The problems occur when one implements the second option. Comparing } } images } } before and after annotation, I found that the annotated images often } } sustained substantial changes in gray or color levels. Case in point, EDS } } maps were so badly affected that the color key was no longer correct. } } } } My solutions follow: (1) Annotate images in Adobe Photoshop before } } importing into Word. Note that the effects of lossy compression on } } annotations (blurred edges) may be pronounced. (2) Use Microsoft } } PowerPoint } } for image presentation. I have encountered no problems with image files } } in } } PowerPoint. } } } } Good luck to you in your endeavors. } } } } Cheers, } } } } "The opinions expressed are those of Gary M. Brown and do not represent } } the } } opinions of ExxonMobil Corporation nor its affiliates." } } } } Gary M. Brown } } ExxonMobil Chemical Company } } Baytown Polymers Center } } 5200 Bayway Drive } } Baytown, Texas 77520-2101 } } phone: (281) 834-2387 } } fax: (281) 834-2395 } } e-mail: Gary.M.Brown-at-ExxonMobil.com } } } } } } "Oakley, Jeff" } } {oakleyj-at-rayovac.c To: {Microscopy-at-sparc5.microscopy.com} } } om} cc: } } Subject: RE: presentation images } } 01/17/03 07:50 AM } } } } } } } } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } This same phenomenon happens with my reports in Microsoft Word. In } } addition to images darkening, they sometimes shift to other pages and/or } } change size and dimension. Adding tables and text boxes to the report } } adds } } to the fun. } } } } The software that we use is networked. Our IS department has told us } } that } } the networked software has a bug that causes these things to happen when } } file sizes increase, and that there is not a patch for it. So we have } } just } } have to deal with it. } } } } Jeff Oakley } } } } } } -----Original Message----- } } } From: Corazon D. Bucana [mailto:bucana-at-audumla.mdacc.tmc.edu] } } Sent: Thursday, January 16, 2003 11:11 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: presentation images } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I created a Power point presentation last November 2002 consisting of } } several fluorescence micrographs. The file which was rather large ( 90 } } Mb) } } } } was left in my laptop all this time. When I opened it again this week I } } find that the images are now too dark and I needed to increase brightness } } by 3-4 clicks on the brightness icon of Power point.I increased } } brightness } } } } of all the micrographs and copied the file to a CD hoping that the image } } will not deteriorate there and then compare it with the one in my laptop } } several weeks from now. Has this happened to anyone else? Is there } } something I should have done to prevent this? } } } } Any suggestions or comments will be greatly appreciated. } } } } Cora Bucana } } } } } } } } } } } } } } } } } } } } } } -- } Ian MacLaren } Technische Universität Darmstadt } Material-und Geowissenschaften } Petersenstr. 23 } 64287 Darmstadt } Germany }
November/December 2002, Vol.10-#6, p.16: Use Adobe Acrobat to Keep Original Resolutions and to Make TIFF Files From Any Program by Jerry Sedgewick. -Matt
Matthew K. Stephenson Analytical Associate Impact Analytical 1910 West Saint Andrews Road Midland, MI 48640 (989) 832-5555 X506 stephenson-at-impactanalytical.com
-----Original Message----- } From: Kevin Frischmann [mailto:kfrisch-at-amnh.org] Sent: Thursday, February 06, 2003 5:31 PM To: Microscopy-at-sparc5.microscopy.com Cc: Dusevich, Vladimir
Sounds like you have lossy image compression enabled in Acrobat, or possibly low output resolution settings.
Lossless or uncompressed options are available in Acrobat, and should give you the high quality you need for publishing. There are quality "presets" as well; "prepress" is the high quality option if I remember correctly (or just manually reduce compression and increase output resolution).
I believe there was an article with tips on using Acrobat for scientific publishing in a recent issue of "Microscopy Today". Anyone remember what issue it was?
-Kevin ------------------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 10:25 AM 2/6/03 -0600, "Dusevich, Vladimir" {dusevichv-at-umkc.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
November/December 2002, Vol.10-#6, p.16: Use Adobe Acrobat to Keep Original Resolutions and to Make TIFF Files From Any Program by Jerry Sedgewick. -Matt
Matthew K. Stephenson Analytical Associate Impact Analytical 1910 West Saint Andrews Road Midland, MI 48640 (989) 832-5555 X506 stephenson-at-impactanalytical.com
Sounds like you have lossy image compression enabled in Acrobat, or possibly low output resolution settings.
Lossless or uncompressed options are available in Acrobat, and should give you the high quality you need for publishing. There are quality "presets" as well; "prepress" is the high quality option if I remember correctly (or just manually reduce compression and increase output resolution).
I believe there was an article with tips on using Acrobat for scientific publishing in a recent issue of "Microscopy Today". Anyone remember what issue it was?
-Kevin ------------------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 10:25 AM 2/6/03 -0600, "Dusevich, Vladimir" {dusevichv-at-umkc.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Randy, If you use molecular seives, you should not just dump them loose into your reagent bottles, but rather eclose them in dialysis tubing or something similar. That would eliminate particles floating free in your ethanols, etc. 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
Randy, I believe that molecular sieves contain alumina that is very detrimental to knives. The fine powder from the sieves takes a very long time to settle and is easily stirred up. Why don't you try using sodium sulfate. We have used that for years without noticeable problems. Of course, we try not to stir up the bottles or use the last 1/3 of the bottle. Rather we pour the remains together with about 1" of fresh sodium sulfate at the bottom of the bottle. Then let the bottle sit a day or two and you should be okay.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
On 2/6/03 10:07 AM, "Tindall, Randy D." {TindallR-at-missouri.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } I'm curious as to how many people have had problems using molecular } sieves in dehydration solvents, with respect to knife damage. We seem } to be going through diamond knives at a uncomfortable rate and we're } wondering if this could be a contributing factor. We are very careful } with our knives and try to minimize contact cleaning of the edges. We } soak the knives often in the recommended solution in a commercial } cleansing unit, and the knife manufacturer has evaluated one of our } knives and confirmed that the edge is chipped, not dirty. } } As a multi-user facility, we do a LOT of ultramicrotomy on a large } variety of samples, so this may just be normal wear and tear, but we } would sure like to minimize this expense. } } Thanks much! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core---We're the Fun Core! } 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/ } } }
Some time ago I have bout a case of 0.5L bottles of 200 alcohol for about $2 ($2.50?) each. No more molecular sieves for me.
Vladimir
} -----Original Message----- } From: Baskin, Tobias } Sent: Thursday, February 06, 2003 1:08 PM } To: Tindall, Randy D.; microscopy-at-sparc5.microscopy.com } Subject: Re: TEM: ultramicrotomy: knife damage } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Randy, } Some people I know keep their molecular sieve in } dialysis tubing. } } Tobias } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } } Dear Listers, } } } } I'm curious as to how many people have had problems using molecular } } sieves in dehydration solvents, with respect to knife } damage. We seem } } to be going through diamond knives at a uncomfortable rate and we're } } wondering if this could be a contributing factor. We are } very careful } } with our knives and try to minimize contact cleaning of the } edges. We } } soak the knives often in the recommended solution in a commercial } } cleansing unit, and the knife manufacturer has evaluated one of our } } knives and confirmed that the edge is chipped, not dirty. } } } } As a multi-user facility, we do a LOT of ultramicrotomy on a large } } variety of samples, so this may just be normal wear and tear, but we } } would sure like to minimize this expense. } } } } Thanks much! } } } } Randy } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core---We're the Fun Core! } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } }
I have used "insert-picture-from file" in Word and a lot of different settings for Distiller, including JPEG and ZIP compression for images, but always line scans and letter on images looked better in Word file, not in PDF.
Anyway, size of the Word 2000 file is close to the sum of image sizes (758K), and if it was up to me, I'd choose Word 2000 (not PDF) files as a standard for publication. It's too bad Microsoft do not advertise as much as Adobe its free viewers, for example this one: http://office.microsoft.com/downloads/2000/wd97vwr32.aspx
Vladimir
} } Set Distiller for printer resolution. } } Were the 4 images imported by reference or } included in the final document? Big difference. } } gary g. } } } At 08:25 AM 2/6/2003, you wrote: } } } Hi Listers, } } } } Preparing my abstract for MM'03 I have found that: } } - Two page document (4 images with superimposed EDS line scans) } } saved in Word 2000 format was 758K size. } } - Saved in Word 6.0 format (as required for uploading by } } submission instructions) it had size of 2.93M (!). } } - Saved in PDF format (Acrobat 5.0) file was just 286K, } } but line scans, still pretty visible, were looking not } } nice. } } } } I believe I should upload PDF files, since Word files } } anyway will be converted in PDF for CD publishing. But are not } } we loosing quality going digital now? } } } } Vladimir } } } } Vladimir M. Dusevich, Ph.D. } } Electron Microscope Lab Manager } } 3127 School of Dentistry } } 650 E. 25th Street } } Kansas City, MO 64108-2784 } } } } Phone: (816) 235-2072 } } Fax: (816) 235-5524 } } Web: http://www.umkc.edu/dentistry/microscopy } } } } } } } } } -----Original Message----- } } } From: Rosemary White [mailto:rosemary.white-at-csiro.au] } } } Sent: Thursday, January 30, 2003 4:00 PM } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: Re: presentation images in Microsoft Word } } } } } } } } } -------------------------------------------------------------- } } } ---------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } } of America } } } To Subscribe/Unsubscribe -- Send Email to } } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } } } ---------. } } } } } } } } } Another trick is to put the images and text into a Word } } } table, if you size } } } the cells in the table to about the size you want, the images } } } will insert } } } to fit the cell size. } } } } } } And if copying and pasting, make sure you do "Paste Special" } } } and unclick } } } the box that says "float over text". That way the images } } } always stay in } } } the same spot with respect to the surrounding text and don't } } } mysteriously } } } jump around when you insert or delete text. } } } cheers, } } } Rosemary } } } } } } } } Thanks Doug- } } } } Great tips- I'm printing them up until I memorize them. } } } } Rgds, } } } } Mike Shaw } } } } Roselle, NJ } } } } } } } } } Gary, } } } } } } } } } } MS Word is such a pain because of the way it works with } } } images. A couple } } } } } of tricks I've discovered over the years. } } } } } } } } } } 1) Place the image within a text box, rather than directly } } } on the page, it } } } } } seems to give you much greater control over the location } } } on the page. It } } } } } also makes it much easier to add text annotations that } } } stay with the image. } } } } } } } } } } } } } } } If you don't want the text box to have a border you can } } } remove it by } } } } } selecting } } } } } the box outline and look for the "paint brush" icon on the } } } Draw toolbar, } } } } } then use the down arrow and select "none". If you'd like } } } the text box to } } } } } be transparent, select the text box outline and look for } } } the "paint bucket" } } } } } icon on the Draw toolbar, then use the down arrow and } } } select "none". If } } } } } you discover its hard to find the text box border once you } } } made the edge } } } } } "invisible", first select the image with a single left } } } click (it should have } } } } } the solid black resizing "handles") and then use the } } } keyboard left or right } } } } } arrow keys and the selection with move out to the textbox } } } outline (with } } } } } black } } } } } bordered resizing "handles"). } } } } } } } } } } 2) Always use the INSERT | PICTURE | FROM FILE option as } } } opposed to copying } } } } } and pasting an image into Word. I find that the images } } } are harder to work } } } } } with if I paste them in. Also, you can insert TIFF or BMP } } } } } images into Word, } } } } } you don't have to use JPEG. } } } } } } } } } } } } } } } } } }
We operate one transmission and a couple of SEMs in one large laboratory. The individual microscopes are each enclosed in a lockable cubicle which makes it easier to control access and rermove the temptation amongst visitors to tamper. Photography and handling of film for the TEM are much easier if you have a lock on the inside - I have even occasionally used the TEM microscope cubicle as an emergency film developing darkroom (not recommended) when the main darkroom was out of commission. You could even use a red light outside linked to a safelight inside the TEM room - it is quite useful.
Our cubicles are ordinary timber painted but I suspect modern fire regulations would require specially treated materials. If the partitions are built around the microscopes then you will need some large plastic sheeting dust-covers, a lot of tape and much careful cleaning after the assembly and painting before you again expose your microscopes. Ideally have the cubicles built first, get everything cleaned a couple of times and the move the microscopes in a few days later when the dust has settled.
One thing, if you do get cubicles try to have either removable panels or very big doors (maybe even extra doors) to allow more visitors, extra equipment and most importantly access for servicing the instrument - discuss this with your service engineer if you can. Before we installed anything we got the floor specifications of each microscope, some little pictures scaled to a graph paper floor plan and could confirm that the rooms were suitable.
Good luck
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
----- Original Message ----- } From: "Malis, Tom" {malis-at-nrcan.gc.ca}
Try putting the molecular sieve inside some dialysis tubing and seal the ends (I just used staples). You end up with a molecular sieve 'sausage' which works quite well. This is not my idea but I can't remember where I got it from. We still get damage to knives but probably from other sources because our absolute ethanol is always crystal clear.
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
----- Original Message ----- } From: "Malis, Tom" {malis-at-nrcan.gc.ca}
On Thursday, February 6, 2003, at 07:06 AM, Lawrence Oakford wrote:
} I was wondering if anyone on the list is familiar with this stain. I } am trying to find a procedure for either making this stain or a } commercial source for the stain. It was listed in a methods section } for negative staining isolated neurofibrillary tangles (Crowther, R. } A. 1991. PNAS 88:2288) but the author did not reference a source for } the stain or a procedure. I would appreciate any help provided. } } Thanks } Dear Lawrence, I would suggest buying phosphotungstic acid and lithium hydroxide and mixing them. I have had success with other negative stains by putting 5 micro-l of the specimen and 5 micro-l of 2% stain on a grid (mixing by repeated pipetting), letting this sit for 1 min, blotting the fluid to near dryness, adding 10 micro-l of 1% stain, letting this sit for 1 min, and blotting to dryness. I would try this, but perhaps someone who has used lithium phosphotungstate has a better protocol. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
On Thursday, February 6, 2003, at 04:01 PM, Malis, Tom wrote:
} Randy, I've not the foggiest as to what molecular sieves are made of. } Are } there any 'hard bits' in them, like fine ceramic particles? } Dear Tom, Molecular sieves are made of zeolites--clays with molecule-sized cavities in their structures. I think they are aluminosilicates, but I'm no expert. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Alternate options are Adobe Framemaker and Quark Express. These are much more industrial strength than Word. For books, dissertations, long articles, etc., these predominate over Word, IMO.
For simple documents, Word is fine. Beyond that, it fails in many ways. The down side of the other options is the steep learning curve. But once this is surmounted, Word is history. This is what has been my experience.
Since Framemaker became an Adobe product, it is tightly integrated to Acrobat. Nevertheless, any program can save or print to Distiller and have really nice results. The trick is to make the settings congruent with your desired results.
gary g.
At 06:25 AM 2/6/2003, you wrote:
} I hate the way Word deals with images. It makes huge files too. I would } recommend Adobe Pagemaker or Deneba Canvas for much more controllable } combination or text and graphics. If you have Acrobat (full version) } installed you can then also export the files to pdf format and share them } easily on the web. Of course, these software packages are not so ideal } for presentations and I have not yet found an alternative to Powerpoint } for that. } } (P.S. This is not just Microsoft bashing, I wish they would do a better } job of Graphics handling in word. In my opinion, the competition is } better at this just now. As a pure Word processor I find MS Word very good). } } Best wishes } } Ian } } On Mon, 20 Jan 2003 09:30:47 -0600, } {"gary.m.brown-at-exxonmobil.com"-at-sparc5.microscopy.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Vladimir I just recently converted MS Word (2000) document with color pictures into PDF using Distiller and was not able to see the difference even the Distiller's settings was not optimal. If I understand correctly, most modern printers used "post-script" format to print. It's actually the same as PDF (yes, ADOBE's job). I mean, PDF is a sequence of command (it's not actually text or graphics), which any modern printer could recognize directly (and print). From another hand any other formats (MS Word, any pictures) should be translated into "post-script" (read PDF) to be printed. Usually it happens when you sent file to printer (printer driver did the job). So, the bottom line here: any image/document you sent to printer converted, actually into PDF (post-script). Therefore, in theory you should be able to have equal quality from PDF and let say TIFF or MS Word when printed. On the screen, they should looks differently (because PDF is actually vector graphics and TIFF for instance is bitmap). I hope it helps. Sergey
At 07:30 AM 2/7/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks for all replies!
I have used "insert-picture-from file" in Word and a lot of different settings for Distiller, including JPEG and ZIP compression for images, but always line scans and letter on images looked better in Word file, not in PDF.
Anyway, size of the Word 2000 file is close to the sum of image sizes (758K), and if it was up to me, I'd choose Word 2000 (not PDF) files as a standard for publication. It's too bad Microsoft do not advertise as much as Adobe its free viewers, for example this one: http://office.microsoft.com/downloads/2000/wd97vwr32.aspx
Vladimir
} } Set Distiller for printer resolution. } } Were the 4 images imported by reference or } included in the final document? Big difference. } } gary g. } } } At 08:25 AM 2/6/2003, you wrote: } } } Hi Listers, } } } } Preparing my abstract for MM'03 I have found that: } } - Two page document (4 images with superimposed EDS line scans) } } saved in Word 2000 format was 758K size. } } - Saved in Word 6.0 format (as required for uploading by } } submission instructions) it had size of 2.93M (!). } } - Saved in PDF format (Acrobat 5.0) file was just 286K, } } but line scans, still pretty visible, were looking not } } nice. } } } } I believe I should upload PDF files, since Word files } } anyway will be converted in PDF for CD publishing. But are not } } we loosing quality going digital now? } } } } Vladimir } } } } Vladimir M. Dusevich, Ph.D. } } Electron Microscope Lab Manager } } 3127 School of Dentistry } } 650 E. 25th Street } } Kansas City, MO 64108-2784 } } } } Phone: (816) 235-2072 } } Fax: (816) 235-5524 } } Web: http://www.umkc.edu/dentistry/microscopy } } } } } } } } } -----Original Message----- } } } From: Rosemary White [mailto:rosemary.white-at-csiro.au] } } } Sent: Thursday, January 30, 2003 4:00 PM } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: Re: presentation images in Microsoft Word } } } } } } } } } -------------------------------------------------------------- } } } ---------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } } of America } } } To Subscribe/Unsubscribe -- Send Email to } } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } } } ---------. } } } } } } } } } Another trick is to put the images and text into a Word } } } table, if you size } } } the cells in the table to about the size you want, the images } } } will insert } } } to fit the cell size. } } } } } } And if copying and pasting, make sure you do "Paste Special" } } } and unclick } } } the box that says "float over text". That way the images } } } always stay in } } } the same spot with respect to the surrounding text and don't } } } mysteriously } } } jump around when you insert or delete text. } } } cheers, } } } Rosemary } } } } } } } } Thanks Doug- } } } } Great tips- I'm printing them up until I memorize them. } } } } Rgds, } } } } Mike Shaw } } } } Roselle, NJ } } } } } } } } } Gary, } } } } } } } } } } MS Word is such a pain because of the way it works with } } } images. A couple } } } } } of tricks I've discovered over the years. } } } } } } } } } } 1) Place the image within a text box, rather than directly } } } on the page, it } } } } } seems to give you much greater control over the location } } } on the page. It } } } } } also makes it much easier to add text annotations that } } } stay with the image. } } } } } } } } } } } } } } } If you don't want the text box to have a border you can } } } remove it by } } } } } selecting } } } } } the box outline and look for the "paint brush" icon on the } } } Draw toolbar, } } } } } then use the down arrow and select "none". If you'd like } } } the text box to } } } } } be transparent, select the text box outline and look for } } } the "paint bucket" } } } } } icon on the Draw toolbar, then use the down arrow and } } } select "none". If } } } } } you discover its hard to find the text box border once you } } } made the edge } } } } } "invisible", first select the image with a single left } } } click (it should have } } } } } the solid black resizing "handles") and then use the } } } keyboard left or right } } } } } arrow keys and the selection with move out to the textbox } } } outline (with } } } } } black } } } } } bordered resizing "handles"). } } } } } } } } } } 2) Always use the INSERT | PICTURE | FROM FILE option as } } } opposed to copying } } } } } and pasting an image into Word. I find that the images } } } are harder to work } } } } } with if I paste them in. Also, you can insert TIFF or BMP } } } } } images into Word, } } } } } you don't have to use JPEG. } } } } } } } } } } } } } } } } } }
I apologise if this topic has gone off the boil but my original message was 'bounced' because my work e-mail software has a nasty habit of adding attachments if I forward a message, although not if I hit reply.
Sorry to clog up the system but I thought I should also add this. I always trim my blocks with glass knives on the ultra microtome and do a final 'facing' (smoothing) of the front of the block with a fresh area of the same or a new glass knife. I never use a diamond knife until I can achieve a smooth finish on the block with a glass knife. If I am desperate then I will either just use a glass knife or an old diamond but it's never been necessary to risk a good diamond on an unknown block this way..
Invariably the biggest problems are found at the tips of pellets where any hard crystalline deposits and broken glass tend to settle and usually deeper material is less troublesome.
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
{snip}
} ------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } AmericaTo Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.ComOn-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------. } } } Dear Listers, } } I'm curious as to how many people have had problems using molecular } sieves in dehydration solvents, with respect to knife damage. We seem } to be going through diamond knives at a uncomfortable rate and we're } wondering if this could be a contributing factor. We are very careful } with our knives and try to minimize contact cleaning of the edges. } We soak the knives often in the recommended solution in a commercial } cleansing unit, and the knife manufacturer has evaluated one of our } knives and confirmed that the edge is chipped, not dirty. } } As a multi-user facility, we do a LOT of ultramicrotomy on a large } variety of samples, so this may just be normal wear and tear, but we } would sure like to minimize this expense. } } Thanks much! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core---We're the Fun Core! } 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/
Just thought maybe this will help someone these abstract submitting days: what I ended up doing for my M&M abstract last year, I made the 1st page (all text) in MS Word, but the 2nd page I made all in Photoshop. Photoshop now allows to add and format text. I then put the two pages together in Acrobat. Yes, I did all that after it came out bad from just inserting the picture into the Word file and distilling the whole thing. To be honest, I don't remember now what was it that I didn't like, but I remember it was unacceptable.
Vlad
-- ------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
Applied Optical Microscopy is just about a month away, in sunny Orlando, Florida. 3.5 days of total immersion in light microscopy, including a special focus session on digital imaging.
Remember.. this course is NOT just for chemists!
Details and registration form available at: http://www.microscopyeducation.com/acs_crse.htm
Sponsor: American Chemical Society Dates: March 7-9 (includes Sat. night "Intro to Digital Imaging") Location: Renaissance Orlando Resort at SeaWorld,Orlando, Fl Tuition, materials, and coffee breaks: $1345 for ACS members, $1445 for non-members
Questions: Call/email me here at the MME offices.
Best regards, Barbara Foster, ACS course coordinator
Microscopy/Microscopy Education, Inc. 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details.
If you are using imaging software, and especially software from Zeiss like KS300 / Axiovision V06.2002, as I am using, you should take care if you use it together with the virusscanner McAfee VirusScan from Network Associates Technology (McAfee).
It seems that with the latest update version (DAT-updatefile 4246) from February 05, 2003, the program recognizes certain DLL-files required for image-capturing as infected with the virus 'HackerDefender'. These file ARE NOT infected, it's a problem of the virusscanner.
Here you will find more information about this so-called virus: http://vil.nai.com/vil/content/v_100035.htm
A part from the website from McAfee:
"Update - Feb 6th 2003: McAfee products using the 4246 DATs are incorrectly reporting certain innocent DLL files as 'trojan or variant HackerDefender'.
The innocent files affected are DLLs related to image analysis software. The following DLLs are known to be incorrectly flagged with the 4246 DATs:
MILCOR.DLL (v6.10.0.186, 626,960 bytes) MILCOR.DLL (v6.01.00.727, 590,096 bytes) MILVGA.DLL (v6.10.00.1618, 348,432 bytes) MILVGA.DLL (v6.01.00.902, 319,760 bytes) MILGEN.DLL (v6.10.00.1257, 426,256 bytes) These DLLs are completely unrelated to the HackerDefender rootkit."
Best regards and good luck,
Sven Terclavers
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Sven Terclavers LM/CLSM Microscopist Center for Transgene Technology and Gene Therapy (CTG) Campus Gasthuisberg K.U.L. O&N Herestraat 49 3000 Leuven Belgium Tel. +32 16 346173 Fax. +32 16 345990 Email: Sven.Terclavers-at-med.kuleuven.ac.be °°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°
We have some problems with a PE-staining (phycoerythrin-group, B-phycoerythrin from Molecular Probes), conjugated with Cd31 to stain bloodvessels. We would like to co-stain with an FITC-conjugate, but it seems that PE is also excited by 488nm (the FITC-excitation wavelenght) instead of only by 568nm, so we cannot trust what we see because of this bleeding-through. Did any of you had the same problem and if you could solve it, how did you do it? Another problem we see is that the PE is bleached really fast. Is this normal and what can be done? Thanks in advance,
Sven Terclavers
PS. Sorry if some of you get this message double because I send it to different mailinglists.
I need retinal tissue, preferably mice. We make a low kV desktop em and some folks are belief-challenged that we image samples without staining and get excellent results.
We want to image the photoreceptors cone and rods in the various scanning modes; TEM SEM and STEM.
We are also interested in expanding our database of images from a broad range of tissue and applications.
So, it's a 2 part email;
1) does anyone know where I can get this retinal tissue ??
2) an invitation to listers who are curious about how their samples will image in our EM to contact me
Thanks,
Ephram
Ephram Shizgal Delong America info-at-delongamerica.com
one of my lab mates was using an edwards auto 306 vaccum coater and a piece of filter paper fell between the plate that seals the bell jar from the diffusion pump. we get a backing pressure fail erroe. we are unable to get into the bell jar to remove the paper. the question is does anyone know how to manulaly get into the coater. we have a service call in but would like to resolve the issue ourselves.
I have run into this in the confocal facility I run. People have brought samples that they have prepped without first consulting me and we have all sorts of strange results. PE is not a stain of choice for fluorescence microscopy. It is very popular with the flow cytometry folks where its rapid bleaching is not an issue, nor apparently is its excitation by multiple wavelengths. I would recommend almost any of the other dyes that excite around 560 in preference to PE. You will also get a stronger (enhanced) signal if you use your Cd31 as a primary Ab then tag that with a fluorescent secondary Ab, although you may have specific reasons for not wanting to do so. Try TRITC, Texas Red, Alexa 546, CY3. They will all fluoresce red and will allow your FITC double stain. 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
out of interest, as a clinical virologist who has a bit of EM background, i would be curious about the unit you are using. i have seen something marketed by one of our canadian distributors, but have no knowledge on how effective the unit might be. sorry, but i forgot the name of the manufacturer.
Does one of you know if allophycocyanin, a member of the phycobiliprotein-group (as PE), has also a high bleeching, just as PE? We have to choose a red label and can only choose between PE, APC and perCP... Thanks,
Many thanks to everyone who replied to my question about possible damage to diamond knives due to use of molecular sieves in the dehydration solvent. There doesn't seem to be a real consensus on this issue, with some folks saying they have never had a problem even after a couple of decades, and others saying they wouldn't use molecular sieves under any circumstances. The latter people advocated using freshly opened absolute ethanol (or whatever) for the final dehydration steps.
One repeated suggestion was that molecular sieves can be safely used if they are put in dialysis tubing to keep particulates safely contained. I think we'll try this.
Another suggestion made by a couple folks was that use of glass pipettes during processing could be the source of glass chips that damage knives. We never use glass pipettes in our sample preps, but interestingly enough, a client who sometimes brings us blocks prepared in his own lab does use them! Mystery solved? We'll see.
Thanks again to everyone. As always, this list is a real resource.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
I have a small problem I hope someone can help with. We have a Lecia DMIL inverted microscope in our facility with a 6V 35W bulb for illumination. It has been using bulbs with a high frequency which surprised me as I have never changed the 100W bulbs in my Zeiss microscopes in 15 years of use. I checked the voltage output at the socket and its maximum is 10.5V. This seems odd for a 6V bulb and Leica cannot tell me if it is abnormal or not, but rates the bulbs at 50 hours which also seems odd (although that is about how long they last). It seems to me that putting a 6V bulb in a 10V socket would certainly lead to a short lifetime, but I don't know if this is our scope, bad design or something is wrong. If someone has a DMIL and can check the socket voltage, that would tell me quickly where the problem lies. Any other suggestions welcome. Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
"Electron energy loss spectroscopy (EELS) is making rapid progress in its ability to provide compositional and chemical information on the nanometre scale. A workshop on the EELS of Steels and Alloys will be held in Bruck an der Mer (Austria) from 12th - 14th June 2003 to discuss the latest developments in this area of application. The deadline for abstracts is 15th May. Details can be found at http://www.cis.tugraz.at/felmi/EELS_of_STEELS_2003.html"
Prof A J Craven Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland, UK
David: I recently went to a bulb website (Topbulb.com) looking for a bulb for my Zeiss Axiophot. They had several brands that were all the right voltage and wattage but to my amazement, the stated lifespan varied from 50 hours for some and 1000 hours for others. I think you could find a longer duration bulb with the same specs. Tom
At 11:05 AM 2/12/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Dear Listservers, Is there anyone who has attempted to compile and use WIEN97/WIEN2k software code on an Apple Macintosh (OSX)? The code is written in FORTRAN90 and runs under Unix on 'practically all platforms'. Your comments and suggestions would be greatly appreciated.
Thanks. Mike Colella Materials & Engineering Science Australian Nuclear Science & Technology Organisation
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The said amount has remained dormant and floating in our Apex Bank. Therefore, we will raise supplementary documents to enable us transfer into an account that is yet to be nominated. We need a foreign partner, because as civil servants, the code of conduct does not allow us to operate foreign bank accounts. Hence we solicit for your assistance, we are requesting you to provide us with the help for the safekeeping of this money until our arrival in your company to collect our share and decide on investment possibilities of this money.
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David You have to check voltage under the load, i.e. bulb should be in place and lit. If the voltage will be still high, it may be a problem. Another things just to keep in mind: does bulb has good electrical contact in the socket? It may happens even on brand new instrument (if somebody put a few drops of water into the socket - terrorist?). The sign of such problem: overheated socket, signs of oxidation/damage on the bulb's "legs" or socket. Sergey
At 08:05 AM 2/12/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I have a colleague who is looking at silica particles in a polymer matrix. He is planning TEM analysis of cross sections to image the particles, determine their distribution and possibly some EDXS mapping. The samples are cast as thin films and the silica could be as small as 5nm. He has found a reference to the use of Phosphotungstic Acid (hope the spelling is correct) to stain the polymer but not the silica, thus improving contrast in the TEM. This was done by floating off the cryo-microtomed sections on a methanol solution of Phosphotungstic Acid. Alas, this is all the information he has.
We don't have cryo microtomy facilities, so we were going to use our microtome at room temp and also attempt to tripod polish.
Does anyone have a more detailed procedure of how to make up the solution and stain the polymer?? Any assistance would be greatly appreciated.
Regards George
George Theodossiou Physicist / Electron Microscopist CSIRO Manufacturing & Infrastructure Technology Private Bag 33 Clayton South MDC Victoria, 3169 tel: +61 3 9545 2012 fax: +61 3 9544 1128
To the extent permitted by law, CSIRO does not represent, warrant and/or guarantee that the integrity of this communication has been maintained or that the communication is free of errors, virus, interception or interference.
The information contained in this e-mail may be confidential or privileged. Any unauthorised use or disclosure is prohibited. If you have received this e-mail in error, please delete it immediately and notify George Theodossiou on +61 3 9545 2012. Thank you.
I am unfamiliar with your particular Lecia, but can offer some general info. Lamp life is significantly shortened under overvoltage conditions, but usually they are much bright brighter and the output spectrum moves to the shorter wavelengths. This could be intended, but not likely due to the very short life.
You need to measure the voltage under load (lamp connected and on). It could be the power supply is poorly regulated and depends on the lamp current to drop the voltage to the correct value. It this is the case, different lamps/currents would result in different loaded voltages which may or may not be correct. If the voltage does not drop, buy lamps by the case (or fix the P.S.)!
Starting incandescent lamps with an overvoltage is just the opposite of what would be the best engineering practice for life extension. Ideally (but rare), a "soft start" would be employed which limits inrush currents when the lamp is turned on. The resistance of a cold filament is very low and rises with temperature to the design value for a particular lamp. Since I=E/R (I=current, E=voltage, R=resistance), the worst time for overvoltage driven excessive current is when the filament is cold.
Another method to limit current would be a magnetically saturating transformer. I have never seen this method of current limiting used in simple lamps, but is almost always used in many welders and everyday microwave oven power supplies.
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: David Knecht [mailto:knecht-at-uconn.edu] Sent: Wednesday, February 12, 2003 11:06 AM To: microscopy-at-sparc5.microscopy.com
I have a small problem I hope someone can help with. We have a Lecia DMIL inverted microscope in our facility with a 6V 35W bulb for illumination. It has been using bulbs with a high frequency which surprised me as I have never changed the 100W bulbs in my Zeiss microscopes in 15 years of use. I checked the voltage output at the socket and its maximum is 10.5V. This seems odd for a 6V bulb and Leica cannot tell me if it is abnormal or not, but rates the bulbs at 50 hours which also seems odd (although that is about how long they last). It seems to me that putting a 6V bulb in a 10V socket would certainly lead to a short lifetime, but I don't know if this is our scope, bad design or something is wrong. If someone has a DMIL and can check the socket voltage, that would tell me quickly where the problem lies. Any other suggestions welcome. Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
We have had the glass in resin blocks problem recently. I only use plastic pipettes. I started suspecting the people who grew the cells but feel there has been a change in some product I use. My paranoid suspicions rest on the type of glass fracture from osmium and accelerator ampoules. Today I found an emnormous sliver 1mm long at the bottom of a block. It looks to big for molecular sieve (a suspect) EDX should indicate if it is glass. (I suppose I could do EDX on the ampoules too!).
I can cope with the problem by using a razor blade to remove the lowest part of the block.
Dave
On Wed, 12 Feb 2003 08:20:09 -0600 "Tindall, Randy D." {TindallR-at-missouri.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Many thanks to everyone who replied to my question about possible damage } to diamond knives due to use of molecular sieves in the dehydration } solvent. There doesn't seem to be a real consensus on this issue, with } some folks saying they have never had a problem even after a couple of } decades, and others saying they wouldn't use molecular sieves under any } circumstances. The latter people advocated using freshly opened } absolute ethanol (or whatever) for the final dehydration steps. } } One repeated suggestion was that molecular sieves can be safely used if } they are put in dialysis tubing to keep particulates safely contained. } I think we'll try this. } } Another suggestion made by a couple folks was that use of glass pipettes } during processing could be the source of glass chips that damage knives. } We never use glass pipettes in our sample preps, but interestingly } enough, a client who sometimes brings us blocks prepared in his own lab } does use them! Mystery solved? We'll see. } } Thanks again to everyone. As always, this list is a real resource. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core---We're the Fun Core! } 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/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
We have received from an other lab a second hand SEM with a Tracor TN 2000 unit for EDX analysis. This Tracor TN 2000 unit does not work anymore. The Tracor supplier in Europe said us that no support can be given for this product anymore. We would like to repair it by ourselves.
Can somebody help us with a copy of operator's manual and, very important for us, with the circuit diagrams? Of course, we will pay the copy and mailing costs.
Thanks a lot in advance. Best regards,
Jean Dr. Jean DILLE Materials Science and Electrochemistry Free University of Brussels, CP 194/03 Avenue F. Roosevelt, 50 1050 Brussels Belgium tel: 32-2-6502723 fax: 32-2-6502786 e-mail: jdille-at-ulb.ac.be
I seem to have missed most of the discussion about the new formulation Kodak 4489 film. Would someone please let me know what's the latest conclusion on that? I have been using the new film for several months without problem, but recently I am experiencing problems with the new film. What alternatives to Kodak film would anyone recommend?
You could try first your specimens without any staining. I have no problem in observation of silica particles and hydroxyapatite crystals (5-50 nm) embedded in adhesive resin. Particles have much greater electron density than resin and staining of the resin will only hide these particles. And for EDS its better do not apply any staining whenever possible.
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: "George.Theodossiou-at-csiro.au"-at-sparc5.microscopy.com } [mailto:"George.Theodossiou-at-csiro.au"-at-sparc5.microscopy.com] } Sent: Thursday, February 13, 2003 1:14 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM Polymer Staining Procedure } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Dear All, } } I have a colleague who is looking at silica particles in a } polymer matrix. } He is planning TEM analysis of cross sections to image the particles, } determine their distribution and possibly some EDXS mapping. } The samples } are cast as thin films and the silica could be as small as 5nm. } He has found a reference to the use of Phosphotungstic Acid (hope the } spelling is correct) to stain the polymer but not the silica, } thus improving } contrast in the TEM. This was done by floating off the } cryo-microtomed } sections on a methanol solution of Phosphotungstic Acid. } Alas, this is all } the information he has. } } We don't have cryo microtomy facilities, so we were going to use our } microtome at room temp and also attempt to tripod polish. } } Does anyone have a more detailed procedure of how to make up } the solution } and stain the polymer?? Any assistance would be greatly } appreciated. } } } Regards } George } } George Theodossiou } Physicist / Electron Microscopist } CSIRO Manufacturing & Infrastructure Technology } Private Bag 33 Clayton South MDC } Victoria, 3169 } tel: +61 3 9545 2012 } fax: +61 3 9544 1128 } } Visit our Web site http://www.cmst.csiro.au } } Shipping address: CSIRO - Manufacturing & Infrastructure } Technology, Gate 4 } Normanby Rd. Clayton, Victoria, } } PLEASE NOTE: } } To the extent permitted by law, CSIRO does not represent, } warrant and/or } guarantee that the integrity of this communication has been } maintained or } that the communication is free of errors, virus, interception or } interference. } } The information contained in this e-mail may be confidential } or privileged. } Any unauthorised use or disclosure is prohibited. If you } have received this } e-mail in error, please delete it immediately and notify } George Theodossiou } on +61 3 9545 2012. Thank you. } } } }
Picking up Woody's point of 'slow-starting' microscope bulbs, a simple way to achieve this with DC bulbs is to connect a hefty capacitor in parallel with the bulb. This gives a gradual switching on and off of the bulb, the speed depending upon the size of the capacitor. We employ this technique for all our small, undergraduate benchtop microscopes as they tend to get switched on and off a lot. This might not be the solution to the problem under discussion but it's very quick, easy and cheap to try and straightforward to undo if it doesn't do the trick.
Justin
} Starting incandescent lamps with an overvoltage is just the opposite of what } would be the best engineering practice for life extension. Ideally (but } rare), a "soft start" would be employed which limits inrush currents when } the lamp is turned on. The resistance of a cold filament is very low and } rises with temperature to the design value for a particular lamp. Since } I=E/R (I=current, E=voltage, R=resistance), the worst time for overvoltage } driven excessive current is when the filament is cold.
---------------------------------- Justin Ritherdon, Materials Science and Engineering, Department of Engineering, University of Liverpool, LIVERPOOL, L69 3GH, United Kingdom
We have had a somewhat similar problem with different make/models of scopes and were told by the scope repair facility that transformers and particularly rheostats anywhere in the electrical system will age and become unreliable. This unreliability results in "spikes" and variations in voltage/power to the bulb and greatly shortens their life. Their suggestion was to rebuild any rheostat when bulb life becomes a issue. On at least one scope with a consistent history of burn out, he was able to demonstrate to us the very brief flickering that the bulb exhibited as it was first turned on and stepped up through the variable power ranges.
bye for now
Tom Parker CSDLAC {tparker-at-lacsd.org}
-----Original Message----- } From: David Knecht [SMTP:knecht-at-uconn.edu] Sent: Wednesday, February 12, 2003 8:06 AM To: microscopy-at-sparc5.microscopy.com
I have a small problem I hope someone can help with. We have a Lecia DMIL inverted microscope in our facility with a 6V 35W bulb for illumination. It has been using bulbs with a high frequency which surprised me as I have never changed the 100W bulbs in my Zeiss microscopes in 15 years of use. I checked the voltage output at the socket and its maximum is 10.5V. This seems odd for a 6V bulb and Leica cannot tell me if it is abnormal or not, but rates the bulbs at 50 hours which also seems odd (although that is about how long they last). It seems to me that putting a 6V bulb in a 10V socket would certainly lead to a short lifetime, but I don't know if this is our scope, bad design or something is wrong. If someone has a DMIL and can check the socket voltage, that would tell me quickly where the problem lies. Any other suggestions welcome. Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
Another good bulb website: bulbman.com. John Mardinly Intel
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Wednesday, February 12, 2003 10:11 AM To: David Knecht Cc: Microscopy-at-sparc5.microscopy.com
David: I recently went to a bulb website (Topbulb.com) looking for a bulb for my Zeiss Axiophot. They had several brands that were all the right voltage and wattage but to my amazement, the stated lifespan varied from 50 hours for some and 1000 hours for others. I think you could find a longer duration bulb with the same specs. Tom
At 11:05 AM 2/12/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Just to give an update. I had been experiencing so called fogging problem with the new formulation of the kodak 4489 film.
Somehow, I can't pinpoint why but my problem has disappeared. I am still using the new formulation film.
Kodak tells me that I was perhaps not agitating enough or properly. I've developed some film without any agitation and I still don't get any fogging (not that I want it back).
Maybe I had a batch before who knows.
Rajesh Patel Robert Wood Johnson Medical School Department of Pathology 675 Hoes Lane Piscataway, NJ 08854
On our 306A there is a bleed valve meant for plasma-glo. When the diffusion pump is cold, open the bleed valve and wait. I think it took about 2hrs to come to atmosphere. Then you can open the bell jar and remove the diffusion pump valve(unscrew like a piano stool). Hopefully your filter paper will be reachable from there.
Ed Hurley
Roswell Park Cancer Institute Corporation
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We also have been struggling with this new formulation for 3 weeks now. The emulsion setting is apparently unchanged [sensitivity to photons], but the film requires much, much more agitation [both more frequent motion, and larger motions help] to get an approximately even development. It is as if the film clings to the developer in a patchy fashion, so that one tends to get very muddy, uneven images. It is almost impossible to get good development near the edge of the film holder, apparently due to poor exchange of solutions, even with lots of agitation. We are also being extra careful to fully drain all developer from the film before going to the 1st rinse, to agitate a little during this rinse, and to agitate a lot in the fix step, again using larger motions, and more frequently.
It is really easy with this film to obtain terrible, unprintable images. But with more concentrated efforts by the user, most of the problem can be dealt with. We've wasted at least one hundred shots getting our methods corrected, and produced some really ugly images for users during January. Hope to be back on track soon. Would love to know how to get an even development all the way to the edge of the film.
We would be interested to hear if people using a nitrogen burst system find any problems? Is that sufficient to deal with the changes in film properties? -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
TO ALL: WE ARE TRYING TO OBTAIN SOME LAB-6 PARTS FOR AN ISI-WB6, SO IF YOU ARE DECOMMISIONING OR SCRAPPING ONE OF THESE MODEL "SEM'S". PLEASE CONTACT US OFF LINE.
Dear Colleagues, The 4th ASEAN Microscopy Conference (ASEANMC4) will be held in Hanoi, VietNam, from October 9 till 10, 2003. This Conference will be a gathering of microscopists from the ASEAN and other regions of the world. An important aim of the Conference is to promote mutual understandings and to develop international collaborative research in life and material sciences. Participants will have the opportunity to review recent work, to discuss recent advances and identify new challenges in the field of electron and light microscopy. The Second announcement are currently in the mail to many microscopist who are interested in this conference. When we receive reply from you then we will send "the Second announcement" to you by post as soon as possible. We would like to extend our warmest welcome to you to participate and look forward to seeing you in Hanoi in October 2003. Chairman, Organizing committee Prof. Nguyen Van Man
*********************************************** Mailing address: Assoc. Prof. Nguyen Kim Giao Electron Microscopy Unit National Institute of Hygiene and Epidemiology N0 1, Yersin street- Hai Ba Trung district - Hanoi - Vietnam Tel: 84.04.9715434 Fax: 84.04.8210853 Email: emlad-at-hn.vnn.vn, or emunihe-at-vol.vnn.vn ***********************************************
Institute of Physics EMAG 2003 Electron Microscopy and Analysis Group Conference 2003 "Towards Objective Oriented Microscopy" The University of Oxford, UK, 3 - 5 September 2003 Call for Papers
Dates for your Diary 21 March 2003 Deadline for Abstract Submission April 2003 Abstract Notification May 2003 Registration Mailing 18 July 2003 Deadline for Registration
General Enquiries: WWW: http://physics.iop.org/IOP/Confs/EMG/ For further information please contact Jasmina Bolfek- Radovani,The Institute of Physics, 76 Portland Place, London W1B 1NT, Tel: +44 (0)20 7470 4800 Fax: +44 (0)20 7470 4900, Email: jasmina.bolfek-radovani-at-iop.org
Exhibition Enquiries: Ron Doole, EMAG03, Department of Materials, University of Oxford, Parks Road, Oxford., OX1 3PH Tel: + 44 (0)1865 273701 Fax: + 44 (0)1865 283333 Email:ron.doole-at-materials.oxford.ac.uk
We're using a nitrogen burst system and haven't had any problems with our 4489 films. Actually I was wondering what all the fuss was about! Maybe that's the solution. Good luck
Marc
On Thursday, February 13, 2003, at 04:33 PM, David Hall wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 also have been struggling with this new formulation for 3 weeks } now. The emulsion setting is apparently unchanged [sensitivity to } photons], but the film requires much, much more agitation [both more } frequent motion, and larger motions help] to get an approximately even } development. It is as if the film clings to the developer in a patchy } fashion, so that one tends to get very muddy, uneven images. It is } almost impossible to get good development near the edge of the film } holder, apparently due to poor exchange of solutions, even with lots } of agitation. We are also being extra careful to fully drain all } developer from the film before going to the 1st rinse, to agitate a } little during this rinse, and to agitate a lot in the fix step, again } using larger motions, and more frequently. } } It is really easy with this film to obtain terrible, unprintable } images. But with more concentrated efforts by the user, most of the } problem can be dealt with. We've wasted at least one hundred shots } getting our methods corrected, and produced some really ugly images } for users during January. Hope to be back on track soon. Would love } to know how to get an even development all the way to the edge of the } film. } } We would be interested to hear if people using a nitrogen burst system } find any problems? Is that sufficient to deal with the changes in } film properties? } -- } David H. Hall, Ph.D. } Center for C. elegans Anatomy } Department of Neuroscience } Albert Einstein College of Medicine } 1410 Pelham Parkway } Bronx, NY 10461 } } www.wormatlas.org } www.aecom.yu.edu/wormem } } phone 718 430-2195 } fax 718 430-8821 } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
I am a hydrophobe when it comes to Araldite and Spurrs. I was taught that sticky blocks were often due to water in the ethanol. I have never tried experiments to check this advice.
Dave
On Fri, 14 Feb 2003 10:26:55 -0500 Geoff McAuliffe {mcauliff-at-UMDNJ.EDU} wrote:
} On the subject of molecular seives and keeping our absolute ethanol } water-free I was always taught that every last molecule of water had to be } removed during processing for EM. Well, it turns out that Epon is miscible } with 70% ethanol! Check out Hayat's Principles and Techniques of EM, } biological applications, vol. 1, page 154. Also, one of my colleagues } routinely goes from 95% ethanol to Epon (ok, an Epon substitute, I don't know } which one) with no problems. I don't know about Araldite, Spurr, etc. } } Geoff } } } On Wed, 12 Feb 2003 08:20:09 -0600 "Tindall, Randy D." } } {TindallR-at-missouri.edu} wrote: } } } } } Many thanks to everyone who replied to my question about possible damage } } } to diamond knives due to use of molecular sieves in the dehydration } } } solvent. There doesn't seem to be a real consensus on this issue, with } } } some folks saying they have never had a problem even after a couple of } } } decades, and others saying they wouldn't use molecular sieves under any } } } circumstances. The latter people advocated using freshly opened } } } absolute ethanol (or whatever) for the final dehydration steps. } } } } } } One repeated suggestion was that molecular sieves can be safely used if } } } they are put in dialysis tubing to keep particulates safely contained. } } } I think we'll try this. } } } } } } Another suggestion made by a couple folks was that use of glass pipettes } } } during processing could be the source of glass chips that damage knives. } } } We never use glass pipettes in our sample preps, but interestingly } } } enough, a client who sometimes brings us blocks prepared in his own lab } } } does use them! Mystery solved? We'll see. } } } } } } Thanks again to everyone. As always, this list is a real resource. } } } } } } Randy } } } } } } Randy Tindall } } } EM Specialist } } } Electron Microscopy Core---We're the Fun Core! } } } 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/ } } } } } -- } ********************************************** } 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 } ********************************************** }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I have a class lecture to give next week on localization, but I don't have an image (nor do I seem to be able to readily locate one) of a GFP molecular model. I was hoping some one out there would have one I could use (digital just good enough for electronic power point) or point me in the right direction.
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
There has been a great deal of discussion about the problems with the newly formatted Kodak type 4489 film. We have been using the Kodak type S-063 for many years without incident. It is more similar to the old Electron Image plates that we used through the late 80¹s. Now I understand that S-063 is faster than 4489 and has slightly larger grain but it also can stand more manipulation regarding exposure and development times. Small film grain is important. However, unless you are doing very critical high magnification imaging, I wonder if the slightly larger grain in S-063 would be a problem.
Has anyone done a comparison between the two films recently? This may be an alternative that you can try.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907 On 2/13/03 9:03 AM, "Eleana Sphicas" {sphicae-at-rockefeller.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } I seem to have missed most of the discussion about the new formulation } Kodak 4489 film. Would someone please let me know what's the latest } conclusion on that? I have been using the new film for several months } without problem, but recently I am experiencing problems with the new film. } What alternatives to Kodak film would anyone recommend? } } Eleana } } }
Hi All: I have been asked by a colleague to ask everyone two things: 1) Is there anyone with a Philips 430 that they are parting out?
2) Is there anyone that has been able to align the Philips 430 without the use of the alignment controls (many of his microscope's electronic alignment controls are non operational). If so, please contact me off-line with ideas/insights or techniques. Thanks in advance, Michael Coviello UT Arlington
Since I haven't had the problem YET, but likely will soon, IF I do see what you have seen, I'll likely try a little PhotoFlo in the developer to help with film wettability.
Fred Monson
-----Original Message----- } From: David Hall [mailto:hall-at-aecom.yu.edu] Sent: Thursday, February 13, 2003 4:33 PM To: Microscopy-at-sparc5.microscopy.com
We also have been struggling with this new formulation for 3 weeks now. The emulsion setting is apparently unchanged [sensitivity to photons], but the film requires much, much more agitation [both more frequent motion, and larger motions help] to get an approximately even development. It is as if the film clings to the developer in a patchy fashion, so that one tends to get very muddy, uneven images. It is almost impossible to get good development near the edge of the film holder, apparently due to poor exchange of solutions, even with lots of agitation. We are also being extra careful to fully drain all developer from the film before going to the 1st rinse, to agitate a little during this rinse, and to agitate a lot in the fix step, again using larger motions, and more frequently.
It is really easy with this film to obtain terrible, unprintable images. But with more concentrated efforts by the user, most of the problem can be dealt with. We've wasted at least one hundred shots getting our methods corrected, and produced some really ugly images for users during January. Hope to be back on track soon. Would love to know how to get an even development all the way to the edge of the film.
We would be interested to hear if people using a nitrogen burst system find any problems? Is that sufficient to deal with the changes in film properties? -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
We have had several reports today that nitrogen burst systems are sufficient to overcome the film's poor wettability, using 2 sec bursts every 8 seconds.
One lab has reported that a pre-rinse in water before the developer also helps to give a more even effect.
We have not had a chance yet to try these ideas [and we don't own a nitrogen burst system], but the problem is serious for anyone lacking nitrogen burst. Clearly the physics at the film surface are quite different to what we are used to.
DHH
} Since I haven't had the problem YET, but likely will soon, IF I do see what } you have seen, I'll likely try a little PhotoFlo in the developer to help } with film wettability. } } Fred Monson
-- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
My response to David Hall's concerns about film agitation during dewvelopment:
Years ago another EM Lab and mine explored this issue. We did all kinds of agitations and came to the conclusion that very minimal agitation produced VERY even development. We exposed Kodak neg emulsions for testing under a darkroom enlarger set up to provide very even illumination and a medium developed density, so did not test using TEM exposed negs of a stained section,for example.
Minimum agitation consisted of the following procedure: at beginning, lower rack of negs into developer (we use full strength Kodak D-19), immediately raise completely out of developer and lower, 2x times. Wait 1 minute, raise out of developer 1x, lower again, wait 1 minute and repeat 1x. Wait 30 sec, raise out, lower. Wait 30 sec, remove and drain 15 sec, place into stop bath. Total time is 3.0 minutes, plus 15 sec drain time.
The 3.0 min development time was empirically determined by running exposure series on TEM (different camera speeds), photographing typical stained biological section (what we look at 90% of time).
Other minimal agitation methods would probably work too, the point is to do just 1 up and down about every minute. Of course the point of agitation is to bring fresh developer to the surface of the emulsion and to remove reaction products from that surface. Perhaps this minimal agitation method means you need a bit more time to overcome slowed development rate due to reaction product buildup between agitations, but one up/down agitation seems sufficient to replenish fresh developer at emulsion surface soon enough to give good development in 3 minutes, full strength D-19, room temp (~22C).
We concluded that the rapid and frequent agitations produce whorls of quick moving developer over the central areas of the emulsions, but create areas near the edges of the film where there is relatively reduced motion of developer and reduced development rates, due to fluid friction in the narrow spacing of about 1/8 inch between films in the rack, developer "boxed in" to the corners between tank sides, edges of films, etc. We would see reduced edge development using those agitations, uneven development across central areas of fillms. For comnparison, we did tray development, few films at a time, with agitation done by moving fingers over the films under development, also got even development that way, as no edge/corner restriction of developer flow in a tray, but who would ever develop 56 filams in a tray! Got to get it to work in a rack in a tank.
Never tried nitrogen burst system, may be fine if you have huge amounts of film to be developed daily, but seems like unnessesary compliction and expense here, given amounts of film we process weekly.
-- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Bera
} We also have been struggling with this new formulation for 3 weeks } now. The emulsion setting is apparently unchanged [sensitivity to } photons], but the film requires much, much more agitation [both more } frequent motion, and larger motions help] to get an approximately } even development. It is as if the film clings to the developer in a } patchy fashion, so that one tends to get very muddy, uneven images. } It is almost impossible to get good development near the edge of the } film holder, apparently due to poor exchange of solutions, even with } lots of agitation. We are also being extra careful to fully drain } all developer from the film before going to the 1st rinse, to agitate } a little during this rinse, and to agitate a lot in the fix step, } again using larger motions, and more frequently. } } It is really easy with this film to obtain terrible, unprintable } images. But with more concentrated efforts by the user, most of the } problem can be dealt with. We've wasted at least one hundred shots } getting our methods corrected, and produced some really ugly images } for users during January. Hope to be back on track soon. Would love } to know how to get an even development all the way to the edge of the } film. } } We would be interested to hear if people using a nitrogen burst } system find any problems? Is that sufficient to deal with the } changes in film properties?
} David H. Hall, Ph.D. } Center for C. elegans Anatomy } Department of Neuroscience } Albert Einstein College of Medicine } 1410 Pelham Parkway } Bronx, NY 10461
Does anyone have a clever easy way to separate new glass slides when you open the box and they are perfectly stuck together. I have heated them in water but that didn't work. I've tapped them on counters, picked at their corners with forceps and fingers....the glass is so clean that they are just "welded" together along their entire surface.
: "You can learn a lot by observation - just by lookin'!" - Yogi Bera : : } We also have been struggling with this new formulation for 3 weeks : } now. The emulsion setting is apparently unchanged [sensitivity to : } photons], but the film requires much, much more agitation [both more : } frequent motion, and larger motions help] to get an approximately : } even development. It is as if the film clings to the developer in a : } patchy fashion, so that one tends to get very muddy, uneven images. : } It is almost impossible to get good development near the edge of the : } film holder, apparently due to poor exchange of solutions, even with : } lots of agitation. We are also being extra careful to fully drain : } all developer from the film before going to the 1st rinse, to agitate : } a little during this rinse, and to agitate a lot in the fix step, : } again using larger motions, and more frequently. : } : } It is really easy with this film to obtain terrible, unprintable : } images. But with more concentrated efforts by the user, most of the : } problem can be dealt with. We've wasted at least one hundred shots : } getting our methods corrected, and produced some really ugly images : } for users during January. Hope to be back on track soon. Would love : } to know how to get an even development all the way to the edge of the : } film. : } : } We would be interested to hear if people using a nitrogen burst : } system find any problems? Is that sufficient to deal with the : } changes in film properties? : : } David H. Hall, Ph.D.
In 40 years of developing film the most consistently even developing of sheet film I ever got was using tubes partially filled with developer constantly agatiteted while floating in water. The commercial version are BZST film tubes http://viewcamerastore.com/catalog/default.php?cPath=27&PHPSESSID=daba0c60b4 7905b73d4ee127ee91c818. I made my own tubes from black sewer pipe.
The most you can do at one time are six negatives and it requires a person full time during the development. You only need to be in the dark while loading the tubes and filling with developer.
I don't know how 4489 would respond to constant agitation but most films can be handled by lowering the developing time by up to a third and or lowering the concentration of the developer. Be careful with dilute developers because you can have more film than you have developer and exhaust the developer before the film is developed.
If you are only doing a limited number of negatives it might be worth looking into. It takes up little space and no dedicated space. Compared to a nitrogen agitations system the price is very low.I would not want to spend 4 hours a day at this. But for one or two negatives it is really great and not bad for a couple of dozen. In a pinch you can load the film and developer in a dark bag. Being completely portable. If you do go probable take you own chemicals. If you don't you will regret it some day.
Expect to do some experiments wiht development.
Good luck
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
No, I don't know a way. Suggest that you contact the manufacturer for any tips. Then, please report them back to us.
Regards,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Tom Parker {tparker-at-lacsd.org To: "microscopy-at-sparc5.microscopy.com" } {microscopy-at-sparc5.microscopy.com} cc: Subject: glass slides 02/14/03 05:54 PM Please respond to "tparker-at-lacsd.org "
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Greetings Microscopers:
Does anyone have a clever easy way to separate new glass slides when you open the box and they are perfectly stuck together. I have heated them in water but that didn't work. I've tapped them on counters, picked at their corners with forceps and fingers....the glass is so clean that they are just "welded" together along their entire surface.
Hello List } } } } I'd like to hear Your opinion/experience about cleaning of the diamond } } indentor like Vickers or Berkovich type used in nano and micro indentation } } instruments. } } They get contaminated mainly by metals or surface deposits. Ultrsonic } } tratment can cause breaks of it or loose the mounting to holder. } } I was thinking about some plasma cleaning method but with some reactive } } media ?? what would be adviced to diamond (holded in steel mount). } } Maybe some places offer repolishing ?? } } } } regards } } } } Krzysztof Herman } } ================================= } } LABSOFT, PL 02-892 Warszawa, ul.Bazancia 45A } } =================================
At 03:31 AM 2/15/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-----Original Message----- } From: Michael O'Keefe [mailto:MAOKeefe-at-lbl.gov] Sent: Thursday, February 06, 2003 7:52 PM To: Anderson, Ron
Sounds like you have lossy image compression enabled in Acrobat, or possibly low output resolution settings.
Lossless or uncompressed options are available in Acrobat, and should give you the high quality you need for publishing. There are quality "presets" as well; "prepress" is the high quality option if I remember correctly (or just manually reduce compression and increase output resolution).
I believe there was an article with tips on using Acrobat for scientific publishing in a recent issue of "Microscopy Today". Anyone remember what issue it was?
-Kevin ------------------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 10:25 AM 2/6/03 -0600, "Dusevich, Vladimir" {dusevichv-at-umkc.edu} wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We use a Knoop hardness tester and check the size of the dent on an SEM. This may sound simple, but scotch tape was the way we cleaned very fine diamond heads at RCA laboratories. Roy Nelson Material Testing Laboratory mtl-at-njcc.com
"Krzysztof M.Herman" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello List } } } } } } I'd like to hear Your opinion/experience about cleaning of the diamond } } } indentor like Vickers or Berkovich type used in nano and micro } indentation } } } instruments. } } } They get contaminated mainly by metals or surface deposits. Ultrsonic } } } tratment can cause breaks of it or loose the mounting to holder. } } } I was thinking about some plasma cleaning method but with some reactive } } } media ?? what would be adviced to diamond (holded in steel mount). } } } Maybe some places offer repolishing ?? } } } } } } regards } } } } } } Krzysztof Herman } } } ================================= } } } LABSOFT, PL 02-892 Warszawa, ul.Bazancia 45A } } } =================================
I've had knife damage from molecular sieves, from glass chips from glass pipets, from cultured cells sscraped off coverslips, and from tissues that had been broken up with ground glass homogenizers. Most of these have been from blocks that were prepared by clients who did not check their protocol with me first!
As do others, we use fairly freshly opened pint bottles of absoute ethanol. We guesstimate when they are no longer close enough to absolute to be useful and dowgrade them to "about 95%". Life's been much easier!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Back in the days when we used to clean glass slides with 99% ethanol we were also able to solve the problem of slide clumps. Just leave them in the alcohol for a while and they should/will slide apart.
Let me know how you get on??
All the best.
Gareth
At 15:54 2003-02-14 -0800, Tom Parker wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Obs/NB New postal/visiting address from July 2002!
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
I use the edge of a straight edge razor blade. This seperates them and you don't have to worry about paper towel residue.
} } } Gareth Morgan {Gareth.Morgan-at-impi.ki.se} 02/17/03 03:52AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi
Back in the days when we used to clean glass slides with 99% ethanol we
were also able to solve the problem of slide clumps. Just leave them in the alcohol for a while and they should/will slide apart.
Let me know how you get on??
All the best.
Gareth
At 15:54 2003-02-14 -0800, Tom Parker wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} water but } that didn't work. I've tapped them on counters, picked at their corners with } forceps and fingers....the glass is so clean that they are just "welded" } together along their entire surface. } } Thanks for any suggestions. } } Tom Parker } {tparker-at-lacsd.org}
Obs/NB New postal/visiting address from July 2002!
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
Dear Tom, I don't know how to separate glass slides stuck together, but the slides we get are "interleaved", which means there is a piece of fine paper between the slides. Mary ----- Original Message ----- } From: "Tom Parker" {tparker-at-lacsd.org} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, February 14, 2003 3:54 PM
Tom Glass cutters use a light oil to keep the split open. Obviously oil would contaminate slides, but you could try immersion in a volatile fluid, such as petroleum spirit, xylene or ethanol. Chris
Dr. Chris Jeffree Inveresk Cottage 26, Carberry Road Inveresk Musselburgh Midlothian EH21 8PR Tel: +44 131 665 6062 FAX +44 131 653 6248 Mobile 07710 585 401 ----- Original Message ----- } From: "Tom Parker" {tparker-at-lacsd.org} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, February 14, 2003 11:54 PM
I just put the on a hotplate for a second and that usually separates them... MC
Mary Mager wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Tom, } I don't know how to separate glass slides stuck together, but the slides we } get are "interleaved", which means there is a piece of fine paper between } the slides. } Mary } ----- Original Message ----- } } } From: "Tom Parker" {tparker-at-lacsd.org} } } To: {microscopy-at-sparc5.microscopy.com} } Sent: Friday, February 14, 2003 3:54 PM } Subject: glass slides } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I need the adapter from a Link detector (c. 1990) to get my good Link detector onto a recently acquired 820 SEM. Has anyone got a dead Link detector from a JEOL 820 SEM. Even one from a good detector that's not mounted would be useful if I can borrow it long enough for my machinist to can make it.
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
A method I have used to separate glass slides that are stuck together is to dip them in liquid nitrogen. They snap apart easily. This is used routinely in the histology lab when pathologists have stacked the slides while still wet with mounting media and the mounting media glues them together. I have not tried this for your application but it won't hurt to try it.
Cheryl Rehfeld Meyer Instruments, Inc. Houston, TX ----- Original Message ----- } From: Tom Parker {tparker-at-lacsd.org} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, February 14, 2003 5:54 PM
Hi all,
We have some OC darkroom filters which are going bad (cracked layers) and I would like to replace them. Unfortunately, our fixtures are 9"x9" and all the filters I've found thus far have been 8x10 or 10x12. Before I get the larger ones and cut them down, is anyone aware of a vendor who sells the 9x9 filters?
TIA, Henk Colijn
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
Try placing slides in warm oven. Perhaps slight expansion will help separate the slides. I haven't tried this approach, so I don't know whether it will work or not.
Gary Gill
-----Original Message----- } From: Tom Parker [mailto:tparker-at-lacsd.org] Sent: Friday, February 14, 2003 6:55 PM To: microscopy-at-sparc5.microscopy.com
Greetings Microscopers:
Does anyone have a clever easy way to separate new glass slides when you open the box and they are perfectly stuck together. I have heated them in water but that didn't work. I've tapped them on counters, picked at their corners with forceps and fingers....the glass is so clean that they are just "welded" together along their entire surface.
This is to inform you that the Call for Abstracts for the forthcoming International Meeting on Applied Physics (APHYS-2003), to be held during October 14-18th 2003 in Badajoz (Spain), is now opened. All the information regarding this interdisciplinary conference can be found at the Conference website
In addition to the regular Scientific Program, several International Workshops will be held as pre-conference events. The following three Workshops are presently confirmed:
1. Workshop on Modern Applied Microscopy in Molecular and Cell Biophysics Research 2. International Interdisciplinar Workshop on Bioengineered Non-crystalline Solids 3. International Workshop on Occupational Radiation Protection
The Conference will be specifically interested in receiving reports on Interdisciplinary researches relating Physics with other Sciences such as Biology, Chemistry, Information Technology, Medicine, etc or relating different Physics areas. In other words, we are specially (but not exclusivelly) interested in reports applying the techniques, the training, and the culture of physics to research areas usually associated with other scientific and engineering disciplines
APHYS-2003 will also serve as a platform to search for partners for transnational collaboration projects, specially for the EU Sixth Framework Program (NETworks of Excellence and Integrated Projects). "Projects Presentations" and "Call for Partners" presentations proposals are therefore encouraged and welcomed. If you are interested in taking part of this Conference feature, please send us the corresponding form available at the website.
In addition to the "traditional" oral contributed and posters presentation, a Virtual Participation modality has been established for those researchers unable to attend it in person. A limited number of works can be presented in this way. Please refer to the Conference website for details.
If you are interested in taking part of APHYS-2003, please send us your PRE-REGISTRATION FORM (at the main website of the conference) as soon as possible. The pre-registration form is also available through the direct URL http://www.formatex.org/aphys2003/preregistration.htm
Deadline for abstracts submission is April 15th 2003 although we highly recommend you to submit your abstracts as soon as possible to avoid saturation during the days before the deadine (more than 800 researchers are expected to attend this large Applied Physics Conference).
Proceedings Accepted and presented papers will be reviewed for publication in special issues of several international Journals such as Journal of Microscopy, Journal of Non-crystalline Solids, Microelectronics Journal, Physica Scripta, Applied Surface Science, Radiation Protection Dosimetry and Applied Physics A (Materials Science & Processing, to be confirmed). Also a book "Advances in Applied Physics" will be published by an international publisher with those papers accepted for presentation but not suitable for the journal issues. For up-to-date information on publications participating at the Conference as publishers, please visit regularly the Conference website (Proceedings sections).
For any question or suggestion, please do not hesitate to contact us at secretariat-at-formatex.org, or visit www.formatex.org/aphys2003/aphys2003.htm (Bookmark the page!!) We would also appreciate if could disseminate this Call for Papers through your Department or Institution.
We hope to meet you at this exciting and interdisciplinar meeting!
----- Original Message ----- } From: "emlad" {emlad-at-hn.vnn.vn} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, February 14, 2003 1:14 PM
Dear colleague
This is to inform you that the Call for Abstracts for the forthcoming International Meeting on Applied Physics (APHYS-2003), to be held during October 14-18th 2003 in Badajoz (Spain), is now opened. All the information regarding this interdisciplinary conference can be found at the Conference website
In addition to the regular Scientific Program, several International Workshops will be held as pre-conference events. The following three Workshops are presently confirmed:
1. Workshop on Modern Applied Microscopy in Molecular and Cell Biophysics Research 2. International Interdisciplinar Workshop on Bioengineered Non- crystalline Solids 3. International Workshop on Occupational Radiation Protection
The Conference will be specifically interested in receiving reports on Interdisciplinary researches relating Physics with other Sciences such as Biology, Chemistry, Information Technology, Medicine, etc or relating different Physics areas. In other words, we are specially (but not exclusivelly) interested in reports applying the techniques, the training, and the culture of physics to research areas usually associated with other scientific and engineering disciplines
APHYS-2003 will also serve as a platform to search for partners for transnational collaboration projects, specially for the EU Sixth Framework Program (NETworks of Excellence and Integrated Projects). "Projects Presentations" and "Call for Partners" presentations proposals are therefore encouraged and welcomed. If you are interested in taking part of this Conference feature, please send us the corresponding form available at the website.
In addition to the "traditional" oral contributed and posters presentation, a Virtual Participation modality has been established for those researchers unable to attend it in person. A limited number of works can be presented in this way. Please refer to the Conference website for details.
If you are interested in taking part of APHYS-2003, please send us your PRE-REGISTRATION FORM (at the main website of the conference) as soon as possible. The pre-registration form is also available through the direct URL http://www.formatex.org/aphys2003/preregistration.htm
Deadline for abstracts submission is April 15th 2003 although we highly recommend you to submit your abstracts as soon as possible to avoid saturation during the days before the deadine (more than 800 researchers are expected to attend this large Applied Physics Conference).
Proceedings Accepted and presented papers will be reviewed for publication in special issues of several international Journals such as Journal of Microscopy, Journal of Non-crystalline Solids, Microelectronics Journal, Physica Scripta, Applied Surface Science, Radiation Protection Dosimetry and Applied Physics A (Materials Science & Processing, to be confirmed). Also a book "Advances in Applied Physics" will be published by an international publisher with those papers accepted for presentation but not suitable for the journal issues. For up-to-date information on publications participating at the Conference as publishers, please visit regularly the Conference website (Proceedings sections).
For any question or suggestion, please do not hesitate to contact us at secretariat-at-formatex.org, or visit www.formatex.org/aphys2003/aphys2003.htm (Bookmark the page!!) We would also appreciate if could disseminate this Call for Papers through your Department or Institution.
We hope to meet you at this exciting and interdisciplinar meeting!
Our lab is converting from sending micrographs manually, to sending scanned pictures over e-mail. What type of programs are you using to do this? Are you finding that you need to make adjustments to the pictures before you send them? Also what type of apparatus are you using to show accurate sizing?
Lloyd Willard, Research Associate Department of Diagnostic Medicine/Pathobiology Phone: (785) 532-4420 Electron Microscopy Laboratory Fax: (785) 532-4039 Kansas State University email: lwillard-at-vet.k-state.edu K-208 Mosier Hall web: www.vet.k-state.edu/depts/dmp/personnel/staff/research.htm Manhattan, Ks 66506-5606
} From: WCRGs-at-aol.com } Date: Mon, 17 Feb 2003 15:34:07 EST } Subject: Joel JSM T-300 } To: ListServer-at-sparc5.microscopy.com } Status: } } Hello Listers; } We have a Joel JSM T-300 SEM. This is an old piece of equipment } that we have been working on for a while. We are at the point where } we can get a filament current. There are two problems that we have } run into - 1) when we turn on the accelerating voltage, the filament } lamp lights up immediately. The instruction manual says that the } lamp should not light up until the filament dial is turned to about } the 3:00 o'clock position. This is even with the gun bias at the } full clockwise position. 2) We seem to hear and see (filament } checker spike), some type of spiking occasionally in the area of the } electron gun. This spiking seemed to show up when we started our } gun alignment procedure. } } If anyone has any suggestions on how we might address these problems } they would be much appreciated. } Thanks, } Ronald Obie } Wood Coatings Research Group, Inc. } 336-841-0264
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ronald.s.najorka-at-intel.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, February 17, 2003 at 15:30:50 ---------------------------------------------------------------------------
Email: ronald.s.najorka-at-intel.com Name: Ron Najorka
Organization: Intel Corp.
Education: Graduate College
Location: Hillsboro,OR,USA
Question: When using the Hitachi s-4700. I was told that if I flashed the tip every time I put a sample in, I would deplete the source to fast. Rather than if I flashed every 8 hours. This is strange because if I flash the tip every 20 minutes for each new sample I get much crisper pictures. Mainly because I am able to work with the aperature and x,y stigs in a much more precise manner.
Hi Tom, I keep a #11 scalpel blade (on the longer small handle) to separate obstreperous slides when I retrieve them from the 70% ethanol into which I immerse them to facilitate the separation. Often, they separate easily after they are wetted.
You could also use 1/2 of a double-edged razor blade in an appropriate holder leaving little of the edge exposed (I recommend a holder lest parts of hands and fingers become mounted along with the section).
Cheers and hope this helps,
Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging
Mail to: Geology, CASI West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383
For help and information only, The CASI houses: An FEI Quanta 400 and Technai 12T, Oxford INCA Energy 400, Tousimis AutoSamdri 815 and Olympus FV-300.
-----Original Message----- } From: Tom Parker [mailto:tparker-at-lacsd.org] Sent: Friday, February 14, 2003 6:55 PM To: microscopy-at-sparc5.microscopy.com
Greetings Microscopers:
Does anyone have a clever easy way to separate new glass slides when you open the box and they are perfectly stuck together. I have heated them in water but that didn't work. I've tapped them on counters, picked at their corners with forceps and fingers....the glass is so clean that they are just "welded" together along their entire surface.
} Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (ronald.s.najorka-at-intel.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, } February 17, 2003 at 15:30:50 } --------------------------------------------------------------------------- } } Email: ronald.s.najorka-at-intel.com } Name: Ron Najorka } } Organization: Intel Corp. } } Education: Graduate College } } Location: Hillsboro,OR,USA } } Question: When using the Hitachi s-4700. I was told that if I flashed } the tip every time I put a sample in, I would deplete the source to } fast. Rather than if I flashed every 8 hours. This is strange because } if I flash the tip every 20 minutes for each new sample I get much } crisper pictures. Mainly because I am able to work with the aperature } and x,y stigs in a much more precise manner.
Both parts of this are true. When you flash the tip you are passing a current through it to heat it up (it's normally a cold cathode tip), driving off contaminating molecules. When clean, you get more electrons off with less of an extraction potential, a better signal-to-noise ratio, and crisper pictures. Also true is that whenever you heat up the tip you drive off more material from that tip and cause it to become duller (less pointy), which translates to less resoution over time, plus shorter tip life. The trick is to find the balance, flashing only when it really needs it, usually whatever Hitachi recommends.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Depending on which e-mail client you use, the most normal way of sending picture files is via an attachment. These attachments can be encoded using several methods. The most common, I believe, is MIME. Outlook and Eudora offer this method.
One thing to look out for is the total packet size. If your outgoing SMTP server limits total size, you will have to keep your encoded message below that number. MIME messages have somewhere between 25% to 50% overhead. The workaround is to have a public ftp site where files can be dumped but not read.
Whatever the pix looks like before sending, it will look the same at the receiving end--if screen gamma is reasonably the same.
gary g.
At 09:05 AM 2/18/2003, you wrote:
} Our lab is converting from sending micrographs manually, to sending } scanned pictures over e-mail. What type of programs are you using to do } this? Are you finding that you need to make adjustments to the pictures } before you send them? Also what type of apparatus are you using to show } accurate sizing? } } Lloyd Willard, Research Associate } Department of Diagnostic Medicine/Pathobiology Phone: (785) } 532-4420 } Electron Microscopy Laboratory } Fax: (785) 532-4039 } Kansas State University email: lwillard-at-vet.k-state.edu } K-208 Mosier Hall web: } www.vet.k-state.edu/depts/dmp/personnel/staff/research.htm } Manhattan, Ks 66506-5606
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (atthom02-at-louisville.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, February 18, 2003 at 22:21:25 ---------------------------------------------------------------------------
Email: atthom02-at-louisville.edu Name: Arun
Organization: Thomas
Education: Graduate College
Location: Louisville, KY
Question: This is Arun Thomas from University of Louisville, Kentucky. We are researching the potential uses of Titanium alloys on medical implant devices. To the Engineering school,a Bausch and Lomb Dynazoom metallograph is donated, but unfortunately it does not have any imaging device with it. So we are interested in getting an attachment to this metallograph. We do not have any contacts for this thing to get done. I would appreciate if you could pass on some contact information regarding the tech support for this machine. Looking forward to hear from you soon, Thanks, Arun
Hi, Some time ago we have solved similar problem by putting the sealed glass slides into common lab ultrasonic cleaner in 70% ethanol. After some time of sonication the glass slides were separated. Maybe it will works for you, too. Best regards Oldrich
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-241062399 Fax: +420-241062347 WEB: http://www.biomed.cas.cz/mbu/lem113/lem.htm
-----Original Message----- } From: Tom Parker [mailto:tparker-at-lacsd.org] Sent: Friday, February 14, 2003 6:55 PM To: microscopy-at-sparc5.microscopy.com
Greetings Microscopers:
Does anyone have a clever easy way to separate new glass slides when you open the box and they are perfectly stuck together. I have heated them in water but that didn't work. I've tapped them on counters, picked at their corners with forceps and fingers....the glass is so clean that they are just "welded" together along their entire surface.
Digital sacanning and transmission of images can be a complex issue, so I am not attempting to be complete here. I will only present some thoughts on digital image scanning and image transmission.
Scanning the images can be done with Adobe Photoshop and several scanners, most of them can be used in combination with a TWAIN driver. Digital images can be sent over the Internet with almost any email program.
Scanning the images at 300 dpi or more is in general good enough for printing purposes afterwards, use about 70 dpi when the images are meant solely for onscreen viewing. Keep in mind that a high resolution of the scanner is not always "hard" but often done by interpolation.
When scanning and transmitting digital images it is important to know what they are meant for. If the images are meant for on-screen viewing, JPEG compression is good enough and a resolution of about 70 dpi (dots per inch) can be used. Care has to be taken if the images are to be used for printing or analysis afterwards. JPEG compression is lossy and introduces artefacts. Tiff with LZW compression is a possible alternative but the file size can be prohibitive.
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Our lab is converting from sending micrographs manually, to sending } scanned pictures over e-mail. What type of programs are you using to do } this? Are you finding that you need to make adjustments to the pictures } before you send them? Also what type of apparatus are you using to show } accurate sizing? } } Lloyd Willard, Research Associate } Department of Diagnostic Medicine/Pathobiology Phone: (785) } 532-4420 } Electron Microscopy Laboratory } Fax: (785) 532-4039 } Kansas State University email: lwillard-at-vet.k-state.edu } K-208 Mosier Hall web: } www.vet.k-state.edu/depts/dmp/personnel/staff/research.htm } Manhattan, Ks 66506-5606
Dear Lloyd, We scan pictures (with contrast/brightness adjusted suitably), do any further necessary contrast/brightness adjustments, label them with a micron bar, and then save them as TIFF. They are uploaded to a company server by ftp. We do the image processing using Photoshop but other packages could also be used.
Remember a micron bar is better than a magnification figure because you don't know if the client will blow up the pictures or something, in which case the magnification number becomes worthless, but the micron bar gets blown up with everything else.
As for image formats, TIFF or EPS preserve all information but can be quite big. JPEG is smaller because it is compressed, but the compression is lossy, i.e. not good for archive quality. Photoshop format is nice because layers (text annotations etc.) are preserved but this format may be large and not readable by all programs. In general we use TIFF for this reason, although I preserve a copy in Photoshop if I want to be able to re-edit the annotations later (perhaps for a conference presentation or such like).
Finally, sending by email is problematic as pictures tend to be large files. If you can set up an ftp server at one end or the other, it is much easier.
If you, however, want to just send someone a couple of pics to look at and not for them to use or edit themselves, then putting them into a pdf document is a good solution. Stick them into some programme that combines images and text reasonably well so that you can put a little label underneath each pic. Some such programmes include MS Powerpoint, Deneba Canvas, Adobe Pagemaker. Install Adobe Acrobat (full version) on your computer. Then when you print, you can print to pdfWriter or Distiller and get a pdf file instead of a printout. With suitable setting of the graphics resolution for pdfWriter or Distiller (eg 150 dpi), you get a nice small pdf file with all your pics in. This is great when writing a paper and you want to show the co-authors at other locations your pics without having to send 10 Mb emails. Instead, the pdf file will weigh in at less than 1 Mb with suitable choices for the Distiller or pdfWriter settings.
So,
for archive quality and editable pics --} TIFF,
for quick showing of pics --} all together in one pdf.
Hope this helps
Ian
On Tue, 18 Feb 2003 11:05:47 -0600, EM Lab {Emlab-at-vet.k-state.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 } -----------------------------------------------------------------------. } } } Our lab is converting from sending micrographs manually, to sending } scanned pictures over e-mail. What type of programs are you using to do } this? Are you finding that you need to make adjustments to the pictures } before you send them? Also what type of apparatus are you using to show } accurate sizing? } } Lloyd Willard, Research Associate } Department of Diagnostic Medicine/Pathobiology Phone: (785) } 532-4420 } Electron Microscopy Laboratory Fax: } (785) 532-4039 } Kansas State University email: lwillard-at-vet.k-state.edu } K-208 Mosier Hall web: www.vet.k- } state.edu/depts/dmp/personnel/staff/research.htm } Manhattan, Ks 66506-5606 } }
-- Ian MacLaren Technische Universität Darmstadt Material-und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany
I agree with these comments, having used FE for 20 years. We flash our SEMs last thing at night and then not again until the following evening. If your IP1 vacuum is good enough, this should be OK. Regular flashing does eventually blunt the tip. We flash on 1 and then on 2 and that's it. Watch IP1 when you flash, this gives a good indication of the vacuum status in the gun. It shouldn't go above 2x10-7. You may need a bake since outgassing samples may put H2O or H2 in the gun chamber. This is difficult for the ion pumps to remove.
Barry Lamb
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: 18 February 2003 22:30 To: by way of MicroscopyListServer Cc: MicroscopyListserver
} Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (ronald.s.najorka-at-intel.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, } February 17, 2003 at 15:30:50 } -------------------------------------------------------------------------- - } } Email: ronald.s.najorka-at-intel.com } Name: Ron Najorka } } Organization: Intel Corp. } } Education: Graduate College } } Location: Hillsboro,OR,USA } } Question: When using the Hitachi s-4700. I was told that if I flashed } the tip every time I put a sample in, I would deplete the source to } fast. Rather than if I flashed every 8 hours. This is strange because } if I flash the tip every 20 minutes for each new sample I get much } crisper pictures. Mainly because I am able to work with the aperature } and x,y stigs in a much more precise manner.
Both parts of this are true. When you flash the tip you are passing a current through it to heat it up (it's normally a cold cathode tip), driving off contaminating molecules. When clean, you get more electrons off with less of an extraction potential, a better signal-to-noise ratio, and crisper pictures. Also true is that whenever you heat up the tip you drive off more material from that tip and cause it to become duller (less pointy), which translates to less resoution over time, plus shorter tip life. The trick is to find the balance, flashing only when it really needs it, usually whatever Hitachi recommends.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} Our lab is converting from sending micrographs manually, to sending } scanned pictures over e-mail. What type of programs are you using to do } this?
Are you referring to the image editing software for converting to formats suitable for e-mail? That is, many e-mail servers will not allow for large attachments, and you therefore need to convert to a compressed format like JPEG. JPEG is however a "lossy" format ... make sure your clients are ok with JPEG artifacts. With respect to "what software", most use Photoshop, but many free softwares are available for conversions ... e.g., search the wwweb for "Irfanview" (Windows).
For other image formats (larger file sizes), you may need to set up an FTP server.
} Are you finding that you need to make adjustments to the pictures } before you send them? Also what type of apparatus are you using to show } accurate sizing?
I don't know how you can ^guarantee^ the final print size. But most image formats, including JPEG, can include the print size definition ... such that if you tell your client "If the image is printed as defined, it will be a specific magnification." Personally, I beleieve all images should include a mag reference in the image itself ... ,e.g., a micron bar which will always reference the correct magnification.
Regarding adjustments, each image should be judged independently ... but you should probably assume they will need something ... even if it's only a micron bar, or the image's print size defined. Again, emphasis should be put on Photoshop, or a quantitative and analytical software (e.g., Image Pro Plus, NIH Image or ImageJ). For editing with respect to presentation, Photoshop offers the best user/peer base and choice of excellent texts, as well as being compatible with quantitative plugins.
People have given some good imaging suggestions, so I won't add to that, but I would suggest that you consider not sending the mages directly at all. Many email systems have arbitrary limits on file size and storage per user. Also many folks still seem unable to handle enclosures (!), or get flustered when they try to read there email form a modem connected computer (while at home).
Because of these issues, I typically share SEM images by posting them on my website. Everybody knows how to use their browser, and from my end there are utilities available that will take an entire folder of images and make HTML thumbnail pages that link to the images. If sombeody wants high res images (usually they don't!) I burn a CD and FEDEX it.
--
Michael J. Herron, U of MN, Dept. of Entomology herro001-at-umn.edu 612-624-3212 (lab) St. Paul, MN 55108
We are interested in purchasing an ultra-thin coater for FESEM work. I would appreciate any opinions on types of materials used ie: chrome, platinum, osmium. Also, any preference on manufacturers. We are imaging semiconductors. Our main application for this coater would be cross-sections of semiconductors. Thanks for your input.
Mark Windland Honeywell Plymouth, Minnesota 763-954-2845 mark.j.windland-at-honeywell.com
I wonder if there are standard (or usual) ways for storing setting data from electron microscopes (magnification, working distance, acceleration V, etc.) into the image file itself, such that they can be automatically imported to a database. Some other devices (like digital cameras) automatically use the IPTC or EXIF fields for this.
Any general idea about how preserve and manage these data together with the images will be very welcome.
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
Question: This is Arun Thomas from University of Louisville, Kentucky. We are researching the potential uses of Titanium alloys on medical implant devices. To the Engineering school,a Bausch and Lomb Dynazoom metallograph is donated, but unfortunately it does not have any imaging device with it. So we are interested in getting an attachment to this metallograph. We do not have any contacts for this thing to get done. I would appreciate if you could pass on some contact information regarding the tech support for this machine. Looking forward to hear from you soon, Thanks, Arun
Arun:
Here's a link that you should investigate:
http://www.diaginc.com/EN/EN.htm
They advertise heavily in Advance Materials & Processes, a magazine for metallurgists. It seems they specialize in digital imaging systems for microscopes.
If you need to get more elaborate in your lab setup, I can pass on some other contacts.
Stu Smalinskas Metallurgist SKF Plymouth, Michigan stu.smalinskas-at-skf.com
__________________________________________________ Do you Yahoo!? Yahoo! Shopping - Send Flowers for Valentine's Day http://shopping.yahoo.com
The first possible explanation is that you have a pretty big leak in the gun which will increase the leakage current from the gun, hence the filament lamp lighting up.
Disassemble the gun column interface and check the "O" rings in that area. Clean the "O" rings by washing in your hands with hot soapy water. When dry gently pull the rings through your fingers, stretching them slightly to check for cracks. These "O" rings have moving surfaces in contact with them so they should be lightly greased.
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "MicroscopyListServer" {zaluzec-at-sparc5.microscopy.com} To: "MicroscopyListserver" {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 18, 2003 7:31 AM
Hi,
We have a Philips 201, lately I have been experiencing lighting problems. Different parts of the negative will be more exposed than others--it is not always in the same spot. Today it happened to be a straight line along the bottom of every negative. About 1/2" or less wide---which causes big problems when printing. Fanning corrected most of the problem. My supervisor has watched me saturate the filament and spread the beam to make sure everything was ok---and it was. I always make sure to center the beam and spread evenly before each shot. Interestingly, my supervisor does not seem to experience these lighting problems on her negatives?
It doesn't happen every time, but most of the time, and I have not experienced this problem when using a different scope (philips 401). If the filament is saturated properly and the beam is centered and spread evenly, where are these shadows on the negatives coming from?
HELP---HELP---HELP
Thank You, Lauren Simmerman Pathology Nebraska Health System-EM Lab 402-502-1811
Does anyone have any unused film cartridges for the JOEL Model #JEM-1200EX? We are looking for Kodak or US made. We have cartridges made in Japan but can not get film in US. Any suggestions would be appreciated.
Lorayne E. Ham Scientific Imaging Specialist
SNBL-USA, Ltd. 6605 Merrill Creek Parkway Everett, WA 98203 (425) 407-0121 ext. 2155 (425) 407-1122 Fax email: lham-at-snblusa.com
Confidentiality Notice: This email, its contents and attachments are confidential and may contain privileged information. It is intended solely for the use of addressee(s) only. Any use, copying or disclosure of this communication or attachments to any other person is expressly prohibited without written permission of SNBL USA, Ltd. If you receive this message in error, please notify the sender at SNBL USA, Ltd. immediately by return e-mail, telephone +1 425 407 0121, or fax +1 425 407 8601. We appreciate your cooperation.
Possibility of ambient light in the room getting to the negatives?
} We have a Philips 201, lately I have been experiencing lighting problems. Different parts of the negative will be more exposed than others--it is not always in the same spot. Today it happened to be a straight line along the bottom of every negative. About 1/2" or less wide---which causes big problems when printing. Fanning corrected most of the problem. } My supervisor has watched me saturate the filament and spread the beam to make sure everything was ok---and it was. I always make sure to center the beam and spread evenly before each shot. Interestingly, my supervisor does not seem to experience these lighting problems on her negatives? } } It doesn't happen every time, but most of the time, and I have not experienced this problem when using a different scope (philips 401). } If the filament is saturated properly and the beam is centered and spread evenly, where are these shadows on the negatives coming from?
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
It seems that you have to open the gun chamber and remove electron gun and then clean a wehnelt , change the filament, if you can see any errosion obviously. To confirm such problem came from dirty gun. 1 After electron gun is removed from the socket, close the gun chamber (without gun) and then evacuate the gun and column until you get ready lamp. Turn on the accelerating voltage, observ that the filament lamp, it should not me light up. 2 you got spike on filament checker, it means that it has a discharge, which is due to dirty wehnelt cap.
I hope that all above will help you to solve the problem Cheers,
Paiboon Nuannin Dept of Physics Faculty of Science Prince of Songkla University Hatyai, Thailand Quoting MicroscopyListServer {zaluzec-at-sparc5.microscopy.com} :
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } From: WCRGs-at-aol.com } } Date: Mon, 17 Feb 2003 15:34:07 EST } } Subject: Joel JSM T-300 } } To: ListServer-at-sparc5.microscopy.com } } Status:ÝÝ } } } } Hello Listers; } } We have a Joel JSM T-300 SEM. This is an old piece of equipment } } that we have been working on for a while. We are at the point where } } we can get a filament current. There are two problems that we have } } run into - 1) when we turn on the accelerating voltage, the filament } } lamp lights up immediately. The instruction manual says that the } } lamp should not light up until the filament dial is turned to about } } the 3:00 o'clock position. This is even with the gun bias at the } } full clockwise position. 2) We seem to hear and see (filament } } checker spike), some type of spiking occasionally in the area of the } } electron gun. This spiking seemed to show up when we started our } } gun alignment procedure. } } } } If anyone has any suggestions on how we might address these problems } } they would be much appreciated. } } Thanks, } } Ronald Obie } } Wood Coatings Research Group, Inc. } } 336-841-0264 } }
------------------------------------------------- This mail sent through IMP: https://chaba.psu.ac.th
You might investigate whether this is uneven development, rather than uneven illumination. We had some similar problems a year or so ago, that turned out to be due to plugged holes in our nitrogen burst system. This created gradients in the density of the film near the edges where there was insufficient mixing. If you are agitating by hand, it may be something different about the way you and your supervisor do the agitation.
Marie
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Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 2242 Storrs, CT 06269-2242 Phone: 860-486-3588 Fax: 860-486-6369
Thank you for your timely responses on slides that stick together. I received suggestions that included:
There is no good way to separate them
Try liquid nitrogen dip
Heat them in oven approx 105 C for period of time
Soak them in solvent such as ethanol
Soak them in ethanol and ultrasonic vibrate them
Pry them apart with razor blade or scalpel blade
I tried both the ethanol soak and oven warming on several batches and the oven warming seemed to separate about half of the clumps (most of my clumps are 2-3 slides per).
Suggestions as to the cause included "water weld" from moist packing of slides to oil adhesion from residue of glass cutter.
It was also suggested by one fairly massive user of slides that manufacturers varied tremendously in this problem.
One person suggested the use of "interleaved slides" that have a thin paper insert between each.
Possibly the list could run a poll on the best slides around?
IrfanView is great tiny but powerful viewer. It has nice editing capacity like brightness, contrast, gamma adjustment. It may work with TWAIN scanners also. I used to associate most image formats with that viewer. It handles even big images very well and it's extremely quick! If you need only to scan image, save it in some format and sent it as an attachment in E.mail, you probably may do it with IrfanView (except sending an E.mail). It's also good idea to embed into the image the scale bar. Personally, I prefer to sent to the customers low-res JPEG images with embedded scale bar for viewing purpose only. If customer satisfied with image, I'll send original high-res TIFF upon request. I also prefer to scan images at the highest possible "optical" scanner's resolution (about 1600 dpi, 16 bit) and save this image as a TIFF untouched (for archival purpose), then in Photoshop I reduce resolution and do some adjustments and save a second copy of the file (TIFF) for working purpose (usually 300 dpi). If I do know that client would be interested to see the image, I also create low-res copy of the image in JPEG (72 dpi) at the same time. I usually use macros to do all these tasks automatically. We used to store archival copy of the image on magneto-optical (MO) disk - 5.2 Gb currently per disk, 10 disks on the shelf...
Recently (with help of my daughter) I discover very nice feature in Photoshop-7 (have no idea does it exist in the earlier versions). It's called 'Adjustable Layer' in the "Layers" Menu. It creates layer with predetermined function like brightness/contrast etc. So, you may change parameters actually not changing the original image. You could create a bunch of such 'Adjustable Layers' over single image with different features. You may re-adjust each layer anytime. So, it's very good if you need to adjust your picture for printing etc. Another things I find in Photoshop-7 that Photoshop files actually smaller (30%) than TIFF. It amused me, but this is true at least in a few cases. It seems to me Photoshop-7 handles memory and other stuff differently (much better) than previous versions. Sergey
At 09:35 AM 2/19/03 -0330, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Lauren, I had a problem like this a number of years ago with our Philips 300. It turned out to be not the scope, but a problem with agitation during development of the negatives. Do you have the problem if someone else develops your negatives and/or do you have the problem if you develop your supervisor's negatives? Dean Abel Biological Sciences 141 BB University of Iowa Iowa City IA 52242-1324
At 08:05 PM 2/19/2003 +0000, you wrote: } ----------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } ----------------------------------------------------------------------- } Hi, } We have a Philips 201, lately I have been experiencing lighting problems. } Different parts of the negative will be more exposed than others--it is } not always in the same spot. Today it happened to be a straight line along } the bottom of every negative. About 1/2" or less wide---which causes big } problems when printing. Fanning corrected most of the problem. } My supervisor has watched me saturate the filament and spread the beam to } make sure everything was ok---and it was. I always make sure to center the } beam and spread evenly before each shot. Interestingly, my supervisor does } not seem to experience these lighting problems on her negatives? } It doesn't happen every time, but most of the time, and I have not } experienced this problem when using a different scope (philips 401). } If the filament is saturated properly and the beam is centered and spread } evenly, where are these shadows on the negatives coming from? } HELP---HELP---HELP } Thank You, } Lauren Simmerman } Pathology } Nebraska Health System-EM Lab } 402-502-1811
I think we have a very good solution for you with our IBS/e Ion Beam Sputter Deposition and Etching system. The IBS/e, is a thin film deposition system which is designed to improve high resolution electron microscopy imaging by depositing ultra-thin, fine grain metal and carbon films on specimens.
Some characteristics of ion beam sputtered films:
* 5 to 8Å Cr, Ta or W films eliminate charging and increase contrast up to 500kX * Film quantity required is proportional to specimen surface roughness * Films hold down fine particles * Ir Films act as cladding on delicate specimens subject to beam damage * 8Å Cr films can be used when doing EDS without producing X-rays above noise * 80Å Cr support substrates can be produced that are cohesive, amorphous, and smooth
Ion beam sputtered material evolves controllably and repeatably with an energy {25eV. There is no heat or radiation artifacts to decorate specimen detail. Properly deposited films are beyond the resolving power of the highest magnification FESEM image!
The dual axes motion of the stage insures uniform specimen coverage in cracks and crevices of small and large specimens up to 50mm diameter - up to 100mm with the large area stage.
If you want to take the fullest advantage of your FESEM, don't settle for a standard "chromium coater" utilizing planar magnetron technology. It is exciting and revealing to see the benefits of ion beam sputtered films on your samples. We can deposit many different metals and carbon or show you examples of contrast enhancement on various types of specimens from our library of micrographs.
Please contact me for more information or visit our website at www.southbaytech.com. I'll look forward to hearing from you.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
"Windland, Mark J (MN14)" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 interested in purchasing an ultra-thin coater for FESEM work. I } would appreciate any opinions on types of materials used ie: chrome, } platinum, osmium. Also, any preference on manufacturers. We are imaging } semiconductors. Our main application for this coater would be } cross-sections of semiconductors. } Thanks for your input. } } Mark Windland } Honeywell } Plymouth, Minnesota } 763-954-2845 } mark.j.windland-at-honeywell.com
-- David Henriks Vice President
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (random-at-pdx.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, February 19, 2003 at 16:22:20 ---------------------------------------------------------------------------
Email: random-at-pdx.edu Name: Random Diessner
Organization: Portland State University
Education: Undergraduate College
Location: Portland, OR, USA
Question: I work in a lab studying the morphology of various Archaeal viruses and virus-like-particles. There seems to be some controversy in the lab as to whether or not one can carbon coat a grid without a pre-existing support film. My understanding was that one needed a support film such as formvar or butvar for the carbon to be deposited on, other vehemently deny this. HELP!! =)
Mark Windland wrote: ================================================================== We are interested in purchasing an ultra-thin coater for FESEM work. I would appreciate any opinions on types of materials used ie: chrome, platinum, osmium. Also, any preference on manufacturers. We are imaging semiconductors. Our main application for this coater would be cross- sections of semiconductors. Thanks for your input. ================================================================ The Osmium Plasma Coaters, such as the OPC-60 shown on URL http://www.2spi.com/catalog/osmi-coat.html
employs a process that is unique and should never be confused with "sputtering". As a result, this is not a matter of small grain size, it it, from all that one can determine, to be **no** grain size since the nucleation of the growth is on an atomic scale (instead of from "active sites"). The coating itself seems to be amorphous at least down to the level one can make such a determination. A good example of the completely structureless and featureless coating (at extreme magnifications) is on URL http://www.2spi.com/catalog/opc-40.html
In order to demonstrate just how really thin of a layer can be deposited and still have conductivity, see URL http://www.2spi.com/catalog/osmium-plasma-coater-demonstration.html The coating thickness is estimated to be 20 nm, but in any case, one would never get that kind of BSE signal through a high Z layer if it was much more than that.
The total lack of grain size, as well the thinness of the layer, when coupled with the inertness relative to chromium, would make the osmium coatings put down using the OPC units something worth considering. We would be happy to run a demo sample for you anytime, contact me off-line for details for the sample submission.
Disclaimer: SPI Supplies is the distributor for the OPC line of Osmium Plasma Coaters made by Nippon Laser and Electronics in Nagoya, Japan. So quite naturally, it would be in our own interest to see more of these systems being sold!
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
Seems a little risky and a waste of liquid N2??????
At 14:34 2003-02-17 -0600, Cheryl Rehfeld - Meyer Instruments, Inc wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Obs/NB New postal/visiting address from July 2002!
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
Two other really nice image-viewers are XnView, a free, explorer-like image viewers with great capabilities (like e.g. converting a serie of images from one type to another, e.g. TIFF to JPEG, very fast, a nice overview via tumbnails,...) Download it for free at: http://www.xnview.com
Another VERY interesting program to adjust your photo's is NeatImage. With this program, you increase your sharpness (e.g. take away pixelation). Take a look at http://www.neatimage.com/examples.html. You can download the program, a free demo, but it has enough capabilities at: http://www.neatimage.com/
Really take some time to take a look, it's great!
Sven Terclavers
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Sven Terclavers LM/CLSM Microscopist Center for Transgene Technology and Gene Therapy (CTG) Campus Gasthuisberg K.U.L. O&N Herestraat 49 3000 Leuven Belgium Tel. +32 16 346173 Fax. +32 16 345990 Email: Sven.Terclavers-at-med.kuleuven.ac.be °°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°
______________ Thursday, February 20, 2003, 1:46:16 AM, you wrote:
SR} ------------------------------------------------------------------------ SR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America SR} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com SR} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html SR} -----------------------------------------------------------------------.
SR} IrfanView is great tiny but powerful viewer. It has nice editing capacity SR} like brightness, contrast, gamma adjustment. It may work with TWAIN SR} scanners also. I used to associate most image formats with that SR} viewer. It handles even big images very well and it's extremely quick! If SR} you need only to scan image, save it in some format and sent it as an SR} attachment in E.mail, you probably may do it with IrfanView (except sending SR} an E.mail). It's also good idea to embed into the image the scale bar. SR} Personally, I prefer to sent to the customers low-res JPEG images with SR} embedded scale bar for viewing purpose only. If customer satisfied with SR} image, I'll send original high-res TIFF upon request. I also prefer to scan SR} images at the highest possible "optical" scanner's resolution (about 1600 SR} dpi, 16 bit) and save this image as a TIFF untouched (for archival SR} purpose), then in Photoshop I reduce resolution and do some adjustments and SR} save a second copy of the file (TIFF) for working purpose (usually 300 SR} dpi). If I do know that client would be interested to see the image, I SR} also create low-res copy of the image in JPEG (72 dpi) at the same time. I SR} usually use macros to do all these tasks automatically. We used to store SR} archival copy of the image on magneto-optical (MO) disk - 5.2 Gb currently SR} per disk, 10 disks on the shelf...
SR} Recently (with help of my daughter) I discover very nice feature in SR} Photoshop-7 (have no idea does it exist in the earlier versions). It's SR} called 'Adjustable Layer' in the "Layers" Menu. It creates layer with SR} predetermined function like brightness/contrast etc. So, you may change SR} parameters actually not changing the original image. You could create a SR} bunch of such 'Adjustable Layers' over single image with different SR} features. You may re-adjust each layer anytime. So, it's very good if you SR} need to adjust your picture for printing etc. Another things I find in SR} Photoshop-7 that Photoshop files actually smaller (30%) than TIFF. It SR} amused me, but this is true at least in a few cases. It seems to me SR} Photoshop-7 handles memory and other stuff differently (much better) than SR} previous versions. Sergey
SR} At 09:35 AM 2/19/03 -0330, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It is possible to evaporate carbon onto a glass slide, float the film onto water and pick the carbon film up on grids much the same as is done with either formvar or butvar. However.... Carbon coated grids prepared in this manner are generally not as strong as formvar or butvar supported films, and thus do not lend themselves to the rigors of negative staining as readily. Preparing such carbon film coated grids is possible. The question is: Why?
Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals -- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (random-at-pdx.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } February 19, 2003 at 16:22:20 } --------------------------------------------------------------------------- } } Email: random-at-pdx.edu } Name: Random Diessner } } Organization: Portland State University } } Education: Undergraduate College } } Location: Portland, OR, USA } } Question: I work in a lab studying the morphology of various Archaeal } viruses and virus-like-particles. There seems to be some controversy } in the lab as to whether or not one can carbon coat a grid without a } pre-existing support film. My understanding was that one needed a } support film such as formvar or butvar for the carbon to be deposited } on, other vehemently deny this. HELP!! =) } } --------------------------------------------------------------------------- }
Dear All, We have a JEOL 5910LV SEM. Recently, I was testing our peltier cryostage to view wet clay samples. We can freeze up to -25 degrees celsius and set the pressure in the sample chamber to 230Pa at maximum. We did not manage to image any ice or wet material. The samples looked freezedried!
Does anyone has experience with this type of work?
Thanks, Ineke Joosten Netherlands Institute for Cultural Heritage Conservation Research Gabriel Metsustraat 8 1072 EA Amsterdam The Netherlands 00 31 (0)20 3054688/728
We do not have holders for JEOL but do have camera boxes and film cassettes for Philips 400 series microscopes. They are free to a good home as long as you pay shipping. Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 2/19/03 3:38 PM, "Lorayne Ham" {Lham-at-snblusa.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have any unused film cartridges for the JOEL Model #JEM-1200EX? } We are looking for Kodak or US made. We have cartridges made in Japan but } can not get film in US. } Any suggestions would be appreciated. } } Lorayne E. Ham } Scientific Imaging Specialist } } SNBL-USA, Ltd. } 6605 Merrill Creek Parkway } Everett, WA 98203 } (425) 407-0121 ext. 2155 } (425) 407-1122 Fax } email: lham-at-snblusa.com } } Confidentiality Notice: This email, its contents and attachments are } confidential and may contain privileged information. It is intended solely } for the use of addressee(s) only. Any use, copying or disclosure of this } communication or attachments to any other person is expressly prohibited } without written permission of SNBL USA, Ltd. If you receive this message in } error, please notify the sender at SNBL USA, Ltd. immediately by return } e-mail, telephone +1 425 407 0121, or fax +1 425 407 8601. We appreciate } your cooperation. } } } }
Lauren, check your film supply....is it the "new" formulation of Kodak 4489? Your problems sound like the ones reported by people using the new film. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Is Os coating durable? I do not use Cr coating because I was getting "disposable" specimens (oxidation was a problem).
Vladimir
} ================================================================ } The Osmium Plasma Coaters, such as the OPC-60 shown on URL } http://www.2spi.com/catalog/osmi-coat.html } } employs a process that is unique and should never be confused with } "sputtering". As a result, this is not a matter of small } grain size, it it, } from all that one can determine, to be **no** grain size since the } nucleation of the growth is on an atomic scale (instead of } from "active } sites"). The coating itself seems to be amorphous at least } down to the } level one can make such a determination. A good example of } the completely } structureless and featureless coating (at extreme } magnifications) is on URL } http://www.2spi.com/catalog/opc-40.html } } In order to demonstrate just how really thin of a layer can } be deposited and } still have conductivity, see URL } http://www.2spi.com/catalog/osmium-plasma-coater-demonstration.html } The coating thickness is estimated to be 20 nm, but in any } case, one would } never get that kind of BSE signal through a high Z layer if } it was much more } than that. } } The total lack of grain size, as well the thinness of the layer, when } coupled with the inertness relative to chromium, would make the osmium } coatings put down using the OPC units something worth } considering. We would } be happy to run a demo sample for you anytime, contact me off-line for } details for the sample submission. } } Disclaimer: SPI Supplies is the distributor for the OPC line } of Osmium } Plasma Coaters made by Nippon Laser and Electronics in } Nagoya, Japan. So } quite naturally, it would be in our own interest to see more of these } systems being sold! } } Chuck } } PS: Remember that we are striving to be 100% paperless, } therefore there } are no paper copies kept of this correspondence. Please be } sure to always } reply by way of "reply" on your software so that the entire string of } correspondence can be kept in one place. } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.2spi.com } ######################## } ============================================ } } } }
We have an older (model E5000) Polaron sputter coater with a broken switch. So far, I've not had any success in finding a replacement. Does anyone know of a source for Polaron parts?
By the way, the switch is a six-position, two-deck design, with two sweeps per deck. Two-deck, six-position, double-pole switches are easy enough to come by, but I haven't seen any with two sweeps per deck.
Thanks
Kevin L. Macke Research Technician Materials Characterization Facility
phone: (215) 898-4555 fax: (215) 573-0620
Department of Materials Science & Engineering University of Pennsylvania 3231 Walnut Street Philadelphia, PA 19104
There is a Phillips EM300, serial #D997, in excellent working condition that is available for the asking. The scope has been under service contract until last December and will no longer be used by DEC. The instrument has a 35mm, 70 mm and standard photographic plate cameras. There is a mount for a digital camera under the column. Various parts, o-rings, filaments, specimen holders, etc. are also available. The water chiller is not included. This TEM is still using the original mercury pumps. Shipping is up to the individual(s) or institution(s) interested.
Parties interested in this TEM will be considered on a first-come, first-serve basis according to the following priorities:
First priority: Any individual/institution willing to accept the TEM as is, accepting the scope with the mercury pumps and lower vacuum system in place.
Second priority: Any individual/institution willing to accept the TEM as is but with the mercury pumps and lower vacuum system removed.
Third priority: The instrument, as a last resort, will be pieced out to those desiring spare parts for their EM300's.
Interested parties are encouraged to contact me offline.
There are two ways to coat a carbon film on a grid. 1. Coat a freshly cleaved mica, float carbon on water and pick up carbon with a grid from below and blot water off. 2. Coat carbon on previously coated grids (with formvar or other plastic film). If the carbon is thick enough, you can dissolve the film with suitable solvent. Carbon is left on the grids. However, you may use the grids without dissolving the plastic film if it is done with a thin layer of carbon to strengthen it.
AnnFook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Room 2091, Bldg. 20, Central Experimental Farm, Ottawa, Ontario Canada K1A 0C6
Tel: 1-613-759-1638 Fax: 1-613-759-1701
e-mail: yanga-at-em.agr.ca
} } } by way of MicroscopyListServer {random-at-pdx.edu} 02/20/03 12:54AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (random-at-pdx.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, February 19, 2003 at 16:22:20 ---------------------------------------------------------------------------
Email: random-at-pdx.edu Name: Random Diessner
Organization: Portland State University
Education: Undergraduate College
Location: Portland, OR, USA
Question: I work in a lab studying the morphology of various Archaeal viruses and virus-like-particles. There seems to be some controversy in the lab as to whether or not one can carbon coat a grid without a pre-existing support film. My understanding was that one needed a support film such as formvar or butvar for the carbon to be deposited on, other vehemently deny this. HELP!! =)
I have tried two variations on the theme. I have used a paper punch to punch out rounds of freshly cleaved mica, stuck one edge of each round onto a clean glass slide with double stick tape, and evaporated carbon onto the slide. Score around the edges of the coated mica, or make a tic-tac-toe grid on each with a needle, leaving the center square large enogh for a grid, then float the films off the mica (one by one) onto water. Place a grid on the film and pick up. My favorite way to pick them up (and which I also use for making Formvar-coated grids) is to come down on top of them with a piece of Parafilm, then lift the Parafilm off. The films seem to float off the mica pieces easier than off a slide, at least in my hands. I've made some pretty sturdy and thin films this way - mostly to image nanoparticles.
Alternatively, I have evaporated carbon onto Formvar-coated grids, stuck them onto a slide as above, then dissolved away the plastic film. With uneven success, I must admit. Right now I can't remember what solvent(s) worked the best, and I often ended up with shreds of Formvar remaining on the grid. However, in these cases I still had enough pure carbon areas that I could easily image proteins and particles.
The pure carbon films do allow much better resolution and contrast than the Formvar or Butvar, but are certainly more hassle!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Brown University announces an opening for a microscopist/manager in the Electron Microscopy Facility in the Center for Advanced Materials Research (CAMR) at Brown University. This central facility serves users in Engineering, Physics, Geology, and Chemistry, as well as visitors from academia and industry. This user-operated facility contains modern electron imaging tools including TEM (JEOL 2010, EM420) and SEM (LEO 1530VP, JEOL 845) and also includes state-of-the-art optical and scanning probe microscopy equipment. The ideal candidate will have experience in the use of transmission electron microscopy for materials research and will teach a graduate level lab course in this area. The facilities manager oversees the daily operation of the facility, trains new users, and works with faculty on sponsored research projects. Other responsibilities of this position include representing the facility in dealings with equipment and service vendors, and troubleshooting sophisticated microscopy and sample preparation equipment. The education and experience of the successful candidate should be equivalent to a Masters level degree in materials science (or a closely related field) and include five to seven years of practical experience. Exceptional candidates with clearly demonstrated expertise in the required areas will also be considered.
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On Wednesday, February 19, 2003, at 09:54 PM, by way of MicroscopyListServer wrote:
} Question: I work in a lab studying the morphology of various Archaeal } viruses and virus-like-particles. There seems to be some controversy } in the lab as to whether or not one can carbon coat a grid without a } pre-existing support film. My understanding was that one needed a } support film such as formvar or butvar for the carbon to be deposited } on, other vehemently deny this. HELP!! =) } Dear Random, Since we cryo-EM folks routinely make holey carbon grids by evaporating carbon onto a plastic film with ~1 - ~10 micrometer holes in it, and, since there is no carbon where there were holes, I would definitely say that you will need a continuous support film on your grid in order to get carbon across the grid openings. If the grid is a high enough mesh, you could dissolve away the formvar, and the carbon would stay intact, but this will not be possible for larger mesh grids. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
You have to have atmosphere of water vapor in a specimen chamber, not air (I do not know if a JEOL 5910LV SEM has this capability).
Even if water vapor is injected in the chamber, it is often not enough during initial pumping, and atmosphere could became too dry. It is recommended to put some additional water droplets into the chamber, preferably close to specimen.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} ---------. } } } Dear All, } We have a JEOL 5910LV SEM. Recently, I was testing our } peltier cryostage to } view wet clay samples. We can freeze up to -25 degrees } celsius and set the } pressure in the sample chamber to 230Pa at maximum. We did } not manage to } image any ice or wet material. The samples looked freezedried! } } Does anyone has experience with this type of work? } } Thanks, } Ineke Joosten } Netherlands Institute for Cultural Heritage } Conservation Research } Gabriel Metsustraat 8 } 1072 EA Amsterdam } The Netherlands } 00 31 (0)20 3054688/728 } } } }
While we're on the subject, does anyone know of a freeware DVD player program?
cheers
rtch
} } } Two other really nice image-viewers are XnView, a free, explorer-like } image viewers with great capabilities (like e.g. converting a serie of } images from one type to another, e.g. TIFF to JPEG, very fast, a nice } overview via tumbnails,...) Download it for free at: } http://www.xnview.com } } Another VERY interesting program to adjust your photo's is NeatImage. } With this program, you increase your sharpness (e.g. take away } pixelation). Take a look at http://www.neatimage.com/examples.html. } You can download the program, a free demo, but it has enough } capabilities at: http://www.neatimage.com/ } } Really take some time to take a look, it's great! } } Sven Terclavers } } } } } } SR} IrfanView is great tiny but powerful viewer. It has nice editing } capacity SR} like brightness, contrast, gamma adjustment. It may work } with TWAIN SR} scanners also. I used to associate most image formats } with that SR} viewer. It handles even big images very well and it's } extremely quick! If SR} you need only to scan image, save it in some } format and sent it as an SR} attachment in E.mail, you probably may do } it with IrfanView (except sending SR} an E.mail). It's also good idea } to embed into the image the scale bar. SR} Personally, I prefer to } sent to the customers low-res JPEG images with SR} embedded scale bar } for viewing purpose only. If customer satisfied with SR} image, I'll } send original high-res TIFF upon request. I also prefer to scan SR} } images at the highest possible "optical" scanner's resolution (about } 1600 SR} dpi, 16 bit) and save this image as a TIFF untouched (for } archival SR} purpose), then in Photoshop I reduce resolution and do } some adjustments and SR} save a second copy of the file (TIFF) for } working purpose (usually 300 SR} dpi). If I do know that client would } be interested to see the image, I SR} also create low-res copy of the } image in JPEG (72 dpi) at the same time. I SR} usually use macros to } do all these tasks automatically. We used to store SR} archival copy } of the image on magneto-optical (MO) disk - 5.2 Gb currently SR} per } disk, 10 disks on the shelf... }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Dear Random, I prepare carbon films by evaporating carbon onto a collodion-covered grid. Then the collodion is dissolved by putting the grid on a chloroform-soaked filter paper stack for 48 hours ("Jaffe washer"). This leaves an amorphous, 20 to 30 nanometer-thick carbon film adhered to the grid. I find these films more robust than the formvar films in my 200kV TEM because they are conductive. I use them to study small particles. Mary ----- Original Message ----- } From: "by way of MicroscopyListServer" {random-at-pdx.edu} To: "MicroscopyListserver" {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, February 19, 2003 9:54 PM
Does any electron microscopist out there in cyber space have a single tilt holder (PW6596) for 3.00 mm grids from a machine they are decommissioning. We would be prepared to pay for shipping and an agreed price.
Hoping someone can help us
Terry Robertson
Dr Terry A Robertson (PhD) Senior Research Fellow School of Surgery and Pathology Division of Pathology University of Western Australia Nedlands Australia 6907 Phone (61) 8 93462935 Mobile 0403025440 Fax (61) 8 93462891 email terryr-at-cyllene.uwa.edu.au
Ineke Joosten wrote: =============================================================== We have a JEOL 5910LV SEM. Recently, I was testing our peltier cryostage to view wet clay samples. We can freeze up to -25 degrees celsius and set the pressure in the sample chamber to 230Pa at maximum. We did not manage to image any ice or wet material. The samples looked freezedried!
Does anyone has experience with this type of work? ================================================================ Once you get above the range of 55-60°C, the sublimation rate of ice becomes considerable. Below that temperature range the rate is very slow. Since you are in the fast sublimation rate range, it would seem that the ice disappeared on you and that is why you are getting the appearance you are seeing.
You would have to be lower in temperature to keep the ice from subliming quickly.
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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My statement should have read: ============================ Once you get above the range of -55 to -60°C, the sublimation rate of ice becomes considerable. Below that temperature range the rate is very slow. Since you are in the fast sublimation rate range, it would seem that the ice disappeared on you and that is why you are getting the appearance you are seeing. ============================= In my original posting I said "55-60°C" . Sorry for not better proof- reading.
My recollection of the discription of elemental osmium in the CRC handbook discribes it as having a discernable oder due to the oxidation of metallic Os to the tetroxide... And OsO4 has enough of a vapor pressure to be used as a heavy-metal stain/fixitive in biological TEM.
Ben Simkin (simkin-at-egr.msu.edu) Michigan State University, dpt. Chemical Engineering and Materials Science
On Thu, 20 Feb 2003, Dusevich, Vladimir wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is Os coating durable? } I do not use Cr coating because I was getting } "disposable" specimens (oxidation was a problem). } } Vladimir } } } ================================================================ } } The Osmium Plasma Coaters, such as the OPC-60 shown on URL } } http://www.2spi.com/catalog/osmi-coat.html } } } } employs a process that is unique and should never be confused with } } "sputtering". As a result, this is not a matter of small } } grain size, it it, } } from all that one can determine, to be **no** grain size since the } } nucleation of the growth is on an atomic scale (instead of } } from "active } } sites"). The coating itself seems to be amorphous at least } } down to the } } level one can make such a determination. A good example of } } the completely } } structureless and featureless coating (at extreme } } magnifications) is on URL } } http://www.2spi.com/catalog/opc-40.html } } } } In order to demonstrate just how really thin of a layer can } } be deposited and } } still have conductivity, see URL } } http://www.2spi.com/catalog/osmium-plasma-coater-demonstration.html } } The coating thickness is estimated to be 20 nm, but in any } } case, one would } } never get that kind of BSE signal through a high Z layer if } } it was much more } } than that. } } } } The total lack of grain size, as well the thinness of the layer, when } } coupled with the inertness relative to chromium, would make the osmium } } coatings put down using the OPC units something worth } } considering. We would } } be happy to run a demo sample for you anytime, contact me off-line for } } details for the sample submission. } } } } Disclaimer: SPI Supplies is the distributor for the OPC line } } of Osmium } } Plasma Coaters made by Nippon Laser and Electronics in } } Nagoya, Japan. So } } quite naturally, it would be in our own interest to see more of these } } systems being sold! } } } } Chuck } } } } PS: Remember that we are striving to be 100% paperless, } } therefore there } } are no paper copies kept of this correspondence. Please be } } sure to always } } reply by way of "reply" on your software so that the entire string of } } correspondence can be kept in one place. } } ============================================ } } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } President 1-800-2424-SPI } } SPI SUPPLIES FAX: 1-610-436-5755 } } PO BOX 656 e-mail:cgarber-at-2spi.com } } West Chester, PA 19381-0656 USA } } Cust.Service: spi2spi-at-2spi.com } } } } Look for us! } } ######################## } } WWW: http://www.2spi.com } } ######################## } } ============================================ } } } } } } } } }
This is in response to your question about carbon films on grids. You received a response involving carbon evaporated onto cleaved mica. Years ago, we used to buy such 10 NM carbon foils on mica from the Arizona Carbon Foil Co., now called ACF Metals. They were beautifully made but not cheap, as I recall. They will essentially make any thickness carbon foil you could ever want. } From the web, I see they are still in business.
http://www.techexpo.com/WWW/acf-metals/page1.html
http://www.techexpo.com/firms/acf-metl.html
This bottom link still says they supply EM substrates.
Disclaimer: I don't work for ACF Co. or ACF-Metals.
I hope this helps.
Paul Beauregard Senior Research Associate PPG Industries Monroeville Technical Center 440 College Park Drive Monroeville, PA 15146 724-325-5131 pabeauregard-at-ppg.com
Chuck - you could subtract 30oC to your figures. It rather depends what you mean by "slow". My rule of thumb over many years of LTSEM with Cambridge 250/Emscope SP2000 and Hitachi 4700/Gatan Alto has been to use -80oC as the standard etching temperature. From the point of view of LTSEM specimens etching is uncontrollably fast at -60oC, conveniently rapid at -80, slower and more controllable at -90, but is observable down to -100oC, probably lower, since water ice has low but measurable vapour pressure beyond -100oC. If ice must be observed at these temperatures the chamber atmosphere must contain water vapour at a partial pressure equilibrated to the vapour pressure of water above the ice. This can probably only be achieved in ESEM.
Ineke - Whatever the temperature of your cryostage, it is a major technical problem, and one we are grappling with currently, to know how to get an ice specimen into a cryoSEM without either removing ice from its surface or adding ice to its surface. Anyone got an answer to this?
Best wishes Chris
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Ineke Joosten wrote: } =============================================================== } We have a JEOL 5910LV SEM. Recently, I was testing our peltier cryostage to } view wet clay samples. We can freeze up to -25 degrees celsius and set the } pressure in the sample chamber to 230Pa at maximum. We did not manage to } image any ice or wet material. The samples looked freezedried! } } Does anyone has experience with this type of work? } ================================================================ } Once you get above the range of 55-60°C, the sublimation rate of ice becomes } considerable. Below that temperature range the rate is very slow. Since } you are in the fast sublimation rate range, it would seem that the ice } disappeared on you and that is why you are getting the appearance you are } seeing. } } You would have to be lower in temperature to keep the ice from subliming } quickly. } } 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 } ######################## } ============================================ } } }
Vladimir Dusevich wrote: ====================================================== Is Os coating durable? I do not use Cr coating because I was getting "disposable" specimens (oxidation was a problem). ======================================================= The osmium metal coating is "durable", indeed relative to chromium, as you suggest, it is inert. After all, it is a precious group metal. Researchers in Japan, where a number of these systems have been installed and used for some years, seem to find that the shelf life for a coated sample is like it would be for gold. Now we have not been able to verify that yet ourselves but specimens coated two years ago by us seem unchanged (when viewed in a conventional non-FESEM instrument, from the way they looked when originally coated.
In order for the Os metal coating to become unstable, it would have to be subjected to some kind of oxidizing agent (and it could be converted back first to the dioxide and then to the tetroxide and that obviously would not be a good thing) but samples sleeping in storage boxes tend to not get exposed to oxidizing agents.......but admittedly, if one was coating particles of sodium periodate, for example, something we have not done, then perhaps one could speculate about its long term stability. But again this is not something we have done.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
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-----Original Message----- } From: Kevin Macke [mailto:macke-at-lrsm.upenn.edu] Sent: Thursday, February 20, 2003 11:56 AM To: Microscopy-at-sparc5.microscopy.com
We have an older (model E5000) Polaron sputter coater with a broken switch. So far, I've not had any success in finding a replacement. Does anyone know of a source for Polaron parts?
By the way, the switch is a six-position, two-deck design, with two sweeps per deck. Two-deck, six-position, double-pole switches are easy enough to come by, but I haven't seen any with two sweeps per deck.
Thanks
Kevin L. Macke Research Technician Materials Characterization Facility
phone: (215) 898-4555 fax: (215) 573-0620
Department of Materials Science & Engineering University of Pennsylvania 3231 Walnut Street Philadelphia, PA 19104
Random - You should be able to coat freshly cleaved mica with carbon, float the carbon off onto water, let the water out so the carbon floats down onto grids that you have placed on a wire mesh. You should be able to use up to 400 mesh grids. The smaller the mesh the less breakage.
Please feel free to contact me off line or by phone if you have any questions.
ML -- Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
We routinely C coat glass slides, float off on water and pick up on 200 mesh grids. The C is coated to a light grey color and works fine for, e.g., 100 Å thick polyethylene single crystals; they may, however, be too thick for some biological samples These will often be dried down on the C coated slides and shadowed before floating. The C is scratched with the point of a tweezers to ca 1/8 in squares before floating. These are then picked up with a grid held in the tweezers, coming up at an angle so the carbon catches on one edge first. If the C layer sticks to the grid, breathing on it after scratching or storing it under an evaporator dish with a small amount of water helps. At times we will also use cover slips as the initial substrate, floating them on ca. 1% HF. There is no way, however, that a C film can be directly deposited on the empty holes of a grid.
--
Phillip H. Geil; Ph. 217-333-0149 Fax 217-333-2736 Department of Materials Science and Engineering University of Illinois 1304 W. Green St. Urbana, IL 61801
Hi all, I'm currently using and Epson 1280 to print photos and it has been adequate for my needs as far as image quality goes but way too slow for printing large numbers of photo quality prints. These past couple of weeks I'm had the need to print large numbers of 8x10 prints FAST! Management wants to purchase a printer that can output a photo quality color print in a minute or less (preferably less!). I just tested a Xerox/Tectronix 6200. The quality was less than the Epson but much faster.
Any recommendations for a fast photo quality printer for digital images? We generally have image files in the 10's of MB and can download to the network 500MB or more worth of photos for a single run.
TIA Damian Neuberger Senior Research Scientist Baxter Healthcare Corp. damian_neuberger-at-baxter.com Tel: 847.270.5888 Fax: 847.270.5897
1) when people want a picture that looks like a photographic print, I use a dye-sub (Kodak). Very slow, software isn;t all that great, and the consumables are very expensive (as was the printer itself, once upon a time). This is used for less than 1% of the work done, and when the printer finally dies it won't be replaced.
2) an Epson outfitted with the Piezographics inks and software, used for about 5% of the printing, produces better-than-photo-quality grey scale prints but is very slow, and also moderately costly per print because of the need for special coated papers. Using grey-scale inks produces fabulous results, but of course this doesn't help much with color (ink jet color prints are so-so; using archival pigmented inks instead of dyes causes some funny color shifts, but the dyes degrade badly with light, heat or humidity).
3) The other 95% is done on a Minolta QMS 3100C laser printer (1200dpi). It is extremely fast, networked to half a dozen computers, uses standard paper, and produces results roughly equivalent to a good magazine or book print. Laser printers have gotten very good for color work (not so wonderful for grey scale). It sounds as though this would fit your needs well. Software is excellent - has built in ICC curves and produces accurate color renditions.
John Russ
======
In a message dated 2/21/03 6:04:30 PM, neuberger1234-at-attbi.com writes:
} I'm currently using and Epson 1280 to print photos and it has been adequate } for my needs as far as image quality goes but way too slow for printing } large numbers of photo quality prints. These past couple of weeks I'm } had } the need to print large numbers of 8x10 prints FAST! Management wants } to } purchase a printer that can output a photo quality color print in a minute } or less (preferably less!). I just tested a Xerox/Tectronix 6200. The } quality was less than the Epson but much faster. } } Any recommendations for a fast photo quality printer for digital images? } We } generally have image files in the 10's of MB and can download to the network } 500MB or more worth of photos for a single run.
There is free open source software: http://gallery.menalto.com/
It allows you to set up image albums very fast and efficient, with differientiated user permissions.
That way, you can avoid e-mailing images altogether, and your customers, clients *or* students can get images at the resolutions they want. You can also ban certain users from downloading full size images for whatever reasons.
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On Mittwoch, Februar 19, 2003, at 02:05 Uhr, michael shaffer wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Lloyd Willard writes ... } } } Our lab is converting from sending micrographs manually, to sending } } scanned pictures over e-mail. What type of programs are you using to } } do } } this? } } Are you referring to the image editing software for converting to } formats } suitable for e-mail? That is, many e-mail servers will not allow for } large } attachments, and you therefore need to convert to a compressed format } like } JPEG. JPEG is however a "lossy" format ... make sure your clients are } ok } with JPEG artifacts. With respect to "what software", most use } Photoshop, } but many free softwares are available for conversions ... e.g., search } the } wwweb for "Irfanview" (Windows). } } For other image formats (larger file sizes), you may need to set up } an FTP } server. } } } Are you finding that you need to make adjustments to the pictures } } before you send them? Also what type of apparatus are you using to } } show } } accurate sizing? } } I don't know how you can ^guarantee^ the final print size. But most } image } formats, including JPEG, can include the print size definition ... } such that } if you tell your client "If the image is printed as defined, it will } be a } specific magnification." Personally, I beleieve all images should } include a } mag reference in the image itself ... ,e.g., a micron bar which will } always } reference the correct magnification. } } Regarding adjustments, each image should be judged independently ... } but } you should probably assume they will need something ... even if it's } only a } micron bar, or the image's print size defined. Again, emphasis should } be } put on Photoshop, or a quantitative and analytical software (e.g., } Image Pro } Plus, NIH Image or ImageJ). For editing with respect to presentation, } Photoshop offers the best user/peer base and choice of excellent } texts, as } well as being compatible with quantitative plugins. } } hth & cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } www.micro-investigations.com (in progress) } }
} } } Hello All: } } } We are contemplating upgrading or trading-in our SEMs } } } including Hitachi S900. Is anyone out there with a } } } functional Hitachi S900 with EDX on it or at least memories of such a } } } system functioning anywhere, who would be willing to discuss its } } } pro-s and con-s ? } } } Marek. } } } } name MAREK MALECKI } building 1052 ANSCI BUILDING } department ANIMAL SCIENCE } division COLLEGE OF AGRICULTURAL & LIFE SCIENCES } email mmalecki-at-wisc.edu } phone (608) 262-0816 } title PROFESSOR } work-email mmalecki-at-wisc.edu } work_address 1675 OBSERVATORY DR MADISON WI 53706 } } } }
I recently looked through a number of recipes for mounting media that were posted a while back. I was looking for something that hardened so that I did not need to use nail polish to seal coverslip edges. Many recipes use Mowiol but I am confused by what role Mowiol plays in mounting media. I thought its job was to harden upon exposure to air so that the coverslip was sealed. However, the Calbiochem catalog lists it as an antifade reagent. If it is not a hardening agent, then what does that job? Also, the protocols for mounting media using polyvinyl alcohol look identical to that for Mowiol. Is Mowiol a trade name of pva? Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
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I was just going to ask at what stage of freezing on the Peltier stage do you actually press the evacuate button and how thick were your specimens. If I am working with anything that I think is likely to lose water easily I wait until the sample has passed about -5 deg C before evacuating to between 70 or ~200 Pa. A thick sample of course may not reach the stage temperature as quickly.
But the use of extra water in the chamber is interesting. Does this not play havoc with the vacuum pumps even on a high pressure system - eg a lot extra gas ballasting or special water traps?
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural & Social Sciences University of Sunderland UK
----- Original Message ----- } From: "Dusevich, Vladimir" {dusevichv-at-umkc.edu}
Hi all, I am considering poster printers and wanted input from the multiple users out there that print large photos on poster printers. I am considering both HP and EPSON. Do you have any comments that I should know about. Thanks in advance, Robert
Dr. Robert K. Pope Indiana University Department of Biology 1700 Mishawaka Avenue South Bend, IN 46634 ropope-at-iusb.edu
I have run into a problem when I set a computer monitor next to my Amray 1830I monitor. The computer monitor shows a moving scan line of the SEM monitor and the scan line is gone when the TV is off. My concern is that the computer monitor would get damaged with time. Did anybody have that kind of problem? Is there a B&W TV monitor that would not cause this kind of effect? Any other suggestions. I would prefer to keep the computer monitor next to the TV.
I would like to hear from any vendors out there who have digital camera systems for mounting on TEM. We're looking at replacing our current system mounted on our Philips 410LS TEM. We will probably be most interested in a side mounted system. My bosses requirements are largest possible viewing field, at highest possible resolution, and a real time viewing of image on the monitor during scope operation. I lack experience in this area so any and all advice, information etc. would be greatly appreciated from anyone. Thanks.
Tom Bargar Electron Microscopy Core Research Facility Dept. of Cell Biology and Anatomy 986395 Nebraska Medical Center Omaha, NE 68198-6395
I have heard John McKenzie make the following suggestion at least two times at his seminars. I haven't tried it yet, but I can't argue with the logic.
He suggested that instead of purchasing a single fast, high-end printer, purchase multiple cheap printers and hook them up in parallel. If the cheap printers are 10x slower than the fast one, just hook up 10x as many on a print server. I haven't tracked the Codonics prices lately, but they were running $10K a few years ago. That would buy twenty Epson 1820s. However, I would hold back some and spend some money on the print server that would distribute jobs to the printers. I believe Windows NT (2000, XP-Pro) has such an ability to distribute jobs among similar printers. I just have never had the occasion to try it.
Regarding image sizes, I would seriously investigate this matter. I rather doubt that an Epson could render a 2000-pixel wide image so that it appeared much better than a 1200-pixel wide image when printed at 8x10 inches. It depends on the capabilities of the printer, but a 1200-dpi printer doesn't render 1200 pixels per inch - it probably can only render a tenth of that or about 120 pixels per inch. If so, all the extra pixels in the image are for naught - at least when it comes to printing.
I suspect (but cannot yet prove) that some print drivers are probably better than others are working with this limitation. There is no sense in sending data to the printer in excess of what can be rendered. However, I think some printer drivers are written to send all the data over anyway and let the printer sort it out. But that takes more time to communicate all that data and takes time at the printer to realize it is superfluous. I think a better driver would know the printer capabilities and only send such a data stream as would be useful.
If someone has experience with these matters, I would appreciate hearing the outcome of their experiments.
Warren
At 04:28 PM 2/21/03 -0600, you wrote: } Hi all, } I'm currently using and Epson 1280 to print photos and it has been adequate } for my needs as far as image quality goes but way too slow for printing } large numbers of photo quality prints. These past couple of weeks I'm had } the need to print large numbers of 8x10 prints FAST! Management wants to } purchase a printer that can output a photo quality color print in a minute } or less (preferably less!). I just tested a Xerox/Tectronix 6200. The } quality was less than the Epson but much faster. } } Any recommendations for a fast photo quality printer for digital images? We } generally have image files in the 10's of MB and can download to the network } 500MB or more worth of photos for a single run. } } TIA } Damian Neuberger } Senior Research Scientist } Baxter Healthcare Corp. } damian_neuberger-at-baxter.com } Tel: 847.270.5888 } Fax: 847.270.5897
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
We have a client looking at individual muscle fibers which need to be fixed and embedded for immunolabeling and which will require longitudinal sections along at least of portion of their miniscule lengths. These things are so tiny that they are barely visible with the naked eye in the fixative. They almost look like little dust motes, if we can see them at all.
Immuno means no osmium, so the problem arises about how we find them once they're in the resin (probably LR White), let alone position them precisely enough to do long sections.
Ideas we're considering include: 1) tying the fibers along a tiny section of dark thread so we can at least see the thread; 2) staining them with an LM stain, such as toluidine blue, before embedding them; and 3) settling them down onto poly-l-lysine coated Thermonox cover slips to stabilize them, then scribing the coverslips and/or using an LM stain.
We are trying to avoid pre-embedding labeling at the client's request, but that will be an option if all else fails. Then we can osmicate following the labeling process.
Questions: Has anyone ever tried adhering muscle fibers to poly lysine coverslips? If so, is the attachment stable enough to get through the processing/embedding process? Is the use of an LM stain practical, or does this introduce a lot of contamination at the EM level? Does toluidine blue penetrate past the fiber membrane? (I've never looked at LM stained materials in a TEM before.)
We're trying to get an idea of what might work before we start, since there is a substantial amount of time-consuming dissection, etc., involved.
Thanks much for any ideas and thoughts.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! 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/
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Damian,
John Mackenzie (North Carolina State) suggested simultaneous printing of images to several Epson printers in parallel. The printers provide great quality images and are dirt-cheap. Several of these in the lab allows overnight printing of a large number of images. I support this approach since it provides (1) the quality, (2) the price, and (3) quantity of images that you need. Remember that what your management wants and what they (you) need are often very different. It is your job to convince them that your approach provides what they want.
Good luck,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Damian Neuberger" {neuberger1234-at-att To: MicroscopyListserver bi.com} (by way of {microscopy-at-sparc5.microscopy.com} MicroscopyListServ cc: er) Subject: Printer recommendation
02/21/03 04:28 PM
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Hi all, I'm currently using and Epson 1280 to print photos and it has been adequate for my needs as far as image quality goes but way too slow for printing large numbers of photo quality prints. These past couple of weeks I'm had the need to print large numbers of 8x10 prints FAST! Management wants to purchase a printer that can output a photo quality color print in a minute or less (preferably less!). I just tested a Xerox/Tectronix 6200. The quality was less than the Epson but much faster.
Any recommendations for a fast photo quality printer for digital images? We generally have image files in the 10's of MB and can download to the network 500MB or more worth of photos for a single run.
TIA Damian Neuberger Senior Research Scientist Baxter Healthcare Corp. damian_neuberger-at-baxter.com Tel: 847.270.5888 Fax: 847.270.5897
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pgan-at-ap.ansell.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, February 24, 2003 at 03:14:57 ---------------------------------------------------------------------------
Email: pgan-at-ap.ansell.com Name: Gan Phay Fang
Organization: Ansell Shah Alam Sdn Bhd
Education: Graduate College
Location: Shah Alam,Selangor, Malaysia
Question: Dear Sir Good day ! I am a beginer as a SEM user. Currently, I notice that the higher the magnification of the SEM , the lesser the penetration of the electron beam as shown by the EDAX spectrum. It would be nice if you could tell me whether there is any correlation between the EDAX and the magnification as well as between the EDAX and the sharpness of the SEM imej.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dha6n-at-virginia.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, February 24, 2003 at 13:27:55 ---------------------------------------------------------------------------
Email: dha6n-at-virginia.edu Name: Dalaver Anjum
Organization: University of Virginia
Education: Graduate College
Location: charlottesville, VA 22904
Question: I'm interested to know the lattest software packages for nano-diffraction in TEM particularly for LACBED/CBED simulations. Will you please provide me some names of these? Thank you very much.
Hello, Has anyone been able to calibrate an ultrasonic bath successfully as described in the ASTM Method D5755-02? If so, what brand of sonicator are you using? Pronda Few
There should be no difference in the depth of penetration as a function of magnification. It will vary according to beam voltage (or effective beam voltage).
I am curious why you think there would be a difference in penetration depth as a function of magnification. What evidence did you see for it? I would guess that you are seeing an underlying layer disappear as you go to higher magnifications.
I would suspect that you might be getting increased charging as you got to higher magnifications and pump the same current into a smaller area. If so, that will reduce the effective beam voltage and you would get less penetration. You should also be getting artifacts in your image. I suggest looking at the high energy limit of your spectra taken at the various magnifications. The background should tail off at the energy of your beam. But say you were using a 10 kV beam but your background tailed off at 8 kV, then your effective beam voltage is only 8 kV because your sample has charged up to 2000 V.
Check it out and let us know what is happening.
Warren
At 02:38 PM 2/24/03 -0600, you wrote: } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (pgan-at-ap.ansell.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, February } 24, 2003 at 03:14:57 } --------------------------------------------------------------------------- } } Email: pgan-at-ap.ansell.com } Name: Gan Phay Fang } } Organization: Ansell Shah Alam Sdn Bhd } } Education: Graduate College } } Location: Shah Alam,Selangor, Malaysia } } Question: Dear Sir } Good day ! I am a beginer as a SEM user. Currently, I notice that the } higher the magnification of the SEM , the lesser the penetration of the } electron beam as shown by the EDAX spectrum. It would be nice if you could } tell me whether there is any correlation between the EDAX and the } magnification as well as between the EDAX and the sharpness of the SEM imej. } } Thanks.
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
It is the magnetic field from the deflection coils in the CRT monitor. Either get a shielded (more costly) computer monitor or an LCD flat panel.
gary g.
At 09:00 AM 2/24/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Randy, I have stained thick epoxy (Polybed 812) sections (1-5 microns) annealed to glass slides with Richardson's Stain (a toluidine blue look-alike) and then re-embedded the sections for thin sectioning and observed no problems as a result of staining. Of course, I stained the sections not the tissue. I am curious as to what you hear from other microscopists. I have worked with non-osmicated tissue and it is difficult to work with when you can't see it! Dean Abel Biological Sciences 141 BB University of Iowa Iowa City IA 52242-1324
At 01:19 PM 2/24/2003 -0600, you wrote:
} Dear Listers, } We have a client looking at individual muscle fibers which need } to be fixed and embedded for immunolabeling and which will require } longitudinal sections along at least of portion of their miniscule } lengths. These things are so tiny that they are barely visible with the } naked eye in the fixative. They almost look like little dust motes, if } we can see them at all. Immuno means no osmium, so the problem arises } about how we find them once they're in the resin (probably LR White), let } alone position them precisely enough to do long sections. } Ideas we're considering include: 1) tying the fibers along a tiny } section of dark thread so we can at least see the thread; 2) staining } them with an LM stain, such as toluidine blue, before embedding them; and } 3) settling them down onto poly-l-lysine coated Thermonox cover slips to } stabilize them, then scribing the coverslips and/or using an LM } stain. We are trying to avoid pre-embedding labeling at the client's } request, but that will be an option if all else fails. Then we can } osmicate following the labeling process. } Questions: Has anyone ever tried adhering muscle fibers to poly } lysine coverslips? If so, is the attachment stable enough to get through } the processing/embedding process? Is the use of an LM stain practical, or } does this introduce a lot of contamination at the EM level? Does } toluidine blue penetrate past the fiber membrane? (I've never looked at } LM stained materials in a TEM before.) We're trying to get an idea of } what might work before we start, since there is a substantial amount of } time-consuming dissection, etc., involved. Thanks much for any ideas and } thoughts. } Randy Tindall } EM Specialist } Electron Microscopy Core---We're the Fun Core! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211
Hi Randy, Try 1%tannic acid, it preserves antigenicity, good for membranes and imparts a pale hue to locate, embed and orient specimen easily. I haven't tried on muscle fibers though, should work.
shashi
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers,
We have a client looking at individual muscle fibers which need to be fixed and embedded for immunolabeling and which will require longitudinal sections along at least of portion of their miniscule lengths. These things are so tiny that they are barely visible with the naked eye in the fixative. They almost look like little dust motes, if we can see them at all.
Immuno means no osmium, so the problem arises about how we find them once they're in the resin (probably LR White), let alone position them precisely enough to do long sections.
Ideas we're considering include: 1) tying the fibers along a tiny section of dark thread so we can at least see the thread; 2) staining them with an LM stain, such as toluidine blue, before embedding them; and 3) settling them down onto poly-l-lysine coated Thermonox cover slips to stabilize them, then scribing the coverslips and/or using an LM stain.
We are trying to avoid pre-embedding labeling at the client's request, but that will be an option if all else fails. Then we can osmicate following the labeling process.
Questions: Has anyone ever tried adhering muscle fibers to poly lysine coverslips? If so, is the attachment stable enough to get through the processing/embedding process? Is the use of an LM stain practical, or does this introduce a lot of contamination at the EM level? Does toluidine blue penetrate past the fiber membrane? (I've never looked at LM stained materials in a TEM before.)
We're trying to get an idea of what might work before we start, since there is a substantial amount of time-consuming dissection, etc., involved.
Thanks much for any ideas and thoughts.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
__________________________________________________ Do you Yahoo!? Yahoo! Tax Center - forms, calculators, tips, more http://taxes.yahoo.com/
Anyone remember Tips & Tricks... Anyway, there are a couple of discussions for locating "invisible" samples in the TEM section at the following url:
http://www.biotech.ufl.edu/EM/tips/tem.html
Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } "Tindall, Randy D." {TindallR-at-missouri.edu} 02/24/03 02:19PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers,
We have a client looking at individual muscle fibers which need to be fixed and embedded for immunolabeling and which will require longitudinal sections along at least of portion of their miniscule lengths. These things are so tiny that they are barely visible with the naked eye in the fixative. They almost look like little dust motes, if we can see them at all.
Immuno means no osmium, so the problem arises about how we find them once they're in the resin (probably LR White), let alone position them precisely enough to do long sections.
Ideas we're considering include: 1) tying the fibers along a tiny section of dark thread so we can at least see the thread; 2) staining them with an LM stain, such as toluidine blue, before embedding them; and 3) settling them down onto poly-l-lysine coated Thermonox cover slips to stabilize them, then scribing the coverslips and/or using an LM stain.
We are trying to avoid pre-embedding labeling at the client's request, but that will be an option if all else fails. Then we can osmicate following the labeling process.
Questions: Has anyone ever tried adhering muscle fibers to poly lysine coverslips? If so, is the attachment stable enough to get through the processing/embedding process? Is the use of an LM stain practical, or does this introduce a lot of contamination at the EM level? Does toluidine blue penetrate past the fiber membrane? (I've never looked at LM stained materials in a TEM before.)
We're trying to get an idea of what might work before we start, since there is a substantial amount of time-consuming dissection, etc., involved.
Thanks much for any ideas and thoughts.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
I agree entirely with Warren. Depth of penetration is independent of magnification. If you are working with a sample that you believe is stoichiometrically homogeneous in the x,y and z axes, then you should not see any difference in spectra. However, if your elemental spacing is such that you do not scan across a region that has one of the elements you are looking for, you won't see it and you may simply not be scanning a large enough area. This is particularly important in quantification. A good example of this is metal alloys wherein you may get regions with only one element in it at high magnifications. Hope that's clear.
Maybe if you state what your sample is the problem will become more clear to everyone.
Regards, Peter Tomic
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Monday, February 24, 2003 4:30 PM To: Microscopy-at-sparc5.microscopy.com Cc: pgan-at-ap.ansell.com
There should be no difference in the depth of penetration as a function of magnification. It will vary according to beam voltage (or effective beam voltage).
I am curious why you think there would be a difference in penetration depth as a function of magnification. What evidence did you see for it? I would guess that you are seeing an underlying layer disappear as you go to higher magnifications.
I would suspect that you might be getting increased charging as you got to higher magnifications and pump the same current into a smaller area. If so, that will reduce the effective beam voltage and you would get less penetration. You should also be getting artifacts in your image. I suggest looking at the high energy limit of your spectra taken at the various magnifications. The background should tail off at the energy of your beam. But say you were using a 10 kV beam but your background tailed off at 8 kV, then your effective beam voltage is only 8 kV because your sample has charged up to 2000 V.
Check it out and let us know what is happening.
Warren
At 02:38 PM 2/24/03 -0600, you wrote: } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (pgan-at-ap.ansell.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, February } 24, 2003 at 03:14:57 } --------------------------------------------------------------------------- } } Email: pgan-at-ap.ansell.com } Name: Gan Phay Fang } } Organization: Ansell Shah Alam Sdn Bhd } } Education: Graduate College } } Location: Shah Alam,Selangor, Malaysia } } Question: Dear Sir } Good day ! I am a beginer as a SEM user. Currently, I notice that the } higher the magnification of the SEM , the lesser the penetration of the } electron beam as shown by the EDAX spectrum. It would be nice if you could } tell me whether there is any correlation between the EDAX and the } magnification as well as between the EDAX and the sharpness of the SEM imej. } } Thanks.
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Randy, We have used the wire loop and formvar film trick for arabidopsis roots that are also extremely small and impossible to see.
Make some small loops out of very thin copper wire leaving a "handle" for future manipulation. Cast a formvar film on a glass slide as is normally done but cut it into squares prior to floating the film off of the slide. Pick up the film squares with the wire loops so that you have a coated loop.
Next lay your fixed muscle fiber on to the film-covered loop. It should adhere fairly nicely.
Final step is to again sandwich the fiber with a formvar film. This takes a bit of practice as you don't want to dislodge your fiber. Just come down from above with your loop and the fiber clinging to the lower surface so it hits the new formvar film piece rather than the water.
The loops + film + fiber can then be carried through all the remaining solutions and even embedded in a flat bottomed capsule. The wire can be dug out o fthe polymerized resin leaving the fiber. The remaining block can then be cut off and reoriented if necessary.
Care does have to be taken so as to keep the film intact. It is sometimes helpful to stick the "handle" down into some wax melted into the bottom of a jar and then gently add and subtract solutions from the jar. In this way you are sure that the formvar film will not touch anything. Since it is a double or triple layer (we sometimes use a rectangular piece of formvar film then applying the final cover so that it actually covers both sides of the wire) it will withstand the s;urface tension changes as you rasie and lower fluid volumes.
Try it...it really works great for very small hard to see tissue.
By the way, I have also used toluidine blue to pre-stain tissue prior to embedding. It seems to work reasonably well without interfering later on although you sometimes loose a fair amount of the stain when dehydrating and infiltrating.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 2/24/03 8:02 PM, "Dean Abel" {dean-abel-at-uiowa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Randy, } I have stained thick epoxy (Polybed 812) sections (1-5 microns) } annealed to glass slides with Richardson's Stain (a toluidine blue } look-alike) and then re-embedded the sections for thin sectioning and } observed no problems as a result of staining. Of course, I stained the } sections not the tissue. I am curious as to what you hear from other } microscopists. I have worked with non-osmicated tissue and it is difficult } to work with when you can't see it! } Dean Abel } Biological Sciences 141 BB } University of Iowa } Iowa City IA 52242-1324 } } At 01:19 PM 2/24/2003 -0600, you wrote: } } } Dear Listers, } } We have a client looking at individual muscle fibers which need } } to be fixed and embedded for immunolabeling and which will require } } longitudinal sections along at least of portion of their miniscule } } lengths. These things are so tiny that they are barely visible with the } } naked eye in the fixative. They almost look like little dust motes, if } } we can see them at all. Immuno means no osmium, so the problem arises } } about how we find them once they're in the resin (probably LR White), let } } alone position them precisely enough to do long sections. } } Ideas we're considering include: 1) tying the fibers along a tiny } } section of dark thread so we can at least see the thread; 2) staining } } them with an LM stain, such as toluidine blue, before embedding them; and } } 3) settling them down onto poly-l-lysine coated Thermonox cover slips to } } stabilize them, then scribing the coverslips and/or using an LM } } stain. We are trying to avoid pre-embedding labeling at the client's } } request, but that will be an option if all else fails. Then we can } } osmicate following the labeling process. } } Questions: Has anyone ever tried adhering muscle fibers to poly } } lysine coverslips? If so, is the attachment stable enough to get through } } the processing/embedding process? Is the use of an LM stain practical, or } } does this introduce a lot of contamination at the EM level? Does } } toluidine blue penetrate past the fiber membrane? (I've never looked at } } LM stained materials in a TEM before.) We're trying to get an idea of } } what might work before we start, since there is a substantial amount of } } time-consuming dissection, etc., involved. Thanks much for any ideas and } } thoughts. } } Randy Tindall } } EM Specialist } } Electron Microscopy Core---We're the Fun Core! } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } }
} Email: dha6n-at-virginia.edu } Name: Dalaver Anjum } } Organization: University of Virginia } } Education: Graduate College } } Location: charlottesville, VA 22904 } } Question: I'm interested to know the lattest software packages for } nano-diffraction in TEM particularly for LACBED/CBED simulations. Will you } please provide me some names of these? Thank you very much.
I favor the plane-wave multislice implementation by Earl Kirkland. In the frozen phonon approximation, it is arguably the most accurate algorithm (see D. A. Muller et al Ultramicroscopy 86, 371 (2001)). Best of all, you get the entire source code and executables for Mac and Windows for the price of Kirkland's book! The book is "Advanced Computing in Electron Microscopy", by Earl J. Kirkland, Plenum 1998, ISBN 0-306-45936-1. It contains a concise summary of electron scattering and image formation, an extensive treatment of the plane-wave multislice image simulation method, and advice for doing accurate simulations with examples.
This package has one drawback for some users: the user-interface is command-line only. There is no slick GUI and no graphical help constructing atomic models.
Good luck! Paul Voyles
Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
I would like to hear from anyone with experience in using digital imaging plates in TEM and how you think this compares to using digital camera systems. Thanks.
Tom Bargar Electron Microscopy Core Research Facility 986395 Nebraska Medical Center Omaha, NE 68198-6395
The 8th Annual RMC Materials Microtomy Course & Workshop is hosted by Boeckeler Instruments in Tucson, Arizona from April 29 - May 2, 2003. Designed specifically for materials scientists needing exposure to advances in specimen preparation for electron microscopy, this is a "hands-on" course catering to all levels of experience. Full details can be seen at www.rmcproducts.com
I have a question for those doing live fluorescent cell imaging in a controlled environment over a period of many hours:
We have a plexiglass incubator installed around the stage of a Nikon inverted microscope, setup for live cell imaging and are looking for a device to both regulate and measure the amount of CO2 inside the chamber. Does anyone know of such a product? Phenol red in culture media can interfere with optical quality, yet we have to maintain a stable pH in the solution. I prefer not to have a probe directly in the dish because often the dish will be closed, therefore, measuring the atmosphere is more practical.
Also, thinking about which gas to use, I thought of a pure CO2 tank but someone said it may make the environment hypoxic if the exhaust is too close to the dish. If the end of the tube (CO2 is bubbled through water) is too distant from the dish, the gas can escape through numerous gaps in the setup and we'll be going through tanks on a daily basis. Would a 5% CO2/air mixture be better?
Thanks for any help Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Queen 30 Bond St. Toronto, ON M5B 1W8 Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
Dear Tom It was huge discussion on ListSerrver about this issue a year or so ago. You may probably find that discussion in the archive. My personal opinion, which I explained in past discussion and may explain again is briefly that image plates are superior in terms of linearity and sensitivity (not necessary better than modern digital cameras). You may load them in the standard film holder and share between the instruments (impossible for digital cameras). The downside of the plates: you need to read them relatively quick after exposure. Scanning will take about 2 min per plate and you have to upload plates from the film holders and load into the scanner and then load them back to film-holder for re-using. So, it's a lot of technical work loading-uploading-scanning etc. I am not sure but it seems to me you have to load-upload in the dark (may be not). In general, image plates may deliver more pixels than moderate digital cameras.
Digital cameras are attached to the instrument, you could not share them. From another hand it always ready: you don't need to load-upload-scan etc as it happening with image plates (if somebody before you uses all plates in the scope for instance). Most moderns digital cameras gives you the chance to keep your film in the scope as well, so you may use film or digital camera without any changes and camera chamber ventilation/vacuuming. The down side of most digital cameras that they do provide less pixels than image plate (not all of them, top models has similar amount of pixels like Ultrascan 4000 from Gatan). Another advantage which appeared to me only when I start using the digital camera on my own is that vacuum in the scope becomes better (there is no frequent ventilation/vacuuming of the camera chamber). Another beauty of the digital cameras (yes, I am voting for digital cameras) is that you have immediate access to the image - you could take the picture and immediately sent it to the printer or to the collaborator. Moreover, you may use Internet and create "video-conference" when people on the opposite side of globe will see exact the same on their screens that you see with digital camera in your microscope room. Personally, I do find that ability to see "live" image on the screen has a great educational potential. It's much easier to teach students how to focus using live image on the screen. I also use that "video-concerning" feature to work with my collaborators on frequent base. Actually, the image from digital camera permanently transmitted to our local network and everyone could see what happening in the microscope.
Personally, I am happy owner of the Gatan's BioScan 600W top mount camera. It's great camera for biological applications. I don't have any commercial interest in Gatan company, just very satisfied user.
Feel free to contact me if you have some further questions. And the very last: do not make final decision unless you will see the stuff in work. Ask for demo, took pictures and compare side-by-side. If company refuse to make demo to you - it looks suspicious to me, I would avoid such company. You may also ask company for references and how many instruments is working in your area. Another things to consider is quality of customer service and avliability (?spell) of it (how many engineers perform service in your area). Godd look.
Sergey
At 08:39 AM 2/25/2003, 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
Has anyone tried compiling the source code from Kirkland's book under Linux? The book says it is standard ANSI C, but I get a number of errors under Caldera OpenLinux Workstation 3.1.1 with gcc 2.95 using the -ansi switch. Primarily these refer to missing trigonometric functions. Reading the documentation for gcc did not help but a look at the include directories revealed a file "tgmath.h" which has the trig-math declarations. Adding #include {tgmath.h} to the source code removed some of the compiler errors, but I still get number of errors relating to sqrtl and other functions esp. from slicelib.c.
If someone has already solved this problem I would like to know the compiler options and changes required to the source code. I do understand that my Linux and gcc are a bit dated. Has it worked directly under a recent version like SuSE Linux 8.1 with gcc 3.0, which is what I am planning to upgrade to?
Thanks for any responses, Divakar
---- Dr R Divakar Physical Metallurgy Section MCG-IGCAR, Kalpakkam 603102, India ----
-----Original Message----- } From: Paul Voyles [SMTP:voyles-at-engr.wisc.edu] Sent: Wednesday, February 26, 2003 10:14 AM To: Microscopy-at-sparc5.microscopy.com Cc: dha6n-at-virginia.edu
} Email: dha6n-at-virginia.edu } Name: Dalaver Anjum } } Organization: University of Virginia } } Education: Graduate College } } Location: charlottesville, VA 22904 } } Question: I'm interested to know the lattest software packages for } nano-diffraction in TEM particularly for LACBED/CBED simulations. Will you } please provide me some names of these? Thank you very much.
I favor the plane-wave multislice implementation by Earl Kirkland. In the frozen phonon approximation, it is arguably the most accurate algorithm (see D. A. Muller et al Ultramicroscopy 86, 371 (2001)). Best of all, you get the entire source code and executables for Mac and Windows for the price of Kirkland's book! The book is "Advanced Computing in Electron Microscopy", by Earl J. Kirkland, Plenum 1998, ISBN 0-306-45936-1. It contains a concise summary of electron scattering and image formation, an extensive treatment of the plane-wave multislice image simulation method, and advice for doing accurate simulations with examples.
This package has one drawback for some users: the user-interface is command-line only. There is no slick GUI and no graphical help constructing atomic models.
Good luck! Paul Voyles
Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
Hello, Has anyone been able to calibrate an ultrasonic bath successfully as described in the ASTM Method D5755-02? If so, what brand of sonicator are you using? Pronda Few
We are beginning to work with samples which contain asbestos. We would like to analyze them by SEM-EDS, but we need help about which criteria to go on in this type of analysis, morphology of the fibers, composition, ...
Can someone help us? Thank you very much in advance Belen -- ************************************************************* Dra. Marìa Belèn Lopez Mosquera Unidade de Microscopìa Servicios Xerais de Apoio á Investigación Universidade da Coruña Edificio Anexo Facultade de Ciencias Campus da Zapateira s/n E-15071 A Coruña
I have been involved in long term multiple time-lapes taking more than 48 hours. For most cell types we used carbogen gas (5% carbon dioxide and 95% oxygen) which we lead into our incubator on the microscope. This 5% carbon dioxide and 95% oxygen mixture is readily available. This way we could keep the cells going in 96 well plates and make several movies in parallel (multiple positions per plate) over the weekend.
================================================= Judy Trogadis wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, Microscopists: } } I have a question for those doing live fluorescent cell imaging in a controlled environment over a period of many hours: } } We have a plexiglass incubator installed around the stage of a Nikon inverted microscope, setup for live cell imaging and are looking for a device to both regulate and measure the amount of CO2 inside the chamber. Does anyone know of such a product? Phenol red in culture media can interfere with optical quality, yet we have to maintain a stable pH in the solution. I prefer not to have a probe directly in the dish because often the dish will be closed, therefore, measuring the atmosphere is more practical. } } Also, thinking about which gas to use, I thought of a pure CO2 tank but someone said it may make the environment hypoxic if the exhaust is too close to the dish. If the end of the tube (CO2 is bubbled through water) is too distant from the dish, the gas can escape through numerous gaps in the setup and we'll be going through tanks on a daily basis. Would a 5% CO2/air mixture be better? } } Thanks for any help } Judy } } Judy Trogadis } Bio-Imaging Coordinator } St. Michael's Hospital, 8Queen } 30 Bond St. } Toronto, ON M5B 1W8 } Canada } ph: 416-864-6060 x6337 } pager: 416-685-9219 } fax: 416-864-6043 } trogadisj-at-smh.toronto.on.ca
Dear Judy, Why not go for live cell chambers available with almost all makes of micrscopes and Confocals, which have a CO2(5%)and temperature controlled stage.
Shashi Singh Scientist CCMB Hyderabad INDIA ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello, Microscopists:
I have a question for those doing live fluorescent cell imaging in a controlled environment over a period of many hours:
We have a plexiglass incubator installed around the stage of a Nikon inverted microscope, setup for live cell imaging and are looking for a device to both regulate and measure the amount of CO2 inside the chamber. Does anyone know of such a product? Phenol red in culture media can interfere with optical quality, yet we have to maintain a stable pH in the solution. I prefer not to have a probe directly in the dish because often the dish will be closed, therefore, measuring the atmosphere is more practical.
Also, thinking about which gas to use, I thought of a pure CO2 tank but someone said it may make the environment hypoxic if the exhaust is too close to the dish. If the end of the tube (CO2 is bubbled through water) is too distant from the dish, the gas can escape through numerous gaps in the setup and we'll be going through tanks on a daily basis. Would a 5% CO2/air mixture be better?
Thanks for any help Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Queen 30 Bond St. Toronto, ON M5B 1W8 Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
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I know that this is somewhat different topic, but I haven't have any idea about this, and I hope you to give me some information.
Spectrometric peaks are often overlapped due to a relatively lower resolution, no matter if the data is from XEDS, XPS, Raman, or XRD. They need to deconvolute or separate the overlapped peaks into individual ones for their precise analysis, and I am one of them.
The question is if you know any software, whether it is commercial, free, share, or not, which could deconvolute or separate the overlapped peaks? What I want to know is the exact energy, intensity, and width of individual peak after deconvolution. I know that professional (expensive) softwares supplied with the spectrometers could handle that, but I want to analyze the data with my computer in my office, after finishing collecting data and getting back to my desk from the instrument analysis center. At the present moment, I am interested in the data from XPS and Raman Spectrometry results.
I was told that Origin graphic software had a function of deconvolution, but I found that the meaning of deconvolution in Origin was quite different from the one used here.
Any comment would be appreciated.
Jondo Yun Division of Advanced Materials Engineering, Kyungnam University Masan, Korea
Depending on the amount and size of the fibers you can use polarized light microscopy or analytical transmission electron microscopy, the SEM is not recommended. This is because you need to measure length, width and aspect ratio in the image, which for small fibers is better resolved in transmission. Also the characteristic hollow tube structure of chrysotile is difficult to see in the SEM. In addition to chemistry that can be obtained in the SEM you need to determine the crystal structure. This can be derived from the light optical properties or from selected area electron diffraction. Morphology and mineralogy are the only two pieces of information required. You need to find out the local regulations for asbestos to decide if your samples need to be regulated. The asbestos literature is huge. I suggest you start with a search on google and read as much as possible. This will give you links to EPA and OSHA regulations in the USA that you can use as a starter.
Dr. Gordon Nord Senior Scientist Environmental Sciences Laboratory Brooklyn College Brooklyn NY 11210
On Wednesday, February 26, 2003, at 04:23 AM, Mª Belén López Mosquera wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 everybody, } } We are beginning to work with samples which contain asbestos. We would } like to analyze them by SEM-EDS, but we need help about which criteria } to go on in this type of analysis, morphology of the fibers, } composition, ... } } Can someone help us? } Thank you very much in advance } Belen } -- } ************************************************************* } Dra. Marìa Belèn Lopez Mosquera } Unidade de Microscopìa } Servicios Xerais de Apoio á Investigación } Universidade da Coruña } Edificio Anexo Facultade de Ciencias } Campus da Zapateira s/n } E-15071 A Coruña } } Teléfono: 34 981 167 000 ext.: 2087 } Telecopia: 34 981 167 068 } Correo eléctrónico: sxaimic-at-udc.es } ************************************************************** } }
I have a currator buddy that has a specimen (frog) that has dried out and he wants to attempt to re-hydrate the specimen for gross morphological examination.
Any sugggestions?
Tim Quinn University of Kansas Ornithology Dept. Histology Lab Director and Program Assistant Natural History Museum and Biodiversity Research Center Dyche Hall Room 414 Lawrence, KS 6604-2454 785-864-4556/785-331-4107 tquinn-at-ku.edu
I NEED TO DRY BACTERY BY SUBLIMATION USING HEXAMETHYLDISILAZANE (HMDS), SOMEONE, COULD HELP ME, BECAUSE ITS THE FIRST TIME I USE IT. SOMEONE COULD TELL ME HOW TO USE HMDS.
Unexposed millipore filters being run in the SEM/EDX as blanks often show particles on them that shouldn't be there, which worries me about potential contamination of particulate samples on filters that have been exposed for investigation. Gently blasting the unexposed filters with canned air helps a lot but not necessarily completely, and in any case would not be advisable for the particulate samples after coating with C or Au/Pd. Can anyone comment or advise?
Many thanks, Dee Breger
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging
Mail to: Geology, CASI West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383
For help and information only, The CASI houses: An FEI Quanta 400 and Technai 12T, Oxford INCA Energy 400, Tousimis AutoSamdri 815 and Olympus FV-300.
-----Original Message----- } From: Mª Belén López Mosquera [mailto:sxaimic-at-udc.es] Sent: Wednesday, February 26, 2003 4:23 AM To: MicroscopyListserver
Hello everybody,
We are beginning to work with samples which contain asbestos. We would like to analyze them by SEM-EDS, but we need help about which criteria to go on in this type of analysis, morphology of the fibers, composition, ...
Can someone help us? Thank you very much in advance Belen -- ************************************************************* Dra. Marìa Belèn Lopez Mosquera Unidade de Microscopìa Servicios Xerais de Apoio á Investigación Universidade da Coruña Edificio Anexo Facultade de Ciencias Campus da Zapateira s/n E-15071 A Coruña
At 10:47 AM 2/26/2003 +0530, you wrote: } Has anyone tried compiling the source code from Kirkland's book under } Linux? The book says it is standard ANSI C, but I get a number of errors } under Caldera OpenLinux Workstation 3.1.1 with gcc 2.95 using the -ansi } switch. Primarily these refer to missing trigonometric functions.
I haven't compiled under Linux, but I have compiled with gcc 3.2 under Cygwin on Win2k and on SunOS 5.8 with gcc 3.1.1. Including {math.h} was sufficient to compile to object code on both. On the Sun, it was necessary to explicitly link the math library with -lm to generate the executable.
If that's not clear, contact me directly - I think we're about the dive off into computerese of limited interest to the rest of the list. :)
Best wishes, Paul
Paul Voyles Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
At 10:03 AM 02/26/2003 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can you email me the source code directly? I can try compiling it here and see what errors pop up.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Wed, 26 Feb 2003, Divakar R wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Has anyone tried compiling the source code from Kirkland's book under Linux? The book says it is standard ANSI C, but I get a number of errors under Caldera OpenLinux Workstation 3.1.1 with gcc 2.95 using the -ansi switch. Primarily these refer to missing trigonometric functions. Reading the documentation for gcc did not help but a look at the include directories revealed a file "tgmath.h" which has the trig-math declarations. Adding #include {tgmath.h} to the source code removed some of the compiler errors, but I still get number of errors relating to sqrtl and other functions esp. from slicelib.c. } } If someone has already solved this problem I would like to know the compiler options and changes required to the source code. I do understand that my Linux and gcc are a bit dated. Has it worked directly under a recent version like SuSE Linux 8.1 with gcc 3.0, which is what I am planning to upgrade to? } } Thanks for any responses, } Divakar } } ---- } Dr R Divakar } Physical Metallurgy Section } MCG-IGCAR, Kalpakkam 603102, India } ---- } } } -----Original Message----- } } From: Paul Voyles [SMTP:voyles-at-engr.wisc.edu] } Sent: Wednesday, February 26, 2003 10:14 AM } To: Microscopy-at-sparc5.microscopy.com } Cc: dha6n-at-virginia.edu } Subject: Re: Ask-A-Microscopist:Diffraction Software for TEM? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Email: dha6n-at-virginia.edu } } Name: Dalaver Anjum } } } } Organization: University of Virginia } } } } Education: Graduate College } } } } Location: charlottesville, VA 22904 } } } } Question: I'm interested to know the lattest software packages for } } nano-diffraction in TEM particularly for LACBED/CBED simulations. Will you } } please provide me some names of these? Thank you very much. } } I favor the plane-wave multislice implementation by Earl Kirkland. In the } frozen phonon approximation, it is arguably the most accurate algorithm } (see D. A. Muller et al Ultramicroscopy 86, 371 (2001)). Best of all, you } get the entire source code and executables for Mac and Windows for the } price of Kirkland's book! The book is "Advanced Computing in Electron } Microscopy", by Earl J. Kirkland, Plenum 1998, ISBN 0-306-45936-1. It } contains a concise summary of electron scattering and image formation, an } extensive treatment of the plane-wave multislice image simulation method, } and advice for doing accurate simulations with examples. } } This package has one drawback for some users: the user-interface is } command-line only. There is no slick GUI and no graphical help } constructing atomic models. } } } } Good luck! } Paul Voyles } } Assistant Professor } Materials Science and Engineering Department } University of Wisconsin - Madison } 1509 University Ave. } Madison, WI 53706-1595 } Voice: (608) 265-6740 } Fax: (608) 262-8353 } voyles-at-engr.wisc.edu } www.engr.wisc.edu/mse/faculty/voyles_paul.html } } } } }
Sandison, A.T. 1955. "The histological examination of mummified material". Stain Technology 30:277-283.
He rehydrated with:
95% ethanol - 30 parts 1% formalin - 50 parts 5% aqueous sodium carbonate - 20 parts
Good luck!
Geoff
"Quinn, Tim Lee" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listies, } } I have a currator buddy that has a specimen (frog) that has dried out and he } wants to attempt to re-hydrate the specimen for gross morphological } examination. } } Any sugggestions? } } Tim Quinn } University of Kansas } Ornithology Dept. } Histology Lab Director and Program Assistant } Natural History Museum and Biodiversity Research Center } Dyche Hall Room 414 } Lawrence, KS 6604-2454 } 785-864-4556/785-331-4107 } tquinn-at-ku.edu } }
-- ********************************************** 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 **********************************************
Emory Neurology Microscopy Core is hosting a week long workshop on Cryo-techniques and immunogold labeling from May 4 through May 9, 2003 in Atlanta, Georgia, USA. The workshop curriculum will include the latest advances in cryo-preparation including cryo-fixation and substitution of biological samples, cryo-ultramicrotomy, and pre- and post-embedding immunogold labeling. Internationally known experts Dr. Kent McDonald (EM Laboratory at the University of California, Berkeley), Dr. Jan Leunissen (Aurion Immunogold Reagents & Accessories) and Mr. Helmut Gnaegi (Diatome) will be the instructors for the workshop. The workshop will include lectures, hands-on training, round table discussions, and presentations on applications. Also, participants in the workshop will be able to work on their own samples. The industrial sponsors for the workshop are Leica Microsystems Inc., Aurion, EMS, and Diatome U.S.
The number of participants in the workshop is limited. The registration deadline is March 31, 2003. If you are interested in attending or need more information about the workshop, please contact the workshop technical coordinator Hong Yi by phone (404-712-8491) or email (hyi-at-emory.edu), or log on www.em-preparation.com, or you may contact the workshop message center at 1-800-248-0665 x 7208.
Thank you all in advance and looking forward to seeing you in Atlanta.
Hong ====================== Hong Yi Emory Neurology Microscopy Core Emory University School of Medicine 6215 Woodruff Memorial Research Building 1639 Pierce Drive Atlanta, GA 30322
I agree with Gordon, especially using polarized light. It is quicker, easier, and in most cases, part of the accepted standard protocol. Chrysotile, for example, exhibits a characteristic "dirty grey" color between crossed polars.
Polarized light can be daunting the first time through, largely because of the complex vocabulary. I've included a simplified but solid introduction to Pol in "Optimizing Light Microscopy" (see the website below for details). Also, I've just finished putting together the workbook for the American Chemical Society short course and can send you a copy of that chapter, if you decide to proceed. Of course, THE definitive authority is the recently deceased Walter McCrone. The Institute has full courses on asbestos analysis. For accessories and written material, you can contact their sales organization: McCrone Microscopes & Accessories in Westmont, IL. Web: www.mccrone.com E-mail address info-at-mccrone.com P: 630-887-7100 F: 630-887-7764.
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 09:33 AM 2/26/03 -0500, Gordon Nord wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As a semi-retired metallurgist I need to make photomicrographs and then publish them digitally with a word processor. I have been using Polaroid film and then digitizing the images on a flatbed scanner - actually three scanners so far - but I'd like to shortcircuit the process and take the digital photomicrographs directly.
I have studied the 2002 archives regarding digital cameras and have in hand a zoom/c-mount interface for a microscope camera for my company's Olympus SZH stereomicroscope, but I'd like also to make digital photomicrographs on a metallograph, where the image quality is a sensitive function of the objective/eyepiece/tube length, which has also been discussed.
It came to mind that I might retrieve the functionality of one of the retired flatbed scanners by removing the light source and glass bed so that the optical element can be presented directly to the imaging area of the metallograph - which is five by seven inches, thereby maximizing the number of pixels by moving the camera to the appropriate position along the optical "bench."
Several questions come to mind ...
1. Will the optical element "see" the light from the projector lens (i.e., the recording eyepiece) if it is coming almost straight on to the original "film" plane ?
2. Given that the answer to (1) is yes, is the light sensitivity of a flatbed scanner good enough to record the image from a metallograph - which might take several seconds to record onto Polaroid type 55 film ?
Thanks for thinking about this.
Best regards, George Langford, Sc.D. Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
Is somebody know a place where we can find parts for a COOLWELL cooling system S-unit, model S-150WCZ. I need the price and part number for a Temperature Board (Loadslave temperature modulator). A complete adress with fax number and email adress would be appreciate. Thank you.
Robert Alain Microscopie lectronique INRS-Instirut Armand-Frappier 531 blvd Des Prairies Laval, Qc, Canada H7V 1B7 Tl: 450-687-5010 poste 4388 Fax: 450-686-5626
-- Hello! I am very happy, because we have a lot of information about analysis of asbestos. Although I know that is a difficult work, I think that this is a very big beginning. Thanks for your information, I have to study and work very much about this. Thanks again. Sincerely, BELEN ************************************************************* Dra. Marìa Belèn Lopez Mosquera Unidade de Microscopìa Servicios Xerais de Apoio á Investigación Universidade da Coruña Edificio Anexo Facultade de Ciencias Campus da Zapateira s/n E-15071 A Coruña
The SEMVision is a defect review tool manufactured by Applied Materials. I acquired an ownership interest in one of these, and seek support to qualify and install my tool. The current version of this tool is described on the Applied Materials web site as follows:
"... an automatic defect review SEM is designed for inline process monitoring and real-time yield enhancement for the sub-100nm generation. Continuing the productivity leadership of the SEMVision product line, the G2 delivers high throughput and automated calibrations offering the most advanced capabilities for production and engineering applications. Field proven applications of HAR (High Aspect Ratio) imaging and Automatic Process Inspection for detection of physical and electrical defects provide customers the ability to control their defects and processes with greater success, leading to higher yields and faster time to market."
On June 26th, 2002, Applied Materials reported that they had shipped over 200 systems to customers worldwide. I believe that this makes the number of tools in the field sufficient to merit third party support.
If you are interested in providing 3rd party support for the Applied Materials SEMVision tool (or know of someone capable) please contact John Edward Moore at telephone 919-434-8457.
Hi Valad: If you are referring to ISI PS-2 (international scientific instruments which became TOPCON) sputter coater it was made by Poalron and named E5000. You can find a manual for this on the internet : http://www.polaron-range.com/Tech%20Support/oldproducts.htm I have had our unit for 20 years and it still works. It doesn't have a cool source so on heat sensitive samples you may run into trouble. I can coat some heat sensitive samples by using low voltage and several short coating times. Call or e-mail if you have any questions. Terry Ellis Hallmark Cards Inc. (yes we do have an scanning electron microscope, think inks, paper, printing plate making, and industrial hygiene concerns) 816-545-6573 e-mail: tellis2-at-hallmark.com
Listers, We have a sample substrate with mammalian cells growing on it that cannot go through ETOH . ETOH is the only transition fluid we use in our particular critical point dryer. I would like to use another drying method such as HMDS but did not know if you can use this compound with other dehydrants.
We have the option of cryo-SEM but it is always more convenient for everyone if we can find a method to dry samples and then image them with more flexibility in our scheduling. Any and all suggestions welcomed.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
To the listserver, I am looking for references on the effects of osmium staining on postembedded immunolabeling, ie reducing the amount and size of gold label on LR Gold/Whites sections. Also has anyone used very low osmium (0.05%?) successfully for IEM?
We have a 1996 model JEOL 6400 and we are looking to upgrade to a stage automation package and looking for information on other people's experiences.
It doesn't have to be real fancy, just something to allow joystick control of the stage (X, Y, rotate) and, if possible, the ability to link to our EDAX Phoenix system (2001 vintage) which is set up to work with stage automation.
Of course, the cheapest model w/ the best performance is preferred. I'm interested in third party versions as well as experiences w/ the JEOL factory version.
Thanks,
John Giles Principal Materials Engineer Honeywell Defense & Space Electronic Systems - Clearwater
Digital Image Capture and Management in Microscopy
May 8, 2003
A course on Digital imaging in light microscopy which will cover the following topics:
Optical Limitations in Light Microscopy...Photographic Imaging Strategies... Digital Imaging Strategies...Selection of Digital Capture (Camera vs. Scanner)...
Image Processing of Captured Images...Image File Formats...Printing Images... Color Management Systems...Database Management Software...Presentation Software for Oral Reports...Website Performance...Integration of Image Data with Sample Information, Calibration, Other Data & Reports...Acrobat and html Software for Written Reports and Archives...Examples of Efficient, Low Cost Image Handling Systems...
Examples of Electronic Microscopy Reports and Databases
The course instructors are Mary and John McCann of McCann Imaging.
WHEN: Thursday, May 8, 2003, from 9 A.M. to 5 P.M.
An advanced course on polarized light microscopy which will cover the following topics:
The nature of polarized light
The origin and interpretation of interference colors
Birefringence and crystal orientation
The Indicatrix
Compensation and variable compensators
Interference figures and their interpretation
The workshop will consist of four Consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of polarized light microscopy. The course instructors include Jan Hinsch of Leica, Inc., Mary McCann of McCann Imaging, John Reffner of SensIR and N.Y.M.S. Instructor Don O'Leary.
WHEN: April 26, May 3, 10 &17, 2003, from 10 A.M. to 4 P.M.
COST: $340 for N.Y.M.S. members, $370 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: advanced course for those who have completed "The Use of the Microscope" or are experienced in microscopy and familiar with the theory of its use.
HOW: Register using the form below. Limited to the first 12 registrants.
Return form to Don O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
FURTHER INFORMATION: Call D. O'Leary (201) 797 -8849 e-mail donoleary-at-att.net
My thanks to everyone who has sent me advice and information on digital image systems for TEM. I am still having trouble understanding why their is a difference in resolution between side mounted cameras and bottom mounted cameras. I see that bottom mounted cameras have a resolution of up to 6 megapixels while the highest I have seen so far for side mounted cameras is 2 megapixels. If anyone out there would be willing to try and explain this to me I would appreciate this and then I could in turn pass that on to my faculty bosses who would also like to know about the difference.
Tom Bargar Electron Microscopy Core Research Facility 986395 Nebraska Medical Center Omaha, NE 68198-6395
George; Check out this website describing building a digital camera from a $100 flatbed scanner! http://www.sentex.net/~mwandel/tech/scanner.html John Mardinly Phone: 408-765-2346 Pager: 877-277-1182
-----Original Message----- } From: George Langford, Sc.D. [mailto:amenex-at-amenex.com] Sent: Thursday, February 27, 2003 8:14 PM To: Microscopy-at-sparc5.microscopy.com
Hello Microscopists !
As a semi-retired metallurgist I need to make photomicrographs and then publish them digitally with a word processor. I have been using Polaroid film and then digitizing the images on a flatbed scanner - actually three scanners so far - but I'd like to shortcircuit the process and take the digital photomicrographs directly.
I have studied the 2002 archives regarding digital cameras and have in hand a zoom/c-mount interface for a microscope camera for my company's Olympus SZH stereomicroscope, but I'd like also to make digital photomicrographs on a metallograph, where the image quality is a sensitive function of the objective/eyepiece/tube length, which has also been discussed.
It came to mind that I might retrieve the functionality of one of the retired flatbed scanners by removing the light source and glass bed so that the optical element can be presented directly to the imaging area of the metallograph - which is five by seven inches, thereby maximizing the number of pixels by moving the camera to the appropriate position along the optical "bench."
Several questions come to mind ...
1. Will the optical element "see" the light from the projector lens (i.e., the recording eyepiece) if it is coming almost straight on to the original "film" plane ?
2. Given that the answer to (1) is yes, is the light sensitivity of a flatbed scanner good enough to record the image from a metallograph - which might take several seconds to record onto Polaroid type 55 film ?
Thanks for thinking about this.
Best regards, George Langford, Sc.D. Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
George; Another option is the "Better Light" camera back, which can create images with over 100 megapixels, and empty your wallet faster than it fill your hard drive. See: http://www.betterlight.com Of course if you are retired and not on a defense department budget, converting the scanner might be the way to go. John Mardinly Intel
-----Original Message----- } From: George Langford, Sc.D. [mailto:amenex-at-amenex.com] Sent: Thursday, February 27, 2003 8:14 PM To: Microscopy-at-sparc5.microscopy.com
Hello Microscopists !
As a semi-retired metallurgist I need to make photomicrographs and then publish them digitally with a word processor. I have been using Polaroid film and then digitizing the images on a flatbed scanner - actually three scanners so far - but I'd like to shortcircuit the process and take the digital photomicrographs directly.
I have studied the 2002 archives regarding digital cameras and have in hand a zoom/c-mount interface for a microscope camera for my company's Olympus SZH stereomicroscope, but I'd like also to make digital photomicrographs on a metallograph, where the image quality is a sensitive function of the objective/eyepiece/tube length, which has also been discussed.
It came to mind that I might retrieve the functionality of one of the retired flatbed scanners by removing the light source and glass bed so that the optical element can be presented directly to the imaging area of the metallograph - which is five by seven inches, thereby maximizing the number of pixels by moving the camera to the appropriate position along the optical "bench."
Several questions come to mind ...
1. Will the optical element "see" the light from the projector lens (i.e., the recording eyepiece) if it is coming almost straight on to the original "film" plane ?
2. Given that the answer to (1) is yes, is the light sensitivity of a flatbed scanner good enough to record the image from a metallograph - which might take several seconds to record onto Polaroid type 55 film ?
Thanks for thinking about this.
Best regards, George Langford, Sc.D. Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
I tried this a couple years ago by projecting a negative (in a photographic enlarger) onto a flatbed scanner. Initially, all I got was an image of the projector bulb. I believe that this is possible if you have a "rear-projection screen" onto which you can project the image.
You would need something like a thin layer of white plastic (that has no structure and is uniform). I set my scanner in the transparency mode and tried different materials. I found that a white garbage bag gave the best images. But they were still unacceptable due to unevenness of the plastic film. I am certain that if one could find the right material that this is doable.
You would need the following:
1. A scanner that can be set to the transparency mode without requiring that the light source (top plate in most scanners) be activated. In some scanners this can be accomplished by simply moving the top plate off of the scan bed but leaving it plugged in (to fool the software).
2. A white, light permeable, rear-projection screen onto which you could project and focus the image.
I belive that if these criteria are met, one would have a kick-ass device to digitize enlarged negatives. With simple modifications to a $400, 300 ppi scanner you would have a device that would outperform a $10-30,000 high end scanner.
JB
} As a semi-retired metallurgist I need to make photomicrographs } and then publish them digitally with a word processor. I have } been using Polaroid film and then digitizing the images on a } flatbed scanner - actually three scanners so far - but I'd like } to shortcircuit the process and take the digital photomicrographs } directly. } } I have studied the 2002 archives regarding digital cameras and } have in hand a zoom/c-mount interface for a microscope camera for } my company's Olympus SZH stereomicroscope, but I'd like also to } make digital photomicrographs on a metallograph, where the image } quality is a sensitive function of the objective/eyepiece/tube } length, which has also been discussed. } } It came to mind that I might retrieve the functionality of one } of the retired flatbed scanners by removing the light source and } glass bed so that the optical element can be presented directly } to the imaging area of the metallograph - which is five by seven } inches, thereby maximizing the number of pixels by moving the } camera to the appropriate position along the optical "bench." } } Several questions come to mind ... } } 1. Will the optical element "see" the light from the projector } lens (i.e., the recording eyepiece) if it is coming almost } straight on to the original "film" plane ? } } 2. Given that the answer to (1) is yes, is the light sensitivity } of a flatbed scanner good enough to record the image from a } metallograph - which might take several seconds to record onto } Polaroid type 55 film ? } } Thanks for thinking about this. } } Best regards, } George Langford, Sc.D. } Amenex Associates, Inc. } amenex-at-amenex.com } http://www.amenex.com/
############################################################## 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 ##############################################################
I'm sure there are references out there, but all I can offer is personal experience and word-of-mouth. I've used 1 (that I can think of off the bat) antibody that gave "normal" staining on sections of tissue with regular osmication, and some that were OK with reduced osmium (osmium mixed with potassium ferrocyanide or ferricyanide - you'll get reams of info on which to use!), and enough that were useless after any osmium to make you cry. Osmium can also mess up your resin polymerization, so you have to be careful - especially if you want to UV cure - dark tissue is a major pain.
What do you mean by osmium reducing the size of gold label? You said you were doing postembedding immuno....I haven't heard of any gold size problems for postembedding (lots with pre-embedding and osmium) - how would that happen?
Do you have some potentially unrescuable blocks that have already been osmicated? You might try a gentle peroxide treatment on the sections, or some EM version of antigen retrieval....
Good luck!
Tamara
On Fri, 28 Feb 2003, Mike Delannoy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To the listserver, } I am looking for references on the effects of osmium staining } on postembedded immunolabeling, ie reducing the amount and size } of gold label on LR Gold/Whites sections. Also has anyone } used very low osmium (0.05%?) successfully for IEM? } } Thank You } Mike D } } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
"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 } -----------------------------------------------------------------------. } } } Hi George, } } I tried this a couple years ago by projecting a negative (in a } photographic enlarger) onto a flatbed scanner. Initially, all I got } was an image of the projector bulb. I believe that this is possible } if you have a "rear-projection screen" onto which you can project the } image. } } You would need something like a thin layer of white plastic (that has } no structure and is uniform). I set my scanner in the transparency } mode and tried different materials. I found that a white garbage bag } gave the best images. But they were still unacceptable due to } unevenness of the plastic film. I am certain that if one could find } the right material that this is doable. } } You would need the following: } } 1. A scanner that can be set to the transparency mode without } requiring that the light source (top plate in most scanners) be } activated. In some scanners this can be accomplished by simply moving } the top plate off of the scan bed but leaving it plugged in (to fool } the software). } } 2. A white, light permeable, rear-projection screen onto which you } could project and focus the image.
Why can't you put the white plastic (or any ol' diffuser, actually) on the sensor? Focus to the depth of the sensor surface. Should keep artifacts from material inconsistencies to a minimum and limit them to horizontal bands, which could then be corrected in processing...? Just my 2 cents, I probably forgot something somewhere.
} I'm acquired an old sputter coater PS-2 and looking for the } manual. Please } let me know if you have one. Thank you in advance.
The ISI PS2 sputter coater is equivalent to a Polaron E5000. Mine (PS2) even has a Polaron sticker on it. I have a Polaron E5000 manual. What do you need to know?
Bruce Girrell Microline Technology Corp. 2397 Traversefield Dr. Traverse City, MI 49686 http://www.microlinetc.com
} There has been some more discussion about imaging plate technology for TEM } and in order to help set the record straight the following is posted with } the permission of the Sysop....
There have been some speculation and misconceptions about the Ditabis imaging plate system...
} When the image storage plate technology, a technology that is widely and } well accepted in other sensitivity critical analytical applications like } radioisotope mapping, was first applied to transmission electron } microscopy it met with only moderate success. This was due in part to } its resolution ( less than film - although even then much higher than } the CCD cameras of the time) and its high cost. Ditabis has addressed } these two shortcomings and has expanded the performance envelop as well . } } In papers published by Dr. Rasmus Schroder of Max-Planck-Institute, the } plates have been used for exposures of 0.1e-/um2 and ongoing experiments } with researchers at the Wadsworth Center in New York are exploring even } lower dosages. The dynamic range of the plates is now 20 bits - orders of } magnitude beyond commercially available CCD cameras for TEM use. The } increased sensitivity means lower exposure damage (cryo) and the extended } dynamic range mean that information that would otherwise be lost is now } available (diffraction - see, Dr. W.-D. Rau, IHP Frankfurt/Oder). } } } Although the images in the plates are certainly not permanent. under } normal exposure setting they are usable for several days. Indeed, we } demonstrate the system by sending plates to prospective customers which } they then expose and send back to us to scan. Except under the very } lowest exposure levels this has worked quite well. We encourage people } to simultaneously shoot film or CCD images of the same fields just for } comparison. } } } The plates are treated much like film - they go into the same film holders } but can be loaded in the light and need to be unloaded in a dim } environment. The same plates can be used in multiple instruments and } upwards of 1000 time before having to be replaced. With 15um pixels and a } field of view the same as film the images can be up to 6000 x 5333 pixels } - almost exactly twice the size of the largest CCD systems (as for } instance, the 4k x 4k Tietz system which, incidentally, we also sell). } Indeed, the plates do not provide the instant gratification of CCD } system - but they do provide a solution for digital images of the full } film field of view of the TEM as well as providing increased sensitivity } and extended dynamic range over other technologies at a cost that is far } below CCD systems with comparable resolution. Like any technology, the } imaging plates are not the solution to all digital TEM imaging problems } but where they are applicable they do provide a high quality, unique and } affordable solution to digitizing the images from transmission electron } microscopes.
I have had a request for a SEM micrograph of a citrus leaf surface. I don't think it matters if it is an orange, grapefruit, lemon, or a lime - they were not specific. Mag. anywhere in the 500X to 2000X range. If anyone has such a photo and they are willing to share (freebie) or sell please let me know off-list. thanks, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) **********************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
In fact, I believe this is how certain, high-end scanning cameras, such as the Leaf Camera, actually work. In my situation, the main stumbling block was the software that would search for the upper light source (in the lid). When I removed the lid, the program would crash or refuse to scan when it failed to detect the light source. I believe that with more time and software/hardware savvy, someone could develop a very inexpensive and high resolution system to digitize enlarger-projected images.
JB
} } } } } Hi George, } } } } I tried this a couple years ago by projecting a negative (in a } } photographic enlarger) onto a flatbed scanner. Initially, all I got } } was an image of the projector bulb. I believe that this is possible } } if you have a "rear-projection screen" onto which you can project the } } image. } } } } You would need something like a thin layer of white plastic (that has } } no structure and is uniform). I set my scanner in the transparency } } mode and tried different materials. I found that a white garbage bag } } gave the best images. But they were still unacceptable due to } } unevenness of the plastic film. I am certain that if one could find } } the right material that this is doable. } } } } You would need the following: } } } } 1. A scanner that can be set to the transparency mode without } } requiring that the light source (top plate in most scanners) be } } activated. In some scanners this can be accomplished by simply moving } } the top plate off of the scan bed but leaving it plugged in (to fool } } the software). } } } } 2. A white, light permeable, rear-projection screen onto which you } } could project and focus the image. } } Why can't you put the white plastic (or any ol' diffuser, actually) on } the sensor? Focus to the depth of the sensor surface. Should keep } artifacts from material inconsistencies to a minimum and limit them to } horizontal bands, which could then be corrected in processing...? Just } my 2 cents, I probably forgot something somewhere.
############################################################## 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 ##############################################################
} Listers, } We have a sample substrate with mammalian cells growing on it that } cannot go through ETOH . ETOH is the only transition fluid we use in our } particular critical point dryer. I would like to use another drying method } such as HMDS but did not know if you can use this compound with other } dehydrants. } } We have the option of cryo-SEM but it is always more convenient for } everyone if we can find a method to dry samples and then image them with } more flexibility in our scheduling. } Any and all suggestions welcomed. } } Debby
Hi Debby We use HMDS in the microwave instead of critical point drying and have found no problem with the final picture. We tried the same cells grown in culture on coverslips through the two methods of drying (conventional cpd and microwave) and got the same sort of picture on the SEM. The filapodia and microvilli were all still intact.
We usually go through ethanol to HMDS but there should be no problem going through acetone. We have the protocol for the microwave processing on our website http://www.emlab.ubc.ca under "Protocols -EM Protocols - Microwave - SEM processing - glutaraldehyde fixation"
If you do not have a microwave, then there may be a problem. I understand that the HMDS works in the same way as liquid CO2 in the critical point dryer in that there is so little surface tension that when it evaporates, there is no damage to the cells. We set the microwave to 45 degrees but we got our protocol from Richard Dearree, California State University, Chico. Richard may have tried several options and have more advice. Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility First Vice President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
For XPS data processing, I can from personal experience recommend a package called CasaXPS, written by Neal Fairley. Runs on a PC only. I believe your university can get a software site license from the author for not-too-much money. See http://www.casaxps.com/
There's a spectroscopic data processing application called SpXZeigR (Spec-tzi-ger), written by Jeff Weimer, that I've heard is good. Runs on a Mac or a PC. See http://lmass.uah.edu/software/
For other possiblities, see Roger Nix's software links site at http://www.chem.qmul.ac.uk/surfaces/#software
or the UK Surface Analysis Forum software links site at http://www.uksaf.org/software.html#1
Regards,
Libby Shaw
} Reply-To: "Jondo Yun XXXXXX" {jdyun-at-kyungnam.ac.kr} } From: "Jondo Yun XXXXXX" {jdyun-at-kyungnam.ac.kr} } To: "MicroscopyListserver" {Microscopy-at-sparc5.microscopy.com} } Subject: How to deconvolute peaks? } Date: Wed, 26 Feb 2003 21:46:25 +0900 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} It came to mind that I might retrieve the functionality of one } of the retired flatbed scanners by removing the light source and } glass bed so that the optical element can be presented directly } to the imaging area of the metallograph - which is five by seven } inches, thereby maximizing the number of pixels by moving the } camera to the appropriate position along the optical "bench." } } Several questions come to mind ... } } 1. Will the optical element "see" the light from the projector } lens (i.e., the recording eyepiece) if it is coming almost } straight on to the original "film" plane ?
.. more snippage of moot points ...
Here's a summary of what was contributed by helpful listers, particularly John Mardinly (the first link):
} http://www.sentex.net/~mwandel/tech/scanner.html This leads to a secondary link which is also quite interesting:
http://www.rit.edu/~andpph/text-demo-scanner-cam.html which in turn leads to: http://www.rit.edu/~andpph/text-better-scanner-cam.html
So then I opened up that IBM scanner (with its green cast) to see what the CCD array looks like ... and ... Arghhhh.
Whatever's there sure isn't a monster array - it's a tiny little lens covering what's gotta be a well collimated CCD chip, effectively about 300 millimeters below the platen, making it essentially a slit camera. The collimation is probably achieved by the narrow width of the CCD array, i.e., one pixel wide.
Which explains how a couple of the workers in the links above managed to make images by panning the scanner while its array moved across the bed. Imagine a light source that illuminates a strip of the field of view like a paintbrush ... now, sweep the brush across the field of view.
The problem in applying this mechanism in a metallograph is that the light is coming from essentially a point source, so the rays make different angles to the platen, depending on where the sensor is along its path. Only some of these rays will make it through the collimation scheme to the CCD chip. On the IBM scanner, that would be in the middle of the field; in the Mustek, it would have been off to one side, because the mirrors that fold the light path aren't perpendicular to the platen.
The aperture of this slit camera at the height of the platen is about six or seven millimeters, so some of the imaging area might still "see" the eyepiece lens. Therefore, in order to apply the cannibalized scanner to a metallograph, I might have to tilt the entire scanner during the scan in order that the slit of collimation always pointed to the eyepiece lens of the metallograph.
Plus which, there are the scanner's lens optics - added to the metallograph's optics. My original intent would be stymied by having to add the scanner's lenses to the optical path. Might as well use a digital camera - as several of you confidently suggested to this doubter.
On the other hand ...
The image in a metallograph's camera is in sharp focus for an extremely long distance on either side of the film plane - a hundred millimeters or so in a 500 millimeter bellows extension - so the projected image would be in focus deep inside the optical path of the scanner. Points of bright reflection on the object plane would become rods of light in the image plane, so they would intersect the slit of collimation at different distances from the original platen height, thereby creating some distortion of the magnification along the direction of motion of the scan. Maybe things aren't so bad after all - I could measure the distortion with a stage micrometer. Hmmmmmmm. Where does the Ewald sphere come into play in all this ...
Thanks for participating in this interesting exercise.
Best regards, George Langford, Sc.D. Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
Here is the table of contents for the March/April issue of Microscopy Today.
New Subscriptions via http://www.microscopy-today.com only, please New subscriptions will close on Tuesday 4 March for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting. Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA. MSA members anywhere have free subscriptions. Non-MSA, non North American subscriptions have been reduced from $80 or $110US to $35US. . More on this later.
Table of Contents:
Membrane Leftovers after Fusion of Vacuoles Stephen W. Carmichael, Mayo Clinic Inspecting Surfaces With a Sharp Stick: Scanning Probe Microscopy - Past, Present, and Future Paul West, Pacific Nanotechnology, Inc. The Life and Death of a Tungsten Hairpin Filament Steve Chapman, Protrain Photoshop Tutorials: Selecting ROIs from Brightfield Images Jerry Sedgewick, University of Minnesota TEM Sample Preparation Using Focused Ion Beam - Capabilities And Limits H.J. Engelmann, B. Volkmann, Y. Ritz, H. Saage, H. Stegmann, Q. de Robillard, E. Zschech; AMD Saxony Beyond the Hype - Is 2-Photon Microscopy Right for You? Jason Kirk, University of Connecticut Creating Multimedia Teaching and Training Tools Steven B. Barlow, San Diego State University Another Way to Implement Diffraction Contrast in SEM Xiaodong Tao and Alwyn Eades, Lehigh University Tardigrades and Microscopes William R. Miller, Chestnut Hill College Choosing a Cantilever for In Situ Atomic Force Microscopy John T. Woodward, National Institute of Standards and Technology Industry News Petrographic Slides Projected in a 35mm Slide Projector Roy Beavers, Southern Methodist University Embedding Media Health Hazards And Medical Documentation E. Ann Ellis, Texas A & M University How to Produce Posters Using PowerPoint John J. Bozzola, Southern Illinois University
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Lots of pixels mean a larger chip. At the side mount position, the diameter of the electron beam is perhaps 1-2 cm, so with a large chip, you would only be illuminating a part of the chip. So, no point in having a chip with lots of pixels at the side mount position.
There are at least two suppliers of 4k by 4k TEM CCD cameras - that is, 16 megapixels, so you might want to do a bit more research on your options.
I would also be careful in use of the word resolution - lots of pixels does not necessarily mean that you get good resolution in the final image. The key factor is the point spread function. To transfer the electron image to the CCD, there is a scintillator and then a coupling mechanism - either lenses/prism or fibre optic. What needs to be considered is how wide an illuminated area is generated at the CCD by a single electron hitting the scintillator. If you have small pixel elements in the CCD, you end up with a single electron illuminating 4 or 9 pixels - in such a case, there would be no advantage to having lots of pixels. Very large pixel elements will result in electrons at different positions in the image illuminating the same CCD pixel element. So, good TEM CCD camera design requires a careful matching of the CCD pixel element size to the point spread function. Given that this match has been achieved, large pixel elements are good - they store more charge, thus giving you greater dynamic range.
I guess what that boils down to is, don't assume more pixels means a better image. If the camera is well designed, it should but it is not an assumption I would make. You also need to think about read-out time - more pixels take longer to read-out to the computer.
My advice would always be to get images from your specimens and compare the results from different cameras.
Here is my protocol for HMDS drying of bacteria. Have fun!
Dave
Preparation of bacteria for SEM using HMDS
Safety Work in fume hood and use gloves Retain ethanol and HMDS (hexamethyldisilazane) for disposal in non-chlorinated waste bottle
Start with a lot of bacteria eg a 3 x1mm cubed pellet
Fixation
Fix in 4% glutaraldehyde in buffer (usually 0.1M )* Leave for 1 hr at room temperature or 24hrs in the fridge Rinse in buffer x3 (total storage time should exceed fixation time by a factor of 3).
SEM Dehydration of bacteria in an Eppendorf
5m in 20% ethanol 5m in 30% ethanol 5m in 50% ethanol 5m in 70% ethanol 5m in 90% ethanol 5m in 100% ethanol 5m in 100% ethanol
5m in 100% ethanol / HMDS (2:1) 5m in 100% ethanol / HMDS (2:2) 5m in 100% ethanol / HMDS (1:2) 5m in 100% HMDS 5m in 100% HMDS
Centrifuge and resuspend in a 2 drops" of HMDS. Put on drop on a clean (use acetone) small round coverslip on a stub. Take rest of sample dilute by a drop and mount one drop. Do this about 4 times to get a range of dilutions.
Sputter with gold and view in SEM.
*For 4% glutaraldehyde in 0.1M buffer Take, say, 50mls of 0.2M buffer, 34ml d water and 16ml of 25% glutaraldehyde.
On Wed, 26 Feb 2003 10:49:26 -0600 (CST) "ggm-at-servidor.unam.mx"-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } hello } } } I NEED TO DRY BACTERY BY SUBLIMATION USING HEXAMETHYLDISILAZANE } (HMDS), SOMEONE, COULD HELP ME, BECAUSE ITS THE FIRST TIME I USE IT. SOMEONE } COULD TELL ME HOW TO USE HMDS. } } THANKS. } } M.C. GUILLERMINA GLEZ. MANCERA } FAC.QUIMICA-UNAM } MEXICO, D.F. } } ------------------------------------------------- } Obtén tu correo en www.correo.unam.mx } UNAMonos Comunicándonos } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
} My thanks to everyone who has sent me advice and information on digital } image systems for TEM. I am still having trouble understanding why their } is a difference in resolution between side mounted cameras and bottom } mounted cameras. I see that bottom mounted cameras have a resolution of up } to 6 megapixels while the highest I have seen so far for side mounted } cameras is 2 megapixels. If anyone out there would be willing to try and } explain this to me I would appreciate this and then I could in turn pass } that on to my faculty bosses who would also like to know about the } difference. } } Tom Bargar } Electron Microscopy Core Research Facility } 986395 Nebraska Medical Center } Omaha, NE 68198-6395 } } phone 402-559-7347 } } tbargar-at-unmc.edu
Lots of pixels mean a larger chip. At the side mount position, the diameter of the electron beam is perhaps 1-2 cm, so with a large chip, you would only be illuminating a part of the chip. So, no point in having a chip with lots of pixels at the side mount position.
There are at least two suppliers of 4k by 4k TEM CCD cameras - that is, 16 megapixels, so you might want to do a bit more research on your options.
I would also be careful in use of the word resolution - lots of pixels does not necessarily mean that you get good resolution in the final image. The key factor is the point spread function. To transfer the electron image to the CCD, there is a scintillator and then a coupling mechanism - either lenses/prism or fibre optic. What needs to be considered is how wide an illuminated area is generated at the CCD by a single electron hitting the scintillator. If you have small pixel elements in the CCD, you end up with a single electron illuminating 4 or 9 pixels - in such a case, there would be no advantage to having lots of pixels. Very large pixel elements will result in electrons at different positions in the image illuminating the same CCD pixel element. So, good TEM CCD camera design requires a careful matching of the CCD pixel element size to the point spread function. Given that this match has been achieved, large pixel elements are good - they store more charge, thus giving you greater dynamic range.
I guess what that boils down to is, don't assume more pixels means a better image. If the camera is well designed, it should but it is not an assumption I would make. You also need to think about read-out time - more pixels take longer to read-out to the computer.
My advice would always be to get images from your specimens and compare the results from different cameras.
The University of Texas Health Science Center and Hamamatsu Photonics KK will co-host A Special Topics Symposium and Workshop on
Fluorescence Lifetime Imaging and Spectral Imaging
Symposium - Academic and Commercial Presentations June 6-7, 2003 Gunter Hotel Student: $ 200.00 (US) Professional/Corporate: $ 250.00 (US) Registration Deadline May 1, 2003
Workshop - Hands-on Experience with Latest Instrumentation June 8-10, 2003 UT Health Science Cnt. Tuition: $700.00 (US) Includes room and board 20 student limit / 4 scholarships available Application Deadline April 7, 2003
For information and forms visit:
http://usa.hamamatsu.com/flim_spectral/default.htm or http://www.uthscsa.edu/csb/imaging-course.html
The -lm switch solved the problem. The math libraries need to be explicitly linked on the gcc command line. It is also not necessary to include tgmath.h. I have successfully compiled atompot, mulslice and image programs and they run successfully on the sample data files provided with the source code. The commands I used were:
My thanks to Paul Voyles and Dimiter Prodanov for their suggestions. The suggestion from Dimiter Prodanov was to upgrade to gcc 3.0 and not to use the -ansi option. I have not used the -ansi option but upgrading to gcc 3.0 is not required for this particular set of programs, IMHO.
With Best Regards, Divakar
---- Dr R Divakar Physical Metallurgy Section MCG-IGCAR, Kalpakkam 603102, India ----
-----Original Message----- } From: Paul Voyles [SMTP:voyles-at-engr.wisc.edu] Sent: Thursday, February 27, 2003 10:02 AM To: Divakar R (Dr) Cc: Microscopy-at-sparc5.Microscopy.Com
Hi everybody,
does anybody know anything about metrizamide embedding of TEM specimens?
Philip
Philip Koeck Södertörns Högskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em
What I remember from an immunogold workshop given by Moise Bendayan, University of Montreal, Canada, is that different antigens react to osmium differently. Some are OK and others are not. If no one had worked on it before you have to find out yourself. You may incubate osmium treated section on a drop of saturarated sodium metaperiodate for 1 hr. before immunostaining. Following reference covers tissue precessing. Bendayan, M. Protein A-gold and Protein G-gold postembedding immunoelectron microscopy. In Colloidal Bold: Principles, Methods and Applications Vol. 1, p33-94. Acacemic Press 1989.
AnnFook yang
} } } Tamara Howard {thoward-at-unm.edu} 02/28/03 01:22PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mike -
I'm sure there are references out there, but all I can offer is personal experience and word-of-mouth. I've used 1 (that I can think of off the bat) antibody that gave "normal" staining on sections of tissue with regular osmication, and some that were OK with reduced osmium (osmium mixed with potassium ferrocyanide or ferricyanide - you'll get reams of info on which to use!), and enough that were useless after any osmium to make you cry. Osmium can also mess up your resin polymerization, so you have to be careful - especially if you want to UV cure - dark tissue is a major pain.
What do you mean by osmium reducing the size of gold label? You said you were doing postembedding immuno....I haven't heard of any gold size problems for postembedding (lots with pre-embedding and osmium) - how would that happen?
Do you have some potentially unrescuable blocks that have already been osmicated? You might try a gentle peroxide treatment on the sections, or some EM version of antigen retrieval....
Good luck!
Tamara
On Fri, 28 Feb 2003, Mike Delannoy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To the listserver, } I am looking for references on the effects of osmium staining } on postembedded immunolabeling, ie reducing the amount and size } of gold label on LR Gold/Whites sections. Also has anyone } used very low osmium (0.05%?) successfully for IEM? } } Thank You } Mike D } } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
Anyone have a manual for a Nikon Labophot Pol? I need to do a little cleaning and probably regreasing for it, but don't have any kind of a manual, basic, service or otherwise. Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
We've lost the operating instructions to our LKB 2178 Knifemaker II. Can someone make us a copy of their's? I'd certainly compensate you for the associated costs. Thanks in advance.
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-487-2002 FAX 906-487-2934 Mailto: opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills.htm
The following party is looking for a outside laboratory to study the structure of DNA in plasmid. This is beyond the scope of our laboratory. I told her that someone in MSA can help her. J. Roy Nelson, Ph.D. Material Testing Laboratory mtl-at-njcc.com
"I am interested in using electron microscopy to look at supercoiled versus open circular DNA in our sample of purified plasmid DNA. I would like to know who might offer this service and their price for the service."
Has anyone done TEM on Armadillidium vulgare (Pillbugs)? We're interested in the structure of the eyes of these pillbugs. We're not very sure about the structures that we are seeing and the preparation of the specimen. If anyone has any literature that they can recommend or experiences that they can share I would greatly appreciate the help.
Khara Scott EM Technician Chicago State University
__________________________________________________ Do you Yahoo!? Yahoo! Tax Center - forms, calculators, tips, more http://taxes.yahoo.com/
Refer to "G. M. Brown and J. H. Butler, "New method for the characterization of domain morphology of polymer blends using ruthenium tetroxide staining and low voltage scanning electron microscopy (LVSEM)", Polymer 38 (15), 3937 (1997)" for a detailed description for preparation of RuO4 solutions.
Regards,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"George.Theodossio u-at-csiro.au" To: Microscopy-at-sparc5.microscopy.com cc: Subject: TEM Polymer Staining Procedure 02/13/03 01:14 AM
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Dear All,
I have a colleague who is looking at silica particles in a polymer matrix. He is planning TEM analysis of cross sections to image the particles, determine their distribution and possibly some EDXS mapping. The samples are cast as thin films and the silica could be as small as 5nm. He has found a reference to the use of Phosphotungstic Acid (hope the spelling is correct) to stain the polymer but not the silica, thus improving contrast in the TEM. This was done by floating off the cryo-microtomed sections on a methanol solution of Phosphotungstic Acid. Alas, this is all the information he has.
We don't have cryo microtomy facilities, so we were going to use our microtome at room temp and also attempt to tripod polish.
Does anyone have a more detailed procedure of how to make up the solution and stain the polymer?? Any assistance would be greatly appreciated.
Regards George
George Theodossiou Physicist / Electron Microscopist CSIRO Manufacturing & Infrastructure Technology Private Bag 33 Clayton South MDC Victoria, 3169 tel: +61 3 9545 2012 fax: +61 3 9544 1128
To the extent permitted by law, CSIRO does not represent, warrant and/or guarantee that the integrity of this communication has been maintained or that the communication is free of errors, virus, interception or interference.
The information contained in this e-mail may be confidential or privileged. Any unauthorised use or disclosure is prohibited. If you have received this e-mail in error, please delete it immediately and notify George Theodossiou on +61 3 9545 2012. Thank you.
I am a book designer with the University of Illinois Press and am searching for a SEM image of a cancer cell(s) to use on the cover of a book we are publishing.
It was suggested that I make my query on this list. The book talks specifically about mantle-cell diffuse lymphoma but the image does not need to be that exact, any malignant lymphoma would do.
We would of course credit any images we use and perhaps be able to pay you or your organization a small fee. Please email me direct if you have any information to pass on.
Thank you,
Paula Newcomb Designer University of Illinois Press 217.244.4706 newcomb1-at-uiuc.edu
Below is the Table of Contents from the upcoming issue of the Journal of Microscopy & Microanalysis. Vol 9, No. 2
Editorial Charles Lyman
Historical Bibliography Key Events in the History of Electron Microscopy F. Haguenau, P.W. Hawkes, J.L. Hutchinson, B. Satiat-Jeunemaître, G.T. Simon, and D.B. Williams
Papers from the Australian Microbeam Analysis Society
Monet's Painting under the Microscope Paula Dredge, Richard Wuhrer, and Matthew R. Phillips
Cathodoluminescence Efficiency Dependence on Excitation Density in n-Type Gallium Nitride Matthew R. Phillips, Hagen Telg, Sergei O. Kucheyev, Olaf Gelhausen, and Milos Toth
Application of Quantitative Electron Microscopy to the Mineral Content of Insect Cuticle Ron Rasch, Bronwen W. Cribb, John Barry, and Christopher M. Palmer
Charge-Related Problems Associated with X-Ray Microanalysis in the Variable Pressure Scanning Electron Microscope at Low Pressures Brendan J. Griffin and Alexandra A. Suvorova
Dear List, I'd like to find a protocol for making colloidal gold in 10 nm and 20 nm sizes. I'm sure someone on the list has either a procedure or a reference to one. TIA for your help. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Dear Colleagues, The 4th ASEAN Microscopy Conference (ASEANMC4) will be held in Hanoi, VietNam, from October 9 till 10, 2003. This Conference will be a gathering of microscopists from the ASEAN and other regions of the world. An important aim of the Conference is to promote mutual understandings and to develop international collaborative research in life and material sciences. Participants will have the opportunity to review recent work, to discuss recent advances and identify new challenges in the field of electron and light microscopy. The Second announcement are currently in the mail to many microscopist who are interested in this conference. When we receive reply from you then we will send "the Second announcement" to you by post as soon as possible. We would like to extend our warmest welcome to you to participate and look forward to seeing you in Hanoi in October 2003.
Chairman, Organizing committee
Prof. Nguyen Van Man
***********************************************
Mailing address:
Assoc. Prof. Nguyen Kim Giao
Electron Microscopy Unit
National Institute of Hygiene and Epidemiology
N0 1, Yersin street- Hai Ba Trung district - Hanoi - Vietnam
Tel: 84.04.9715434 Fax: 84.04.8210853
Email: {mailto:emlad-at-hn.vnn.vn} emlad-at-hn.vnn.vn, or {mailto:emunihe-at-vol.vnn.vn} emunihe-at-vol.vnn.vn
Why make your own gold, so much easier to buy and store premade?
Anyway, way back when there was a review article in the EMSA Bulletin entitled Colloidal Gold as a Marker and Tracer in Light and Electron Microscopy. Published around 1986??? Author was J. Dewey This is a nice review article on most aspects of colloidal gold labeling..
Other references to look up are:
1). Mulpferdt, H. Experientia, 1982 (38) pg 1127-1128 2). Roth, J. 1983b Techniques in Immunocytochemistry, Vol. II (Academic Press) pg 217-284.
best of luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
Note to list: This E-mail included a rather lengthy attachment that cannot be sent via the list. Please E-mail me directly if you want the procedure for making colloidal gold conjugates.
Bill, I have used the enclosed procedure many times and it works great. Although I am lazy these days and usually buy gold conjugates, I still have some prepared by this method that worked great 8-10 years after being made. I made primarily 5 and 10nm gold so cannot vouch for the 20nm results.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 3/3/03 5:52 PM, "Bill Tivol" {tivol-at-caltech.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 List, } I'd like to find a protocol for making colloidal gold in 10 nm and 20 } nm sizes. I'm sure someone on the list has either a procedure or a } reference to one. TIA for your help. } Yours, } Bill Tivol } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 } tivol-at-caltech.edu } } }
I kindly ask you for your advice concerning inverted microscope. We need to count tiny mites in alcohol samples. Mites were washed from plants, so some debris is in samples, too. As mites are very small - adults are 0.2 times 0.05 mm, white soft bodies, observing them using dissection microscope is very difficult, determination developmental stages and sex impossible.
Therefore, we consider to purchase an inverted microscope but we are not sure if to ask for that one which is equipped with relief contrasting method. If yes, which would be the best? Hoffman modulation contrast usually needs special condenser and lenses and is therefore more expensive. Zeiss microscope has so called Variable relief (VAREL) contrast which allows unilateral darkfield, perhaps convenient for our white mites.
As we do not have a possibility to try inverted microscope with relief contrast, I would very appreciate your opinion/advice.
Dear Colleagues: Tungsten bulbs change as they age (e.g., move toward more yellow wavelengths). Can anyone tell me if the mercury bulbs used in fluorescence microscopes also change with age? If so, is it possible that an old bulb might result in weakened fluorescence? --Many thanks, Jan Factor
--------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Division of Natural Sciences Purchase College State University of New York Purchase, NY 10577 --------------------------------------- Ofc. Tel: 914-251-6659 Ofc. Fax: 914-251-6635 E-mail: jfactor-at-purvid.ns.purchase.edu ---------------------------------------
Whether your specialty involves fundamental research or practical applications of surface treatments and coatings, we encourage you to make a technical presentation at the 2nd ASM International Surface Engineering Congress & Exposition, to be held Sept. 15-18, 2003 in Indianapolis.
This premier event uniquely focuses on the practical engineering as well as the science of surface treatments and coatings. It's your best opportunity for networking with industrial engineers and scientists, industrial managers, and academic and government researchers.
To further enhance the technological and economic relevance of this year's technical sessions, our Congress will be co-located with America's largest heat treating event, the 22nd ASM Heat Treating Society Conference & Exposition, as well as the annual meeting of the North American Die Casters Association.
Abstracts are due by March 10 for presentations (platform and poster sessions) in the following areas:
Corrosion resistant surfaces PVD/CVD processes and applications Laser processes Thermal barrier coatings Computer modeling of deposition processes and applications Characterization of coatings and thin films Surface finishing effects and technologies Surface metrology Tribological coatings Residual stresses development and effect on properties Biomedical applications of coatings New deposition equipments and manufacturing Thermally sprayed coatings and applications Plasma enhanced processes Scanning probe microscopy for nanosciences
Please note that while "paper" submissions are encouraged for publishing in our proceedings volume, they are NOT required. To submit your abstract by March 10, please click on the following link: http://www.asminternational.org/absubmit_surface.cfm
Sincerely,
Dr. Oludele Popoola, General Chair Consultant Novi, MI popoola1-at-yahoo.com
Dr. Sudipta Seal, Program Chair University of Central Florida Orlando, FL sseal-at-pegasus.cc.ucf.edu
Dr. Arnold Deutchman, Expositions Chair Worthington BeamAlloy Corp., a Division of Worthington Industries, Inc. Columbus, OH ahdeutch-at-worthingtonindustries.com
Professor Narendra Dahotre, Proceedings Editor University of Tennessee-Center for Laser Applications Knoxville, TN ndahotre-at-utk.edu
Ask the vendor to bring the instrument you are considering to your lab for a demonstration. We always do this for expensive items. Any vendor who will not comply is not worth dealing with.
Geoff
Rostislav Zemek wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 users of ListServer, } } I kindly ask you for your advice concerning inverted microscope. We need } to count tiny mites in alcohol samples. Mites were washed from plants, so } some debris is in samples, too. As mites are very small - adults are 0.2 } times 0.05 mm, white soft bodies, observing them using dissection } microscope is very difficult, determination developmental stages and sex } impossible. } } Therefore, we consider to purchase an inverted microscope but we are not } sure if to ask for that one which is equipped with relief contrasting } method. If yes, which would be the best? Hoffman modulation contrast } usually needs special condenser and lenses and is therefore more } expensive. Zeiss microscope has so called Variable relief (VAREL) contrast } which allows unilateral darkfield, perhaps convenient for our white } mites. } } As we do not have a possibility to try inverted microscope with relief } contrast, I would very appreciate your opinion/advice. } } Thank you. } } Rostislav Zemek } } ------------------------------------------------------------------------------ } Rostislav Zemek Phone : 00420-387775227 } Institute of Entomology } Branisovska 31 Fax : 00420-385310354 } 370 05 Ceske Budejovice } Czech Republic E-mail: rosta-at-acarus.entu.cas.cz } ------------------------------------------------------------------------------ }
-- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
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Eighth Annual INTERNATIONAL 12-Day Short Course on
3D Microscopy of Living Cells June 15 - 26, 2003 (Pre-course: afternoon, June 14)
Seventh, Post-course Workshop on
3D Image Processing, June 28 - July 30, 2003
http:// www.3dcourse.ubc.ca/
Organized by Prof. James Pawley, (University of Wisconsin-Madison)
in association with
The Departments of Pharmacology and Physiology and the Brain Research Centre, University of British Columbia Vancouver, BC, Canada
DATES
Applications must be received by March 15, 2003 Deposit due April 15, 2003 Registration 5:00 - 7:00 PM Saturday, June 14, 2003 First Lecture 7:30 PM Saturday, June 14, 2003 Live-cell Course ends, noon Thursday, June 26, 2003 3D Image Processing Course, June 28 - 30, 2003
APPLICATIONS
Applicants must complete a questionnaire on the web to assess knowledge level, field of interest and proposed personal, live-cell, project. www.3dcourse.ubc.ca/application.htm
Enrollment will be limited to about 24-32 participants (exact number depends on number of 3D Systems available). Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with access to basic texts to read before the course begins. Application forms can be obtained from:
Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU
Additional information is available from: www.3dcourse.ubc.ca/brochure.htm
We expect to have at least 11, 3D microscope workstations for student use during 14 3D-Lab session and there will be an international faculty of 20.
Application deadlines:
Application forms must be received for screening by March 15, 2003. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2003. In general, refunds of the deposit will only be possible if someone on the waiting list can take the place. The remainder is due before Registration.
Pre-Course Tuition (1/2 day Basic Optical principles) $120 (US) 3D Live-cell Course Tuition (includes lunches and snacks): $2450 (US) 3D Image Processing Workshop Tuition (includes lunches and snacks): $900 (US)
Room/board about $40/day (US) depending on room type.
I hope that this includes all of the information that you need, but if not, please get back to me.
Jim Pawley --
-- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU 3D Microscopy of Living Cells Course, June 14 - 26, 2003, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2003
I'm in the market for a new critical point dryer, and trying to get a feel for the pros and cons of the various units that are available. If you have purchased a CPD in the last few years, would you please contact me (off-line)? I would appreciate hearing what you have and how well it works.
Thanks!
Leslie Eibest SEM lab manger Dept. of Biology Duke University
JiaMing manufacturing company, a world class forged iron, stained class, wood furniture, bamboo floor, stone product manufacturing company. Currently, is looking for European distributor. Please let us know if you're interested. You can also visit our web site www.jmwired.com
The article was by J. DeMay from Janssen Pharm in Beerse, Belgium and it's on pg 54 of EMSA Bulletin 14, 1 Spring 1984. It has a detailed appendix on how to make colloidal gold. The article is an extensive work with a zillion references.
I could either a.) photocopy the article and send it to Bill, b.) scan it in to PDFs and post their availability on the listserver, or c.) seek permission to reprint a shorter version in Microscopy Today. That said, Ed's suggestion to just go out and buy the stuff is a good one.
What does the community want?
Ron Anderson, Editor Microscopy Today and Editor of the EMSA Bulletin in 1984
-----Original Message----- } From: Edward Calomeni [mailto:calomeni-1-at-medctr.osu.edu] Sent: Tuesday, March 04, 2003 7:48 AM To: Microscopy-at-sparc5.microscopy.com
Morning Bill,
Why make your own gold, so much easier to buy and store premade?
Anyway, way back when there was a review article in the EMSA Bulletin entitled Colloidal Gold as a Marker and Tracer in Light and Electron Microscopy. Published around 1986??? Author was J. Dewey This is a nice review article on most aspects of colloidal gold labeling..
Other references to look up are:
1). Mulpferdt, H. Experientia, 1982 (38) pg 1127-1128 2). Roth, J. 1983b Techniques in Immunocytochemistry, Vol. II (Academic Press) pg 217-284.
best of luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
Looking for some lab in the Dallas area ( will consider other areas ) that could take some silicate powder material and heat it to 1100-1200C in a vacuum. Want to melt the material into a sample that can be sectioned and polished for microprobe analysis.
Thanks
Roy Beavers Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, Tx. 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
Greetings, Arc lamps probably do not change in spectrum as they age, but for sure they lose intensity. Sometimes when you change an old one, you will see a cloud of metalic deposit on the glass, presumably lowering the output intensity. In my experience the loss depends on the individual bulb, some lose more than others. But I have bulbs that are clearly and obviously less bright at the end of their life. Much rejoicing when the bulb gets changed.
Hope this helps, Tobias Baskin
} } } Dear Colleagues: Tungsten bulbs change as they age (e.g., move toward } more yellow wavelengths). Can anyone tell me if the mercury bulbs used } in fluorescence microscopes also change with age? If so, is it possible } that an old bulb might result in weakened fluorescence? } --Many thanks, Jan Factor } } --------------------------------------- } Jan Robert Factor, Ph.D. } Professor of Biology } --------------------------------------- } Division of Natural Sciences } Purchase College } State University of New York } Purchase, NY 10577 } --------------------------------------- } Ofc. Tel: 914-251-6659 } Ofc. Fax: 914-251-6635 } E-mail: jfactor-at-purvid.ns.purchase.edu } ---------------------------------------
Dear List, Thank you for the quick and informative responses. I have two protocols in hand and several promising references. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Jan I think mercury lamp may emit less light over the years for so many reasons (decreasing pressure or just dust cover). I would suspect more the filters: they may fade or change the properties under extended exposure to the UV light. In general, all optical components with age became dirtier and UV light may just make case the worse. Finally you may see funny sometimes colored layers of decomposed (or what?) material covered optical surfaces. It definitely will not increase the amount of light you have. Sergey
At 08:33 AM 3/4/2003, 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 can appreciate that anyone would prefer to have a vendor deliver an instrument to their door to do an on-site demonstration, but you also must realize that it is simply not possible in every instance. To say that "Any vendor who will not comply is not worth dealing with" is quite simplistic and offensive. I have nothing to do with this particular customer, but I can certainly understand how several excellent microscopy vendors may have difficulty justifying an on-site demo in the Czech Republic. I personally have been to the Czech Republic several times to demonstrate our systems, however, it can certainly not be justified in every instance. To dismiss vendors so capriciously would be doing a disservice to Dr. Zemek. Perhaps the company offering the best solution at the best price has a better method of demonstrating its performance than through an on-site demo. Technology these days is pretty impressive.
Speaking as a family business owner and microscopy vendor for nearly 40 years, I guess I take it a bit personally when vendors are treated with such disrespect. Scientists and vendors working together rather than as adversaries is much more productive and serves the scientific community much more effectively.
Best regards-
David
Geoff McAuliffe wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Ask the vendor to bring the instrument you are considering to your lab for a } demonstration. We always do this for expensive items. Any vendor who will not } comply is not worth dealing with. } } Geoff } } Rostislav Zemek wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 users of ListServer, } } } } I kindly ask you for your advice concerning inverted microscope. We need } } to count tiny mites in alcohol samples. Mites were washed from plants, so } } some debris is in samples, too. As mites are very small - adults are 0.2 } } times 0.05 mm, white soft bodies, observing them using dissection } } microscope is very difficult, determination developmental stages and sex } } impossible. } } } } Therefore, we consider to purchase an inverted microscope but we are not } } sure if to ask for that one which is equipped with relief contrasting } } method. If yes, which would be the best? Hoffman modulation contrast } } usually needs special condenser and lenses and is therefore more } } expensive. Zeiss microscope has so called Variable relief (VAREL) contrast } } which allows unilateral darkfield, perhaps convenient for our white } } mites. } } } } As we do not have a possibility to try inverted microscope with relief } } contrast, I would very appreciate your opinion/advice. } } } } Thank you. } } } } Rostislav Zemek } } } } ------------------------------------------------------------------------------ } } Rostislav Zemek Phone : 00420-387775227 } } Institute of Entomology } } Branisovska 31 Fax : 00420-385310354 } } 370 05 Ceske Budejovice } } Czech Republic E-mail: rosta-at-acarus.entu.cas.cz } } ------------------------------------------------------------------------------ } } } } -- } ********************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane, Piscataway, NJ 08854 } voice: (732)-235-4583; fax: -4029 } mcauliff-at-umdnj.edu } **********************************************
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Can anyone tell me if the mercury bulbs used } in fluorescence microscopes also change with age? If so, is it possible } that an old bulb might result in weakened fluorescence? } --Many thanks, Jan Factor
Mercury lamps lose their output as a function of time. At the end of the rated life of the lamp the output is diminished by 30%, per the manufacturer. Not letting the lamp warm up before shutting it off can also cause the lamp to lose output, by darkening the envelope. It may also ruin the lamp completely. These lamps must be on for a minimum of 15 minutes after being lit, per the manufacturer. I suggest 20 minutes as a rule.
Alignment of the lamp is critical and can have a dramatic effect on the light delivered to the specimen. It is very common for the lamp to be installed incorrectly, or to have someone try and "adjust" the lamp for you and throw the alignment off. The adjusting mechanism in the lamphouse of many of these systems freezes up over time as the lubricants are dried up by the intense heat of the lamp and this also contributes to an inability to make the proper adjustments...
Good luck,
Try Lamp Technology or Bulbman as a resource for replacement lamps. Great prices, good service.
Std disclaimer, I have purchased thousands of lamps from both but have no other relationship with them.
David Burton Optical Specialist, University of Washington dbuw-at-u.washington.edu
We might be in the market for a new digital camera for microscopy/image analysis. I am currently using a Nikon DXM 1200 (12 magapixels) but wonder what else is out there as the development of such devices is going so quickly.
Contacts from users with likes/dislikes and also from the trade would be welcome.
Obs/NB New postal/visiting address from July 2002!
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
We are trying to make small optical cells for use under the optical microsccope, by building up square glass "walls" on a microscope slide, or mounting a metal washer on the same. The cavity is to contain some rather agressive solvent mixtures, also sometimes to be heated, so sticking with superglue or Aralidite does not always make a sound cell. Could anyone tell us of a tough adhesive for these purposes?
(School essay, AD3000: "The greatest playwrite of the Early Computer Age was William Shakespeare, who wrote the Comedy of Errors.")
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
Ron et al. I vote for creating Protocol Bank on the MSA WWW site and having Debby's and Jan's protocols there as pdf files. Marek.
At 03:17 PM 3/4/03 -0500 Tuesday, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I went through all the SEM tips in the archives but I still couldn't find answers to two specific problems that I am facing:
1. Mouse Embryos: After OTO processing and CPD the samples became very fragile and some are even damaged during the run. How does one get the embryos onto the stub and orient them without damaging them? Or should the sample be attached on to something before CPD?
2. Insects: In preparing whole insects for SEM, the problem I found was that after CPD they also become very brittle, easily damaged. Some suggest to stick a pin through the insect, but I don't know at what stage should this be done. Before fixation? before CPD?
Eleana Sphicas Bio-imaging Resource Center Rockefeller university (212)-327-8125
} Geoff: } } I can appreciate that anyone would prefer to have a vendor deliver an } instrument to their door to do an on-site demonstration, but you also } must realize that it is simply not possible in every instance.
That may be true.
} To say } that "Any vendor who will not comply is not worth dealing with" is quite } simplistic and offensive.
Hey, times are very tight here in University research land. We spend our money the way we want to. We can't afford to buy equipment we have not tested. If this offends you, so be it. In 30 years in university science, I have never had a vendor refuse to demo a product, even an inexpensive one.
} I have nothing to do with this particular } customer, but I can certainly understand how several excellent } microscopy vendors may have difficulty justifying an on-site demo in the } Czech Republic.
If that is the nature of their business, then that is the nature of their business. The vendor runs his business the way he wants to, the customer spends his money the way he wants to. Capitalism.
} I personally have been to the Czech Republic several } times to demonstrate our systems, however, it can certainly not be } justified in every instance. To dismiss vendors so capriciously would } be doing a disservice to Dr. Zemek.
No, Dr Zemek can make his own decisions. I was suggesting one course of action for his consideration.
} Perhaps the company offering the } best solution at the best price has a better method of demonstrating its } performance than through an on-site demo. Technology these days is } pretty impressive.Speaking as a family business owner and microscopy vendor for } nearly 40
} years, I guess I take it a bit personally when vendors are treated with } such disrespect.
Not disrespect, it is just the way things operate in my world. How is 'not buying a product without a demo to see if it fits your lab's needs' disrespectful??
} Scientists and vendors working together rather than as } adversaries is much more productive and serves the scientific community } much more effectively.
That is a very nice, general statement that no one would argue with. It is also 180 degrees opposite of buying a product without knowing if it will solve your problem.
Geoff
} Best regards- } } David } } Geoff McAuliffe wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Ask the vendor to bring the instrument you are considering to your lab for a } } demonstration. We always do this for expensive items. Any vendor who will not } } comply is not worth dealing with. } } } } Geoff } } } } Rostislav Zemek wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 users of ListServer, } } } } } } I kindly ask you for your advice concerning inverted microscope. We need } } } to count tiny mites in alcohol samples. Mites were washed from plants, so } } } some debris is in samples, too. As mites are very small - adults are 0.2 } } } times 0.05 mm, white soft bodies, observing them using dissection } } } microscope is very difficult, determination developmental stages and sex } } } impossible. } } } } } } Therefore, we consider to purchase an inverted microscope but we are not } } } sure if to ask for that one which is equipped with relief contrasting } } } method. If yes, which would be the best? Hoffman modulation contrast } } } usually needs special condenser and lenses and is therefore more } } } expensive. Zeiss microscope has so called Variable relief (VAREL) contrast } } } which allows unilateral darkfield, perhaps convenient for our white } } } mites. } } } } } } As we do not have a possibility to try inverted microscope with relief } } } contrast, I would very appreciate your opinion/advice. } } } } } } Thank you. } } } } } } Rostislav Zemek } } } } } } ------------------------------------------------------------------------------ } } } Rostislav Zemek Phone : 00420-387775227 } } } Institute of Entomology } } } Branisovska 31 Fax : 00420-385310354 } } } 370 05 Ceske Budejovice } } } Czech Republic E-mail: rosta-at-acarus.entu.cas.cz } } } ------------------------------------------------------------------------------ } } } } } } } -- } } ********************************************** } } 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 } } ********************************************** } } -- } David Henriks } Vice President } } South Bay Technology, Inc. } 1120 Via Callejon } San Clemente, CA 92673 USA } } TEL: +1-949-492-2600 } Toll-free in the USA: +1-800-728-2233 } FAX: +1-949-492-1499 } } email: henriks-at-southbaytech.com } } Celebrating 38 years of providing Materials Processing Solutions for } Metallogaphy, } Crystallography and Electron Microscopy. } } Please visit us online at www.southbaytech.com. } } The information contained in this message and any attachments is } privileged and } confidential. This message is intended for the individual or entity } addressed. } If you are not the intended recipient, please do not read, copy or } disclose this } communication. Notify the sender of the mistake by calling } +1-949-492-2600 and } delete this message from your system.
-- ********************************************** 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 **********************************************
Hello everyone, I've been trying to enlarge a "virtual museum" of older, significant scopes and equipment used in electron microscopy. If you have a picture of older equipment (out of use or still being used) I would like to include it on the site. I would also like to include some information about the equipment (dates, usage, location, any interesting features, etc..) and cite the provider of the image as a contributor. This could also be a link to an existing image at your site. At present it represents equipment collected by the late Dr. Jerry Paulin. I would like to add a "Contributed Collection" addition to this. I think it would be great to show EM students the variety and provide them an appreciation for the changes in equipment.
The museum, as it stands now, can be seen at www.uga.edu/caur/museum.htm
Thanks in advance for your participation. John Shields EM Lab University of Georgia Athens, GA 30602 jshields-at-cb.uga.edu
What I've done for such delicate samples as you describe and with some success is to take a pin (like an insect mounting pin, or push pin) cut to about 1/4 inch in length, file a flat end, dip end in colloidal carbon paint, gently touch that end to "underside" (or side opposite the one you want to look at) of insect or embryo. The sample will stick to the paint so then you can lift it up. Hold the pin by its side with a clamping type fine tipped forceps.
Next step is to mount it onto an SEM sample stub. You can drill a small hole into the stub surface that will take the diameter of the pin, or use a clamping type SEM stub, both of which I have here. After mounting/clamping the pin onto the stub, paint carbon paint over the stub surface, then you will have a nice dark background behind the sample, which may also be conveniently out of focus, due to height of sample above stub surface, which helps to give dark featureless background for those full body glamour shots!
Good luck!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Bera
} Hi list members, } } I went through all the SEM tips in the archives but I still couldn't find } answers to two specific } problems that I am facing: } } 1. Mouse Embryos: After OTO processing and CPD the samples became very } fragile and some are even damaged during the run. How does one get the } embryos onto the stub and orient them without damaging them? Or should the } sample be attached on to something before CPD? } } 2. Insects: In preparing whole insects for SEM, the problem I found was } that after CPD they also become very brittle, easily damaged. Some suggest } to stick a pin through the insect, but I don't know at what stage should } this be done. Before fixation? before CPD? } } Eleana Sphicas } Bio-imaging Resource Center } Rockefeller university } (212)-327-8125
A lot of the problem with arc sources is the electrodes. As the arc files over a long period of time, the electrodes erode, decreasing intensity. This problem adds to the deposition you have mentioned.
OptiQuip has a new stabilizer (LTS) and power supply (model 1600) for their Nobska fluorescence illuminators which automatically tests the lamp intensity 2000 times/s, compares it to a stored intensity, then signals the power supply to adjust the voltage to maintain the intensity. Normal stabilized power supplies control just the flicker. It's really great for everything from measurement to montage to deconvolution.
We ran an article last November in American Lab which discusses this new approach in detail: Foster, B. "Optimizing illumination for Microscopy and Measurement", Am Lab, Nov 2002, Pp13-17.
Caveat: MME has no financial interest in this product.
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
"Why didn't they teach us that sooner?" ... probably because no one thought to call MME about customized, on-site courses. Offered in all areas of microscopy, sample prep,and image analysis, they make an immediate impact on your productivity. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
At 03:32 PM 3/4/03 -0600, Tobias Baskin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I use a 5/0 Red Sable artist's brush. You can buy these from the EM suppliers like Ted Pella for a couple bucks. I use them on embryos and insects, just like you. Just gently touch the sample and it will usually stick to the hairs. Then touch it down to your adhesive substrate. If the sample won't stick to the brush, touch the brush to your tongue (or touch your finger to your tongue then touch the brush to your finger). Saliva is a wonderful thing.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
At 09:26 AM 3/5/2003 -0500, Eleana Sphicas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Silicone sealant will probably work(something like Silastic (TM)). It's both heat and solvent resistant. You can try the hardware store variety, or contact Dow, GE, or one of the other manufacturers if you need something more specialized.
On Wednesday, Mar 5, 2003, at 05:38 America/New_York, Robert H. Olley wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 trying to make small optical cells for use under the optical } microsccope, by building up square glass "walls" on a microscope slide, } or mounting a metal washer on the same. The cavity is to contain some } rather agressive solvent mixtures, also sometimes to be heated, so } sticking with superglue or Aralidite does not always make a sound cell. } Could anyone tell us of a tough adhesive for these purposes? } } (School essay, AD3000: "The greatest playwrite of the Early Computer } Age was William Shakespeare, who wrote the Comedy of Errors.") } } +-----------------------------------------+ } Robert H.Olley } J.J.Thomson Physical Laboratory } University of Reading } Whiteknights } Reading RG6 6AF } England } +-----------------------------------------+ } Phone: } {direct line +44 (0) 118 9318572 } {University internal extension 7867 } Fax: +44 (0) 118 9750203 } Email: R.H.Olley-at-reading.ac.uk } URL: http://www.reading.ac.uk/~spsolley } +-----------------------------------------+ } } } } All the best, Andy Buechele
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Eleana Sphicas {sphicae-at-rockefeller.edu} 03/05/03 09:26AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi list members,
I went through all the SEM tips in the archives but I still couldn't find answers to two specific problems that I am facing:
1. Mouse Embryos: After OTO processing and CPD the samples became very fragile and some are even damaged during the run. How does one get the embryos onto the stub and orient them without damaging them? Or should the
sample be attached on to something before CPD?
Fixed and dehydrated tissues will become brittle. Process them adhered to something. Amazingly SuperGlue does a nice job on fresh tissue. Use very sparingly and do not cover or the fumes will deposit on the surfaces. There are also ways to bind to glass coverslips (aminosilane), and then just process the coverslip itself.
2. Insects: In preparing whole insects for SEM, the problem I found was that after CPD they also become very brittle, easily damaged. Some suggest
to stick a pin through the insect, but I don't know at what stage should this be done. Before fixation? before CPD?
Ha, the bane of my recent existence. How to keep insects looking lively. Already dried specimens may be placed in a steam bath to absorb moisture and then CAREFULLY arranged (Some of my researchers just plunk in warm water overnight with a little surfactant). I prefer the steam because it doesn't require CPD or HMDS so long as you control it. If samples are in alcohol or fixative , you may pin mount them, but not through the body. Splay out the critter, and place a pin to either side of the body, angled so that they cross just over the structure. It takes time and lots of practice especially to pin down all the legs, antenna, wings, body... but the results are well worth it. I use silicone mats or filter paper underneath. If you don't have the mini pins, a cactus works really well (my preference). From this point dry in whatever manner suits you.
Fresh samples are most easily done by getting them to land on a damp piece of filter paper. When happy, set on a liquid nitrogen chilled block to snap freeze. I have also had success getting creepers to walk across tape and then freezing in a lab freezer to kill. Freeze drying would be the preferred method from this point.
Method three- Use an ESEM or cryo stage equipped instrument. Getting them to sit still under the beam is the major drawback with ESEM
Good luck!!
Eleana Sphicas Bio-imaging Resource Center Rockefeller university (212)-327-8125
Someone just dropped off a handful of "ancient" coins and wants to know if there is anything an SEM could tell him about their authenticity. The person dropping them off is not a crackpot treasure hunter type. He is a careful and knowledgeable professor who is really stumped.
They come from Syria, where they were bought from a street vendor. They are supposed to be silver and appear to be the real thing. If fake, they are really good fakes. If I got the story right, the vendor did not claim that they were authentic, but couildn't, wouldn't say where they came from. The price was about right for reproductions, but on closer inspection, they do not look like obvious forgeries.
We suspect they may be cast rather than struck and were wondering if there are any characteristic features that would tell us how the coins were made. We will do a little EDS to check on the metal, are there any trace metals that might be a give away, either way?
If you know anything about coins or anyone who we could contact, we would appreciate it very much if you could help us out.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
On Tuesday, March 4, 2003, at 12:17 PM, MicroscopyToday wrote:
} Way to go Ed--you've got a great memory! } } The article was by J. DeMay from Janssen Pharm in Beerse, Belgium and } it's on pg 54 of EMSA Bulletin 14, 1 Spring 1984. It has a detailed } appendix on how to make colloidal gold. The article is an extensive } work } with a zillion references. } } I could either a.) photocopy the article and send it to Bill, b.) scan } it in to PDFs and post their availability on the listserver, or c.) } seek } permission to reprint a shorter version in Microscopy Today. That said, } Ed's suggestion to just go out and buy the stuff is a good one. } } What does the community want? } Dear Ron, I can print out the article from PDF files, so that would be my preference; however, either of the other options is acceptable. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Jonathan: I've had a fair amount of experience using the SEM in the authentication of ancient coins. Based on your description of the origin of these coins from a street vendor, the chances of their being modern fakes are about 99.99% The individuals in the countries where ancient coins are found know exactly what genuine ancient coins are worth, especially silver ones, and they know where to sell them, which is not a vendor on a street corner.
You are correct in your assumption that they are most likely a cast fakes; most inexpensive reproductions are. Look carefully for a joining line at the edge where the two halves were glued together, or look for an edge that has been filed and then roughened to hide the joining line. Also, look for small casting bubbles in the metal.
In terms of the SEM, do quantitative analysis of the metal. If the coin is covered with toning, you may have to scrap a minute area clean to get an accurate analysis. If the metal is too pure, i.e. greater that 92% silver, be very suspicious. Look for trace metals. In genuine coins you may see small amounts of lead (~2 wt%), gold (less than 1 wt% and often below minimum detectable concentration), as well as iron. If it is cast, you are more likely to see some common casting metal alloy that just looks like silver. Look at the toning using EDS. Silver fakes are often toned with paint (titanium), iodine, etc. Genuine toning will usually show lead, iron, or chlorine from silver chloride (horn silver as it is know to ancient numismatists). If the composition of the toning is identical all over the coin, then be suspicious. Genuine ancients usually have considerable variation in chemical composition of the toning.
I hope this helps. Stanley L. Flegler, Director Center for Advanced Microscopy Michigan State University
I could put you in touch with one of the authors of that study, if you wish to pursue it further.
"To achieve results never before accomplished, we must employ methods never before attempted." -Sir Francis Bacon
Jeffrey A Fortner Argonne National Laboratory CMT/205 9700 S. Cass Avenue Argonne, IL 60439 (630) 252-5594 FAX: (630) 972-4438
} ---------- } From: jmkrupp-at-cats.ucsc.edu } Sent: Wednesday, March 5, 2003 1:40 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM (?) of ancient coins } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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: } } Someone just dropped off a handful of "ancient" coins and wants to know if } there is anything an SEM could tell him about their authenticity. The } person dropping them off is not a crackpot treasure hunter type. He is a } careful and knowledgeable professor who is really stumped. } } They come from Syria, where they were bought from a street vendor. They } are } supposed to be silver and appear to be the real thing. If fake, they are } really good fakes. If I got the story right, the vendor did not claim that } they were authentic, but couildn't, wouldn't say where they came from. The } price was about right for reproductions, but on closer inspection, they do } not look like obvious forgeries. } } We suspect they may be cast rather than struck and were wondering if there } are any characteristic features that would tell us how the coins were } made. } We will do a little EDS to check on the metal, are there any trace metals } that might be a give away, either way? } } If you know anything about coins or anyone who we could contact, we would } appreciate it very much if you could help us out. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
Barbara, we are using two sets of OptiQuip 1600 Power supply and LTS, which runs two of our Mercury lamp housings. We have shipped (Currently packing one) 7 times as of today in the year starting from January 1, 2002 (excluding this time) to New York for repair. That means an average of repair on either one of the system in every two months. Most of the time the Power supply blows the main fuses and burns the cord, which is connecting the LTS. It will take a two weeks to get the system back. If you have used this system for a long time, do you had any problems like these? We appreciate your comments.
We have now asked the company's President to put a full stop on this saga through our local Olympus Representative Mr. David Kinast. Thanks
Shiv
Mayandi Sivaguru, PhD Associate Director Molecular Cytology Core Facility 2, Tucker hall University of Missouri Columbia, MO 65211-7400
Voice: 1-573-882-4895 Fax: 1-573-882-0123
www.biotech.missouri.edu/mcc/
At 11:42 AM 3/5/03 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
There is a large body of information on this in the literature of the fields of archaeometry and art conservation science. The criteria that are important depend on the culture represented by the coins, their age, the burial conditions, ore types exploited for the metals and the treatment or mishandling that they have received since they were found.
Compositional information is often not sufficient by itself to make a determination. Phase analysis (by X-ray diffraction) or microstructural analysis by very limited metallographic testing (to minimize destructive action)may be required.
I've never heard this term "toning" before. Perhaps you are referring to the corrosion layer, oxide or patina that metal surfaces typically acquire and that may be falsified in the case of a forgery.
Unfortunately the criteria that you site could be highly misleading to someone unfamiliar with numismatics or with the methods that have been used over the years to treat ancient metals. Applying these criteria in the way that you suggest could result in a large proportion of legitimate ancient pieces being rejected.
On the other hand (other side of the coin?) in antiquity there were often large numbers of counterfeit coins in circulation. This is particularly true in an area like Syria that was a cross roads for many cultures and often outside close control of distant capitols. These ancient counterfeits are often debased but extremely valuable historical indicators of conditions at the time. Your criteria seem to assume that to be an antiquity a coin must be of high quality and from the official mint.
I would urge you to run any coins of potential value by a knowledgable numismatist before engaging in destructive tests or attempting to clean them. And not to base a decision on broad assumptions such as those regarding the purity of the metal.
John Twilley Art Conservation Scientist
-------Original Message------- } From: "Stanley L. Flegler" {flegler-at-pilot.msu.edu} Sent: 03/05/03 03:38 PM To: Microscopy-at-sparc5.microscopy.com, Jon Krupp {jmkrupp-at-cats.ucsc.edu}
Jonathan: I've had a fair amount of experience using the SEM in the authentication of ancient coins. Based on your description of the origin of these coins from a street vendor, the chances of their being modern fakes are about 99.99% The individuals in the countries where ancient coins are found know exactly what genuine ancient coins are worth, especially silver ones, and they know where to sell them, which is not a vendor on a street corner.
You are correct in your assumption that they are most likely a cast fakes; most inexpensive reproductions are. Look carefully for a joining line at the edge where the two halves were glued together, or look for an edge that has been filed and then roughened to hide the joining line. Also, look for small casting bubbles in the metal.
In terms of the SEM, do quantitative analysis of the metal. If the coin is covered with toning, you may have to scrap a minute area clean to get an accurate analysis. If the metal is too pure, i.e. greater that 92% silver, be very suspicious. Look for trace metals. In genuine coins you may see small amounts of lead (~2 wt%), gold (less than 1 wt% and often below minimum detectable concentration), as well as iron. If it is cast, you are more likely to see some common casting metal alloy that just looks like silver. Look at the toning using EDS. Silver fakes are often toned with paint (titanium), iodine, etc. Genuine toning will usually show lead, iron, or chlorine from silver chloride (horn silver as it is know to ancient numismatists). If the composition of the toning is identical all over the coin, then be suspicious. Genuine ancients usually have considerable variation in chemical composition of the toning.
I hope this helps. Stanley L. Flegler, Director Center for Advanced Microscopy Michigan State University
I'm not sure if your cells would be compatible, but we often used a non-CO2 dependent HEPES buffer for relatively long term atomic force microscopy/calcium imaging/electrophysiology experiments with primary hippocampal neuron and glial cultures. Might be worth looking at.
All the best,
Raj
Dr. Raj Lartius Novascan Technologies, Inc. Iowa State University Research Park 2501 North Loop Drive Ames, Iowa 50010 USA
At 01:14 PM 2/25/03, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Novascan Technologies, Inc. Iowa State University Research Park 2501 North Loop Drive Ames, Iowa 50010 USA
is anyone having any information on the lens design of the old ISI SMS 2 SEM. I'd require data on the geometry of the pole pieces as well as the number of A-t. But for the meantime typical values for the lens currents might already be helpful.
Besides the replies already given, I have used "live insect handling forceps" for both types of samples mentioned (and many other fragile, driec samples). These are blunt or fine-pointed forceps made of thin, flexible stainless steel that allow a sure grip with minimal force. As the name implies, they were originally developed by entomologists, but I believe not for live insects, but minute, dried insects in collections. I got mine from Fine Science Tools, cat #26029-10 & 26030-10 http://www.finescience.com/ . There should be other sources. The other trick is to put a pointy bit of latex rubber on the end of a dissecting needle and rub it on silk or wool or something to give it a charge. The static electricity can then be used to pick up and move fragile specimens. (My apologies to whoever first did this long ago, but I don't remember whom to credit.)
Phil
} Hi list members, } } I went through all the SEM tips in the archives but I still couldn't } find answers to two specific } problems that I am facing: } } 1. Mouse Embryos: After OTO processing and CPD the samples became } very fragile and some are even damaged during the run. How does one } get the embryos onto the stub and orient them without damaging them? } Or should the sample be attached on to something before CPD? } } 2. Insects: In preparing whole insects for SEM, the problem I found } was that after CPD they also become very brittle, easily damaged. } Some suggest to stick a pin through the insect, but I don't know at } what stage should this be done. Before fixation? before CPD? } } Eleana Sphicas } Bio-imaging Resource Center } Rockefeller university } (212)-327-8125
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Other than finding a joining line and/or casting bubbles, there is no one single factor that can be used. Instead look for "mistakes" that a modern counterfeiter may have made.
Doing an EDS check on the metal is an excellent first method of determining authenticity. A high proportion of these tourist fakes are cast and can be eliminated immediately by their alloys. A surprising number do not even contain silver, but instead a casting alloy such as Bismith, Lead, Tin, and Cadmium. A surprising number are cast using solder!
Among the struck (as opposed to cast) fakes, the single most common mistake of modern counterfeiters is to use either four nines pure silver or silver from common silver coins (for example 90% Ag, 10% Cu) without any trace metals. The ancients were not able to produce silver of ultra high purity and trace amounts of other metals (lead, iron, and gold) are present. Athens produced some of the highest silver content ancient coins, but even then, they are perhaps 92% to 94% silver with other trace elements. An excellent source of information on this area is {underline} Metallurgy in Numismatics {/underline} , vol 1 and 2, published by the Royal Numismatic Society, {underline} Analysis of Ancient Metals {/underline} , by E. Caley, and {underline} The New Style Silver Coinage of Athens {/underline} , by Margaret Thompson.
The term toning is often used by numismatists rather than corrosion layer, oxide, etc. It is possible that a genuine ancient coin has been cleaned and artificially retoned. Therefore, if you find titanium or iodine on the surface, it still could be ancient.
There were counterfeit coins in ancient times. The most common example is known as a "fourre." A copper core or blank was used with a thin layer of silver bonded to it. These were then struck using dies that attempted to copy the genuine coins in circulation. In ancient times, silver coins were often tested by making a test cut on the edge to see if a copper core was present. A number of these "tested" coins have survived and are in collections, etc. The ancient counterfeiter made a profit by passing these coins off as high silver coins, when in fact they were mainly copper. Most of these coins can be identified by the breaks in the silver layer that developed over the centuries. If there is any doubt, the coin can be placed in the SEM and if a suspicious area proves to copper, then that is a good indication that it is a fourre. If further proof is needed, specific density testing can be done, short of total destructive analysis.
Recently, we just installed a new JEOL 2010F TEM. The TEM has a 2k*2K ultrascan CCD (not yet installed), EDS, Enfina PEELs and Digiscan STEM system. We also have both Emispec and Gatan systems. This is a user based facility. The TEM is booked everyday.
We are trying to figure out a way for data storage, to let users save data for a period of time and let them transfer data through Internet.
I appreciate for the following informations:
what is the data storage system in your Lab? how much data in a period of time (a day, a week) the users save? how frequently they save and transfer the data?
Any other suggestion on internet security is also welcome
Thank you very much
Sincerely,
Jinguo Wang
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, (814) 863-0637 email: jqw11-at-psu.edu
A histology question for the light microscopists out there: Are Haversian systems present in the flat bones formed by intramembranous ossification or are they only in long bones? Thanks, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I have the Cressinton MTM10 thickness monitor. It has stopped working. Does anyone know anything about this monitor and what I can check other than the fuse which seems to be fine? OR does anyone know how to get in touch with Cressington. I have left phone messages and sent them email from their web site and have gotten no replies.
Thanks in advance for any suggestions.
Mei Lie -- Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
I installed a Windows NT server 6 years ago. I made each microscope with a computer interface a client. There is a Philips TEM with Gatan CCD, Hitachi SEM, Leica confocal, 2 graphics workstations, and a backup server. All the systems run Windows based software so they can be networked as clients of the server (we include one Mac workstation for poster printing). There is also a UNIX based deconvolution scope that shares some parts of the facility. The server has about 60 gig of RAID 5 storage. Images acquired from any client are saved directly to the server. Images are never saved to the client workstations so the workstations stay more or less as delivered. No extraneous software is written to the workstations so the microscope application that is installed at setup remains unchanged. Users get an account on the server and this allows them space on the server. Once the image is written to the server it can be downloaded on the LAN, on the Internet via HTTP, or via FTP. Even with the confocal on the network the 60 gig is plenty of space for now. The server is considered a "transfer station" for information. We do not store images. My policy is that images over 30 days old may be removed at any time without notification. In actual practice, I do not remove anything less than 6 months old. If you want to provide storage, you may need more space. We have over 200 accounts so we leave storage and archiving to the individual but we do provide CD writing and DVD writing hardware in the facility. We do not set quotas on data storage. That feature is not native to Windows NT4.0 but can be added from another vendor. That feature is available in Windows2000 Server. Storage is cheap so quotas are not needed here yet. Security is managed by keeping aware of patches and fixes to the operating system. A firewall is being considered. In the 6 years that the system has been running there has been no security breach and no viruses.
Contact me directly if you have questions.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
At 11:39 AM 3/6/2003 -0500, Jinguo Wang wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
We may need to buy a table to put our Zeiss Axiovert on. I am aware of the various vibration isolation systems discussed here previously, and have used a heavy slab on sorbothane feet on a standard table for years with good success. However the tables that we received with our new building seem to have alot of flex in teh legs, so that even with an isolated slab on top, we are concerned about stage vibration. Can anyone suggest a good solid table to use as a base, or should I just go for a TMC/Kinetic systems table. Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
We have been very happy with our Snap server otherwise known as an NAS server. Basically just a bunch of hard drives and ethernet card. I have the Quantum Snap with 240G of space mirrored. They are scalable and you can add a bunch together to really serve some data. Users access their data in a variety of ways, and it took about as long to set up as to unpack. It also serves as the backup dump for the lab PC's.
We serve roughly 70 researchers and their associates annually generating 15-20k images. Our Molecular group uses a bunch of these as well for their data sets (get big quick) and the users write data to CD's as they fill up their quotas. Again authentication is done on a separate server and it all sits behind the SI firewall keeping the riffraff out. Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Jinguo Wang {jqw11-at-psu.edu} 03/06/03 11:39AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers
Recently, we just installed a new JEOL 2010F TEM. The TEM has a 2k*2K ultrascan CCD (not yet installed), EDS, Enfina PEELs and Digiscan STEM system. We also have both Emispec and Gatan systems. This is a user based facility. The TEM is booked everyday.
We are trying to figure out a way for data storage, to let users save data
for a period of time and let them transfer data through Internet.
I appreciate for the following informations:
what is the data storage system in your Lab? how much data in a period of time (a day, a week) the users save? how frequently they save and transfer the data?
Any other suggestion on internet security is also welcome
Thank you very much
Sincerely,
Jinguo Wang
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, (814) 863-0637 email: jqw11-at-psu.edu
I am trying to get our kevex system running for fully quantitative analysis and have been saving reference standards. When I try to use the standards in deconvolution some are accepted while others are rejected with the message "not a standard for this element and line". Elements up to Al seem to be accepted while everything after Ca is rejected. We thought it may have something to do with the Kb line. I've tried everything I can think of, re-saving etc, I know I have used the same process for all the standards. Any suggestions?
Kind Regards,
Jodi Reynolds. Onesteel Wire Mills Newcastle Australia
Does anyone have any recommndations on laminar flow hoods for optical microscopy? We are looking for one to create a clean space for sample prep and optical micrscopy.
Joe Neilly, Research Investigator Abbott Laboratories R45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Hi folks - does anyone have a confocal with a Coherent Enterprise II 651 UV argon laser or similar, operating close to high resolution TEMs and SEMs - are there likely to be any problems with electrical interference, either in the power supply or radiated?
cheers
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University, Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 http://www.anu.edu.au/EMU
Hi All, I'd like to hear opinions about people's impression of the OEM (original equipment manufacturer sponsored) service they receive for their TEM's. Is it timely (average time from request to completion)? Are issues resolved the first time (call backs for same problem)? Do you feel it is cost effective? Is your downtime excessive? Minimal? Service personnel to service area ratio? Feel free to add any topic I might have missed. We are looking at acquiring new instrumentation, and thus are trying to evaluate every aspect of such a purchase.
All responses will remain confidential.
Thanks in advance, Randy Nessler University of Iowa Central Microscopy Research Facility Iowa City, Iowa 52242 Phone 319-335-8142
Listers, An immediate opening for an experienced (T)EM tech is available here at the Carolinas Medical Center in Charlotte, NC. Please see the listing and apply on-line at {http://www.carolinashealthcare.org} www.carolinashealthcare.org.
Go to Find A Job, then Job Search. Complete Step 1 with "Research Services," Step 2 with "Cannon Research Center," leave Step 3 blank, and click Find (Step 4.) Page down to Res Tech II, Position number 74366.
After viewing the listing (shown below and at the MSA job website,) if interested go to "Apply On Line," and submit your cva/resume indicating the position number.
Hope to hear from you soon!
Res Tech II POSITION NUMBER: 74366 / LOCATION: Cannon Research Center / CATEGORY: Research Services FT,M-F,days. Prepare chemical reagent's, follow chemical protocols, fixation through embedding of specimen, photo darkroom techniques with print processing & related duties. Bio/Chem/Med laboratory, must be able to stand & walk for extended periods. BA/BS in Biology/ Chemistry /or related.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor Cannon Electron Microscopy Lab Carolinas Medical Center P.O. Box 32861; Charlotte, NC 28232-2861 Ship to: 1000 Blythe Blvd; Charlotte, NC 28203 Ofc:704-355-1267 ; Lab:704-355-7220 ; Fax 704-355-0589 E-mail: {mailto:WWiggins-at-Carolinas.org} Winston.Wiggins-at-CarolinasHealthCare.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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PLEASE CIRCULATE THIS TO ALL THAT MAY BE INTERESTED
The Midwest Microscopy and Microanalysis Society (MMMS) is co-sponsoring the following upcoming meetings:
March 27th at Northwestern University (with the Biological Imaging Facility atNorthwestern) Topics include Tomography, Cryo FESEM in addition to others (see details below)
Location: Norris University Center, Lewis Room, Evanston,IL
May 13th - Joint meeting with the Milwaukee ASM Chapter
Topic - Color Metallography presented by George F. Vander Voort, Buehler Ltd.
Location: Klemmer's Banquet Center,10401 W. Oklahoma Ave.Milwaukee, WI 53227(414) 541-0401, 5:00pm Social 6:00pm Dinner 7:00pm Program
For additional information on either program contact Robb Mierzwa (mierzwa-at-jeol.com) or Jim DiOrio (jim_diorio-at-baxter.com)
Northwestern Program: (3/27/03)
9:00 am : Registration, 9:45 Welcome and introduction
10:00 Brad Marsh,Univeristy of Colorado at Boulder:3D Structure studies of the pancreatic beta cell by high resolution EM tomography
11:00 Valerie Kilman,Northwestern University:A new role for an old kinase in the Drosophila circadian clock
11:30 Veronica Ledoux,Northwestern University:Serial EM study of inhibitory hippocampal synapses
12:00 Lunch
1:00 Stan Erlandsen,Univeristy of Minnesota Medical School:Visualizing the glycocalyx:correlative microscopy on cell interaction and detecting individual molecules with immunogold by cryoSEM
2:00 Kendrick Boardman,Northwestern University:Lymphangiogenesis:roles of interstitial fluid flow and fluid channel formation
2:30 Greg Beitel,Northwestern University:Unexpected roles for the Na K ATPase in epithelial tube-size control and septate (tight)junctions organization
3:00 Reception:
4:00 Close
Driving directions to Northwestern University Evanston Campus available on the Web. at http://www.northwestern.edu/campus/directions/
RSVP Required
Please send RSVP via email,phone,or fax to: Robb Mierzwa,MMMS Program Coordinator c/o JEOL USA,Inc. 3906 Lisa Avenue Sheboygan,WI 53083 phone:(920)803-8945;FAX:(920)803-8946 email:mierzwa-at-jeol.com Admission:Free to MMMS Members MMMS Membership:$10.00 MMMS membership available at registration
Mei Lie, of course we've already contacted you, but for clarification to the rest of the list: Ted Pella, Inc. provides sales & service for Cressington vacuum equipment (except the CFE-60) in North, Central and South America. You can reach us at (800) 237-3526.
Tom Pella Ted Pella, Inc. http://www.tedpella.com
-----Original Message----- } From: Mei Lie Wong [mailto:wong-at-msg.ucsf.edu] Sent: Thursday, March 06, 2003 10:23 AM To: Microscopy-at-sparc5.microscopy.com
I have the Cressinton MTM10 thickness monitor. It has stopped working. Does anyone know anything about this monitor and what I can check other than the fuse which seems to be fine? OR does anyone know how to get in touch with Cressington. I have left phone messages and sent them email from their web site and have gotten no replies.
Thanks in advance for any suggestions.
Mei Lie -- Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
} We have been very happy with our Snap server otherwise known as an NAS } server. Basically just a bunch of hard drives and ethernet card. I have the } Quantum Snap with 240G of space mirrored. They are scalable and you can add } a bunch together to really serve some data. Users access their data in a } variety of ways, and it took about as long to set up as to unpack. It also } serves as the backup dump for the lab PC's.
I second the vote for the Snap server. Easy to set up and administer, plus our main unit network guru has ours set up to backup on another one in another building, while that one backs up on ours each night in case one building burns down...
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Hello, I bought an EDAX model JSM-6100 detector for our lab through ebay at a good price. The information on the detector is as follows: 132-01, 10 mm2 active area, ser# 7455-57440ME, amp model#194-SUTW, PV9757/44ME.
It is the detector, connecting support arm, and dewar. It looks like it has an amplifier or electronic box on it as well. I am however missing the support electronics which go from the detector to the computer - wiring, software, computer card, and computer. If anyone has any experience with these detectors, or with installing them with homebuilt computers - let me know. It's a long term project I have to upgrade an Electronscan E3 ESEM to have analysis capability.
The detector looks like this one from EDAX: http://www.edax.com/products/Microanalysis/detectors/standard_EDS/10_liter.html
Thanks everyone.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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
A couple of days ago I embedded some samples for microtomy, using Procure 812. Today when I went to remove the blocks from the mould I found that the mould material had adhered to the blocks and came away with the blocks, thus destroying two mould blocks. I have never experienced this problem in the past to such a degree.
The resin was measured and mixed properly and cured at 60 deg celcius. When embedding I half fill the mould and cure it in the oven at 60 deg celcius for an hour, then position the sample, fill the rest of the mould and back in the oven for final curing. This has worked in the past for me.
Does anybody have a better method of removing the cured blocks and not destroying the moulds, use a release agent etc etc??
Your help would be greatly appreciated.
Regards George
George Theodossiou Physicist / Electron Microscopist CSIRO Manufacturing & Infrastructure Technology Private Bag 33 Clayton South MDC Victoria, 3169 tel: +61 3 9545 2012 fax: +61 3 9544 1128
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George: My experience has been that the moulds age and that with repeated usage tend to start adhering to the cured blocks. At that point it's time to purrchase new moulds. I have had no success with release agents, etc. Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals Ridgefield, CT -- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } } A couple of days ago I embedded some samples for microtomy, using Procure } 812. Today when I went to remove the blocks from the mould I found that the } mould material had adhered to the blocks and came away with the blocks, thus } destroying two mould blocks. I have never experienced this problem in the } past to such a degree. } } The resin was measured and mixed properly and cured at 60 deg celcius. When } embedding I half fill the mould and cure it in the oven at 60 deg celcius } for an hour, then position the sample, fill the rest of the mould and back
} in the oven for final curing. This has worked in the past for me. } } Does anybody have a better method of removing the cured blocks and not } destroying the moulds, use a release agent etc etc?? } } Your help would be greatly appreciated. } } Regards } George } } George Theodossiou } Physicist / Electron Microscopist } CSIRO Manufacturing & Infrastructure Technology } Private Bag 33 Clayton South MDC } Victoria, 3169 } tel: +61 3 9545 2012 } fax: +61 3 9544 1128 } } Visit our Web site http://www.cmst.csiro.au } } Shipping address: CSIRO - Manufacturing & Infrastructure Technology, Gate 4 } Normanby Rd. Clayton, Victoria, } } PLEASE NOTE: } } To the extent permitted by law, CSIRO does not represent, warrant and/or } guarantee that the integrity of this communication has been maintained or } that the communication is free of errors, virus, interception or } interference. } } The information contained in this e-mail may be confidential or privileged.
} Any unauthorised use or disclosure is prohibited. If you have received this } e-mail in error, please delete it immediately and notify George Theodossiou } on +61 3 9545 2012. Thank you. } } }
Does anybody knows what is the acceptable variation in the extractor current on a LEO 1500 FE-SEM? Is it expected to observe a variation of the current for a Schottky FE-tip? Or might it be some external field? I am setting up a monitoring log now to see if this variation is periodic.
Our observed variation is about 3%.
Thanks in advance Sara
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} A histology question for the light microscopists out there: Are Haversian } systems present in the flat bones formed by intramembranous ossification or } are they only in long bones? Thanks, Tom
Tom:
Haversian systems are present in bone formed both ways. Haversian systems may be absent in very thin trabeculae of bone. In that case nourishment by diffusion through canaliculi is sufficient since the thickness of the trabecula is not greater than the diameter of a Haversian system.
Geoff
} Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I have a geriatric JEOL 100C TEMSCAN that's older than dirt. It is under service contract with JEOL and the JEOL crew here in the Houston area have been phenomenally good about keeping it operating and operating well. They respond very quickly, at least within 24 hours. Now, that may be because we are in a large medical school in a large medical center and they have a lot of instruments in the area and are generally close by. It could also be because they just wander by my lab in a pavlovian reflex. I think they also understand that I run a diagnostic lab and patients don't stop getting sick just because Cartwright's scope is down. I am very happy with the service from JEOL.
Joiner Cartwright, Jr., Ph.D. Baylor College of Medicine Dept. of Pathology Houston, Texas U.S.A.
P.S. I have no financial ties to JEOL America, nor do I own stock in that company. However my car is parked on the 5th floor of Garage #6 today with the window down just enough for a plain brown envelope to slide through.
************************************************
At 06:03 PM 03/06/2003 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have any thoughts about the advantages/disadvantages of the use of various types of embedding molds (I'm referring to the substances from which they're made). If there's a preference for a certain type of material, then why?
We use Durcupan and EMbed/Araldite resins. I know that glycol methacrylate will not polymerize correctly in silicone molds, but does well in polyethylene. Other than that example, are there any problems in which the resin reacts with the mold material in any way?
Our "clear" silicone molds eventually turn opaque, with it happening faster around cavities that are used more often. Is this due to oven heat or the resin?
Thank you,
J.M. Lett Harold W. Siebens Hearing Research Center Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
My bosses are interested in sending me to a short course or workshop in digital imaging and image analysis. In particular they are looking for a workshop or short course which would be using Image-Pro Plus and Photoshop. Also geared toward biological EM applications. Anyone out there offering a short course or workshop along these lines?
Tom Bargar Core Electron Microscopy Research Facility 986395 Nebraska Medical Center Omaha, Ne 68198-6395
Hello George, When I was a graduate student at the University of Kansas, I was taught to spray the embedding molds with a teflon spray as one would do to a frying pan (I forget the brand name of the stuff). But for 15 years at the University of Iowa I have not done this and have had no problems removing blocks from "good" molds. When the molds begin to crack and the blocks become difficult to remove, I discard the mold. I have never experienced a sudden or overnight problem as you describe. Good luck.
Dean Abel Biological Sciences 141 BB University of Iowa Iowa City IA 52242-1324 USA
At 05:21 PM 3/7/2003 +1100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
AFter three Zr/O-W Schottky guns, my experience suggests that 3% variation is OK. I log Iext twice per month and perform gun brightness measurment every two months. I also record IPG and IPC pump currents.
These types (my type?) tend to rise in Iext after install and then stabilize at some certain current. At 92uA, mine varies by no more than +- 2uA. When the Iext starts to change more than this, that is an indicator that the gun is dying. Along with higher or lower Iext, gun chamber vacuum will deteriorate IPG will increase). When this happens, it's about two weeks to a totally dead gun. Typical life is about two years. My last one lasted exactly two years two months. some folks report half that lifetime. Keeping good vacuum and proper protocol for protecting the gun saves the cost of a new gun....or at least, puts it off longer.
gary g.
At 04:41 AM 3/7/2003, you wrote:
} Hello All } } Does anybody knows what is the acceptable variation in the extractor } current on a LEO 1500 FE-SEM? Is it expected to observe a variation of } the current for a Schottky FE-tip? Or might it be some external field? I } am setting up a monitoring log now to see if this variation is } periodic. } } Our observed variation is about 3%. } } Thanks in advance } Sara } } } -- } This message has been scanned for viruses and dangerous content by } MailScanner, and is believed to be clean. } } "The CSIR exercises no editorial control over E-mail messages and/or } attachments thereto/links referred to therein originating in the } organisation and the views in this message/attachments thereto are } therefore not necessarily those of the CSIR and/or its employees. } The sender of this e-mail is, moreover, in terms of the CSIR's Conditions } of Service, subject to compliance with the CSIR's internal E-mail and } Internet Policy."
I never used to have the problem with molds cracking or sticking to the resin but about 3 years ago it became a common occurence in my lab. I thought at first the supplier had changed the formulation but it was true from more than one supplier. Teflon sprays - available at your hardware store - help some but not much. I think using BDMA rather than DMP-30 in your epoxy resin mix makes it worse but that is not something I have carefully examined. Recently I switch to polyethylene "disposal" molds and they work pretty well - I re-use them but have only done this 3-4 times so I don't know what their lifespan will be.
} Hello George, } When I was a graduate student at the University of Kansas, I was } taught to spray the embedding molds with a teflon spray as one would do } to a frying pan (I forget the brand name of the stuff). But for 15 years } at the University of Iowa I have not done this and have had no problems } removing blocks from "good" molds. When the molds begin to crack and the } blocks become difficult to remove, I discard the mold. I have never } experienced a sudden or overnight problem as you describe. Good luck. } } Dean Abel } Biological Sciences 141 BB } University of Iowa } Iowa City IA 52242-1324 } USA } } } At 05:21 PM 3/7/2003 +1100, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
George, It sounds like your rubber mold has had one to many cures-the same happened to me with old rubber molds and Eponate 12.
Mike D ----- Original Message ----- } From: "George.Theodossiou-at-csiro.au"-at-sparc5.microscopy.com
Perfect timing. I was just about to post the following on this list. I think it exactly corresponds to your needs. We use Photoshop as a platform for the course because of its familiarity to a wide user base, but all of the plugins and techniques supplied are also fully compatible with Image Pro Plus. typically about 40% of the attendees are from the biological research community (other disciplines represented include materials science, medicine, forensic applications, geology, industrial imaging, etc.).
John Russ ===
The three-day intensive hands-on workshop on Image Processing and Measurement presented by John Russ (author of "The Image Processing Handbook" and "Practical Stereology") through the North Carolina State University Department of Continuing and Professional Education is now in its 21st year. The course dates for 2003 are May 21 - 23 in Raleigh, NC. This course has generated highly favorable reviews from the thousands of previous students. The primary focus is on images from various types of microscopy, with practical guidance in correcting imaging defects, enhancing the images for presentation and measurement, and performing stereological meaningful measurements on them. Textbooks and computer software are provided to attendees. Lab sessions with an opportunity to bring your own images makes this course immediately useful and highly productive.
For full information on the course, including outlines, faculty information, a downloadable brochure, and on-line registration, go to
http://members.aol.com/ipcourse
Class size is limited to maintain a high ratio of instructors to students, so make your reservation now. You may also contact Cindy Allen at NCSU Continuing Education, at 919-515-8171 ====
In a message dated 3/7/03 12:46:15 PM, tbargar-at-unmc.edu-at-sparc5.microscopy.com writes:
} My bosses are interested in sending me to a short course or workshop in } digital imaging and image analysis. In particular they are looking for } a } workshop or short course which would be using Image-Pro Plus and Photoshop. } Also geared toward biological EM applications. Anyone out there offering } a } short course or workshop along these lines?
George, You can use a TFE release agent to extend the life of your moulds, but the repeated heating, over time, causes the material to breakdown...what you experienced is pretty much inevitable. Moulds don't last forever. 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
Joiner, I don't know who you are, but I like your sense of humor. I'm thinking of trying the plain brown envelope trick with Leica and Philips. Let me know if it works. Mary Gail Engle
At 09:39 AM 3/7/03 -0600, Joiner Cartwright, Jr., Ph.D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
I would like to hear opinions or experiences using a Peltier cooled cryo-stage for SEM. Is this appropriate for high vacuum FEGSEM?
Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295
Dear George, I always use a silicon release agent (a spray) when I embed samples. Sometimes they release, sometimes they don't or they take chunks of the mold with them. The molds have a life, but I make my own out of silicon rubber, so I just make some more. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {"George.Theodossiou-at-csiro.au"-at-sparc5.microscopy.c om} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 06, 2003 10:21 PM
We are looking for a technician with experience in clinical electron microscopy. Position Available immediately. Please contact me if you are interested know anyone who might be.
Lois Anderson Johns Hopkins University Dept. of Pathology Laboratory Manager Electron Microscopy/Immunofluorescence 600 N. Wolfe Street/Pathology 709 A Baltimore, MD 21287 (410) 955-2861/fax (410) 614-7110 landers-at-jhmi.edu
I was taught that going to Spot Size 1 in TEM gives more light at the expense of more beam damage to the specimen and lower resolution. In addition, I was told that one uses Spot Size 2 or 3 for high resolution work. Can one of the TEM experts out there expand on these simple statements. At what magnification does going to Spot Size 1 really begin to hurt resolution - if i am working at 20K, should it make a difference? I don't see much difference except in the amount of light. Comments and personal prejudices will be warmly accepted. If it makes any difference, I am working with conventional epon thin sections of biological material that was osmicated and UA and Pb counterstained; most photos are 20K or less but can sneak up to 40K or 50K. Thanks.
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Hi, I have two questions for image processing experts. A) how to change a gray scale image into a false color image. If the intensity of a gray scale image is in the range of 0 - 255, red, orange and blue color pixels in the corresponding color image should represent the pixels with intensity in the range of 0-100, 101 - 200, and 201-255 in the gray scale image, respectively. B) How to emerge three gray scale images into a color image. If the intensity of gray scale images is in the range of 0 - 255, red, orange and blue pixels in the color image should represent the pixels with intensity of 201-255 in the three gray scale images, respectively. Could you please tell me which software can solve the problems and how to do them? Your help is highly appreciated. Thanks, Jian-Guo
*********************************** Jian-Guo Zheng Research Assistant Professor Materials Science & Engineering Manager, NUANCE-EPIC http://www.nuance.northwestern.edu/epic/
Northwestern University 2220 Campus Drive, 1156 Cook Hall Evanston, IL 60208-3108, USA Phone: (847) 491-7807, Fax: (847) 491-7820 E-mail: j-zheng3-at-northwestern.edu http://www.nuance.northwestern.edu/epic/zheng/index.htm ***********************************
The problem of blocks sticking to the moulds might be the mould itself. They vary greatly in quality. We had to give up on one supplier because the moulds would start to deteriorate after only two or three usages. We are quite happy with the moulds supplied by Ted Pella (I have no financial interest in the company), but even these vary from batch to batch.
Ralph Common Michigan State University Division of Human Pathology
-----------------
Dear Colleagues,
A couple of days ago I embedded some samples for microtomy, using Procure 812. Today when I went to remove the blocks from the mould I found that the mould material had adhered to the blocks and came away with the blocks, thus destroying two mould blocks. I have never experienced this problem in the past to such a degree.
The resin was measured and mixed properly and cured at 60 deg celcius. When embedding I half fill the mould and cure it in the oven at 60 deg celcius for an hour, then position the sample, fill the rest of the mould and back in the oven for final curing. This has worked in the past for me.
Does anybody have a better method of removing the cured blocks and not destroying the moulds, use a release agent etc etc??
Your help would be greatly appreciated.
Regards George
George Theodossiou Physicist / Electron Microscopist CSIRO Manufacturing & Infrastructure Technology Private Bag 33 Clayton South MDC Victoria, 3169 tel: +61 3 9545 2012 fax: +61 3 9544 1128
Dear Colleagues, Most of grids are made of metals. Can anyone tell me if there are ceramic grids/slots/holes? Many thanks, Rong Yu
-- ************************************* Rong Yu Ph.D. Materials Science Division Lawrence Berkeley National Laboratory University of California Berkeley, CA94720, USA Phone: 1-510-486-6809 Fax: 1-510-486-7768 *************************************
The larger spot, the worse spatial coherence, and thus lower resolution for "High-Resolution" imaging. But such difference in resolution is hardly be detected at lower magnifications, e.g. 20k-50k.
Tom Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I was taught that going to Spot Size 1 in TEM gives more light at the } expense of more beam damage to the specimen and lower resolution. In } addition, I was told that one uses Spot Size 2 or 3 for high resolution } work. Can one of the TEM experts out there expand on these simple } statements. At what magnification does going to Spot Size 1 really begin to } hurt resolution - if i am working at 20K, should it make a difference? I } don't see much difference except in the amount of light. Comments and } personal prejudices will be warmly accepted. If it makes any difference, I } am working with conventional epon thin sections of biological material that } was osmicated and UA and Pb counterstained; most photos are 20K or less but } can sneak up to 40K or 50K. Thanks. } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- ************************************* Rong Yu Ph.D. Materials Science Division Lawrence Berkeley National Laboratory University of California Berkeley, CA94720, USA Phone: 1-510-486-6809 Fax: 1-510-486-7768 *************************************
In Photoshop, convert your image to indexed color:
Image, Mode, Indexed Color
In the Indexed Color dialogue box, choose:
Palette, Custom
You will see a grid of 256 squares. Click and drag over the first 85 squares and a color picker box will appear. Set those squares to the color you want by setting both the beginning and end square of the 85 squares to the same color. Do this three times for the three colors you want. Think about saving the indexed color profile so you don't have to do it again.
For your second question, I believe the Channels, Merge feature should be used. No time for details now but review the PhotoShop manual for this feature. We routinely use it to reconstruct 3 channel false color images from the confocal.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
} Hi, } I have two questions for image processing experts. A) how to change a } gray scale image into a false color image. If the intensity of a gray } scale image is in the range of 0 - 255, red, orange and blue color } pixels in the corresponding color image should represent the pixels with } intensity in the range of 0-100, 101 - 200, and 201-255 in the gray } scale image, respectively. B) How to emerge three gray scale images into } a color image. If the intensity of gray scale images is in the range of } 0 - 255, red, orange and blue pixels in the color image should represent } the pixels with intensity of 201-255 in the three gray scale images, } respectively. } Could you please tell me which software can solve the problems and how } to do them? Your help is highly appreciated. Thanks, } Jian-Guo } } } *********************************** } Jian-Guo Zheng } Research Assistant Professor } Materials Science & Engineering } Manager, NUANCE-EPIC } http://www.nuance.northwestern.edu/epic/ } } Northwestern University } 2220 Campus Drive, 1156 Cook Hall } Evanston, IL 60208-3108, USA } Phone: (847) 491-7807, Fax: (847) 491-7820 } E-mail: j-zheng3-at-northwestern.edu } http://www.nuance.northwestern.edu/epic/zheng/index.htm } ***********************************
I totally agree with Roger. You said, "etc. etc." I took this to mean you wanted a lot of help and info.
I use EPONATE 12 clone epoxy and the stuff releases fine when new PE molds are used. I have used flat silicone? embedding molds colored green and blue. This gets to be expensive when they give out. When they get old, the silicone rubber sticks as streaks on the blocks and wrecks the mold for any further use. They work fine for a time, just like the PE does. I have used my PE disposable molds, below, for over 8 years.
I use cheap Wheaton Bottle PE caps that can be purchased separately in different size diameters. (see below) I like a flat mold over the 'vertical' BEEM capsules that you can buy because of the types of samples I do and control of sample geometry. Anyway, after 5-10 uses, the epoxy sticks so bad that the polyethelene has to be trashed to get the round block out. I just throw the cheap mold away and use a new one. Hint: Just before the PE is going to stick, some crazing and slight sticking seems to take place. Also a thin or thick white area in the PE after removal signals the next embedding will be very hard to remove.
I believe the PE has some plasticizer on or near the surface that acts as a release agent. I could be wrong but there is something on the surface, in any case.
I use this large low profile mold because I do a lot of automotive coatings that contain TiO2 crystals and the larger molds are easier to pop the block out of. TiO2 trashes a diamond knife faster than most other things I run. This costs money and uses up diamond knife edge needlessly. So once I release a coil coat, 5 µm soda can ink or thicker 'mil' coating, I pin it down flat with straight pins that stick into the PE bottle caps and that hold the corners of the coating. Why use the pins? Sometimes the heat cure can cause the samples to curl during the curing and that also translates into excessive diamond knife edge useage and other problems in the TEM. I don't have to worry about a small sized sample being submitted. (The metal panels I get can be 10's of sq. inches in size.) If you only put a small amount of epoxy in at first, the raised number in the cap mold allows a thin film of epoxy to wick under the film even after pinning. After wetting, you can add the rest of the epoxy. This totally surrounds the embedded film or coating with epoxy. I saw the cured block with a X-ACTO RAZOR saw. It's faster but messy. Anyway, this flatness allows me to orient the straight and planar sample in the microtome so the smallest amount or profile is presented to the diamond knife edge used to thin section the coating for the TEM or for staining.
So why don't I use the small silicone molds? What I want to do is select the best flat area in the block that has the plane of the coating co-planar with the bottom of the mold. This also makes the coating co-planar with the jaws of the microtome vise mount on my RMC 6000 XL. This ensures that the diamond thin section cuts will be rectangular and not skewed. This is very critcal to the EDS examination of anti-reflective coatings on polymers.
These caps are listed in the regular Wheaton Catalog and can be ordered from Fisher Scientific using the Wheaton cap number.
I have never used Procure 812 but most clones of EPON 812 are the same. I have tested the ORIGINAL EPON 812 against Medcast (no longer available from Pella) and their Eponate 12 by having the NMR spectra run. Yes I still have a bottle of real EPON 812 from Shell ! NMR is not that sensitive as an analytical assay technique but the spectra match very close. The original stuff is thicker than Eponate 812 or the old Medcast. Obviously the molecular weight is now different in these newer clones. One other embedding kit I tested had minor levels of aromatics, FYI.
For cutting or trimming epoxy, use extra long Weck SS Razor Blades. They cost more but the pressure needed to be applied to cut the epoxy is 3-5 times less than a regular razor blade. This makes them safer to use from this stand point only. THESE blades CUT TISSUE like your FINGERS a lot easier also! They are extremely sharp and dangerous to use. They make a new regular razor seem dull. Take heed!!!! Never place your fingers or anything else below the edge of these blades.
I hope this helps give you some perspective and an overview of some hints on how others use Epon 812 clone epoxy and about silicone versus PE molds.
Paul Beauregard Senior Research Associate PPG Industries Monroeville Chemicals Technical Center 440 College Park Drive Monroeville, PA 15146 724-325-5131
} } George: } My experience has been that the moulds age and that with repeated usage tend to } start adhering to the cured blocks. At that point it's time to purrchase new } moulds. I have had no success with release agents, etc. } Roger Moretz, Ph.D. } Dept of Toxicology } BI Pharmaceuticals } Ridgefield, CT } -- } } } Dear Colleagues, } } } } A couple of days ago I embedded some samples for microtomy, using Procure } } 812. Today when I went to remove the blocks from the mould I found that the } } mould material had adhered to the blocks and came away with the blocks, thus } } destroying two mould blocks. I have never experienced this problem in the } } past to such a degree. } } } } The resin was measured and mixed properly and cured at 60 deg celcius. When } } embedding I half fill the mould and cure it in the oven at 60 deg celcius } } for an hour, then position the sample, fill the rest of the mould and back } } } in the oven for final curing. This has worked in the past for me. } } } } Does anybody have a better method of removing the cured blocks and not } } destroying the moulds, use a release agent etc etc?? } } } } Your help would be greatly appreciated. } } } } Regards } } George } } } } George Theodossiou } } Physicist / Electron Microscopist } } CSIRO Manufacturing & Infrastructure Technology } } Private Bag 33 Clayton South MDC } } Victoria, 3169 } } tel: +61 3 9545 2012 } } fax: +61 3 9544 1128
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mwb142-at-psu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, March 7, 2003 at 12:42:42 ---------------------------------------------------------------------------
Email: mwb142-at-psu.edu Name: Matt Bell
Organization: Penn State
Education: Graduate College
Location: State College, PA
Question: I know that there is a Sparrow approximation for resolution of an optical wave. Does that apply for acoustic waves? And if so, what is the formula for the Sparrow approximation for lateral and vertical resolution?
What you are suggesting is not the only way. Of course you can create a false color image, but you can just as easily create a full color image by assigning each of the three colors to one color channel of the full color (24-bit) image. If you really do need a false color image, you can simply convert this image. The results are likely to be better as any reasonable software will optimize the color palette.
As for software, look for software that supports Fluorescence imaging (such as, but not limited to, our analySIS software). Your question is a standard problem in fluorescence, as often a b/w camera is used (because of higher sensitivity) to acquire several fluorescence channels and merge them into a color image. Often it is more than 3 colors, which makes it more complicated, but it is still a standard technique (multiple Fluorescence) for many software packages.
Please contact me offline if you want more information.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Jian-Guo Zheng [mailto:j-zheng3-at-northwestern.edu] Sent: Friday, March 07, 2003 4:56 PM To: Microscopy-at-sparc5.microscopy.com Cc: 'Jian-Guo Zheng'
Hi,
We have 3 SEM's and 2 TEM's in our facility with varying costs and amounts of use associated with each. Using the Cost/Hours of Use=Rate leaves us with scopes having unappealingly high rates simply because they aren't used much or have higher annual costs. Your heads are nodding in agreement now, right?
What can't I use an "average" rate for SEM's and TEM's that (somewhat) smooths out those irregular use and costs patterns? Of course I can, right? Well, in truth it would help me to be able to tell Research Accounting that others are doing it too :} ).
Is anyone else doing this or something like it?
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Thank you for all the replies and useful hints, for release agents and different types of moulds.
I understand that moulds have a finite life but what really annoyed me about this instance was that the moulds were white silicone rubber, had only been used 3-4 times previously and had very few cracks. I thought they had a bit of life left in them. Oh well, it just makes work more fun.
Regards George
George Theodossiou Physicist / Electron Microscopist CSIRO Manufacturing & Infrastructure Technology Private Bag 33 Clayton South MDC Victoria, 3169 tel: +61 3 9545 2012 fax: +61 3 9544 1128
To the extent permitted by law, CSIRO does not represent, warrant and/or guarantee that the integrity of this communication has been maintained or that the communication is free of errors, virus, interception or interference.
The information contained in this e-mail may be confidential or privileged. Any unauthorised use or disclosure is prohibited. If you have received this e-mail in error, please delete it immediately and notify George Theodossiou on +61 3 9545 2012. Thank you.
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what is the timescale of these variations? In case seconds, then there is something wrong with your filament ... this may be fixed with an bakeout and a "reconditioning": trying to find a new extractor voltage and filament current is usually the last measure before exchanging the filament I think the DENKA's behave a bit different compared to the one's from FEI ...
There is also some further reading which may be helpful for understanding: F. Zhou:"Handbook of charged particles optics", pp.77-102 "A Review of the ZrO/W Schottky Cathode", CRC Press
But the main point to check the status of your GEMINI is the probe current: With a 30µm aperture -at- 1kV our's have been at 90..130pA (...as long as the aperture is o.k.). The DENKA we are using right now has about 5500h and has been at more than 180µA extractor current (...EHT on) more or less form the beginning!!! On the second to minute timescale the variations are definitely below 0,5%(!!!) ... o.k.: we are doing only LV-morphology work (...no EDX/WDX) ... and we do not change the extractor voltage from the set working point of 6KV for more than +/- 400V for longer than 4h ... IGP vac always better than 2x10^-9Žmbar
If the variation (... always talking about the time after having reached the saturation, that means 4..6h after switching on the gun!!!) is periodically on a minute to hour scale, there is something wrong with the ZrO droplet which should wet the tip of the W single crystal ...
Hope this helps a bit?
Gunnar
Date sent: Fri, 07 Mar 2003 10:02:19 -0800 To: "Sara Prins" {SPrins-at-csir.co.za} } From: Gary Gaugler {gary-at-gaugler.com}
Owen, I ran into the same problem when we set up our rate structure last year. Accounting wanted each scope treated separately based on it's use and service contract costs. This amounted to vastly different rates for our three scopes. We intend to go back and argue for the need to treat all as one unit and combine maintenance costs plus use hours to come up with a reasonable hourly rate for all. Our internal people are fortunate in that the rate will be heavily subsidized for most of them, make it very affordable. It will affect external users and some unsubsidized university users the most but should end up being more fair overall than the way it is now. Hopefully this will be accepted by the university accounting office.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 3/9/03 4:35 PM, "Owen P. Mills" {opmills-at-mtu.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, } } We have 3 SEM's and 2 TEM's in our facility with varying costs and amounts } of use associated with each. Using the Cost/Hours of Use=Rate leaves us } with scopes having unappealingly high rates simply because they aren't } used much or have higher annual costs. Your heads are nodding in } agreement now, right? } } What can't I use an "average" rate for SEM's and TEM's that (somewhat) } smooths out those irregular use and costs patterns? Of course I can, } right? Well, in truth it would help me to be able to tell Research } Accounting that others are doing it too :} ). } } Is anyone else doing this or something like it? } } Owen } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills } } } }
I can not answer your specific question about whether others are using average cost. However, as a recent graduate from business school, and one who three times attended summer hockey camp at your university (Go Huskies!) I offer some input that I hope you find helpful.
Accounting costs are an approximation of opportunity cost, and to the extent that they reflect opportunity cost they serve as a fair measure for transfer pricing (of services purchased within the organization) or a useful measure for external pricing (of services sold outside the organization).
Opportunity cost indicates the value that you might extract from your tool given the options that you have. Options may include performing research, charging an external customer, and selling the tool. In an extreme, if you choose one option, others are foregone. If you strictly perform research, for example, your opportunity cost is the difference between the present value of performing that research and the greater of charging external customers or selling the tool. You might have reason to argue that you are doing the right thing and to do so by stating alternatives, and noting that those alternatives provide less value than your desired use.
Opportunity cost may be the marginal cost of operating a tool. Marginal costs would be those costs that occur only when you turn the tool on, excluding those costs which are otherwise committed to, such as purchase price of the tool and cost to maintain the building. Considering another extreme, it may be that your highest and best use of this tool and the facility would be to rent it out at a fee merely greater than the cost of electricity to run the tool.
In contrast to opportunity costs of today, your organization's accounting costs are probably what is required to recover dollars already spent to purchase the unit and to operate the facility that it houses it. Indeed, this would reflect the opportunity cost at the time the tool was purchased. However, such accounting may reflect little of today's economic situation and in the worst case it leads to failure to extract the most possible value from the tool.
While you may find standard practices to support use of "average pricing", I hope that you might also consider opportunity cost as a method for both decision making as well as persuasion. Finally, I recommend an easy reading novel that provides an understanding of standard accounting practices and suggests a framework for extracting maximum value from an organization's assets, "The Goal", by Elijahu M. Goldratt.
Sincerely,
John Moore Montara Industries New Hill, North Carolina 919-434-8457
"Assembling a team to provide 3rd party support for the Applied Materials SEMVison."
----- Original Message ----- } From: "Owen P. Mills" {opmills-at-mtu.edu} To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, March 09, 2003 4:35 PM
Hi,
We have 3 SEM's and 2 TEM's in our facility with varying costs and amounts of use associated with each. Using the Cost/Hours of Use=Rate leaves us with scopes having unappealingly high rates simply because they aren't used much or have higher annual costs. Your heads are nodding in agreement now, right?
What can't I use an "average" rate for SEM's and TEM's that (somewhat) smooths out those irregular use and costs patterns? Of course I can, right? Well, in truth it would help me to be able to tell Research Accounting that others are doing it too :} ).
Is anyone else doing this or something like it?
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
The Connecticut Microscopy Society is pleased to present Dr. Pietro De Camilli on Thursday, March 27 at the Peabody Museum at Yale University. Dr. De Camilli will describe his work on synaptic transmission. A reception and dinner precede the lecture. Information is on our website, http://www.connms.org We hope to see you there!
Elizabeth Cowles, President of the Connecticut Microscopy Society
Elizabeth A. Cowles, Ph.D. Associate Professor, Department of Biology 220 Goddard Hall Eastern Connecticut State University 83 Windham St. Willimantic, CT 06226 voice: 860-465-4385 fax: 860-465-5213 email: cowlese-at-easternct.edu home page: http://www.easternct.edu/personal/faculty/cowlese/index.html http://biology.easternct.edu
To amplify what Geoff has said slightly, I would only add that "FIRST" dermal bone is often called "woven bone" (non-lamellar in its initial structure) and may even become known as "spicular or trabecular" bone thereafter before it becomes remodeled as lamellar bone. The reason I mention this is that if one is looking at embryonic/fetal bone, then one may not see haversian systems or periosteal or endosteal lamellar bone at the time of sampling. Furthermore, there are even some small fish whose bone is actually acellular in the adult (as I recall dimly from coursework taken years ago). Some of the smaller sesamoid bones may also lack haversian and/or lamellar bone, although without looking I would bet that either or both of my patellas do. The haversian system is considered the fundamental unit of bone remodeling and is often referred to as an 'osteon'.
It would be relatively easy to distinguish between the lamellae of periosteal and endosteal bone from those in haversian bone. Indeed, one can often observe between periosteum and endosteum the arcs of remnant older haversian systems between the newer, uninterrupted osteons.
It would also be worthwhile mentioning that the axes of collagen fibrils are parallel within a single lamella and opposing in adjacent lamellae [mindful of micelles of cellulose in adjacent layers of wood growth] and not described as being paraxial with the long axis of a long bone. Thus, perfect transections of long bone viewed with the TEM will never show transverse sections of collagen micells without some tilt.
The most recent really good basic histology book is the 12th edition of Bloom and Fawcett, published in 1994 (ISBN: 0-412-04691-1). There should be some of this latest in a classic series of higher-end biology books available on the used book market. I got mine as new just last year.
Hope this helps too,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging
Mail to: Geology, CASI West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383
For help and information only, The CASI houses: An FEI Quanta 400 and Technai 12T, Oxford INCA Energy 400, Tousimis AutoSamdri 815 and Olympus FV-300.
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Friday, March 07, 2003 10:00 AM To: Tom Phillips Cc: Microscopy-at-sparc5.microscopy.com
Tom Phillips wrote:
} A histology question for the light microscopists out there: Are Haversian } systems present in the flat bones formed by intramembranous ossification or } are they only in long bones? Thanks, Tom
Tom:
Haversian systems are present in bone formed both ways. Haversian systems may be absent in very thin trabeculae of bone. In that case nourishment by diffusion through canaliculi is sufficient since the thickness of the trabecula is not greater than the diameter of a Haversian system.
Geoff
} Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- ********************************************** 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 **********************************************
On Friday, March 7, 2003, at 04:55 PM, Rong Yu wrote:
} The larger spot, the worse spatial coherence, and thus lower } resolution for } "High-Resolution" imaging. } But such difference in resolution is hardly be detected at lower } magnifications, e.g. 20k-50k. } } Tom Phillips wrote: } } } I was taught that going to Spot Size 1 in TEM gives more light at the } } expense of more beam damage to the specimen and lower resolution. In } } addition, I was told that one uses Spot Size 2 or 3 for high } } resolution } } work. Can one of the TEM experts out there expand on these simple } } statements. At what magnification does going to Spot Size 1 really } } begin to } } hurt resolution - if i am working at 20K, should it make a } } difference? I } } don't see much difference except in the amount of light. Comments and } } personal prejudices will be warmly accepted. If it makes any } } difference, I } } am working with conventional epon thin sections of biological } } material that } } was osmicated and UA and Pb counterstained; most photos are 20K or } } less but } } can sneak up to 40K or 50K. Thanks. } } Dear Tom and Rong, Another consideration for very high mags is that the closer the beam is to crossover, the poorer the resolution, so one may want to use a smaller spot size number (actually, bigger spot) further from crossover rather than a bigger spot size number at crossover. Also, your specimen may be better behaved vis-à-vis charging/heating induced beam movement if a larger area is illuminated. As you said, Rong, all this is irrelevant at 20k-50k. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Just some last minute comments from a long-time mold manufacturer:
Standard blue silicone rubber molds - These are quite pliable. We've found the life span is mainly dependent upon the number of cures and components of the epoxy mixture. Over-curing at high temps can also limit lifespan.
Clear silicone rubber molds - These require a different silicone formulation. They do tend to crack when bent. Obviously you have to balance this with the bottom illumination advantage.
We've looked at other rubber molds manufactured in the U.S. and most use pretty good material. Our labs do use our TFE spray which helps in certain situations and it is now environmentally-friendly.
We also do a lot of custom silicone molds and some are quite large. These may require modification of curing procedures.
John Arnott Disclaimer: Ladd sells EM supplies and accessories
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
Does anyone know of an opaque white resin that can be used for tissue infiltration? We want to progressively section and photograph the block face
Thanks Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University, Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 http://www.anu.edu.au/EMU
Is anyone using an automatic negative/film developer for TEM negatives that they are happy with? Our EM Lab is looking for an auto developer that would develope Technical Pan Film TP-120 (a roll of approx 8 negatives that is taken with a Zeiss TEM 109). Manufacturer and price will be most helpful. Any imput will be appreciated. Many thanks to all who respond or take precious time to read. Teresa
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rajmeister-at-msn.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, March 10, 2003 at 22:49:28 ---------------------------------------------------------------------------
Email: rajmeister-at-msn.com Name: Raj
Organization: Mary Washington College
Education: Undergraduate College
Location: F'brg, VA, USA
Question: Hello, My question is, what do you think about the Observer IV microscope overall for its price($265)? And also what is the procedure for preparing an oil immersion slide for use at 1000x? Thanks
We are the latest folks to get caught by the "New Formulation" TEM films. We had a couple of batches come out of our nitrogen-burst processor looking like they'd been tossed into a vat of developer and left untouched for a couple minutes. The negs were way light and mottled with streaks and patches indicative of poor agitation. Unfortunately, I didn't even realize we had any of this film in our inventory, since we hadn't bought any since before the controversy hit the listserver, so our poor student lab assistant got the blame.
When we finally hit on the cause of this, we were able to remedy it by increasing our agitation cycle to 2 seconds every 10 seconds, instead of 2 seconds every 30 seconds (which worked beautifully with the old film). We use a 4-minute development in d-19 at 68-70 degrees F, and we may move to 5 minutes. The agitation marks have been eliminated and density is back into the good range, although still a bit lighter than before. We had also started to get a "thumbprint" mark on the bottom of the negatives which puzzled us greatly until our faculty coordinator pointed out that it coincided exactly with the round plastic rod that the negatives rest on at the bottom of the rack. It was causing increased local agitation in a half-oval pattern as the nitrogen bubbles swirled around it. Lifting the negatives up slightly seems to have cured this. For some reason we had never noticed it before.
In case you were curious, the student lab assistant has been duly apologized to and was very gracious in his victory.
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! 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/
Consult a professional photographer who is familiar with the type of job you are doing. Proper lighting coupled with a specific film and processing may allow tissue in a paraffin or epoxy block to stand out from the background. You might also try mixing something into your embedding material to make it opaque.
Geoff
Sally Stowe wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone know of an opaque white resin that can be used for tissue } infiltration? We want to progressively section and photograph the block } face } } Thanks } Sally } } Dr Sally Stowe } Facility Coordinator, ANU Electron Microscopy Unit } Research School of Biological Sciences } Australian National University, Canberra ACT0200 } AUSTRALIA } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } http://www.anu.edu.au/EMU
-- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I am looking for a statement of acceptable water quality for TEM usage. This may be familiar to those of us who are *blessed* to be inspected by CAP. All I know about water quality is when it's NOT! Does anyone have a statement that they use that they would be willing to share?!
Thanks,
Karen Bovard EM Lab Creighton University Medical Center Omaha, Nebraska
The Oklahoma Microscopy Society and the Central States Microscopy and Microanalysis Society will be holding a joint meeting in Tulsa, Oklahoma on April 10-11, 2003. The theme of tis meeting is "The Art of the Science Image."
Thusday, April 10 Session I: Image Creation 9:00-9:50 Opening Address: "Images and Human Understanding", John Russ, NCSU
10:00-10:45 "A Review of X-Ray Mapping", John Friel, PGT
10:45-11:00 BREAK
11:00-12:00 "Spectrum Imaging: The Next Step in Microanalysis", Paul Kotula, Sandia Labs
12:00-1:00 LUNCH
1:00-1:30 "Winning the Depth of Field Battle in Light Microscopy", Jim DeMian, 3M Co.
1:30-2:30 "Specialized Methods in Light Microscopy", Robert Weaver, McCrone Inst.
Evenong Reception - IMAX Theatre 6:00-7:00 Buffet Dinner 7:00-8:30 "Four Million House Guests" a.k.a. "The Hidden Dimension", 3D movie and Keynote Addrerss: "SEM Stop-Frame, Color, 3D Animation for Motion Pictures", David Scharf, Scarf Photography
Friday April 11 Session II: Image Processing and Analysis 8:30-12:30 "Guide to Image Analysis Workshop", John Russ, NCSU
12:30-1:30 LUNCH
Session III: Image Presentation 1:30-2:15 "Basic Principles of Image Composition for Scientists", TU Art Dept.
2:15-2:30 BREAK
2:30-3:30 "Exploring the Invisible: the Cultural Impact of Microscopy", Lynn Gamwell, SUNY
Special Features Corporate Exhibitors Demonstrations of image processing software Region-wide "Art of the Science Image" competition Art Gallery displays "Images from Science", Rochester Inst. of Tech and from David Scharf
Hotel Accommodations Renaissance Tulsa, the city's newest hotel, located across from the IMAX theatre, special meeting rate of $84 per night for single or double; complimentary pool and fitness facilities; complimentary shuttle from airport and to nearby shopping mall.
Registration In advance ( {3/31), $40 for members, $50 for non-members, $20 for students On-site (} 3/31), $50 for members, $60 for non-members, $25 for students Thursday night reception: $20 note: non-member registration includes a 2003 membership to either OMS or CSMMS
For more information or to receive a registration packet, contact Lou Ross at rosslm-at-missouri.edu or at (573) 882-4777.
-- Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211-5120 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.biotech.missouri.edu/emc
Leslie, We have been in the very fortunate position to have extensive university support to cover costs associated with this microscopy facility. The administration, primarily the School of Agriculture, has provided sufficient funds to cover service contracts on 3 EMs and additional funds for other operating costs. In addition salaries of two full-time professional level positions are covered by schools of Agriculture, Science, and Veterinary Medicine since this core facility caters primarily to researchers within those schools. This means that for many years we had no charges for use of the facility other than to cover consumables such as film. The facility encourages multi-user access but also does provide full service (there were always charges for service).
Under current financial stress, we have instituted a rate system to try to cover future increases in costs as well as to try to reclaim some of the current operating costs. Salaries are not figured into the recharge except for the service option. Administrators here understand that EM facilities do not make money...they are fortunate to recover most of their costs but even this is difficult if you want the facility used by the greatest possible number of researchers.
Now, that is the brief history behind the facility funding. It is true that you must charge identical amounts to all users of a facility if any are paying for costs using grant funds. You cannot preferentially exempt some people and supplement others. We use the subsidies to keep our rates low for all users. By doing this there is also the commitment by the administrators to continue the subsidies and not expect the facility to cover all associated costs. Everyone within the university who are doing sponsored research or unfunded research pay the same amount.
However, in some cases researchers do proprietary work on contract with private companies. In this case, they are not able to share the results with the research community so are not entitled to subsidized rates. At the present time they pay the full amount based on actual costs divided by use. Since the use is by university staff, no indirect charges are applicable. This also applies to companies who have direct ties to the university such as those who are industrial affiliates of departments or programs (in which case they are paying a fee for access to university staff and facilities).
Those companies who do not have formal ties to the university pay the unsubsidized rates plus indirect costs to the university to cover infrastructure costs. They also would pay consultant costs for assistance by university staff.
We have found so far that keeping rates low through the subsidies increases the number of users. Since the equipment does no good if unused, this is very desirable. In return, as the number of users increases, so does the revenue. The revenue then goes to "pay back" funds used for the subsidies. I would guess that we generate more overall revenue with the lower rates and more users benefiting from the facility than if we charged double the cost and user numbers fell accordingly. Its like anything else...if users perceive a bargain they will come...and often spend more than they originally intended because they feel they are getting value for their $.
Now, some equipment has been purchased by asking university faculty to contribute required funds. This is the case for some of our light microscopes and computer equipment. In this case, when these researchers agree to help fund facility equipment, they understand that access is on a first-come basis. In return the equipment is maintained and students are trained in the use by facility staff. The researchers individually put in much less money than if they were purchasing the entire instrument so we usually end up getting higher end equipment. There are no fees associated with using this jointly funded equipment.
Major equipment is funded almost entirely through instrumentation grants with required matches from the university. Researchers who assist in writing these grants understand that they are doing this partially as a service to the university community. They do not get preferential treatment if the instrument resides in the common core facility but usually end up using the equipment most since they had the research justification for it in the first place.
Devising a fair rate system is a complicated business and we have found that we need to clarify and revise as the system reveals the need.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 3/10/03 11:53 AM, "Leslie Eibest" {leibest-at-duke.edu} wrote:
} Hi Debby, } } I'm very interested in the method used there to subsidize some } users. Currently, we have a couple of departments that subsidize my salary } and the service contract, and those department members have free use of the } equipment and technical assistance. Outsiders were charged reasonable } rates. Everyone loved the system, and it provided a lot of stability for } the SEM lab. However, Sponsored Programs has come down on our necks, saying } that this unfairly subsidizes some grant holders and not others. All } federal grant holders should be charged the lowest rate. We will have to } raise more funds from user fees, so now we'll have to charge } everyone. Researchers and students without funding won't be able to use } the equipment. Adding to the fun, they won't let me keep reserves for } equipment repairs and upgrades. It's ironic, since NSF was pleased that } our lab was accessible to so many users. } } Have you had to deal with these issues yet? If so, how do you get around } the problems? } } Leslie } } } } } We intend to go back and argue for the need to treat all as one unit and } } combine maintenance costs plus use hours to come up with a reasonable hourly } } rate for all. Our internal people are fortunate in that the rate will be } } heavily subsidized for most of them, make it very affordable. It will } } affect external users and some unsubsidized university users the most but } } should end up being more fair overall than the way it is now. Hopefully } } this will be accepted by the university accounting office. } } } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-052 Whistler Building } } 170 S. University Street } } West Lafayette, IN 47907 } } } } } } On 3/9/03 4:35 PM, "Owen P. Mills" {opmills-at-mtu.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, } } } } } } We have 3 SEM's and 2 TEM's in our facility with varying costs and amounts } } } of use associated with each. Using the Cost/Hours of Use=Rate leaves us } } } with scopes having unappealingly high rates simply because they aren't } } } used much or have higher annual costs. Your heads are nodding in } } } agreement now, right? } } } } } } What can't I use an "average" rate for SEM's and TEM's that (somewhat) } } } smooths out those irregular use and costs patterns? Of course I can, } } } right? Well, in truth it would help me to be able to tell Research } } } Accounting that others are doing it too :} ). } } } } } } Is anyone else doing this or something like it? } } } } } } Owen } } } } } } Owen P. Mills } } } Electron Optics Engineer } } } Materials Science & Engineering } } } Michigan Technological University } } } Rm 512 M&M Bldg. } } } Houghton, MI 49931 } } } PH 906-369-1875 } } } FAX 906-487-2934 } } } mailto:opmills-at-mtu.edu } } } http://www.mm.mtu.edu/~opmills } } } } } } } } } } } } } }
Oops, sorry. My posting was about the 4469 film. Thanks for the reminder, Mike.
Randy
-----Original Message----- } From: Mike Coviello [mailto:coviello-at-mae.uta.edu] Sent: Tuesday, March 11, 2003 12:52 PM To: Tindall, Randy D.
To all, The Core Facility Management session at the annual Microscopy and Microanalysis meeting is a forum to discuss topics relating to the management issues associated with core facilities in both the academic and private sector. The format has been to have a presenter to introduce the topic and then having an interactive discussion session. Many of the proceedings have been taped, transcribed and published in Microscopy Today so that those who could not attend the meetings still had the benefit of the discussions. Some of the topics discussed in past sessions include:
Managing Users Justification of costs/cost recovery Equipment maintenance issues Training Users Calibration of Ems Scientific Ethics and responsibilities of facility managers.
I must finalize the topic(s) for M&M 2003. As we want to make this session timely, I would like input from readers on topics of current interest. This year we will receive no support from M&M 2003 but they have promised us a room and audio-visual equipment.
Please respond immediately with topic suggestions.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
First, let me assure you I know how to spell cryostat! The listserver spam filter blocks messages with CRY (as in Cry for Help) in them and Nestor is working on it. In the meantime I thought I would try this work around. I have several questions for the cryostat gurus out there.
First, how do you decide whether to infiltrate with sucrose? I will be fixing my tissue with 2% paraformaldehyde and some protocols call for direct freezing and others for infiltration with 30% sucrose in PBS first. I know the sucrose will cryo-protect and suspect it will also improve the plasticity but is there a disadvantage? A minor side question is whether the 30% sucrose can be simply made up in 1x PBS or do you need to account for the change in volume that the sucrose will probably cause. Presumably osmolarity is no longer a big issue if you are using 30% sucrose.
Second, what is the feeling on knives - high profile vs low profile? Is it worth paying extra for the heavy duty ones to minimize chatter or is that only needed for hard tissues. I will be cutting lymph nodes.
Thanks for any tips. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
} First, how do you decide whether to infiltrate with sucrose?
If you are going to freeze the tissue cryoprotection is a good idea. If your samples are small and you freeze quickly, you may be able to get away w/o cryoprotection.
} I will be } fixing my tissue with 2% paraformaldehyde and some protocols call for } direct freezing and others for infiltration with 30% sucrose in PBS } first. I know the sucrose will cryo-protect and suspect it will also } improve the plasticity but is there a disadvantage?
Not that I know of.
} A minor side question } is whether the 30% sucrose can be simply made up in 1x PBS or do you need } to account for the change in volume that the sucrose will probably } cause.
I don't. I have used sucrose concentrations from 20% to 30% with no apparent differences so if you are off a bit, no problem. What is important is the speed of freezing, the faster the better.
} Presumably osmolarity is no longer a big issue if you are using 30% } sucrose.
Nope. At least not in my experience.
} Second, what is the feeling on knives - high profile vs low profile? Is it } worth paying extra for the heavy duty ones to minimize chatter or is that } only needed for hard tissues. I will be cutting lymph nodes.
I can't help you there, I usually cut rodent brains with plain old microtome knives.
} Thanks for any tips. Tom } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
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 **********************************************
This is perhaps a bit unusual of a topic but it is centered on EDS viability for foiling double blind testing.
Question: Can EDS be used to analyze a double blind tablet to ascertain whether it is placebo or a study medication?
It would seem that the inability to see light element H might not be a limiting factor. Are there signatures of placebos that would strongly differentiate them from study meds? How might one guard against educated analysis of study meds? If the basic elements are C, H and O, are there distinct signatures or quantities that one could use to distinguish between placebo and study meds?
Randy, I'm glad you apologized to your student...its very frustrating to be lamed for someting that is totally out of your control! Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
HI Tom, The preference in our facility is to cryoprotect with sucrose whenever possible. We even freeze our tissues in a 1:1 mixture of sucrose (20% in PBS) and OCT. It makes a softer block, and you may have to drop your chamber temp to -20, but it cuts very smoothly and yields very nice structure. The only time we don't use sucrose is when the primary Ab's to be used won't tolerate any fixation, then we snap freeze and pray. Our protocol is a modification of that published by Barthel & Raymond in J Histochem Cytochem 3899) 1383-1388 1990. They were looking at eyes. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I am not sure what is the film thickness you need to cross-section, if it's over 10 microns it might be quite difficult. However, we have successfully cross-sectioned individual multi-walled carbon nanotubes and bundles of those (on SiO2/Si substrates) using combination of FIB 'lift-out' and broad beam ion milling in Gatan Ion Mill. At the first stage, you use the FIB 'lift-out' technique to make a sample with the thickness 0.8-1.0 microns. The sample is then mounted with the glue on the edge (on a bar) of the Cu half-grid and subjected to ion milling (4 kV, followed by 2 kV) in Gatan Ion Mill. Typical milling time in Gatan: 4-6 minutes. Keep checking the sample thickness in TEM and continue milling as necessary. The sample quality is good enough for high resolution TEM and EELS.
Let me know if you are interested in more detail, I can send you our recent paper.
Regards, Katharine ******************************* Katharine Dovidenko Assistant Professor School of NanoSciences and NanoEngineering & UAlbany Institute for Materials University at Albany-SUNY 251 Fuller Rd., Albany, NY 12203 Phone: (518) 437-8781
-----Original Message----- } From: yimin yao [mailto:yimin-at-fy.chalmers.se] Sent: Wednesday, March 12, 2003 3:49 AM To: Microscopy-at-sparc5.microscopy.com
Dear colleagues,
Does anyone have experience to use FIB to make cross-section TEM samples for a carbon nanotube film on substrate?
Best regards,
Yiming
------------------------------------ Dr. Yiming Yao
Microscopy and Microanalysis Department of Experimental Physics Chalmers University of Technology SE-41296, Göteborg Sweden
UVM Practical Course on Three-dimensional Cryo Electron Microscopy of Single Particles
August 11-17, 2003 Burlington, Vermont
The course will teach the principles of three-dimensional reconstruction of single particles from electron micrographs.
It includes demonstrations of the experimental aspects, teaching of the basic theoretical principles and six hours per day of hands on experience in processing data sets from cryo-electron microscopy images.
Participants will work in groups of two and six instructors will be available to guide everyone step by step through the complete reconstruction process. Four participants will have the opportunity to carry out practical microscopy work the two days following the end of the course.
For further information and application visit our web site: http://physioweb.med.uvm.edu/Cryo_Practical/
Organizers and Teachers:
Michael Radermacher Teresa Ruiz Montserrat Barcena Jean Francois Menetret Montserrat Samso T.B.A.
__________________________________________________ Michael Radermacher, Assoc. Prof. University of Vermont College of Medicine Department of Molecular Physiology and Biophysics HSRF Building Burlington, VT 05405
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Study medications would most likely contain detectable amounts of nitrogen. There might also be Na or K.
-Ken Gaugler
Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This is perhaps a bit unusual of a topic } but it is centered on EDS viability for } foiling double blind testing. } } Question: Can EDS be used to analyze } a double blind tablet to ascertain whether } it is placebo or a study medication? } } It would seem that the inability to see } light element H might not be a limiting } factor. Are there signatures of placebos } that would strongly differentiate them } from study meds? How might one guard } against educated analysis of study meds? } If the basic elements are C, H and O, are } there distinct signatures or quantities } that one could use to distinguish between } placebo and study meds? } } gary g. } } }
On Tuesday, March 11, 2003, at 06:16 PM, Gary Gaugler wrote:
} This is perhaps a bit unusual of a topic } but it is centered on EDS viability for } foiling double blind testing. } } Question: Can EDS be used to analyze } a double blind tablet to ascertain whether } it is placebo or a study medication? } } It would seem that the inability to see } light element H might not be a limiting } factor. Are there signatures of placebos } that would strongly differentiate them } from study meds? How might one guard } against educated analysis of study meds? } If the basic elements are C, H and O, are } there distinct signatures or quantities } that one could use to distinguish between } placebo and study meds? } Dear Gary, Most medications consist of small amounts of the active ingredient mixed with a much larger amount of matrix (which provides sufficient volume to handle a tablet, gets the active ingredient into the stomach--and sometimes further along the digestive tract--dissolves at a rate appropriate to deliver the drug as desired, etc.). The placebo in a properly designed trial consists of matrix identical to that for the drug, so EDS analysis would almost certainly give the same result for both. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
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In your message you only mention elements you believed would be present in the placebo tablet. However as Ken Gaugler alluded to, if you had information regarding the chemical composition of the active ingredient you might be able to ascertain if it were absent in a placebo tablet. For example, if the active contained sulfur EDS would (likely) be able to detect the sulfur in a tablet assuming the excipients/binders (see response from Bill Tivol) did not contain sulfur. However, unless you can verify this on a tablet containing the active ingredient I would be very careful in coming to a conclusion based upon a negative finding by EDS.
FYI Polarized light microscopy can also be helpful with this type of investigation.
Regards, jr
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Wednesday, March 12, 2003 8:51 PM To: microscopy-at-sparc5.microscopy.com
On Tuesday, March 11, 2003, at 06:16 PM, Gary Gaugler wrote:
} This is perhaps a bit unusual of a topic } but it is centered on EDS viability for } foiling double blind testing. } } Question: Can EDS be used to analyze } a double blind tablet to ascertain whether } it is placebo or a study medication? } } It would seem that the inability to see } light element H might not be a limiting } factor. Are there signatures of placebos } that would strongly differentiate them } from study meds? How might one guard } against educated analysis of study meds? } If the basic elements are C, H and O, are } there distinct signatures or quantities } that one could use to distinguish between } placebo and study meds? } Dear Gary, Most medications consist of small amounts of the active ingredient mixed with a much larger amount of matrix (which provides sufficient volume to handle a tablet, gets the active ingredient into the stomach--and sometimes further along the digestive tract--dissolves at a rate appropriate to deliver the drug as desired, etc.). The placebo in a properly designed trial consists of matrix identical to that for the drug, so EDS analysis would almost certainly give the same result for both. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
We have a pleasure to announce the final circular for the:
12th Croatian Slovenian Crystallographic Meeting to be held in Hotel Jezero, National Park Plitvice Lakes, Croatia (June 19-22, 2003). All additional information with Instructions for Abstract, Registration Form and Hotel Accommodation is contained on the web site:
http://www.chem.pmf.hr/~hkz/plitvice
Looking forward to seeing you on Plitvice Lakes Organizing Committee
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (didier.goux-at-unicaen.fr) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, March 13, 2003 at 08:01:06 ---------------------------------------------------------------------------
We use a balzers CPD 020 for critical point drying. When the engineer show me how to use it, I noticed that the temperature were set to 42 degres celcius (314 K) but the critical temperature is 31 degres celcius (304 K).
Dear All This is one of those "odd ones". Seems like administration love there admin tools. Here is one they would like a realistic answer of. "How many publications can the University expect per year from a EMU with one TEM, one SEM, and one CLSM ?" The EMU is staffed by 3 people. The size of the University: Overall total 12,286 students
Full-time 9977 Part-time 2309
Male - 6328 Female - 5958
Undergraduate - 11,336 Post graduate - 950
After a nice laugh, please help me on this one. I anticipate the normal "next" question in the near future: Cost recovery through consultancy!
Stephan H Coetzee Department of Physics EMU Private Bag 0704 Abalone, Botswana
It is good to know that administrators the world over all ask for wacky things. It helps to show there are no real differences between us all. This is a silly exercise but here is how I would answer it. A realistic level of production for a faculty member is 2 refereed papers a year (clearly 5-6 would be an excellent but not impossible goal but on average, 2 would be good for cell biologists). I would suggest that you estimate 2 / year for every faculty member whose research publications would always include at least 1 micrograph and a lesser fraction for the heathen biochemist and molecular biologists who only occasionally dabble in the most difficult science of microscopy. The real flaw with this exercise is that it will be taken as a measure of productivity of the EMU when, unless your staff are terribly incompetent (an unlikely prospect), it is really a measure of faculty productivity and quality. You could have a great EMU but if you don't have aggressive faculty, it won't get used.
At 05:28 PM 3/13/2003 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
(1) Thanks to all those who replied to my 'adhesive' question. I also now remember the correspondence on this listerver concerning the sawing up of M-Bond 610 chunks. I've found that M-Bond 610 is supplied in something like 25 ml bottles - does one have to use the whole of one pair of bottles once it's opened?
(2) Something completely different - has anyone recently purchased a new vacuum coating unit for TEM? How much did it cost, and do you like it? Trying to get a price from a manufacturer directly can be very wearing - they seem to want to interrogate you about what you want to do with it, rather than letting you know if the thing is within your budget.
LAB TALK (Caption searching for a cartoon):
"Call me an airhead? There's a Penning gauge on yours!"
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
Hi All, Please allow me to pick your collective brains.... A client just requested a TEM study of mouse ankle joints. I have looked at many things in the scope over the years, but I've never had to process & cut bone. Mice have tiny bones, and this will be the ankle...even tinier: Do I need to decalcify? How should we fix (I usually use a modified Karnovsky's (2.5% GA, 4% PFA + 0,02% picric acid) as my primary fix.) How & when do I decalcify? Any preferred resin? I feel like a babe in the woods. Thanks, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I am in the market for an osmometer. Does anyone have experience with these instruments to share? I am also interested in literature from vendors. Thank you in advance.
I have grown weary of snagging digital images with a camera held in my bare hand over the eyepiece or camera port of a stereomicroscope while manipulating an object with the other hand, which actually does work passably well because of the effectively high "film" speed of the digital camera. The images have vignetting and there's usually way too much light, though. It's not a good optical match, because the digital camera's lens gets in the way; there's a need for a transfer lens ... which would be too dear and too specialized.
Therefore, I bought a suitable video adapter with C mount for my company's stereomicroscope and then bought a Kodak MDS 100 camera, both on eBay, in my price range (less than $500 in it so far). While waiting for the camera to arrive (hopefully complete in its original wrapping as advertised by the eBay seller, as Kodak has disowned the thing) I'm anticipating with the following questions:
1. Have any of you light microscopists got experiences to share ? 2. Has anyone tried to mate the camera with a PC running Linux ? 3. Are there any wavelength issues, such as poor image quality due to infrared seeping through all that glass ?
I do have experience operating a flatbed scanner that uses the USB port as well as the TWAIN interface - and that is all favorable so far. My digital camera uses floppy disks, so image transfer via that route isn't very fast, but it's pretty reliable except when I make images on very hot days, whereupon some of the floppy files are DOA. And no, I'm not trying to get it to work from a Linux PC right from the get-go. The main PC is using W98SE.
And for the time being I have given up all hope of making one of the old, defunct flatbed scanners work like a film-plane scanner. Too many mechanical and optical issues. Thanks for your help on that.
Best regards, George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
*From: "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} *To: "Listserver Microscopy (E-mail)" {Microscopy-at-sparc5.microscopy.com} *Subject: Number of publications supported by the EMU per year? *Date sent: Thu, 13 Mar 2003 17:28:58 +0200
*------------------------------------------------------------------------ *The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Best regards,
Witold Zielinski
*Dear All *This is one of those "odd ones". Seems like administration love there admin *tools. Here is one they would like a realistic answer of. *"How many publications can the University expect per year from a EMU with *one TEM, one SEM, and one CLSM ?" *The EMU is staffed by 3 people. *The size of the University: *Overall total 12,286 students * *Full-time 9977 *Part-time 2309 * *Male - 6328 *Female - 5958 * *Undergraduate - 11,336 *Post graduate - 950 * *After a nice laugh, please help me on this one. I anticipate the normal *"next" question in the near future: Cost recovery through consultancy! * * *Stephan H Coetzee *Department of Physics *EMU *Private Bag 0704 *Abalone, *Botswana * *Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} * *Telephone: (+267) 355 2462 *Fax: (+267) 3185 097 *Telephone: (+267) 355 0000 Switchboard * * * * :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) ;)
Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
I will preface my remarks by stating that I have not worked with osmometers since the 1980's. That said, there are two different types of osmometers/two different ways of measuring osmolarity. 1. Freezing point depression 2. Vapor pressure. How #1 works is obvious, how #2 works I have no clue. I prefered a Wescor Vapor pressuer osmometer because I could use a sample as small as 7 microliters. The freezing point depression osmometers I had access to needed a much larger volume for accurate results. That may have changed in subsequent years. Most renal physiologists have an osmometer for measuring urinc concentration, perhaps someone in the physiology dept can help.
Geoff
kwalters wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am in the market for an osmometer. Does anyone have experience with these } instruments to share? I am also interested in literature from vendors. } Thank you in advance. } } Kathy } } Kathy Walters // } Central Microscopy Research Facility / / } 85 EMRB / /\ } University of Iowa / /\ \ } Iowa City, Iowa 52242 / / \ \ } Phone #: (319) 335-8142 / / \ \ } Fax #: (319) 384-4469 ______ ((0)) } email: Katherine-Walters-at-uiowa.edu |__| / / } || / / } -------------- } ------------------
-- ********************************************** 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 **********************************************
} Does anyone have experience with these instruments to share?
There are two basic types commonly used: vapor pressure osmometers and freezing point osmometers. The major manufacturer of freezing point osmometers went out of business several years ago. I am not aware of other manufactures. As a cell physiologist who works with osmoregulation, I use the vapor pressure osmometer regularly. Every colleague that I know uses the Wescor brand (Utah). I am not familiar with any other brands. I am absolutley satisfied with my Wescor, but have no means for making a comparison with other brands. These are not that cheap--I think one could cost $4-5K.
Best,
Don ____________________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
Greetings: I have not heard much lately about Holographic TEM lately.
From what I read, even though unstained polymer sections provide little modulation of the electron wave amplitude, there are variations in the wave phase. Holographic imaging techniques can be used to recover these phase modulations and thereby create contrast between multi-phase polymers without the use of stains. Can anyone provide an update on the state-of-the-art of this technique?
regards,
Anthony J. Ribaudo Staff Scientist Analytical Sciences Microscopy Facility Honeywell International Specialty Materials Division
Following the sudden death of my husband General Sani Abacha the former head of state of Nigeria in August 1998, I have been thrown into a state of utter confusion, frustration and hopelessness by the present civilian administration, I have been subjected to physical and psychological torture by the security agents in the country. My son is still under detention arraigned before the federal high courtof Nigeria for an offence he did not commit even though the government might release him soon. As a widow that is so traumatized,I have lost confidence with anybody within the country. You must have heard over the media reports and the internet on the recovery of various huge sums of money deposited by my husband in different security firms abroad, Some companies willingly gave up their secrets and disclosed our money confidently lodged there or many outright blackmail. In fact the total sum discovered by the Government so far is in the tune of $700. Million dollars. And they are not relenting to make me poor for life. I got your contacts through my personal research,and out of desperation decided to reach you through this medium.I will give you more information as to this regard as soon as you reply. I repose great confidence in you hence my approachto you due to security network placed on my day to day affairs I cannot afford to visit the embassy so that is why I decided to contact you and Ihope you will not betray my confidence in you. I have deposited the sumof 40 million dollars with a security firm abroad whose name is witheld for now until we open communication.shall be grateful if you could receive this fund into your account for safe keeping. This arrangement is known to you and my son Mustapha alone, so my son will deal directly with you as security is up my whole being. I am seriously considering to settle down abroad in a friendly atmosphere like yours as soon as this fund get into your account so that I can start all over again if only you wish, but if it is impossible,just help me in diverting this fund into your account which will accrue you 30% of this fund . Please honesty is the watch word in this transaction. I will require your telephone and fax numbers so that we can commence communication immediately and I will give you a more detailed picture of things. In case you dont accept please do not let me out to the security as I am giving you this information in total trust and confidence I will greatly appreciate if you accept my proposal in good faith. Please expedite action.
Sincerely yours,
Hajia Mariam Abacha.
N/B: Please copy your response:hajiamahaaba-at-netscape.net
Following the sudden death of my husband General Sani Abacha the former head of state of Nigeria in August 1998, I have been thrown into a state of utter confusion, frustration and hopelessness by the present civilian administration, I have been subjected to physical and psychological torture by the security agents in the country. My son is still under detention arraigned before the federal high courtof Nigeria for an offence he did not commit even though the government might release him soon. As a widow that is so traumatized,I have lost confidence with anybody within the country. You must have heard over the media reports and the internet on the recovery of various huge sums of money deposited by my husband in different security firms abroad, Some companies willingly gave up their secrets and disclosed our money confidently lodged there or many outright blackmail. In fact the total sum discovered by the Government so far is in the tune of $700. Million dollars. And they are not relenting to make me poor for life. I got your contacts through my personal research,and out of desperation decided to reach you through this medium.I will give you more information as to this regard as soon as you reply. I repose great confidence in you hence my approachto you due to security network placed on my day to day affairs I cannot afford to visit the embassy so that is why I decided to contact you and Ihope you will not betray my confidence in you. I have deposited the sumof 40 million dollars with a security firm abroad whose name is witheld for now until we open communication.shall be grateful if you could receive this fund into your account for safe keeping. This arrangement is known to you and my son Mustapha alone, so my son will deal directly with you as security is up my whole being. I am seriously considering to settle down abroad in a friendly atmosphere like yours as soon as this fund get into your account so that I can start all over again if only you wish, but if it is impossible,just help me in diverting this fund into your account which will accrue you 30% of this fund . Please honesty is the watch word in this transaction. I will require your telephone and fax numbers so that we can commence communication immediately and I will give you a more detailed picture of things. In case you dont accept please do not let me out to the security as I am giving you this information in total trust and confidence I will greatly appreciate if you accept my proposal in good faith. Please expedite action.
Sincerely yours,
Hajia Mariam Abacha.
N/B: Please copy your response:hajiamahaaba-at-netscape.net
On Thursday, March 13, 2003, at 08:55 AM, Robert H. Olley wrote:
} (2) Something completely different - has anyone recently purchased a } new } vacuum coating unit for TEM? How much did it cost, and do you like it? } Trying to get a price from a manufacturer directly can be very wearing } - } they seem to want to interrogate you about what you want to do with it, } rather than letting you know if the thing is within your budget. } } Dear Robert, I recently priced out several vacuum evaporators, and the one we ordered was slightly under $20k. We wanted a desk-top unit, and we wanted a turbo pump for the vacuum. It has not yet arrived, so I can't tell you whether we'll like it, but I've had good experiences with similar units. If your requirements are different, you could possibly get away with a much cheaper unit, ~$5k, with a mechanical pump for the vacuum. We need to evaporate both carbon and metals, but we don't need sputtering capability. Since the uses to which the unit will be put make a big difference in the price, I think the manufacturer or supplier is justified in asking rather than just selling the fanciest unit to each customer. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Hi Stephan This is a question of particular interest to us as we have to be accountable too and I am looking forward to the replies We use:
Utilization of our facility is monitored by 1) The numbers of researchers using the facility from individual departments. 2) The numbers of researchers using the facility from off campus and from industry. 3) The numbers of participants in the in-house workshops. 4) The numbers of graduate students taking credit courses in electron and light microscopy. 5) The numbers of students in various UBC courses visiting the facility. 6) The numbers of research papers produced with the assistance of the facility. 7) The numbers of projects handled by the facility's staff. 8) The number of research theses produced with support from the facility.
The numbers of users we mostly generate from the billing. For the numbers of papers and theses we have to remind users constantly that we need those numbers. There is a note on the billing when it is sent out. Does anyone have any other ideas how to generate these numbers? Elaine
} Dear All } This is one of those "odd ones". Seems like administration love there admin } tools. Here is one they would like a realistic answer of. } "How many publications can the University expect per year from a EMU with } one TEM, one SEM, and one CLSM ?" } The EMU is staffed by 3 people. } The size of the University: } Overall total 12,286 students } } Full-time 9977 } Part-time 2309 } } Male - 6328 } Female - 5958 } } Undergraduate - 11,336 } Post graduate - 950 } } After a nice laugh, please help me on this one. I anticipate the normal } "next" question in the near future: Cost recovery through consultancy! } } } Stephan H Coetzee } Department of Physics } EMU } Private Bag 0704 } Abalone, } Botswana } } Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} } } Telephone: (+267) 355 2462 } Fax: (+267) 3185 097 } Telephone: (+267) 355 0000 Switchboard
-- Dr. Elaine Humphrey Director, BioImaging Facility First Vice President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
In brief, one usually sets the temperature slightly higher than the critical point to insure that one has passed it.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging
Mail to: Geology, CASI West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383
For help and information only, The CASI houses: An FEI Quanta 400 and Technai 12T, Oxford INCA Energy 400, Tousimis AutoSamdri 815 and Olympus FV-300.
-----Original Message----- } From: didier.goux-at-unicaen.fr [mailto:didier.goux-at-unicaen.fr] Sent: Thursday, March 13, 2003 10:23 AM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (didier.goux-at-unicaen.fr) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, March 13, 2003 at 08:01:06 ---------------------------------------------------------------------------
We use a balzers CPD 020 for critical point drying. When the engineer show me how to use it, I noticed that the temperature were set to 42 degres celcius (314 K) but the critical temperature is 31 degres celcius (304 K).
I am not sure if my first posting got through, but, the definitive answer to this question comes from either Raman microscopy or FT-IR microscopy. No guessing, just good chemical fingerprinting.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 08:53 AM 3/13/03 -0500, jrobson-at-rdg.boehringer-ingelheim.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Near IR (NIR) focal plane array systems provide direct visual differentiation of active and placebo tablets, as well as the areal distribution of components within the tablet. For an example, see http://www.spectraldimensions.com/products/index.html.
James Martin Orion Analytical, LLC www.orionanalytical.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
I am not sure if my first posting got through, but, the definitive answer to this question comes from either Raman microscopy or FT-IR microscopy. No guessing, just good chemical fingerprinting.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 08:53 AM 3/13/03 -0500, jrobson-at-rdg.boehringer-ingelheim.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (henryp-at-bhphoto.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, March 14, 2003 at 12:11:56 ---------------------------------------------------------------------------
Email: henryp-at-bhphoto.com Name: Henry Posner
Organization: B&H
Education: Undergraduate College
Location: New York, NY USA
Question: I am in possession of my father's WW II era Leica microscope. It's missing the chrome clips which hold a slide in place. Where can I get a pair and where can I learn more about this instrument and its current value? TIA
the company "Knauer" in Berlin / Germany builds among other scientific instruments osmometers for over 40 years. I have a freezing point one and am very contented with it. they have represantatives all over the world, just check their website http://www.knauer.net/ peter -- ********************************** peter.heimann-at-uni-bielefeld.de Dr. Peter Heimann Developmental Biology & Molecular Pathology; W7-107 University of Bielefeld D 33501 Bielefeld / Germany phone: xx49(0)521-106-5628 / 5627 FAX : " " - 5654 www.uni-bielefeld.de/biologie/Entwicklungsbiologie/ www.uni-bielefeld.de/SFB549 ***********************************
In the past I have been working on Calcium ratio measurements in microscopy. It has been a while since I was actively involved and I am curious about new developments in the field.
People have been using ultrafast camera's for Calcium ratio imaging, which puzzles me a bit. From what I remember, at 20 deg. Celsius, the Calcium sensitive probe Fura-2 needs 5-10 ms to reach equilibrium in a solution of about 140 mM Calcium. We used regular PAL video cameras (25 fps, 40 msec./frame) with a frame splitter device, which enabled us to monitor intracellular Calcium changes and relocation at 1/4 of the frame rate, at 10 msec in individual cells. Taking the Nyquist sampling theorem into account, this is sufficient to monitor most phenomena we were interested in.
We used Fura-2, as the use of Calcium ratio imaging which has some advances compared to non-ratio Calcium measurement (i.e. concentration changes of the probe). What are good alternatives to Fura-2 for intracellular Calcium measurements. For Calcium ratio measurements, a Mercury arc lamp provides the best result I guess, with its high emission of light in the near-UV range (I do not like Mercury arcs for other fluorescent work, due to their "spiky" spectrum, but prefer Xenon arcs instead, as their emission is more evenly spread through the visible spectrum).
For the optics we used Fluorite lenses, as they transmit better in the near-UV range, than glass objectives. What is your opinion of the optics for Calcium ratio measurements ?
We used mechanical filter changers for switching between the excitation wavelengths, what about elctronic devices ? From what I remember, their transmission efficiency is much lower than "traditional" filters ?
Some data : The concentration of intracellular Calcium lies between 10 nM and 10 uM or even higher I believe. The excitation wavelength optimum changes when Fura-2 binds Calcium. Fura-2 has a Kd of about 145 nM and is about saturated above 1 uM Calcium (this may change depending on the conditions ?).
When Fura-2 binds Calcium, the excitation optimum shifts between 300 and 400 nm, 363 (ion free) and 335 nm, in practice 340 nm and 380 nm are taken for the two excitation wavelengths (Mercury arc peaks). The more Calcium is present, the higher the absorption shift towards 340 nm. One can detect this shift by monitoring the emission at 510 nm (~green light), the change in emission intensity will reflect the absorption shift between 340 and 380 nm. The Kd (dissociation constant) for Calcium of Fura-2 is ~135 nM in Mg2+ free Calcium buffers and ~224 nM in the presence of 1 mM Mg2+. Although Fura-2 may accurately indicate peaks of Calcium up to 500 nM in cells, Fura-2 exhibits limited sensitivity above 500 nM Calcium.
There is no Calcium dependent change in absorption at about 360 nm., which is called the isosbestic point of Fura-2. Measurement of the 510 nm emission when exciting at 360 nm can be used as a measure for fotobleaching of Fura-2.
Dear all, I am sure I remember that someone had produced a routine for the rotational averaging of SAD ring patterns to produce an intensity versus line plot analagous to a XRD diffractogram. Could anyone give me any hints where I could get such a thing? A plug-in for Gatan's digital micrograph would be especially good, since we use this program often. A Photoshop plug-in would be another good solution.
Thanks in advance for any help you can give.
-- Ian MacLaren Technische Universität Darmstadt Material-und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany
In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes:
} Dear all, } } I am sure I remember that someone had produced a routine for the rotational } averaging of SAD ring patterns to produce an intensity versus line plot } analagous to a XRD diffractogram. Could anyone give me any hints where } I could get such a thing? A plug-in for Gatan's digital micrograph would } be especially good, since we use this program often. A Photoshop plug-in } would be another good solution. } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit images) both include that routine, as a Photoshop-compatible plugin. see http://ReindeerGraphics.com
---------------------------------------------------------------------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes:
} Dear all, } } I am sure I remember that someone had produced a routine for the rotational } averaging of SAD ring patterns to produce an intensity versus line plot } analagous to a XRD diffractogram. Could anyone give me any hints where } I could get such a thing? A plug-in for Gatan's digital micrograph would } be especially good, since we use this program often. A Photoshop plug-in } would be another good solution. } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit images) both include that routine, as a Photoshop-compatible plugin. see http://ReindeerGraphics.com
A one-year postdoctoral position (with a possible one-year extension) is opened at the CNRS in Strasbourg, France (CEPE: Centre dEcologie et de Physiologie Energétiques). The aim of the project is to look at the morphological and functional flexibility of the intestinal mucosa in vertebrates through light, transmission and scanning electron microscopy. An experienced microscopist is required with skills in immunohistochemistry (immunofluorescence) and transmission and scanning electron microscopy (immunogold). Skills in Environmental Scanning Electron Microscopy (ESEM) -and possibly in western blotting- would be an advantage. If interested, please send a resume to Dr JH Lignot by fax or by email.
Dr Jean-Hervé Lignot, Université Louis Pasteur / CNRS CEPE 23 rue Becquerel, 67087 Strasbourg, Cedex 2, France Fax: 00 33 0388106906 Tel: 00 33 0388106938
A one-year postdoctoral position (with a possible one-year extension) is opened at the CNRS in Strasbourg, France (CEPE: Centre dEcologie et de Physiologie Energétiques). The aim of the project is to look at the morphological and functional flexibility of the intestinal mucosa in vertebrates through light, transmission and scanning electron microscopy. A good microscopist is required with skills immunohistochemistry (immunofluorescence), transmission and scanning electron microscopy (immunogold). Skills in Environmental Scanning Electron Microscopy (ESEM) -and possibly in western blotting- would be an advantage. If interested, please send a resume to Dr JH Lignot by fax (0033 388106 906) or by email (J-H.Lignot-at-c-strasbourg.fr).
Dr Jean-Hervé Lignot, Université Louis Pasteur / CNRS CEPE 23 rue Becquerel, 67087 Strasbourg, Cedex 2, France Fax: 00 33 0388106906 Tel: 00 33 0388106938
I'm building a humidified/heated chamber to do live cell imaging of mammalian cells. iIm looking for a vendor that sells warm air blowers that would be useful.
Thanks in advance for the input.
Best,
Gary
Gary S. Laevsky, Ph.D. Research Associate The Scripps Research Institute 10550 N. Torrey Pines Road/IMM-24 La Jolla, CA 92037 (858) 784-9372
I am in the market for a electropolisher for TEM sample preparation & was wondering if anyone had any suggestions. Preferably a twin jet electropolisher for amorphous metal sample preparation.
Any feedback would be greatly appreciated,
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
Does anyone know where I could get another rubber drive belt for the Reichert OMU3 ultramicrotome? Ours broke, and when we try to glue it, it just keeps breaking again.
Does this rotational averaging routine from Reindeergraphics take into account the ellipticity of the (imperfect) SAD ring patterns? If so, does it automatically find the major and minor axes or does that require manual adjustment?
Thanks,
Tom Schamp
On 3/17/03 7:03 AM, ""DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com" {"DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes: } } } Dear all, } } } } I am sure I remember that someone had produced a routine for the rotational } } averaging of SAD ring patterns to produce an intensity versus line plot } } analagous to a XRD diffractogram. Could anyone give me any hints where } } I could get such a thing? A plug-in for Gatan's digital micrograph would } } be especially good, since we use this program often. A Photoshop plug-in } } would be another good solution. } } } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit } images) both include that routine, as a Photoshop-compatible plugin. see } http://ReindeerGraphics.com } } } }
---------------- Tom Schamp ph: (434) 982-4595 University of Virginia fx: (434) 982-5660 Dept. of Materials Science and Engineering email: cts2v-at-virginia.edu 116 Engineer's Way Charlottesville, VA 22904
I am interested in looking at the general Laminin distribution within a basement membrane. Does anybody know of, or have, an antibody that will cross react with a number of Laminin subclasses, rather than just a specific Laminin subclass ?
I am more than happy to receive answers from antibody suppliers.
Thanks in advance
Allan
-- ------------------------------------------------- Allan Mitchell Technical Manager Otago Centre for Electron Microscopy C/-Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
On Monday, March 17, 2003, at 01:40 AM, Ian MacLaren wrote:
} I am sure I remember that someone had produced a routine for the } rotational averaging of SAD ring patterns to produce an intensity } versus line plot analagous to a XRD diffractogram. Could anyone give } me any hints where I could get such a thing? A plug-in for Gatan's } digital micrograph would be especially good, since we use this program } often. A Photoshop plug-in would be another good solution. } } Thanks in advance for any help you can give. } Dear Ian, I wrote a routine both for SPIDER and as a stand-alone, which determines the center and radius of a ring from an input of between 3 and 20 points on the ring. SPIDER can then produce a rotational average by padding your original diffractogram into a larger image such that the center of the pattern is the center of the larger image and using the rotational averaging module in SPIDER. This does not qualify as a plug-in for DM, but it could do the job. Since I have moved from Albany, I no longer have access to the code, but perhaps someone there could get it to you, if you decide you need it. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Ian, You may want to look at the program ProcessDiffraction http://www.mfa.kfki.hu/~labar/ProcDif.htm by János L. Lábár. It will do what you are asking and is a stand alone program. I have used it a handful of times, but never very intensely.
-Ray
Ray D. Twesten, PhD. Center for Microanalysis of Materials Seitz Materials Research Laboratory 104 S. Goodwin Ave., Urbana, IL 61801 +1 217 244-6177 (Fax -2278)
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: Monday, March 17, 2003 3:41 AM To: Microscopy Listserver Cc: Ralf Theissmann
Dear all, I am sure I remember that someone had produced a routine for the rotational averaging of SAD ring patterns to produce an intensity versus line plot analagous to a XRD diffractogram. Could anyone give me any hints where I could get such a thing? A plug-in for Gatan's digital micrograph would be especially good, since we use this program often. A Photoshop plug-in would be another good solution.
Thanks in advance for any help you can give.
-- Ian MacLaren Technische Universität Darmstadt Material-und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany
Rotational averaging is included in a software package for Digital Micrograph written at the NCEM. It's available for download at http://ncem.lbl.gov/frames/software.htm.
Larry Thomas Pacific Northwest National Laboratory P.O. Box 999 Mail Stop P8-16 Richland, WA, USA
} ---------- } From: Tom Schamp } Sent: Monday, March 17, 2003 11:13 AM } To: "DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com; ian.maclaren-at-physics.org; Microscopy-at-sparc5.microscopy.com } Subject: Re: Rotational averaging } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dr. Russ, } } Does this rotational averaging routine from Reindeergraphics take into } account the ellipticity of the (imperfect) SAD ring patterns? If so, does } it automatically find the major and minor axes or does that require manual } adjustment? } } Thanks, } } Tom Schamp } } } On 3/17/03 7:03 AM, ""DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com" } {"DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes: } } } } } Dear all, } } } } } } I am sure I remember that someone had produced a routine for the rotational } } } averaging of SAD ring patterns to produce an intensity versus line plot } } } analagous to a XRD diffractogram. Could anyone give me any hints where } } } I could get such a thing? A plug-in for Gatan's digital micrograph would } } } be especially good, since we use this program often. A Photoshop plug-in } } } would be another good solution. } } } } } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit } } images) both include that routine, as a Photoshop-compatible plugin. see } } http://ReindeerGraphics.com } } } } } } } } } } } } } ---------------- } Tom Schamp ph: (434) 982-4595 } University of Virginia fx: (434) 982-5660 } Dept. of Materials Science and Engineering email: cts2v-at-virginia.edu } 116 Engineer's Way } Charlottesville, VA 22904 } } } } }
This is an open reply to the listserver for your posting, but my questions are directed to you.
I tried the radial profile option in Fovea Pro on a ring pattern file that I have that has the pattern centered. I got a nice radial profile. I then offset the image and put a black background where the image was offset. If you take that radial profile, you get something quite different. Could you tell us the general algorithm that you use to obtain this?
I would also like to know whether the circular Hough Transform that you discussed with me a few years ago would take into account this offset or must the pattern be centered?
Again with the circular Hough Transform, could that address an elliptical pattern?
On another note:
I helped MDI, Inc. modify their electron diffraction module in WinJade several years ago. Here are some of the problems that had to be addressed with their system. 1) some diffraction patterns can have an aspect ratio that is not 1:1 either due to the microscope or the digitizing process. This needs to be taken care of because it can be as much as 2%. For the microscope itself, the presence of an elliptical pattern can be checked quite easily by taking two identical diffraction patterns of polycrystalline gold and overlaying one over the other, but rotated 90 degrees. 2) patterns are often off-center and must be centered for the radial profile 3) Their original algorithm simply took the number of pixels in a ring and divided by the number of pixels in the ring for an average. This had the effect of decreasing the intensities quickly. In effect, they were taking the integral over dxdy, not the integral over d(theta)rdr, i.e. not taking the average in polar coordinates. For spotty patterns, this drastically cut down the intensities at larger radii in the pattern because of the large number of black pixels in the rings bring the intensities down. We developed four algorithms for using on patterns and which was best depended on the type of pattern e.g. completer rings, spotty rings, etc. The four algorithms were:
J1: Average All Pixels in a Ring --The radius for each pixel in the image is calculated, the intensity at each radius is summed, divided by the number of pixels at that radius, and the pattern is normalized to 10,000 counts.
J2: Average Above-Average Pixels --The average of each radius is found and only pixels at each radius that have intensities above the average for the radius is used. The intensity is calculated for each radius by averaging these pixels and the pattern is normalized.
J3: Sum of Above-Average Pixels --The sum of the above-average pixels is used instead of the average and the pattern is normalized.
J4: Take Maximum Pixel in Ring --The highest intensity pixel in each ring is used as the intensity.
Here are the conclusions that I found with WinJade to real world diffraction patterns:
Care must be taken when choosing the mode of data reduction. J1 and J2 give very good results with complete rings. J2 works very well with incomplete but readily apparent rings. J2 gives an improved peak/bkg ratio because it excludes pixels below the average intensity value for a given ring. J3 is noisy. It works well when there are discrete spots on a black background. J4 works well for spotty patterns and finds all of the d-spacings present. It provides good values of d-spacings at large radii.
This program was really great for working with SAD patterns. You could compare the radial profile with the powder diffraction database, compare with X-ray diffraction data, overlay either experimental XRD data or PDF patterns on the SAD pattern, and easily calibrate the camera constant easily for the program. Unfortunately, in the new version of WinJade, they did not include the electron diffraction module. They have it in another program that they sell, but I already bought the upgrade to the new program and can't use the old version on Windows 2000. -Bummer.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: "DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com [mailto:"DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com] Sent: Monday, March 17, 2003 7:04 AM To: ian.maclaren-at-physics.org; Microscopy-at-sparc5.microscopy.com
In a message dated 3/17/03 4:48:38 AM, maclaren-at-tu-darmstadt.de writes:
} Dear all, } } I am sure I remember that someone had produced a routine for the rotational } averaging of SAD ring patterns to produce an intensity versus line plot } analagous to a XRD diffractogram. Could anyone give me any hints where } I could get such a thing? A plug-in for Gatan's digital micrograph would } be especially good, since we use this program often. A Photoshop plug-in } would be another good solution. } The Image Processing Tool Kit (for 8 bit images) and Fovea Pro (for 16 bit images) both include that routine, as a Photoshop-compatible plugin. see http://ReindeerGraphics.com
Scott (and anyone else interested in this conversation)
The plugin uses a circular hough to get the radially averaged intensities. It expects the center of the pattern to be "near" the center of the image, as it does not store the full 3D accumulator space for a Hough (the axes are x-center, y-center and radius). I suppose that with computer memories getting larger it would be possible to store a larger array, if someone has a pressing need. The method works by adding up the grey scale brightnesses of each pixel in the original image into the points in the Hough array that define a possible circle through those points - in other words a cone in the Hough space. Then the central axis of the pattern is found by locating the brightest column in the space, and the intensities along that axis form a plot of the circularly averaged intensities. The routine does NOT perform an elliptical hough transform, which would require a 5 dimensional array (x-center, y-center, minor axis, major axis, angle). It is certainly possible to write something to do that, but it simply hasn't come up before. What I usually do with images that have some distortion (e.g., non-round SAED patterns) is fix that first, before the measurement. But writing an elliptical Hough wouldn't be hard (given enough memory) and if someone wants to agitate for that, I'll consider doing it. Might even give it away, as this seems like a pretty specialized application.
John Russ
In a message dated 3/17/03 8:05:29 PM, walck-at-ppg.com writes:
I tried the radial profile option in Fovea Pro on a ring pattern file that I have that has the pattern centered. I got a nice radial profile. I then offset the image and put a black background where the image was offset. If you take that radial profile, you get something quite different. Could you tell us the general algorithm that you use to obtain this?
I would also like to know whether the circular Hough Transform that you discussed with me a few years ago would take into account this offset or must the pattern be centered?
Again with the circular Hough Transform, could that address an elliptical pattern?
No question it can be done. All of the math is pretty trivial, but there is a modest amount of programming involved and before embarking on it I'd like to see how much interest there is. Apparently someone has written something that does circular averaging in DM, and if that meets peoples' needs there is no point in my writing a Photoshop plugin as well. If a groundswell of interest develops (i.e., more than just you), I'll try to design a fairly simple but adequate user interface to get the camera constant, so that the output plot and Excel file have all of the 1/d, d, etc values included. If the ellipticity of the pattern is an instrument constant, it is still easier to fix it before measurement rather than add the extra parameters to the Hough.
John Russ
=====
In a message dated 3/17/03 8:48:57 PM, walck-at-ppg.com writes:
} John, } } I think that you would be surprised how many elliptically patterns are } out there that people don't even know they have until someone points it } out. The microscope manufacturers don't go out of their way to tell you. } } } } One more question for you. How do you calibrate the output. The data } is given (in my case) with a maximum radius as something like 4.6 um. } I then went in and made the um/pixel value =1 and so now even though it } is saying that the max radius is in um, I know that it is in pixels. Still, } is there anyway you could have a diffraction pattern calibration in the } program similar to the magnification calibration for images? this way, } d-spacing, 1/d spacings, 2theta, values, and 2theta(XRD Cu-Ka) values could } be calculated. } } } } I could see an elliptical check with the camera constant calibration. } Two vertical lines that you can adjust in and out so that they are tangent } to opposite sides of the same ring and then two horizontal lines to do } the same thing. The value of the separations of the lines would agree } for a circular pattern as a check. This could also check the centering } of the pattern when the two lines are over top of each other and their } separation is zero. } } } } -Scott
I was wondering if anyone can tell me where I might find demographics for research and clinical labs in the US. Any site's you can refer me to would be greatly appreciated.
*Date sent: Mon, 17 Mar 2003 10:53:06 -0600 *From: William Stratton {wgstratton-at-wisc.edu} *Subject: Recommendations for Electropolisher? *To: Microscopy-at-sparc5.microscopy.com
*------------------------------------------------------------------------ *The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
We're using double jet polisher Tenupol made by Struers for many years and may say that it's good maschine for metallic TEM specimen preparation. However such requirment as electropolishing of amporhous metal is maybe too specific. Good performence of this method depends rather on the metal than on its state.
Best regards,
Witold Zielinski
*Greetings all, * *I am in the market for a electropolisher for TEM sample preparation & was *wondering if anyone had any suggestions. Preferably a twin jet *electropolisher for amorphous metal sample preparation. * *Any feedback would be greatly appreciated, * *William Stratton * *------------------- *William G. Stratton *Research Assistant *University of Wisconsin - Madison * *1509 University Avenue *Madison, WI 53706 *Office: 608-265-6391 *Fax: 608-262-8353 *wgstratton-at-wisc.edu * * * :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) ;)
Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
The company Solent Scientific manufactures full enclosure incubation chambers for Research Inverted Microscopes, Confocal Microscopes and Multi Photon Microscopes. Maybe they can help with the necessary equipment ?
Their website: http://www.solentsci.com/
Note: I have no commercial relation with this company. At my previous employer they used to build their own fully equipped incubator chambers for long-term time-lapse studies of mammalian cells (} 24 h.).
==================== Gary Laevsky wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear all, } } I'm building a humidified/heated chamber to do live cell imaging of } mammalian cells. iIm looking for a vendor that sells warm air blowers that } would be useful. } } Thanks in advance for the input. } } Best, } } Gary } } Gary S. Laevsky, Ph.D. } Research Associate } The Scripps Research Institute } 10550 N. Torrey Pines Road/IMM-24 } La Jolla, CA 92037 } (858) 784-9372
Dear Garry, You can get it stitched neatly, That is what I did for my Reichert ultracutE, It is working for 3 years like that. Shashi
Does anyone know where I could get another rubber drive belt for the Reichert OMU3 ultramicrotome? Ours broke, and when we try to glue it, it just keeps breaking again.
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Does anyone have a source for black or darkly colored membrane filters. I am setting up to do an OM particle counting / sizing study and need a filter with a pore size of {0.45 micrometers that will be resistant to cyclohexane.
Thanks, John A. Robson Boehringer Ingelheim Pharmaceuticals, Inc. PO Box 368 900 Ridgebury Rd Ridgefield, CT 06877
The University of Texas Health Science Center at San Antonio will host a Symposium and Workshop sponsored by Hamamatsu Photonics KK on
Fluorescence Lifetime Imaging and Spectral Imaging
Academic Participants: Philippe Bastiaens (GER)*Robert Clegg (USA)*Mary Dickinson (USA)*Paul French (UK) Hans Gerritsen (Neth)*Brian Herman (USA)*Ammasi Periasamy (USA) Herbert Schneckenburger (GER)*Ton Visser (Neth)*Timo Zimmermann (GER)
Symposium Registration Deadline: May 1, 2003 June 6-7, 2003 The Sheraton Gunter Hotel 205 E. Houston St. San Antonio, TX Student: $200 ($250 after May 1) Professional: $250 ($300 after May 1)
Workshop Application Deadline: April 7, 2003 June 8-10, 2003 UT Health Science Center San Antonio, TX Tuition: $700 (includes room and board) 20 student limit/4 scholarships
For Information and Forms Visit: http://usa.hamamatsu.com/flim_spectral/default.htm or http://www.uthscsa.edu/csb/imaging-course.html
Rushing around South Africa at the moment I have just seen the request for data on Spot Size in a TEM, perhaps I can help?
The Spot Size control adjusts the first condenser lens and in combination with the second condenser lens they demagnify the virtual source to provide a certain beam "spot size" on the specimen. This spot size may be measured by setting the magnification at 10,000X where 1um = 1cm. When trying to produce a quality image I would always suggest one uses a spot size smaller than 3um.
Whilst the Spot Size is used by most people at its largest setting (bright) it is not the way the designers expect the instrument to be. Good operating techniques should always take the spot size into account, even at lower magnifications better quality images will be attained at smaller spot sizes.
Since papers were produced in 1944 about transmission images (LM and EM) we have known that parallel beams are very important if you are chasing the best image quality, biology or materials we always desire high coherence. There are some misunderstandings on how to obtain a parallel beam or high coherence, for example setting the final condenser under focus is incorrect. The procedure for high coherence would be to use the smallest spot size you could tolerate (this probably means you must up the emission current, use at least 20 to 30 uA). Once in this condition over focus the final condenser (clockwise from crossover) the spot, whatever size it is now, becomes your new virtual source. The further over focus you go, the greater the distance between the specimen and the crossover the more parallel the beam and therefore it attains a higher coherence, which is what we are after. You will deduce the smaller the condenser aperture the sharper the spot and the smaller the spot the greater the coherence for a given degree of over focus.
Work with a design team and they expect everyone to over focus, they expect everyone to use small condenser apertures and they expect everyone to use small spot sizes. They do not expect everyone to use too low an emission current because this makes the task too difficult! Unfortunately almost everyone does use too low a current, I have talked before about filament life being the most important feature of many laboratories and it is always to the determent of image quality.
So in short you should always use a smallish spot size when taking micrographs and you should always run with the second condenser over focus. With sheet film photography try to use 3 to 4 seconds exposure, as this will give you better coherence too. Also remember that the denser the negative the more contrast you will build in your final print.
Hope this helps, spot size IS so important in the production of a quality image.
Steve Chapman
Senior Consultant EM
Protrain for Electron Microscopy Courses World Wide
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alvarobq-at-fcien.edu.uy) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, March 18, 2003 at 11:42:39 ---------------------------------------------------------------------------
Question: may i adding potassium ferrocyanide to 1%OsO4 postfixation to enhance contrast-staining cellular components in a routin coloidal-gold immunocytochemistry method? how do you prepare this ferrocyanide solution? what rate do you use? and if i use Lowicril(absence of osmium treatment), may i adding to the glutaraldehyde solution? how do you prepare and what rate?
Can anyone fill me in on some of the issues surrounding doing high res cryoEM and/or the need for something like a PhotoScan 2002 high res scanner from ZI Imaging.
I need to get some of the basics down so I can have a reasonable chance of understanding a proposal being made here.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have a guy who wants to look at the interior of some wood blocks using our conventional SEM. He is trying to see bacteria and anything else that might be hiding in there.
His blocks are pretty small, about 5mm cubes. I thought about CPD, but had second thoughts about how long to soak in CO2 etc. He did some experiments and it took a couple of hours for some dye to make it all the way into the middle of the block. Plus he has about 30 of these things to look at. That could add up to a lot of time and CO2.
I got an idea that HMDS might work for this project, but have no experience with it. I think that being able to soak the blocks for a long time in HMDS would be good, as would the chance to let them sublime to dryness over a long time without too much attention.
Do you think I am on the right track or is there something I am missing.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
A journal editor requested me to provide a reference regarding how to measure electron dose. I looked over some textbooks and experimental sections of some papers on electron beam damage. However, I could not find. If someone knows a reference, please advise.
Hiromi Konishi University of New Mexico hkonishi-at-unm.edu
Steve, thanks for the very good comment on spot size issue.
Without knowing all these details I was told many years ago to use smaller possible spot size (yes, I do have some idea what coherence is) and owerfocus second condenser. Working in Russia and here in US I always set all my TEMs for spot size 3 (JEM12000EX) surviving the battle with previous "spot size 1" lovers. If microscope aligned correctly, I never ever had a problem with illumination even with regular tungsten filament in magnification range x5-80K. My standard exposure time is 1.5-1.7 sec for 4489 film. In order to insure dynamic range, negatives should be very grayish. If it's too dark - you will loose the film's dynamic range. If you need extra contract - use more contrasty paper. In the nowadays, when we switched mostly for digital imaging, low illumination is just not an issue. My digital camera is exactly 10x more sensitive than 4489 film, so I have exposure time 0.1-0.2 sec with tungsten filament, spot size 3. Another reason to use smaller spot size - it's more easy to focus the images, at least to me: I do see focus changes more clear with smaller spot size. As for illumination, we have to keep in mind that e-beam itself damage our fragile biological samples, so less light - better for your sample. Thanks again for such detailed comment. Sergey
At 08:20 AM 3/18/2003, 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 saw a black (Whatman) Nuclepore polycarbonate membrane filter (0.2 and 0.4 um pore) in VWR. You'd have to check the chemical resistance charts if polycarbonate is resistant to cyclohexane.
Lizette Tuason
-----Original Message----- } From: "jrobson-at-rdg.boehringer-ingelheim.com"-at-sparc5.microscopy.com [mailto:"jrobson-at-rdg.boehringer-ingelheim.com"-at-sparc5.microscopy.com] Sent: Tuesday, March 18, 2003 5:28 AM To: microscopy-at-sparc5.microscopy.com
Jonathan,
We recently looked at wood blocks in our FESEM and the only preparation we did was to freeze them in liquid nitrogen, then fracture them with pliars to expose the internal structure, followed by several days in a vacuum chamber to get as much moisture and air out as possible. We glued them down with copius amounts of conductive paint, then sputtered the bejesus out of them.
The structure was well preserved and we did find bacteria.
The only problem was breaking the harder pieces of wood. Some of the wood was ancient and very brittle, but the new pieces were tough.
Hope this is applicable to your problem.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Jon Krupp [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Tuesday, March 18, 2003 3:03 PM To: Microscopy-at-sparc5.microscopy.com
Hi again:
I have a guy who wants to look at the interior of some wood blocks using our conventional SEM. He is trying to see bacteria and anything else that might be hiding in there.
His blocks are pretty small, about 5mm cubes. I thought about CPD, but had second thoughts about how long to soak in CO2 etc. He did some experiments and it took a couple of hours for some dye to make it all the way into the middle of the block. Plus he has about 30 of these things to look at. That could add up to a lot of time and CO2.
I got an idea that HMDS might work for this project, but have no experience with it. I think that being able to soak the blocks for a long time in HMDS would be good, as would the chance to let them sublime to dryness over a long time without too much attention.
Do you think I am on the right track or is there something I am missing.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Has anybody experience in praparation of weak milk foams? We prefer freeze fracture technique. The bubbles size is at maximum 1 mm. Of special interest is the air-milk interface.
The next meeting of the Metropolitan Microscopy Society will be held on April 9th, 2003 at the EDAX facility in Mahwah, NJ. EDAX has graciously agreed to host our meeting and to provide complimentary coffee and lunch to all the attendees.
The forthcoming meeting comprises five presentations offering a blend of microscopic, spectroscopic and image processing applications. Mike Marko will describe the cutting edge methods for tomographic reconstruction of biological organelles. Paul Kotula and Tom Hancewicz will discuss methods to extract maximum value from the data by analyzing spectral images. Matt Libera will illustrate ways to form novel surface patterns on polymeric materials, using electrons, for controlling biological activity and Del Redfern will highlight the advances in high-resolution imaging in projection x-ray microscopy.
We invite all members to attend and to inform their colleagues and fellow workers who may also wish to attend the meeting. This will not only offer an opportunity to listen to some of the experts in their respective fields but also a venue to meet colleagues and exchange ideas as well as to discuss some of the new items that are in the pipeline for this year.
It is important that members should pre-register as it will help us plan for lunches. The pre-registration deadline is April 4 and can be accomplished electronically. Please respond via email or fax to Evan Slow directly. For all attendees, the meeting fee, which includes lunch, will be $20.00.
For any additional information about the meeting, please contact any of the officers. We hope to see you on April 9th at the meeting in Mahwah!
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Hi. I am looking for a reference regarding electron dose measurement. Journal editor requested me to provide a reference on dose measurement in microscope. I looked over some textbooks and papers on electron damages, but I could not find. If someone knows a reference (textbook or experimental section in papers), please advise. Thank you,
Hiromi Konishi, Ph.D. University of New Mexico hkonishi-at-unm.edu
I was wondering if anyone can tell me where I might find demographics for research and clinical labs in the US. Any site's you can refer me to would be greatly appreciated.
I'm not sure that the requirements are clearly stated but I'll try my best to respond. If the person already has the blocks of wood cut, and they are only 5mm square, then they most likely are dried out as are any bacteria within the blocks and there is no point in rehydrating and then dehydrating with HMDS or CPD - the damage has already been done. On the other hand, if the blocks of wood are in a buffer solution, then perhaps HMDS would work but you will have to soak for quite a while and then it will take some time to dry. Five mm blocks could be dehydrated in a graded ethanol series (10, 20, 30, 50, 70, 80, 95, 3x100, 3:1 ethanol:HMDS, 1:1, 1:3 and then 100% HMDS. I'm guessing on the times as it would depend on the type of wood. I'd try 30min or so for each step.
How does he plan to look at anything in the interior of the block with an SEM? Will he section them? Depending on the wood, he might need a sliding microtome and a good sharp microtome blade (I've never had much success with razor blades and always preferred a good heavy, sharp blade. If he is going to look at only the surface, then whatever is there is already dried by the above process. I really don't know how the xylem parenchyma and the xylem rays are going to look (I'm assuming secondary xylem as you wrote "wood blocks". And depending on the material and what he wants to look at he might have to embed in some type of removable embedment.
If he is planning on sectioning the wood then the blocks will have to be soaked in warm water or sometimes I've had to use steam to soften the lignified secondary walls immediately before and during sectioning. This would create sections that you could then run up into HMDS; they would dehydrate much faster.
Just a few ideas and thoughts, Damian Neuberger
} Hi again: } } I have a guy who wants to look at the interior of some wood blocks using } our conventional SEM. He is trying to see bacteria and anything else that } might be hiding in there. } } His blocks are pretty small, about 5mm cubes. I thought about CPD, but had } second thoughts about how long to soak in CO2 etc. He did some experiments } and it took a couple of hours for some dye to make it all the way into the } middle of the block. Plus he has about 30 of these things to look at. That } could add up to a lot of time and CO2. } } I got an idea that HMDS might work for this project, but have no experience } with it. I think that being able to soak the blocks for a long time in HMDS } would be good, as would the chance to let them sublime to dryness over a } long time without too much attention. } } Do you think I am on the right track or is there something I am missing. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
I would propose to measure the beam current by using a Faraday cage and, by integration in time, you can get an acceptable approximation of the electron dosis of irradiation of your sample. The Faraday cups/cages are commercially available; you may have some doubts about the precision of electron beam current measurement, but it will work.
Sincerely,
Corneliu Sarbu
At 19/03/03 13:40, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Corneliu Sarbu, PhD Department of Metallurgy and Applied Materials Science (MTM Dept.) Catholic University of Leuven (KULeuven) Kasteelpark ARENBERG nr. 44 B-3001 Heverlee-Leuven, Belgium **************************************************************** Phone: +32-16-32.1241 - office +32-16-32.1264 - secretary of department Fax: +32-16-32.1992 or +32-16-32.1270 e-mail: Corneliu.Sarbu-at-mtm.kuleuven.ac.be web-site: www.mtm.kuleuven.ac.be ****************************************************************
I would propose to measure the beam current by using a Faraday cage and, by integration in time, you can get an acceptable approximation of the electron dosis of irradiation of your sample. The Faraday cups/cages are commercially available; you may have some doubts about the precision of electron beam current measurement, but it will work.
Sincerely,
Corneliu Sarbu
At 19/03/03 13:40, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Corneliu Sarbu, PhD Department of Metallurgy and Applied Materials Science (MTM Dept.) Catholic University of Leuven (KULeuven) Kasteelpark ARENBERG nr. 44 B-3001 Heverlee-Leuven, Belgium **************************************************************** Phone: +32-16-32.1241 - office +32-16-32.1264 - secretary of department Fax: +32-16-32.1992 or +32-16-32.1270 e-mail: Corneliu.Sarbu-at-mtm.kuleuven.ac.be web-site: www.mtm.kuleuven.ac.be ****************************************************************
SEM -cryo. need info on conduction by Leit-C CCC at low temperatures
I am looking at samples of ice (glacier & artificial chemical doped) using SEM with cryo-stage.
The samples are stuck to stubs using Leit-C conductive carbon cement. A co-worker has suggested to me that carbon has greatly reduced electrical conduction at liquid nitrogen temperatures but could not remember where he had seen it. Does anyone have any information on this? Thanks,
David Mallard Department Earth Sciences Bristol University, UK
Please take note of the upcoming School on Electron Microscopy of Powdered Nanostructured Materials
which will be hold in Wroc?aw (Poland), 19-20 September 2003. For complete information, please look at: http://celtam.int.pan.wroc.pl/ISEM/ Or Email: kepinski-at-int.pan.wroc.pl
Looking forward to seeing you in Wroc?aw Organizing Committee
Currently I am working on a project which need label cytokeratin in skin sample. we use NCK-CK15 from Novocastra Lab as primary antibody. we got nice result on paraffin section at LM level. But we couldn't get any K15 gold labeling on LR white sections. The sample was fixed with 4% paraformaldehyde and embedded in LR White at 50C for 1 day. We did get gold labeling of filaggrin on the LR white sections from same block. I was wondering if the cytokeratin antigen survived after sample preparation. Here are my questions. 1) Has anyone done this kind of labeling on resin embedded sections? If anyone did similar experiment, can you share the reference or protocol? 2) Immunohistochemistry always need perform antigen retrieval on sections before labeling, what about EM immunolabeling? 3) I have some references about antigen retrieval of resin sections. I tried high pressure heat and microwave heat with sodium citrate buffer. But I lost almost all sections from grids. Is there any tricks to keep sections stick on the coated or bare nickel grids.
Any suggestions are really appreciated!
Shanling
Shanling Shi Advanced Imaging & Measurement Unilever Research & Development -Edgewater 45 River Road Edgewater, NJ 07020 201-840-2340 Shanling.Shi-at-unilever.com
The U.S. Post Offices new Merlin (?) system has rejected the following addresses from the mailing of the March issue of Microscopy Today.
These subscribers were not sent magazines. I noticed several instances of 4-digit ZIP codes, other than that they look OK to me.
If you see your name here will you please resubscribe at http://www.microscopy-today.com
Thanks,
Ron Anderson, MT Editor
Ms. Becca Hoffman Triquint Semiconductor Dr. Stephen Giovannoni Oregon State Univ Dr. Andrew Gabor Evergreen Solar Inc Dr. Emil Adamec McLean Hospital Dr. Arthur Coates Microtherm Mr. Vincenzo Lordi Princeton University Dean Face DuPont Ms. Hermina Borgerink Wake Forest Univ Medical School Ms. M.L. Henson Usacil - Conus Mr. Larry L. Flinn US Army Criminal Inv Lab-Conus Larry Flynn Usacil-Conus[[same person]] Dr. Irene Kokkala North Georgia College Heike Gabrisch Cal Tech Ms. Vina R. Diderrich Univ Of Tennessee Ms. Judith A. Mescher Desc - Eltf Dr. Lewis D. Stegink Univ Of Iowa Mr. Wayne Engleman Monsanto BB3G Dr. John Longlet Chevron Chemical Co Daryl L Goad Texas A & M University Mr. John Curulli University of Southern California Dr. David Carter Center for Plant Cell Biology Ms. Susan DeMaggio Univ Of California Matthias Rief Stanford Univ Victoria Doyle-Jones Stanford University
Actually, this is a fairly short list for a database of over 16,500 subscribers!
Just a reminder about the exciting and innovative joint meeting of the Oklahoma Microscopy and Central States Microscopy and Microanalysis Societies that will be held at the University of Tulsa on April 10-11. Entitled "The Art of the Scientific Image", the two-day event will focus on the creation of scientific images, their digital processing and analysis, and their effective presentation. Connections between scientific imaging, human vision, and art will be explored throughout the meeting, which will feature an art exhibit and a region-wide contest for scientific images presented for aesthetic appeal rather than scientific content. Renowned imaging expert Dr. John Russ, author of The Image Processing Handbook, will present a workshop on the basics of image analysis, and Dr. Rob Weaver from the McCrone Research Institute will demonstrate techniques for forming beautiful images using specialized light microscopy techniques. A highlight of the meeting will be the keynote address by David Scharf, called by Time magazine "an Ansel Adams of inner space," whose scanning electron microscope images are regularly published in science journals, as well as in popular magazines such as Time, Nature, and Discovery. His address will accompany a viewing of the IMAX film "Hidden Dimensions" for which his contributions received an Emmy award. The meeting will close with an address by Dr. Lynn Gamwell, whose recent book Exploring the Invisible documents two centuries of connections between science, art, architecture, and culture.
The program and a registration form can be found on our website, see below
We are looking into a chartering a bus for those wishing to attend the meeting. The cost would be ~$50 per person and need to reserve it by Wednesday March 26 so please let us know asap if you are interested.
For more information contact Randy (tindallr-at-missouri.edu) or Lou (rosslm-at-missouri.edu).
Thanks, Lou Central States Microscopy and Microanalysis Society http://treefrog.cvm.uiuc.edu/~lam/csmms/
We have immunolabeled human skin for filaggrin and keratins at both light and electron microscopic levels. We have used mostly Zamboni's fixed tissue embedded in LRWhite with good success for filaggrin, K10 and Pan Keratin. However, I haven't tried K15. Two of my favorite tricks for unmasking are, 1) 0.3% sodium dodecyl sulphate (SDS) in 0.01M tris pH 7.6 incubated for 5-10 min, then rinse and continue as normal. 2) incubate the grid on 0.01% trypsin in PBS for 5-10 min., rinse and continue as normal.
Robert Underwood U of Washington Div. of Dermatolgy
On Thu, 20 Mar 2003, Shanling Shi wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi. there, } } Currently I am working on a project which need label cytokeratin in skin } sample. we use NCK-CK15 from Novocastra Lab as primary antibody. we got nice } result on paraffin section at LM level. But we couldn't get any K15 gold } labeling on LR white sections. The sample was fixed with 4% paraformaldehyde } and embedded in LR White at 50C for 1 day. We did get gold labeling of } filaggrin on the LR white sections from same block. I was wondering if the } cytokeratin antigen survived after sample preparation. Here are my questions. } 1) Has anyone done this kind of labeling on resin embedded sections? If anyone } did similar experiment, can you share the reference or protocol? } 2) Immunohistochemistry always need perform antigen retrieval on sections } before labeling, what about EM immunolabeling? } 3) I have some references about antigen retrieval of resin sections. I tried } high pressure heat and microwave heat with sodium citrate buffer. But I lost } almost all sections from grids. Is there any tricks to keep sections stick on } the coated or bare nickel grids. } } Any suggestions are really appreciated! } } Shanling } } Shanling Shi } Advanced Imaging & Measurement } Unilever Research & Development -Edgewater } 45 River Road } Edgewater, NJ 07020 } 201-840-2340 } Shanling.Shi-at-unilever.com } } } }
Could you use OCT compound? The Oxford reps say it can be mixed with a graphite powder. I have never got round to trying it.
Dave
On Thu, 20 Mar 2003 11:03:04 +0000 "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } I am looking at samples of ice (glacier & artificial chemical doped) } using SEM with cryo-stage. } } The samples are stuck to stubs using Leit-C conductive carbon cement. A } co-worker has suggested to me that carbon has greatly reduced } electrical conduction at liquid nitrogen temperatures but could not } remember where he had seen it. Does anyone have any information on this? } Thanks, } } David Mallard } Department Earth Sciences } Bristol University, UK }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
On Thu, 20 Mar 2003 11:03:04 +0000 "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } I am looking at samples of ice (glacier & artificial chemical doped) } using SEM with cryo-stage. } } The samples are stuck to stubs using Leit-C conductive carbon cement. A } co-worker has suggested to me that carbon has greatly reduced } electrical conduction at liquid nitrogen temperatures but could not } remember where he had seen it. Does anyone have any information on this? } Thanks, } } David Mallard } Department Earth Sciences } Bristol University, UK }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
The 2003 Spring Meeting of THE TEXAS SOCIETY FOR MICROSCOPY Embracing All Forms of Microscopy
April 3, 4, 5, 2003
Meeting will be held in Hubbard Hall on the campus of Texas Womans University, Denton, TX.
THURSDAY WORKSHOP Microwave Techniques Presented by Rick Giberson, Research and Development Manager, Ted Pella, Inc. Workshop sponsored by Ted Pella, Inc. and TSM
MSA LAS-SPONSORED SPEAKER FRIDAY, APRIL 4, 2003 HUBBARD HALL TWU - DENTON CAMPUS Dr. Lucille A. Giannuzzi, Ph.D., Associate Professor, Mechanical, Materials & Aerospace Engineering and Director, UCF/Cirent Materials Characterization Facility, University of Central Florida, Orlando, FL. Topic: "Focused Ion Beam Specimen Preparation for Everything". Focused ion beam techniques have been developed
to prepare site-specific specimens in a reproducible and rapid manner for SEM and TEM analysis. The conventional and the lift-out methods of specimen preparation will be discussed and FIB applications on a wide range of materials will be presented. The usefulness for using the FIB for solving scientific research problems will be emphasized.
Registration forms, hotel information, and author's instructions can be found on our website, http://www.texasmicroscopy.org/ . Questions about the program can be directed to Jo Taylor, Program Chair, at jtaylor-at-sfasu.edu.
I have a group looking at very early germinated Arabidopsis sp. They are having problems with good infiltration of the Epon-Araldite epoxy. While they are out searching the literature for solutions, I thought I'd drop a note to the list. Any suggestions for an infiltration protocol for these tiny plants? Spurr's? Dice up those little seeds?
TIA
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
On Thursday, March 20, 2003, at 03:03 AM, "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote: } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } I am looking at samples of ice (glacier & artificial chemical doped) } using SEM with cryo-stage. } } The samples are stuck to stubs using Leit-C conductive carbon cement. A } co-worker has suggested to me that carbon has greatly reduced } electrical conduction at liquid nitrogen temperatures but could not } remember where he had seen it. Does anyone have any information on } this? } Thanks, } Dear David, I do not know the properties of the cement you mention, nor do I have at hand any quantitative data on the conductivities of carbon at 293 K vs 77 K; however, the conductivity of carbon evaporated films at 77 K is sufficient for TEM observations at that temp. The conductivity of carbon at 4.2 K is not adequate for observations, and maybe that is what your co-worker remembers. On the other hand, SEM and TEM are sufficiently different that carbon could also be inadequate for SEM observations at 77 K. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
After 24 years of managing our small facility as a collaborative effort with interested faculty, I now have a new supervisor with little technical expertise. Our facility consists of an older TEM, a new SEM, a new deconvolution scope, and a Lieca SP1 confocal. I don't want to air our dirty laundry but I need a little help in justifying our equipment to the new boss. Now, she is proud of the fact that she still uses PhotoShop 3.0 and states that the best way to get a good picture is to take a good picture and then don't mess with it. While there is certainly some truth in the second fact, it is also very naive. And the first fact classifies her as a Luddite. Her research centers around the confocal and the deconvolution scope. So the move is on to dump the TEM and give the Digital Darkroom to the department Network Admin. This will free me up to tackle confocal projects for her and spend more time dusting. Yes, after 24 years of managing the facility on my own my new boss has actually given me a dusting schedule. However, she is quick to point out that this is not micro-managing. Alright, I digress. I need to justify having a couple graphics workstations, a slide scanner, a flat bed scanner with transparency tray, and a photo-quality printer. She would give this equipment to the Network Admin who would integrate it with the common computer room. I treat this equipment like I do optical equipment. I am concerned that once relocated to a common room, with no supervision, it will rapidly deteriorate. I also take exception with the apparently wide-spread belief that the computer support staff is the best source of information on digital imaging. I guess because it is digital?
So how do I keep those pieces of equipment I deem necessary for the facility? I think we should have a photo-quality printer. I think we need the scanner with tray for those increasingly rare occasions that we need to scan in a TEM neg or an illustration for a PowerPoint slide or publication. I find that I am not good at making an argument for what seems blindingly obvious to me. I just sputter and fume and ask, "... how can anyone have an Imaging Facility without a couple workstations and printers?". Or a TEM.
And then there is always the possibility she is right. Maybe we don't need to do this stuff to the standards I embrace. Should I convert to her mantra: "Do more, less well."?
Any good arguments out there that have worked for any of you?
I have an ageing Jeol JSM840 SEM (~20 years old) with a rapidly fading viewing screen and with no framestore. I cannot get a replacement viewing screen so am looking at other options such as getting a third part image grabbing system that would allow tv rate viewing of a reasonable sized image on a pc screen for focussing etc. A framestore for averaging would also be useful. I don't need the system for capturing digital images, I can already do that via my EDS system, it's the viewing screen that is the problem. The EDS capture system does not allow tv rate viewing.
The Deben Pixie 3000 system appears to do what I want but I'm interested to know of any other similar systems available. I'd also like to hear comments from users of the Deben system.
Cheers
Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
David Carbon has very useful conductivity at 77K, though 100K would be more typical of an LTSEM system cooled by LN2. This is easy to confirm in the lab with a multimeter. My measurements indicate that a 3mm dia. graphite rod 70mm long has a resistance of 1.2 ohms at room temperature and 2.1 ohms at 77K. No dramatic conductivity loss, therefore.
Leit-C is solvent-based paint, and expensive too. Is it really the best choice for your purpose, I wonder. I use TissueTek or aqueous colloidal graphite (Aquadag) to attach ice samples to stubs. Effective and much cheaper. Best wishes Chris
On 20 Mar 03, at 15:00, Bill Tivol wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Thursday, March 20, 2003, at 03:03 AM, } "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote: } } } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } } } I am looking at samples of ice (glacier & artificial chemical doped) } } using SEM with cryo-stage. } } } } The samples are stuck to stubs using Leit-C conductive carbon } } cement. A co-worker has suggested to me that carbon has greatly } } reduced electrical conduction at liquid nitrogen temperatures but } } could not remember where he had seen it. Does anyone have any } } information on this? Thanks, } } } Dear David, } I do not know the properties of the cement you mention, nor do I have } } at hand any quantitative data on the conductivities of carbon at 293 K } vs 77 K; however, the conductivity of carbon evaporated films at 77 K } is sufficient for TEM observations at that temp. The conductivity of } carbon at 4.2 K is not adequate for observations, and maybe that is } what your co-worker remembers. On the other hand, SEM and TEM are } sufficiently different that carbon could also be inadequate for SEM } observations at 77 K. Yours, Bill Tivol EM Scientist and Manager } Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 } California Institute of Technology Pasadena CA 91125 (626) 395-8833 } tivol-at-caltech.edu } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 /8669 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
This is a rough one. We have just stated a new EM lab (less than two years operational) Our TEM (Which is new!) is not functioning due to the SIS camera being away for repairs under warranty now for some time. The EDS has been sent back twice now and I hope this time around the repairs will be successful. Being in Africa does have it pro's to like the weather, the birds and wildlife. We also have a lot more freedom in research. I am seeing forward to a prosperous year full of publications with EM input and well trained motivated users! (my whish list and also the units goal)
Thus if your TEM is running this is already a blessing. This alone (service contract cost depending) should be enough to motivate for keeping it. We are in a process to monitoring all publications on all instruments as a justification to keep it running or to replace it should the kneed arise.
My wife is brilliant at management and she helps me a lot. Her motto is always: "if you motivate something well enough you can cell ice to an Eskimo."
This is the most important point of all: Since she is now directing the facility, please go ask her to give the Objectives, Goals and mission statement for the facility to you in writing (Her view). If possible ask her also for a 5, and 10 Year plan for the facility. Before attempting to motivate anything think things over carefully. You two must be able to work in peace since strive will destroy the facility quickly and drive users away! With unity much can be achieved since her signature is required most likely on all purchase requests, motivations and budget expenditure.
When you do the motivation, link your instruments to the objectives and goals of your institute as well as the ones you got from her. Let her see that you are on her side and you would like to see these goals achieved.
If you can provide a list of publications where each instrument was involved in over its lifespan as a percentage of the total number of publications published in your institution it should help to explain the kneed of each instrument, scanner, SG's etc...
You can probably get a letter from the IT guys to state that your application, software, set-up and configuration is unique to the institution and it serve the best to keep it out the hands of ordinary villains to prevent expensive reconfiguration and repair. I had to fight to keep them out without driving them away since I kneed there help but not there software upgrades etc!
If you have a good relationship with your research office ask them for a opinion in writing.
It is worth it to do a survey in the institute to all relevant users, past users and possible users including head of departments. Things worth getting a answer on are questions like: What do you expect from the facility, What instruments would you like to be present in the unit, Past usage, usefulness of input in their research. Possible future utilisation of your unit. Suggestions to improve the service as well as suggestions wishes for instruments/capability's currently not present. If you can get postgads promised to instruments and research programs including your facility the better. I know that is not always possible but if you do not ask you will not know. We did that at a previous institute where I was employed. This was a very useful exercise. This helped to direct the unit and improve on our service. These documents are also a very useful source for equipment motivation when it is combined into a format top management can understand.
Hopefully from this information the wise path to follow will be much more clearer to all.
Good luck.
-----Original Message----- } From: Rick Harris [mailto:raharris-at-ucdavis.edu] Sent: Friday, March 21, 2003 1:08 AM To: microscopy-at-sparc5.microscopy.com
After 24 years of managing our small facility as a collaborative effort with interested faculty, I now have a new supervisor with little technical expertise. Our facility consists of an older TEM, a new SEM, a new deconvolution scope, and a Lieca SP1 confocal. I don't want to air our dirty laundry but I need a little help in justifying our equipment to the new boss. Now, she is proud of the fact that she still uses PhotoShop 3.0 and states that the best way to get a good picture is to take a good picture and then don't mess with it. While there is certainly some truth in the second fact, it is also very naive. And the first fact classifies her as a Luddite. Her research centers around the confocal and the deconvolution scope. So the move is on to dump the TEM and give the Digital Darkroom to the department Network Admin. This will free me up to tackle confocal projects for her and spend more time dusting. Yes, after 24 years of managing the facility on my own my new boss has actually given me a dusting schedule. However, she is quick to point out that this is not micro-managing. Alright, I digress. I need to justify having a couple graphics workstations, a slide scanner, a flat bed scanner with transparency tray, and a photo-quality printer. She would give this equipment to the Network Admin who would integrate it with the common computer room. I treat this equipment like I do optical equipment. I am concerned that once relocated to a common room, with no supervision, it will rapidly deteriorate. I also take exception with the apparently wide-spread belief that the computer support staff is the best source of information on digital imaging. I guess because it is digital?
So how do I keep those pieces of equipment I deem necessary for the facility? I think we should have a photo-quality printer. I think we need the scanner with tray for those increasingly rare occasions that we need to scan in a TEM neg or an illustration for a PowerPoint slide or publication. I find that I am not good at making an argument for what seems blindingly obvious to me. I just sputter and fume and ask, "... how can anyone have an Imaging Facility without a couple workstations and printers?". Or a TEM.
And then there is always the possibility she is right. Maybe we don't need to do this stuff to the standards I embrace. Should I convert to her mantra: "Do more, less well."?
Any good arguments out there that have worked for any of you?
Dear Rick, Definitely spurr's is the best. I do not how hard is the seed coat, You could try araldite-DDSA-DMP mixture. after fixation and dehydration, PO etc, Infilterate with 20% resin mix in PO and leave your sample vial open overnight. Next day(~20h) change it to fresh 100% resin mix for about 2hrs before embedding. curing, usual at 60 deg C for 72hrs. Shashi Singh Scientist CCMB Hyderabad-India
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I have a group looking at very early germinated Arabidopsis sp. They are having problems with good infiltration of the Epon-Araldite epoxy. While they are out searching the literature for solutions, I thought I'd drop a note to the list. Any suggestions for an infiltration protocol for these tiny plants? Spurr's? Dice up those little seeds?
TIA
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
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Peter No, I did a two-point measurement with the rod connected at both ends. The rod was immersed in liquid nitrogen while remaining connected, so the resistances are comparable. The figures I quote are the resistance of the specified length of the 3mm carbon rod, not the resistivity of carbon, which is usually quoted as ohms per square. Chris
} David; } } In doing this measurement of electrical resistivity, did you use a } "four point Kelvin" measurement to determine sheet resistance? It's } the only way I know of to know and compare resistances unless all } things in the comparison are held the same, i.e. thickness, width and } length. } } Peter Tomic } } } ----- Original Message ----- } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } To: {David.Mallard-at-bristol.ac.uk} ; "Bill Tivol" {tivol-at-caltech.edu} } Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, March 21, 2003 } 3:45 AM Subject: Re: SEM -cryo. need info on conduction by Leit-C CCC } at low temperatures } } } } -------------------------------------------------------------------- } } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } } ---. } } } } } } David } } Carbon has very useful conductivity at 77K, though 100K would be } } more typical of an LTSEM system cooled by LN2. This is easy to } } confirm in the lab with a multimeter. My measurements indicate that } } a 3mm dia. graphite rod 70mm long has a resistance of 1.2 ohms at } } room temperature and 2.1 ohms at 77K. No dramatic conductivity loss, } } therefore. } } } } Leit-C is solvent-based paint, and expensive too. Is it really the } } best choice for your purpose, I wonder. I use TissueTek or aqueous } } colloidal graphite (Aquadag) to attach ice samples to stubs. } } Effective and much cheaper. Best wishes Chris } } } } On 20 Mar 03, at 15:00, Bill Tivol wrote: } } } } } ------------------------------------------------------------------ } } } ---- -- The Microscopy ListServer -- Sponsor: The Microscopy } } } Society 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 Thursday, March 20, 2003, at 03:03 AM, } } } "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote: } } } } } } } } SEM -cryo. need info on conduction by Leit-C CCC at low } } } } temperatures } } } } } } } } I am looking at samples of ice (glacier & artificial chemical } } } } doped) using SEM with cryo-stage. } } } } } } } } The samples are stuck to stubs using Leit-C conductive carbon } } } } cement. A co-worker has suggested to me that carbon has greatly } } } } reduced electrical conduction at liquid nitrogen temperatures } } } } but could not remember where he had seen it. Does anyone have } } } } any information on this? Thanks, } } } } } } } Dear David, } } } I do not know the properties of the cement you mention, nor do I } } } have } } } } } } at hand any quantitative data on the conductivities of carbon at } } } 293 K vs 77 K; however, the conductivity of carbon evaporated } } } films at 77 K is sufficient for TEM observations at that temp. } } } The conductivity of carbon at 4.2 K is not adequate for } } } observations, and maybe that is what your co-worker remembers. On } } } the other hand, SEM and TEM are sufficiently different that carbon } } } could also be inadequate for SEM observations at 77 K. Yours, Bill } } } Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility } } } Broad Center, Mail Code 114-96 California Institute of Technology } } } Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu } } } } } } } } } } } } ========================================== } } Dr. Chris Jeffree } } University of Edinburgh } } BIOSEM - Biological Sciences Electron Microscope Facility } } Institute of Cell and Molecular Biology } } Waddington Building } } King's Buildings, Mayfield Road } } EDINBURGH, EH9 3JN, Scotland, UK } } Tel. #44 (0) 131 650 5554 /8669 } } FAX. #44 (0) 131 650 6563 } } Mobile 07710 585 401 } } email c.jeffree-at-ed.ac.uk } } ========================================= } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 /8669 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
More questions! How do you decide whether to use TissueTek or aqueous colloidal graphite (Aquadag) for an application? Where do get the latter?
Dave
On Fri, 21 Mar 2003 08:45:18 -0000 Chris Jeffree {cjeffree-at-srv0.bio.ed.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } David } Carbon has very useful conductivity at 77K, though 100K would be } more typical of an LTSEM system cooled by LN2. This is easy to } confirm in the lab with a multimeter. My measurements indicate } that a 3mm dia. graphite rod 70mm long has a resistance of 1.2 } ohms at room temperature and 2.1 ohms at 77K. No dramatic } conductivity loss, therefore. } } Leit-C is solvent-based paint, and expensive too. Is it really the } best choice for your purpose, I wonder. I use TissueTek or aqueous } colloidal graphite (Aquadag) to attach ice samples to stubs. } Effective and much cheaper. } Best wishes } Chris } } On 20 Mar 03, at 15:00, Bill Tivol wrote: } } } ---------------------------------------------------------------------- } } -- The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Thursday, March 20, 2003, at 03:03 AM, } } "David.Mallard-at-bristol.ac.uk"-at-sparc5.microscopy.com wrote: } } } } } } SEM -cryo. need info on conduction by Leit-C CCC at low temperatures } } } } } } I am looking at samples of ice (glacier & artificial chemical doped) } } } using SEM with cryo-stage. } } } } } } The samples are stuck to stubs using Leit-C conductive carbon } } } cement. A co-worker has suggested to me that carbon has greatly } } } reduced electrical conduction at liquid nitrogen temperatures but } } } could not remember where he had seen it. Does anyone have any } } } information on this? Thanks, } } } } } Dear David, } } I do not know the properties of the cement you mention, nor do I have } } } } at hand any quantitative data on the conductivities of carbon at 293 K } } vs 77 K; however, the conductivity of carbon evaporated films at 77 K } } is sufficient for TEM observations at that temp. The conductivity of } } carbon at 4.2 K is not adequate for observations, and maybe that is } } what your co-worker remembers. On the other hand, SEM and TEM are } } sufficiently different that carbon could also be inadequate for SEM } } observations at 77 K. Yours, Bill Tivol EM Scientist and Manager } } Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 } } California Institute of Technology Pasadena CA 91125 (626) 395-8833 } } tivol-at-caltech.edu } } } } } } } ========================================== } Dr. Chris Jeffree } University of Edinburgh } BIOSEM - Biological Sciences Electron Microscope Facility } Institute of Cell and Molecular Biology } Waddington Building } King's Buildings, Mayfield Road } EDINBURGH, EH9 3JN, Scotland, UK } Tel. #44 (0) 131 650 5554 /8669 } FAX. #44 (0) 131 650 6563 } Mobile 07710 585 401 } email c.jeffree-at-ed.ac.uk } ========================================= }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
If you just want to see the interior of the wood, it should be easy enough
to split 5mm cubes with a single edge razor blade, knife or wood chisel along the longitudinal axis. You have your choice of exposing surfaces with orientations that are radial, tangential or something in between. (If
those terms don't mean anything to you, take a look at the Soc. for Wood Sci. & Tech. at http://www.swst.org/teach/set2/struct1.html for an excellent introduction to wood structure and terminology).
If you need to expose a cross-section, I'd echo the LN2 idea with a couple
of suggestions. For my wood science MS research, I had to break numerous 5mm square by 50mm "matchsticks" with as brash (perpendicular to the long axis and not jagged) a failure as possible for an SEM study. What worked best for me was to lightly score the wood on the tension side where I wanted the break and insert it into a small jig that clamped the matchstick on both sides of the score line. I immersed the jig w/ wood in LN2 until the boiling slowed significantly (may take a while, wood is a good insulator) and broke the sample immediately and energetically on removal from the LN2. If your pieces are only 5mm cubes, you may not be able to use such a jig, but perhaps you don't need the brash failure I did. Another factor I found important was the moisture content of the wood
- too low was a problem. No matter what technique you use, you'll likely encounter the variability of wood. Count on lots of replications!
Pete Eastman Microscopy Rohm and Haas Company 215 619-5229
"Tindall, Randy D." {TindallR-at-missouri.edu} 03/19/2003 09:13 AM
To: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} cc: {microscopy-at-sparc5.microscopy.com} bcc: Subject: RE: CPD or HMDS for wood blocks?
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Jonathan,
We recently looked at wood blocks in our FESEM and the only preparation we did was to freeze them in liquid nitrogen, then fracture them with pliars to expose the internal structure, followed by several days in a vacuum chamber to get as much moisture and air out as possible. We glued them down with copius amounts of conductive paint, then sputtered the bejesus out of them.
The structure was well preserved and we did find bacteria.
The only problem was breaking the harder pieces of wood. Some of the wood was ancient and very brittle, but the new pieces were tough.
Hope this is applicable to your problem.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Jon Krupp [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Tuesday, March 18, 2003 3:03 PM To: Microscopy-at-sparc5.microscopy.com
Hi again:
I have a guy who wants to look at the interior of some wood blocks using our conventional SEM. He is trying to see bacteria and anything else that might be hiding in there.
His blocks are pretty small, about 5mm cubes. I thought about CPD, but had second thoughts about how long to soak in CO2 etc. He did some experiments and it took a couple of hours for some dye to make it all the way into the middle of the block. Plus he has about 30 of these things to look at. That could add up to a lot of time and CO2.
I got an idea that HMDS might work for this project, but have no experience with it. I think that being able to soak the blocks for a long time in HMDS would be good, as would the chance to let them sublime to dryness over a long time without too much attention.
Do you think I am on the right track or is there something I am missing.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
On Thursday, March 20, 2003, at 07:12 PM, Dave Phelan wrote:
} I have an ageing Jeol JSM840 SEM (~20 years old) with a rapidly fading } viewing screen and with no framestore. I cannot get a replacement } viewing } screen so am looking at other options such as getting a third part } image } grabbing system that would allow tv rate viewing of a reasonable sized } image on a pc screen for focussing etc. A framestore for averaging } would } also be useful. I don't need the system for capturing digital images, } I can } already do that via my EDS system, it's the viewing screen that is the } problem. The EDS capture system does not allow tv rate viewing. } Dear Dave, If you have old screens or screen blanks, they can be recoated with phosphor. There are companies that do this; I don't have access to their names or addresses, but I am pretty sure I have posted at least one to the list in the past. You can also recoat the screen blank yourself, and I think I posted a procedure to the list also. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Dave Phelan wrote: ============================================================================ ======== I have an ageing Jeol JSM840 SEM (~20 years old) with a rapidly fading viewing screen and with no framestore. I cannot get a replacement viewing screen so am looking at other options such as getting a third part image grabbing system that would allow tv rate viewing of a reasonable sized image on a pc screen for focussing etc. A framestore for averaging would also be useful. I don't need the system for capturing digital images, I can already do that via my EDS system, it's the viewing screen that is the problem. The EDS capture system does not allow tv rate viewing.
The Deben Pixie 3000 system appears to do what I want but I'm interested to know of any other similar systems available. I'd also like to hear comments from users of the Deben system. ============================================================================ ========= Another alternative is the passive image capture software called Spectrum Mono, as described on URL http://www.2spi.com/catalog/software/spectrum.shtml
It does what you want it to do, and, it is perhaps the lowest cost alternative for a product of this type. On most older SEMs it is easily installed, either by the user or the preferred service engineer. We have had the software running on one of our own Model 840 JEOL SEMs and it seems to perform up to expectations.
Disclaimer: SPI Supplies has distributed this "passive" image capture software for some time and we have a vested interest in its promotion.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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Can anyone recommend a source of 5-10L LN2 portable dewars? These are for filling a 10L EDS detector. The LN2 will be transported by car.
Before I introduce myself, I wish to inform you that this letter is not a hoax mail and I urge you to treat it serious. I am Dr. Muhammad Khalil Hasan, a banker with the Mashreq Bank Plc Dubai, UAE. I am the Accounts officer late Mr. Robert Chapman the African Area Director of SIL International, who unfortunately died in the crash of Kenya Airways Flight 431 in Abidjan, Ivory Coast, January 30 2000. You will read more stories about the crash on visiting this website, news.airwise.com/airlines/archive/2000/kenya2000.html and also in this website, where Chapman's company talked about his death in the Kenya crash. You shall as well find the pictures of Chapman and his wife there. Mr. Chapman was from Hamilton, Ontario Canada.
Since the death of Chapman, I as his accounts officer in the bank, have made several enquiries to locate his only surviving relation, without any success. I came across your name and contact, on the course of my personal searching for Chapman's relations, so I decided to contact you for this project.
I am contacting you to assist in repatriating and securing the wealth left behind in a fixed deposit account by Robert, before they get confiscated or declared unserviceable by my bank. The board of my bank, has issued a notice that after 2 months from now and no next of kin shown up for the claim, the funds will be confiscated and declared unserviceable. Since I and my team have been unsuccessful in locating Robert's relatives for sometime now, I seek your consent to present you as the Next of Kin of the deceased since you are a foreigner, so that the proceed of this deposit valued at $USD7.5 Million Dollars can be released to you.
The bank will release the funds to any foreigner who has all related information/documents to the bank account. I am in charge of this matter in my bank, because I am his accounts officer. Your application will be directed to my department for verification and approval. Everything is under my control.
I shall provide you all the information and copy of the certificate of deposit issued to Robert when he deposited the funds. I shall also involve a good attorney who shall represent you in all the appropriate offices for the claim. Please, find in the attachment my work ID Card. This is just to proof to you that my proposal is genuine. I also have all necessary information we need for the claim and once the money is transferred to you, I shall destroy all the documents used for the claim and leave no traces. After everything, you shall have 40% of the total sum,while 60% for me.
All requires is your honest cooperation to enable us see this business successful. I guarantee that this will be executed under a legitimate arrangement that will protect you from any breach of the law. Robert was a very good man and it is not wise to allow his hard earned wealth to be stolen by the greedy directors of my bank.
Further details awaits your response by email. PLEASE, TREAT THIS PROPOSAL AS TOP SECRET. Please, confirm the receipt of my Id Card in the attachment, as it was difficult to attach.
Tell her NO! Make no consigns not afoot to compromise or be her friend just tell her no.
When you end up in her supervisors offices explaining the dusting duties, her ludilite ways and designs to enslave you. If he tells you to go along to get along say no. Let her find some other domestic servant to do her work. and call a head hunter.
Unless dusting is what you should be doing I would think if you are any good you can find a better paying job down the hall and let here dust her own lab.
The only thing is you have to mean no you can say it and not mean it shows.
One of the reasons women get less pay is they will put up with a lot more abuse on the job than most a,m/
No is an extremely powerful word that is not used near enough. If the whole lab will tell her no se will be gone by the end of the moth unless she is sleeping wiht the boss. Then it will take another month or two. In the mean time a match stem propping the valve on a tire open will annoy here. do it to two tires cost here 45 minutes that day.A 10 dollar bill will get the doomster sat where her car can't get over night. A 40 dollar tip will get a pitcher of ice water down the front of her dress as she gets up to speak at a meeting..
Even the densest manager gets the clue some where along the line.
Bad management is on of the most embedded instructions in our country and to letters NO can bring them to their knees if everyone cooperates and be rid of them.
They may be a great tech and like the work give them power over other can turn Dr. Jeckel into Mr. Hyde.
Being a bad manger is only a third the mangers fault. A third or more rests with their boss and third rests with the people that work for them. You were looking for a job when you found this one and the next one should pay better with new challenges and new manure piles to afford.
good luck
Gordon Couger Stillwater, OK www.couger.com/gcouger ----- Original Message ----- } From: "Rick Harris" {raharris-at-ucdavis.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 20, 2003 5:07 PM
I am in need of a scoring wheel cartridge for a Leica Reichert Knifemaker II. Please reply with source contact info.
Thanks.
William C. Hamlett, Ph.D. Professor of Anatomy & Cell Biology Indiana University School of Medicine Notre Dame, Indiana 46556, USA e-mail whamlett-at-nd.edu Telephone 574-631-7194 FAX 574-631-7821 http://www.nd.edu/~whamlett
This is to inform you that the Call for Abstracts for the forthcoming International Meeting on Applied Physics (APHYS-2003), to be held during October 14-18th 2003 in Badajoz (Spain), is open. All the information regarding this interdisciplinary conference can be found at the Conference website
www.formatex.org/aphys2003/aphys2003.htm
Some of the topics to be covered will be
- Surfaces, Interfaces and Colloids - Imaging Techniques, Microscopy - Nano-sciences and Technologies - Biophysics and Biophysical Chemistry - Materials Science & Engineering, Applied Solid State Physics/Chemistry - Advanced and Fuctional Materials - Biomedical Engineering and Biomaterials - Biological & Medical Physics - Computational Physics - Radiation Physics, Applied Nuclear Physics/chemistry, Radiation Protection
In addition to the regular Scientific Program, several International Workshops will be held as pre-conference events. The following Workshops are presently confirmed
1. Workshop on Modern Applied Microscopy in Molecular and Cell Biophysics Research 2. International Interdisciplinar Workshop on Bioengineered Non-crystalline Solids 3. Workshop on Interfaces in Colloidal and Particulate Systems 4. International Workshop on Radiation Protection and Dosimetry
The Conference will be specifically interested in receiving reports on Interdisciplinary researches relating Physics with other Sciences such as Biology, Chemistry, Information Technology, Medicine, etc or relating different Physics areas. In other words, we are specially (but not exclusivelly) interested in reports applying the techniques, the training, and the culture of physics to research areas usually associated with other scientific and engineering disciplines
APHYS-2003 will also serve as a platform to search for partners for transnational collaboration projects, specially for the EU Sixth Framework Program (NETworks of Excellence and Integrated Projects). "Projects Presentations" and "Call for Partners" presentations proposals are therefore encouraged and welcomed. If you are interested in taking part of this Conference feature, please send us the corresponding form available at the website.
In addition to the "traditional" oral contributed and posters presentation, a Virtual Participation modality has been established for those researchers unable to attend it in person. A limited number of works can be presented in this way. Please refer to the Conference website for details.
If you are interested in taking part of APHYS-2003, please send us your PRE-REGISTRATION FORM (at the main website of the conference) as soon as possible. The pre-registration form is also available through the direct URL http://www.formatex.org/aphys2003/preregistration.htm
Deadline for abstracts submission is April 30th 2003 (for oral presentations proposals; for posters additional time will be provided) although we highly recommend you to submit your abstract as soon as possible to avoid saturation during the days before the deadline.
Proceedings Accepted and presented papers will be reviewed for publication in special issues of several international Journals: - Journal of Microscopy - Journal of Non-crystalline Solids - Applied Surface Science - Colloids and Surfaces A - Powder Technology (to be confirmed) - Microelectronics Journal - Physica Scripta - Radiation Protection Dosimetry - Applied Physics A (Materials Science & Processing, to be confirmed). - Biomedial Materials and Engineering
Also a book "Advances in Applied Physics" will be published by an international publisher (Kluwer or the American Institute of Physics, with those papers accepted for presentation but not suitable for the journal issues.For up-to-date information on publications participating at the Conference as publishers, please visit regularly the Conference website (Proceedings sections).
For any question or suggestion, please do not hesitate to contact us at secretariat-at-formatex.org, or visit www.formatex.org/aphys2003/aphys2003.htm (Bookmark the page!!) We would also appreciate if could disseminate this Call for Papers through your Department or Institution.
We hope to meet you at this exciting and interdisciplinar international meeting!
SUV would be fine versus car. I thought that there were Dewars that could be filled and had pressure relief vents. Then, comes the problem of getting the LN2 out of the Dewar.
This is a tough area to tackle for a low use system. I suspect EDS would be used a half a day per week, perhaps less. Total of around 100 hours per year. For the non-use periods, the detector would be empty. This is suppose to be OK.
gary g.
At 09:42 AM 3/22/2003, you wrote: } Gary } There is really only one way to carry liquid nitrogen safely by car } and that is outside it. A pick-up truck, or SUV with a rear transport } cage for a dewar would be ideal, but if liquid nitrogen spills inside } the car you, your passengers and other road users are in mortal } danger. Taylor-Wharton manufacture dry shippers that can safely } transported inside cars and aircraft, but unfortunately they are } designed as refrigerators, and will not release liquid nitrogen when } inverted. They are therefore useless for refilling your EDS } detector. } Chris } } Dr. Chris Jeffree } Inveresk Cottage } 26, Carberry Road } Inveresk } Musselburgh } Midlothian } EH21 8PR } Tel: +44 131 665 6062 } FAX +44 131 653 6248 } Mobile 07710 585 401 } ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} } To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} } Sent: Saturday, March 22, 2003 2:20 AM } Subject: LN2 dewar } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 recommend a source of 5-10L } } LN2 portable dewars? These are for filling } } a 10L EDS detector. The LN2 will be } } transported by car. } } } } Any ideas? } } } } tnx, } } gary g. } } } }
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Gary Gaugler wrote: ============================================================================ SUV would be fine versus car. I thought that there were Dewars that could be filled and had pressure relief vents. Then, comes the problem of getting the LN2 out of the Dewar.
This is a tough area to tackle for a low use system. I suspect EDS would be used a half a day per week, perhaps less. Total of around 100 hours per year. For the non-use periods, the detector would be empty. This is suppose to be OK. ============================================================================ == I thought that in the USA at least, since liquid nitrogen is considered to be a "dangerous goods" material, in order to transport it, one would need to be a licensed "dangerous goods" transporter. For example those who deliver liquid nitrogen have their trucks placarded with the green flammable gas placard. This is all highly regulated by the US DOT.
Is it not similar to the case for transporting hazardous waste, one needs to be licensed to carry it in a vehicle?
I am not the last word on this, however, and there could be some exemptions, perhaps for personal use, but if there are, I have not heard of any. We all have to make our own decisions about such things, but one should be sure that laws and/or regulations are not being violated. In the event of an accident, enormous legal liability for someone could be the result (not a legal opinion, just a lay presumption).
Chuck
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Rick, There are a number of factors to consider. It would help to know what parts are not infiltrated well to determine the cause of the problems.
Spurr's is definitely easier to get into plant material. If the seeds are still intact it will be hard to get any resin in. You stated that they are early germinants. Does this mean that they have broken through the seed coat? If they haven't, you may have to abrade the seeds in some way to get fixation and infiltration. Larger seeds can be shaken over sandpaper, but this may not work with the smaller seeds. (Dicing the seeds may work but could smash the cells.) Consider extending the infiltration time to several days to a week. I have had to infiltrate for more than a week to get successful embedding of some woody material. You will have to make regular changes of resin, but it does make a difference.
I don't know your protocol, but another factor may be fixation and especially dehydration. You have to use extended fixation and dehydration times to allow hydrophobic resin to penetrate plant material. Often the woody parts retain water and the resin won't penetrate.
If you have access to a lab microwave then you should consider trying infiltration in there. It really helps!
Good luck,
Kim {} {} {} {} {} Kim Rensing PhD Department of Botany, UBC 6270 University Blvd. Vancouver BC, Canada V6T 1Z4
On Thursday, March 20, 2003, at 02:02 PM, Rick Harris wrote: } } I have a group looking at very early germinated Arabidopsis sp. They } are having problems with good infiltration of the Epon-Araldite epoxy. } While they are out searching the literature for solutions, I thought } I'd drop a note to the list. Any suggestions for an infiltration } protocol for these tiny plants? Spurr's? Dice up those little seeds? } } TIA } } } Rick A. Harris, Director } Microscopy and Imaging Facility
Rick, There are a number of factors to consider. It would help to know what parts are not infiltrated well to determine the cause of the problems.
Spurr's is definitely easier to get into plant material. If the seeds are still intact it will be hard to get any resin in. You stated that they are early germinants. Does this mean that they have broken through the seed coat? If they haven't, you may have to abrade the seeds in some way to get fixation and infiltration. Larger seeds can be shaken over sandpaper, but this may not work with the smaller seeds. (Dicing the seeds may work but could smash the cells.) Consider extending the infiltration time to several days to a week. I have had to infiltrate for more than a week to get successful embedding of some woody material. You will have to make regular changes of resin, but it does make a difference.
I don't know your protocol, but another factor may be fixation and especially dehydration. You have to use extended fixation and dehydration times to allow hydrophobic resin to penetrate plant material. Often the woody parts retain water and the resin won't penetrate.
If you have access to a lab microwave then you should consider trying infiltration in there. It really helps!
Good luck,
Kim {} {} {} {} {} Kim Rensing PhD Department of Botany, UBC 6270 University Blvd. Vancouver BC, Canada V6T 1Z4
On Thursday, March 20, 2003, at 02:02 PM, Rick Harris wrote: } } I have a group looking at very early germinated Arabidopsis sp. They } are having problems with good infiltration of the Epon-Araldite epoxy. } While they are out searching the literature for solutions, I thought } I'd drop a note to the list. Any suggestions for an infiltration } protocol for these tiny plants? Spurr's? Dice up those little seeds? } } TIA } } } Rick A. Harris, Director } Microscopy and Imaging Facility
JEOL will have your picture tube (CRT) for a replacement. If they don't by some miracle, then contact Richardson Electronics at www.rell.com or (630)208-2200. Use information from your SEM CRT label (monitor pulls out, label is on the side of a CRT). You can get new CRT form Richardson for couple-three hundred $. Make sure they are aware that CRT phosphor must have longer decay time (phosphor type is also stated on the CRT label). Longer phosphor decay time is not critical, but helpful.
If you in the mood, get a monochrome video monitor (such as security monitor), and connect TV video from SEM to it.
Countless video frame grabbers with frame averaging function are available- too many to list. What features and what price range are you looking for?
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
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----- Original Message ----- } From: "Dave Phelan" {emudp-at-mail.newcastle.edu.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 20, 2003 10:12 PM
Hi, Bill
Could you re-post the directions for a home-cooking procedure? I didn't think it was possible to recoat CRTs.
cheers
rtch
} } } On Thursday, March 20, 2003, at 07:12 PM, Dave Phelan wrote: } } } I have an ageing Jeol JSM840 SEM (~20 years old) with a rapidly } } fading viewing screen and with no framestore. I cannot get a } } replacement viewing screen so am looking at other options such as } } getting a third part image grabbing system that would allow tv rate } } viewing of a reasonable sized image on a pc screen for focussing } } etc. A framestore for averaging would also be useful. I don't need } } the system for capturing digital images, I can already do that via } } my EDS system, it's the viewing screen that is the problem. The EDS } } capture system does not allow tv rate viewing. } } } Dear Dave, } If you have old screens or screen blanks, they can be recoated with } phosphor. There are companies that do this; I don't have access to } their names or addresses, but I am pretty sure I have posted at least } one to the list in the past. You can also recoat the screen blank } yourself, and I think I posted a procedure to the list also. Good } luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron } Microscopy Facility Broad Center, Mail Code 114-96 California } Institute of Technology Pasadena CA 91125 (626) 395-8833 } tivol-at-caltech.edu } }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
} From: "Garber, Charles A." {cgarber-at-2spi.com} : : Gary Gaugler wrote: : ============================================================================ : SUV would be fine versus car. I thought that there were Dewars that could : be filled and had pressure relief vents. Then, comes the problem of getting : the LN2 out of the Dewar. : : This is a tough area to tackle for a low use system. I suspect EDS would be : used a half a day per week, perhaps less. Total of around 100 hours per : year. For the non-use periods, the detector would be empty. This is : suppose to be OK. : ============================================================================ : == : I thought that in the USA at least, since liquid nitrogen is considered to : be a "dangerous goods" material, in order to transport it, one would need to : be a licensed "dangerous goods" transporter. For example those who deliver : liquid nitrogen have their trucks placarded with the green flammable gas : placard. This is all highly regulated by the US DOT. : : Is it not similar to the case for transporting hazardous waste, one needs to : be licensed to carry it in a vehicle? : : I am not the last word on this, however, and there could be some exemptions, : perhaps for personal use, but if there are, I have not heard of any. We : all have to make our own decisions about such things, but one should be sure : that laws and/or regulations are not being violated. In the event of an : accident, enormous legal liability for someone could be the result (not a : legal opinion, just a lay presumption). : : Chuck : Liquid nitrogen should be much less dangerous the liquid oxygen and I see trucks that haul live fish with LOX tanks on them. They may be hauling the LOX without the proper permits. The people in the bait fish business are not particular careful about obeying the law. Artificial insemination technicians carry LN2 dewers all over the country as well.
To pump the LN2 put a pipe that reaches the bottom of the tank. Just close the vents and put a little current through a heater submerged in the nitrogen and the pressure will pump the LN2 out the pipe. Four psi should pump about 10 gallons a minutes or more though a 3/4 inch pipe to a container the same height as the LN2 tank.
If the temperature of boiling propane is low enough you could vaporize propane and dispose ot it in a flame and greatly reduce your storage and procurement problems. A propane tank has to be in sealed compartment in vehicle or mounted out it and the vehicle should not be parked in a building. It can legally be installed in vehicles.
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (spohnheimer-at-dalsemi.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, March 22, 2003 at 19:57:07 ---------------------------------------------------------------------------
Email: spohnheimer-at-dalsemi.com Name: John Spohnheimer
Organization: Dallas Semiconductor (Commercial)
Education: Graduate College
Location: Dallas, TX
Question: I am an engineer/physicist working with semiconductors & have used high-quality optical/SEM/TEM microscopes for ~25 years. Recently, a friend (also a physicist) asked why we can see sub-micron geometries (sometimes even one-two tenth micron diameter particles are visible) using optical microscopes with wavelengths nominally in the 1/2 micron range.
I argued that what we're really seeing is simply a phase interference pattern generated by the sub-micron particle, not the real particle itself. That's why most sub-resolution particles appear spherical in an optical microscope and often-times look completely different under SEM examination.
I'm curious if a professional in this field has a better/more accurate explanation.
I have learned your name from the Internet. We are the manufacturer&exporter in China, specializing in producing Bathroom Products. Please visit our website www.china-bathroom.com for more information. OEM is available. Please let me know if I may be of further assistance. ( E-mail: jimmy-at-china-genesis.com )
Yours sincerely, Mr. Jimmy Sky
Ningbo Genesis Industry Co., Ltd. Tel: 86-574-87330333 Fax: 86-574-87725245 Location: Ningbo city, Zhejiang province, China ( a port city, 150km from Shanghai )
If you think this is spam, please send email Subject "remove" to jimmy197809-at-hotmail.com.
I have learned your name from the Internet. We are the manufacturer&exporter in China, specializing in producing Bathroom Products. Please visit our website www.china-bathroom.com for more information. OEM is available. Please let me know if I may be of further assistance. ( E-mail: jimmy-at-china-genesis.com )
Yours sincerely, Mr. Jimmy Sky
Ningbo Genesis Industry Co., Ltd. Tel: 86-574-87330333 Fax: 86-574-87725245 Location: Ningbo city, Zhejiang province, China ( a port city, 150km from Shanghai )
If you think this is spam, please send email Subject "remove" to jimmy197809-at-hotmail.com.
To upgrade our old microscope (a double condenser lens VG HB501) we are looking for a second-hand parallel electron energy loss spectrometer (PEELS) or a second-hand Gatan Image Filter (GIF). Please e-mail any information to me if you have one. Thanks marie
----------------------------------------------------------------- Marie-Claude Cheynet Groupe Physique du métal LTPCM/ENSEEG/CNRS(UMR-5614) Domaine Universitaire BP75 Saint-Martin d'Hères 38402 e-mail mcheynet-at-ltpcm.inpg.fr tel : 33 (0)4.76.82.66.14 http://www.inpg.fr/LTPCM/ -------------------------------------------------------------------------------- -----------------
Greetings, This reflects a common misconception about the quantitative definition of 'resolution' in microscopy. For light microscopy, one is quite used to hearing that the 'resolution limit' is about 0.25 microns, or thereabouts. So it is natural to wonder how come it is pretty easy to see things like microtubules, which have a diameter around 1/10th of that limit. The answer lies in the definitition of resolution: that limit defines how close two objects can approach and still be resolved as two objects. When they get closer than the limit, then they are seen as one single object. However, the definition says nothing about the smallest object that can give rise to an image. That is limited only by the sensitivity of the detector. So objects like microtubules many times smaller than the resolution limit can be detected. If you measure their diameter, you will find that they measure up to whatever the diffraction limit is in your particular scope. When two of them happen to be closer than that limit, then they are detected as one object.
Hope this helps, Tobias } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (spohnheimer-at-dalsemi.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, } March 22, 2003 at 19:57:07 } --------------------------------------------------------------------------- } } Email: spohnheimer-at-dalsemi.com } Name: John Spohnheimer } } Organization: Dallas Semiconductor (Commercial) } } Education: Graduate College } } Location: Dallas, TX } } Question: I am an engineer/physicist working with semiconductors & } have used high-quality optical/SEM/TEM microscopes for ~25 years. } Recently, a friend (also a physicist) asked why we can see } sub-micron geometries (sometimes even one-two tenth micron diameter } particles are visible) using optical microscopes with wavelengths } nominally in the 1/2 micron range. } } I argued that what we're really seeing is simply a phase } interference pattern generated by the sub-micron particle, not the } real particle itself. That's why most sub-resolution particles } appear spherical in an optical microscope and often-times look } completely different under SEM examination. } } I'm curious if a professional in this field has a better/more } accurate explanation. } } Thank you. } } } ---------------------------------------------------------------------------
I looked around on the US DOT web site, and was a little surprised to find that it seems there is not, in the United States at least, a class of hazardous material called "Cryogen". The most useful document I found in my (brief) search was http://hazmat.dot.gov/files/registration/0203/regbrch2002.pdf which clearly lists the classes of materials that require placarding of vehicles and registration of carriers. My lay reading of this leads me to the conclusion that liq. N2 would be placarded as "Non-flammable gas" which is only regulated in quantities over 1000 pounds. It may, naturally, be different in other countries. Nevertheless, common sense must prevail, and that tells us that one must be thoughtful about transporting any quantity of the material. There will be other applicable safety regulations that apply to liquid nitrogen at all times, whether during transportation or not, (for example, occupational safety regulations), which I have not even begun to address here.
Once you add the "W" word to any material, though, the issue is **VERY** different. Even a microgram of waste is highly regulated (or so it seems!).
Tony.
At 04:07 PM 3/23/2003 -0600, Gordon Couger wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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When I was working in a lab with low-overhead - basically owned by someone with shallow pockets - I successfully transported LN2 in my car using a styrofoam cooler. I wasn't too keen on it and I don't recommend doing it, but I thought this may be worth mentioning. LN2 thermoses are somewhat expensive... I believe $200-$500, but worth it for a lab committed to doing EDS work.
If you must transport by car, keep the windows open or blower fan on if it's in the passenger cabin. I doubt that there are DOT issues involved. LN2 is not a substance one would consider highly hazardous, plus I believe DOT has jurisdiction only for amounts above a certain minimum. Only practical issues apply.
On the matter of occasional EDS use, one can simply fill LN2 when needed, allowing for proper detector cooldown (roughly 2-4 hours for my particular model). This method has to weighed against the thermal cycling that occurs, which is not good and potentially may - with time - damage the detector by delamination.
Stu Smalinskas Metallurgist SKF NATC Plymouth, Michigan
Gordon cougar wrote:
Liquid nitrogen should be much less dangerous the liquid oxygen and I see trucks that haul live fish with LOX tanks on them. They may be hauling the LOX without the proper permits. The people in the bait fish business are not particular careful about obeying the law. Artificial insemination technicians carry LN2 dewers all over the country as well.
To pump the LN2 put a pipe that reaches the bottom of the tank. Just close the vents and put a little current through a heater submerged in the nitrogen and the pressure will pump the LN2 out the pipe. Four psi should pump about 10 gallons a minutes or more though a 3/4 inch pipe to a container the same height as the LN2 tank.
If the temperature of boiling propane is low enough you could vaporize propane and dispose ot it in a flame and greatly reduce your storage and procurement problems. A propane tank has to be in sealed compartment in vehicle or mounted out it and the vehicle should not be parked in a building. It can legally be installed in vehicles.
Gordon Couger gcouger-at-couger.com
Dr. Chris Jeffree wrote:
} There is really only one way to carry liquid nitrogen safely by car } and that is outside it. A pick-up truck, or SUV with a rear transport } cage for a dewar would be ideal, but if liquid nitrogen spills inside } the car you, your passengers and other road users are in mortal } danger. Taylor-Wharton manufacture dry shippers that can safely } transported inside cars and aircraft, but unfortunately they are } designed as refrigerators, and will not release liquid nitrogen when } inverted. They are therefore useless for refilling your EDS } detector. } Chris } } Dr. Chris Jeffree } Inveresk Cottage } 26, Carberry Road } Inveresk } Musselburgh } Midlothian } EH21 8PR } Tel: +44 131 665 6062 } FAX +44 131 653 6248 } Mobile 07710 585 401
Gary Gaugler wrote:
} } Can anyone recommend a source of 5-10L } } LN2 portable dewars? These are for filling } } a 10L EDS detector. The LN2 will be } } transported by car. } } } } Any ideas? } } } } tnx, } } gary g.
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There's a long answer involving such great party conversation pieces as Rayleigh's criterion and Ernst Abbe's formula for resolution in any optical system by diffraction which is R=0.61 lambda/ n sine alpha.
The important bit in this context is n sine alpha. Where n is refractive index of the medium through which light/electrons pass from the specimen into the lens and sine alpha is the sine of the semi angle of the objective lens This combined figure is generally called the Numeric Aperture (NA) of the lens and even our old teaching microscopes have an NA of 1.28. In other words to make resolution smaller or better you need a high refractive index (eg immersion oil 1.4 or greater) and a big lens as close as possible (eg an oil immersion lens) which give a NA as large as possible.
Some quick maths tells you that a basic light microscope is capable of a resolution of 0.61/1.28 lambda which is less than half the wavelength of light. But in the e.m. with an aperture of 100um diam and focal length of ~3mm you are lucky to get a NA of better than about 0.01. Why the e.m. has such a poor numerical aperture is because of lens aberrations especially spherical which are only corrected by using a small central objective lens aperture.
Basically light microscope lens design is so good that resolution is only limited by diffraction effects whereas electromagnetic lenses are a compromise.
My apologies if I've oversimplified.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: spohnheimer-at-dalsemi.com (by way of MicroscopyListServer)
Thanks to many who replied to my post about LN2 Dewars and transporting them. It is possible to do this. however, several have pointed out the issues surrounding temperature cycling of the EDS Dewar. The makers claim that this is not a problem--even with Si(Li) detectors. And the systems will power down the amplifier if LN2 is low or out. And, they will not allow the unit to power up if LN2 is not correct. But there seems to be issues surrounding the window. The makers claim that this is not so.
OK. Plan B. So what about the closed loop refrigerated EDS units like EDAX makes? Does anyone have any direct experience with these? Good or bad. I need to detect down to N.
I had heard that these Kleemenko cycle cryocoolers are notoriously unreliable. The pumps fail frequently. Perhaps also, there are sensitivity issues and low Z detection limits. I had looked at closed loop WDS systems before and got the same reliability feedback from users. How about EDAX EDS units?
This is a rough one. We have just stated a new EM lab (less than two years operational) Our TEM (Which is new!) is not functioning due to the SIS camera being away for repairs under warranty now for some time. The EDS has been sent back twice now and I hope this time around the repairs will be successful. Being in Africa does have it pro's to like the weather, the birds and wildlife. We also have a lot more freedom in research. I am seeing forward to a prosperous year full of publications with EM input and well trained motivated users! (my whish list and also the units goal)
Thus if your TEM is running this is already a blessing. This alone (service contract cost depending) should be enough to motivate for keeping it. We are in a process to monitoring all publications on all instruments as a justification to keep it running or to replace it should the kneed arise.
My wife is brilliant at management and she helps me a lot. Her motto is always: "if you motivate something well enough you can cell ice to an Eskimo."
This is the most important point of all: Since she is now directing the facility, please go ask her to give the Objectives, Goals and mission statement for the facility to you in writing (Her view). If possible ask her also for a 5, and 10 Year plan for the facility. Before attempting to motivate anything think things over carefully. You two must be able to work in peace since strive will destroy the facility quickly and drive users away! With unity much can be achieved since her signature is required most likely on all purchase requests, motivations and budget expenditure.
When you do the motivation, link your instruments to the objectives and goals of your institute as well as the ones you got from her. Let her see that you are on her side and you would like to see these goals achieved.
If you can provide a list of publications where each instrument was involved in over its lifespan as a percentage of the total number of publications published in your institution it should help to explain the kneed of each instrument, scanner, SG's etc...
You can probably get a letter from the IT guys to state that your application, software, set-up and configuration is unique to the institution and it serve the best to keep it out the hands of ordinary villains to prevent expensive reconfiguration and repair. I had to fight to keep them out without driving them away since I kneed there help but not there software upgrades etc!
If you have a good relationship with your research office ask them for a opinion in writing.
It is worth it to do a survey in the institute to all relevant users, past users and possible users including head of departments. Things worth getting a answer on are questions like: What do you expect from the facility, What instruments would you like to be present in the unit, Past usage, usefulness of input in their research. Possible future utilisation of your unit. Suggestions to improve the service as well as suggestions wishes for instruments/capability's currently not present. If you can get postgads promised to instruments and research programs including your facility the better. I know that is not always possible but if you do not ask you will not know. We did that at a previous institute where I was employed. This was a very useful exercise. This helped to direct the unit and improve on our service. These documents are also a very useful source for equipment motivation when it is combined into a format top management can understand.
Hopefully from this information the wise path to follow will be much more clearer to all.
Good luck.
-----Original Message----- } From: Rick Harris [mailto:raharris-at-ucdavis.edu] Sent: Friday, March 21, 2003 1:08 AM To: microscopy-at-sparc5.microscopy.com
After 24 years of managing our small facility as a collaborative effort with interested faculty, I now have a new supervisor with little technical expertise. Our facility consists of an older TEM, a new SEM, a new deconvolution scope, and a Lieca SP1 confocal. I don't want to air our dirty laundry but I need a little help in justifying our equipment to the new boss. Now, she is proud of the fact that she still uses PhotoShop 3.0 and states that the best way to get a good picture is to take a good picture and then don't mess with it. While there is certainly some truth in the second fact, it is also very naive. And the first fact classifies her as a Luddite. Her research centers around the confocal and the deconvolution scope. So the move is on to dump the TEM and give the Digital Darkroom to the department Network Admin. This will free me up to tackle confocal projects for her and spend more time dusting. Yes, after 24 years of managing the facility on my own my new boss has actually given me a dusting schedule. However, she is quick to point out that this is not micro-managing. Alright, I digress. I need to justify having a couple graphics workstations, a slide scanner, a flat bed scanner with transparency tray, and a photo-quality printer. She would give this equipment to the Network Admin who would integrate it with the common computer room. I treat this equipment like I do optical equipment. I am concerned that once relocated to a common room, with no supervision, it will rapidly deteriorate. I also take exception with the apparently wide-spread belief that the computer support staff is the best source of information on digital imaging. I guess because it is digital?
So how do I keep those pieces of equipment I deem necessary for the facility? I think we should have a photo-quality printer. I think we need the scanner with tray for those increasingly rare occasions that we need to scan in a TEM neg or an illustration for a PowerPoint slide or publication. I find that I am not good at making an argument for what seems blindingly obvious to me. I just sputter and fume and ask, "... how can anyone have an Imaging Facility without a couple workstations and printers?". Or a TEM.
And then there is always the possibility she is right. Maybe we don't need to do this stuff to the standards I embrace. Should I convert to her mantra: "Do more, less well."?
Any good arguments out there that have worked for any of you?
In the past I have thermally cycled several detectors without ill effect, but not enough times to be absolutely confident. Several vendors have told me that modern detectors should be far more stable to thermal cycling than older ones.
I have a detector from EDAX with a small cryostat designed for cooling on demand. This works fine, but we find the 1-2hr cool-down time is a nuisance in our facility for users who see a feature, then decide it would be good to do EDX analysis on it. We still tend to keep the detector cool except during the weekend. If you were more sure of when you would and wouldn't be using the EDX (and if the occasions when you would be using it are relatively few) then I would say this would be a good way to go. I have had one window failure (in 3.5 years), but then I have had window failures at about this rate with other detectors that are not cycled regularly.
If you are thinking of new technology, you might consider the silicon drift detector from Rontec. To the casual user this works just like a conventional EDX, but it only needs cooling to -15C or thereabouts; this is done easily with a thermoelectric cooler. Since the temperature change is small, it also cools fast if it is not already on - it can be used 15 mins or less after being switched on. On standby, the amplifiers are left on but the detector is allowed to warm up. I don't know if I got a particularly good one, but its resolution is comparable to that of a premium Si(Li) of a few years ago, and it is sensitive to x-rays down to C. It also will count happily at 160,000 counts/sec. It does have a few minor disadvantages, so it may not be for everyone, but I was attracted by the lack of need for Liq. N2, and I have been a happy customer. (I have no connection with Rontec except as a customer, but I should mention that the terms of the purchase allow Rontec to use my installation for demonstration purposes).
Cheers,
Tony.
At 01:51 PM 3/24/2003 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** Manager, CMSE Shared Experimental Facilities ** MIT Rooms 13-1027 or 13-3090 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
dear sir or madam, i was wondering whether you could inform me of the modern developments in microscopy and their advantages over traditional methods and machines
-- ******************************* This message was sent from an ISS public cluster at the University of Leeds using Netscape Communicator. Its authenticity cannot be guaranteed.
It would seem this request has reached the ultimate in vague and general questions. I suppose the only improvement on it would be to avoid the mention of microscopy altogether.
At 02:41 PM 3/25/03 +0000, you wrote: } Date: Tue, 25 Mar 2003 14:41:04 +0000 } From: "R.M. Carr" {BMB2RMC-at-leeds.ac.uk} } Reply-To: noreply-at-leeds.ac.uk } Organization: University of Leeds } To: Microscopy-at-sparc5.microscopy.com } Subject: modern microscopy } Content-Type: text/plain; charset=us-ascii } Content-Transfer-Encoding: 7bit } } dear sir or madam, } i was wondering whether you could inform me of the modern developments } in microscopy and their advantages over traditional methods and machines } } -- } ******************************* } This message was sent from an ISS public cluster at the University of } Leeds using Netscape Communicator. Its authenticity cannot be } guaranteed.
} From: "Richard Edelmann" {edelmare-at-MUOHIO.EDU} } Date: Tue Mar 25, 2003 5:15:32 AM US/Pacific } To: Kim Rensing {krensing-at-interchange.ubc.ca} } Subject: Re: Arabidopsis infiltration } Reply-To: edelmare-at-MUOHIO.EDU } } Rick: } } Arabidopsis is evil - pure and simple. Having worked with a variety } of } plants, arabidopsis (the tiny parking lot weed) takes 4 - 8x longer } for ALL } fixation steps than normal plants 10-20x the size - But short answer is } LONGER IS BETTER. I can find you a reference for our protocols if you } want but basically (yes these numbers are right): } } 1) aldehyde fix 8-10hrs -at- rt } } 2) rinses 4x -at-20-30min each } } 3) Osmium fix 2-4 hr -at-rt (this one is short who knows why . . ) } } 4) rinse } } [ U acetate en bloc if wanted - over night] } } 5) dehydrate 25-75% -at-30-60 mins, 95, 100, 100, 100, 100 60-120 + mins } } 6) Infiltration (with Spurr's, Quetol 651, or LR White) on rotator at } RT. } } 1/2 day steps: 3:1, 1:1, 1:3, 100%, {- no catalyst } } 1 -day 100% {- no catalyst } } 100% 1/2-day to overnight with catalyst } } Molds w/catalyst } } } What age plants? Seedlings 1-6 days old mostly, but works with } mature/maturing plants (including infloresence) as well. } } On Thursday, March 20, 2003, at 02:02 PM, Rick Harris wrote: } } } } } } I have a group looking at very early germinated Arabidopsis sp. They } } } are having problems with good infiltration of the Epon-Araldite } } } epoxy. } } } While they are out searching the literature for solutions, I thought } } } I'd drop a note to the list. Any suggestions for an infiltration } } } protocol for these tiny plants? Spurr's? Dice up those little } } } seeds? } } } } } } TIA } } } } } } } } } Rick A. Harris, Director } } } Microscopy and Imaging Facility } } } } } } } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 350 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } http://www.emf.muohio.edu } } "RAM disk is NOT an installation procedure." }
I have just started using the JEMS image and diffraction simulation software. Is there a users group / listserver dedicated to JEMS? If so how do I join? Cheers,
Mark Blackford ANSTO, Materials and Engineering Science PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
Over the years we have had visitors use our Liquid Nitrogen and found it very convenient to purchase a refill for our tank to replace what they used and the convenience of having the Liq N2 on site. This avoided the problem of transporting the nitrogen accross the city etc. Pat Connelly Dept. of Biology Univ. of Pennsylvania Philadelphia, PA 19104-6018
I'm in need of some new specimen block chucks for an old Sorval MT-2B ultramicrotome. Can any one give me a source for such items new or used. I'm also looking for some extra stainless steel camera plate film (31/4 X 4) carriers for a Hitachi 7000 TEM. TIA
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Have you ever been in this situation? You are representing a client who is supposed to receive a monthly payment. On some months, that payment has been made on time, but on other months it has been paid late and often not at all. Your client is entitled to interest on amounts owing, but figuring out how much interest precisely is another matter.
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Dear listers, Could some one give me tips on how to process plated cells for immuno gold using LG gold or white. and also I have been processing some rabbit aortas with S Steel stent in it for semi thick sections using glycol-methacrylate. For some reason some blocks have turned out be verysoft. Is it true that they cannot be re processed and reembedded. Can I do some thing to get sections of the region containing the stent without disturbing the structure. Shashi Singh CCMB Hyderabad INDIA
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I would like to remind you of our international post-graduate short-courses on histological techniques. Please note that you can still register for the Multiple Staining in Immunohistochemistry course, starting April 2nd.
We offer these international post-graduate short-courses on the histological techniques: - FISH Techniques in Molecular Pathology (16-20 June 2003) - Confocal Light Microscopy: fundamentals and biological applications (12-16 May 2003) - Tissue micro-array (18-20 June 2003) - Multiple Staining in Immunohistochemistry (2-4 April 2003) All these courses are organized in close collaboration with experts in the field of interest.
Please visit our website http://www.novaknowledge.nl/english.htm for detailed information about these courses and other courses that we offer (e.g. quantitative PCR and strategic protein purification).
Best regards, Hans van Hirtum
Ing. J.P. van Hirtum Hogeschool Brabant Nova Knowledge P.O.box 5690 4801 EB Breda, the Netherlands T: +31 (0) 76 572 2644 F: +31 (0) 76 572 2640 E: hvanhirtum-at-hsbb.nl W: http://www.novaknowledge.nl
Maybe so, but why shouldn't someone feel able to post on a microscopy forum in order to gain information about microscopy?
Perhaps it would be more helpful for those well versed in the subject to suggest literature which covers the field of microscopy more gererally than is practicable here, or to offer advice on how the question might be made more specific.
If there's one thing that puts people off from beginning to learn a subject, it's being sneered at by those already in the know.
----- Original Message ----- } From: Warren E Straszheim {wesaia-at-iastate.edu} To: {Microscopy-at-sparc5.microscopy.com} Cc: R.M. Carr {BMB2RMC-at-leeds.ac.uk} Sent: Tuesday, March 25, 2003 4:18 PM
I find the contrast between "modern" and "traditional" somewhat amusing.
Who determines when something changes from "modern" to "traditional".
For example, a brand new Olympus, Nikon or other "high end" optical microscope with all the trimmings and a digital camera could certainly be called a "modern" microscope, but I suspect it would be classified as "traditional" (in some cases "obsolete"?).
Sorry, I couldn't resist spending the bandwidth to share my amusement.
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Tuesday, March 25, 2003 10:19 AM To: Microscopy-at-sparc5.microscopy.com Cc: R.M. Carr
I agree with Justin. We need to be careful how we word our comments on e-mail. E-mail is especially harsh, as we don't have the help of voice inflection or gestures to help us communicate the sentiment we really mean. Having said that, in order to answer the question, you do need to clarify what you mean by modern microscopy.
As for modern microscopy, do you mean techniques for biology that use immunohistochemistry, confocal microscopy that allows for imaging of live cultured cells with incorporation of dyes and markers, scanning tunneling microscopes and field emission microscopes that allow imaging beyond the standard electron microscopes? Some on this stuff isn't that "new", but it is all more modern than standard histologic techniques, SEM of carbon or platinum coated materials and TEM of heavy metal stained, epoxy embedded tissues. Are you more interested in nonbiological (materials) microscopy? Please clarify and you may get more responses.
Karen Pawlowski
Justin Ritherdon wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 so, but why shouldn't someone feel able to post on a microscopy forum } in order to gain information about microscopy? } } Perhaps it would be more helpful for those well versed in the subject to } suggest literature which covers the field of microscopy more gererally than } is practicable here, or to offer advice on how the question might be made } more specific. } } If there's one thing that puts people off from beginning to learn a subject, } it's being sneered at by those already in the know. } } ----- Original Message ----- } } From: Warren E Straszheim {wesaia-at-iastate.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Cc: R.M. Carr {BMB2RMC-at-leeds.ac.uk} } Sent: Tuesday, March 25, 2003 4:18 PM } Subject: Re: modern 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 } } -----------------------------------------------------------------------. } } } } } } It would seem this request has reached the ultimate in vague and general } } questions. I suppose the only improvement on it would be to avoid the } } mention of microscopy altogether. } } } } At 02:41 PM 3/25/03 +0000, you wrote: } } } Date: Tue, 25 Mar 2003 14:41:04 +0000 } } } From: "R.M. Carr" {BMB2RMC-at-leeds.ac.uk} } } } Reply-To: noreply-at-leeds.ac.uk } } } Organization: University of Leeds } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: modern microscopy } } } Content-Type: text/plain; charset=us-ascii } } } Content-Transfer-Encoding: 7bit } } } } } } dear sir or madam, } } } i was wondering whether you could inform me of the modern developments } } } in microscopy and their advantages over traditional methods and machines } } } } } } -- } } } ******************************* } } } This message was sent from an ISS public cluster at the University of } } } Leeds using Netscape Communicator. Its authenticity cannot be } } } guaranteed. } } } } } }
There was a thread like this a while ago, where the merits of answering this type of questions was discussed. The result was in favor of what you are saying, however, I think it was assumed that the questions were of a certain quality. Questions like "Can you tell me about microscopy" or similar will not provoke many answers, because frankly, most people can use their time better than answering these basic questions. I am sure, nobody on the list would object if you answered with a few pointers to books, but I wouldn't hold my breath for a large response.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Justin Ritherdon [mailto:J.Ritherdon-at-liverpool.ac.uk] Sent: Wednesday, March 26, 2003 3:28 AM To: Microscopy-at-sparc5.microscopy.com
Maybe so, but why shouldn't someone feel able to post on a microscopy forum in order to gain information about microscopy?
Perhaps it would be more helpful for those well versed in the subject to suggest literature which covers the field of microscopy more gererally than is practicable here, or to offer advice on how the question might be made more specific.
If there's one thing that puts people off from beginning to learn a subject, it's being sneered at by those already in the know.
----- Original Message ----- } From: Warren E Straszheim {wesaia-at-iastate.edu} To: {Microscopy-at-sparc5.microscopy.com} Cc: R.M. Carr {BMB2RMC-at-leeds.ac.uk} Sent: Tuesday, March 25, 2003 4:18 PM
Is there anybody out there who has an epon embedding of isolated mammalian centrosomes/centrioles (or basal bodies) and might be willing to let me have a few of sections?
A great deal has been published by many companies and individuals over the years in American Lab. The Focus on Microscopy column, appearing over the past 8 years (typically appears in April and November, occasionally in July/August issues), emphasizes new directions (www.iscpubs.com). See also "Microbrew" column which appeared periodically in Advanced Imaging (a Cygnus publication).
Other key publications: Microscopy Today, BioPhotonics, R&D (ww.rdmag.com), Semiconductor International, Advanced Materials and Practices (published by the Am. Soc. for Materials).
Our firm tracks new develops. From our perspective, the hottest new areas include -the convergence of microscopy and spectroscopy (especially FT-IR and Raman, but there are interesting new techniques in X-ray emerging), -evolution of new chip technologies for cameras -push into nanotechnology from multiple directions (especially EM, interferometry).
Your question is worthy of a MAJOR publication... far beyond the scope of this response. Please feel free to contact me off-line if you are interested in something specific.
Best regards, Barbara Foster Microscopy/Microscopy Education, Inc 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
"Why didn't they teach us that sooner?" ... probably because no one thought to call MME about customized, on-site courses. Offered in all areas of microscopy, sample prep,and image analysis, they make an immediate impact on your productivity. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
At 02:41 PM 3/25/03 +0000, R.M. Carr wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I agree with Warren Straszheim. The question as posed sounds like an essay topic and would require a several-thousand word reply once one did define terms. There was no evidence that the person solicitng aid had done any research that might have allowed them to pose the question in a more easy-to-answer form. I think people whose time and energy are being solicited deserve some respect.
One clear technical revolution in microscopy occurred in the 1960s with the advent of scanning electron microscopy. Up to that point, all microscopes used lenses, either optical or electron-optical, to generate a magnified image of an object. The scanning electron microscope was the first type of microscope to generate magnified images without the use of lenses. It was a hugely important advance in the characterization of surface morphology of materials. More recently the optical analogue of that technology has appeared in the form of the laser scanning confocal microscope, which again has had a major impact, particularly in the life sciences. Yes, both these microscopes use lenses, but the purpose of the lenses is solely to generate a probe of electrons or light radiation, and not to generate the image itself. Since the principle of scanning or raster microscopy was introduced in the SEM, pure non-optical scanning microscopes have appeared in the forms of the scanning tunnelling microscope, the atomic force microscope and their relatives.
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Facility
----- Original Message ----- } From: "Karen Pawlowski" {kpawlow-at-swbell.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 26, 2003 2:45 PM
Oh, come on!
rtch
} } } Maybe so, but why shouldn't someone feel able to post on a microscopy } forum in order to gain information about microscopy? } } Perhaps it would be more helpful for those well versed in the subject } to suggest literature which covers the field of microscopy more } gererally than is practicable here, or to offer advice on how the } question might be made more specific. } } If there's one thing that puts people off from beginning to learn a } subject, it's being sneered at by those already in the know. }
} } } } } It would seem this request has reached the ultimate in vague and } } general questions. I suppose the only improvement on it would be to } } avoid the mention of microscopy altogether. } }
} } } } } } dear sir or madam, } } } i was wondering whether you could inform me of the modern } } } developments in microscopy and their advantages over traditional } } } methods and machines } } } } } } --
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
-----Original Message----- } From: Justin Ritherdon [mailto:J.Ritherdon-at-liverpool.ac.uk] Sent: Wednesday, March 26, 2003 2:28 AM To: Microscopy-at-sparc5.microscopy.com
Maybe so, but why shouldn't someone feel able to post on a microscopy forum in order to gain information about microscopy?
Perhaps it would be more helpful for those well versed in the subject to suggest literature which covers the field of microscopy more gererally than is practicable here, or to offer advice on how the question might be made more specific.
If there's one thing that puts people off from beginning to learn a subject, it's being sneered at by those already in the know.
----- Original Message ----- } From: Warren E Straszheim {wesaia-at-iastate.edu} To: {Microscopy-at-sparc5.microscopy.com} Cc: R.M. Carr {BMB2RMC-at-leeds.ac.uk} Sent: Tuesday, March 25, 2003 4:18 PM
Posted on behalf of Ken Taylor, chair of the 2003 Gordon Conference on Three-Dimensional Electron Microscopy of Macromolecules
This is a heads up about application to this years Gordon Conference on 3-D Electron Microscopy of Macromolecules. The conference will be closed to new applications at the end of March. Attendance to the conference is by invitation only based on your application. Application does not guarantee acceptance but if you want to attend, you have to apply.
Application takes only a few minutes and can be done on-line at the following URL:
On Tuesday, March 25, 2003, at 06:41 AM, R.M. Carr wrote:
} i was wondering whether you could inform me of the modern developments } in microscopy and their advantages over traditional methods and } machines } Dear RM, Two modern developments are the scanning probe microscopies, which can look at signals previously unseen, and cryoscopic methods. The first development includes atomic force microscopy (with its several modes), near-field optical microscopy, Hall probe microscopy, and many more--some of which I'm sure I've not heard of. The advantages are the ability to measure the rigidity of surfaces on the atomic level, the ability to get much better resolution than a wavelength of light, and the ability to measure magnetic field variations on the micrometer scale. The second development enables the observation of biological materials without the necessity of chemical fixation and staining, so the unperturbed biological molecules themselves can be measured. As others on the list have suggested, you should find the appropriate materials in your library or on the web to get the latest details. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wang-at-verrillon.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 26, 2003 at 14:54:52 ---------------------------------------------------------------------------
Email: wang-at-verrillon.com Name: Chih-Hao Wang
Organization: Verrillon
Education: Graduate College
Location: Grafton, MA, 01721
Question: I want to measure the thickness for a very thin layer (20nm) of coating on the glass surface. I tried SEM but the resolution is not good enough for this.
Mr, Wang, A few suggestions I have are using ellipsometry, profilometry, and/or X-ray reflectivity. We have obtained best results using X-ray reflectivity (GIXR) for polymer thin films. I hope this helps.
Roberto Olayo-Valles Department of Chemical Engineering and Materials Science University of Minnesota
-----Original Message----- } From: by way of MicroscopyListServer [mailto:wang-at-verrillon.com] Sent: Wednesday, March 26, 2003 6:39 PM To: MicroscopyListserver
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wang-at-verrillon.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 26, 2003 at 14:54:52 ---------------------------------------------------------------------------
Email: wang-at-verrillon.com Name: Chih-Hao Wang
Organization: Verrillon
Education: Graduate College
Location: Grafton, MA, 01721
Question: I want to measure the thickness for a very thin layer (20nm) of coating on the glass surface. I tried SEM but the resolution is not good enough for this.
There is actually an inexpensive book that might answer the question: Light and Electron Microscopy by Elizabeth and Henry Slayter, Cambridge University Press 1992 ISBN 0-521-33948-0
It contains a few chapters describing microscopy using about anything you can imagine, light, electrons, neutrons, X-rays, ions, sound or nothing at all. I think that should give an overview, although it's strongest side is light microscopy in my opinion.
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em
The phrase 'We have always done things this way.' is as much a reason to change as a reason not to. - Dartwill Aquila _______________________________________ } dear sir or madam, } i was wondering whether you could inform me of the modern developments } in microscopy and their advantages over traditional methods and machines
You don't tell us what the composition is of your very thin layer. But if the average Z value is not too low, you can use the X-ray spectrometer on the SEM to measure the thickness. There are today optional programs for that. If you don't possess such a program, you can follow the procedure described by Bishop and Poole in J. Phys. D: Appl. Phys., vol. 6 (1973) 1142. Using this procedure I have once measured the thickness of gold layers in the nm range on holmium.
-----Original Message----- } From: wang-at-verrillon.com [mailto:wang-at-verrillon.com] Sent: 27. marts 2003 01:39 To: MicroscopyListserver
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wang-at-verrillon.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 26, 2003 at 14:54:52 ---------------------------------------------------------------------------
Email: wang-at-verrillon.com Name: Chih-Hao Wang
Organization: Verrillon
Education: Graduate College
Location: Grafton, MA, 01721
Question: I want to measure the thickness for a very thin layer (20nm) of coating on the glass surface. I tried SEM but the resolution is not good enough for this.
One of the easiest ways to do this is with optical modeling. If you can find someone with an ellipsometer, it would be relatively straightforward. Making cross section on glass that can measure films that thin in the TEM are not real easy, but can be done. You can use the normal dimple and ion milling approach or the Tripod polish approach. Phil Swab published an article on how to prepare a sample with ultramicrotomy that is fairly straitforward. I dug out a response to a previous question that he answered here and have copied it below.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
____From Phil Swab:
Glass is readily microtomed with a diamond knife and may be a suitable inexpensive technique for you to consider. Particularly if the materials of your sample are sufficiently dissimilar in reaction to the chemical and ion etching of some techniques. I've been embedding and sectioning coated glass, optics, and other hard materials for 18 years (even diamond coated silicon in Microscopy Research and Technique, vol. 31, p. 308, 1995). The imaging and analysis of nano-structures in micron sized areas near the surface of glass is routine, fast, and inexpensive for physical microstructure and chemistry. Mechanical artifacts generated in ultramicrotomy tend to be quite large, readily visible, and easily ignored but may interfere with the analysis of naturally occurring deformation features (i.e. twinning, slip, etc.). 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.
The critical elements for microtomy of hard, non-porous materials include: 1. 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 may be further broken to form very pointed thin samples. [Time = ~20 minutes] 2. Optimize sample orientation for sectioning and preferred orientation. Some physical microstructures are anisotropic and are difficult to interpret when viewed in the wrong orientation. [Time = ~10 minutes] 3. Maximize adhesion to the resin through the selection of an appropriate resin (low viscosity and non-reactive with your sample), meticulous and contamination-free sample prep, and the addition of adhesion promoters (such as Dow Corning Z-6040). [Time = ~1 hour, with an over night epoxy cure] 4. Section using standard procedures, but minimize the sectioning speed (optimize cutting speed). [Time = ~1 hour]
These times are approximate for 1 sample, and there could be economy in numbers. As always, each case will require individual attention. Cheers,
Phil Swab Engineering Development Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com
-----Original Message----- } From: wang-at-verrillon.com [mailto:wang-at-verrillon.com] Sent: Wednesday, March 26, 2003 7:39 PM To: MicroscopyListserver
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wang-at-verrillon.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 26, 2003 at 14:54:52 ---------------------------------------------------------------------------
Email: wang-at-verrillon.com Name: Chih-Hao Wang
Organization: Verrillon
Education: Graduate College
Location: Grafton, MA, 01721
Question: I want to measure the thickness for a very thin layer (20nm) of coating on the glass surface. I tried SEM but the resolution is not good enough for this.
Here is a method I have used for processing cell culture monolayers into LRWhite for subsequent immunolabeling:
LRWhite Cell monolayer Processing Cells ar grown in thermanox petri dishes.
1. Fload cells with Zambonies fixative 4 degrees: 5 min- 24hr...test. For 50 ml...Heat and stir 16.5ml water to 55 degrees C, add 4 drops 2M NAOH. Add 1g paraformaldehyde (prills) for 2% or 2g for 4%...It will take about 5 minutes to clear. Do not let the temperature exceed 60 degrees. Cool on ice. Add 25 ml 2x Sorensons. Add 7.5 ml Picric acid. pH to 7.4 using the pH indicator strips. Store at 4 degrees. Good for 2 weeks.
2. Rinse in PBS 3x 5 minutes. 3. 35% EtOH 1 min. 4. 50% EtOH 1 min. 5. 70% EtOH 2 min. 6. 95% EtOH 2 min. 7. 2:1 100% EtOH/LRWhite 5 min. (Rotator). 8. 1:1 100% EtOH/LRWhite 10 min. (Rotator). 9. 1:2 100% EtOH/LRWhite 20 min. (Rotator). 10. Pure LRWhite (overnight). (Rotator). 11. Change into fresh LRWhite. 1 hr (Rotator) 12. Change into fresh LRWhite. 13. Place into Vacuum Oven 50-55 degrees, flushed several times with dry nitrogen and allow to polymerize for 24-48 hr. I have about 3-5 lbs. of vacuum to keep the door sealed. Note: Steps from fixation to 70% can be carried out at 4 degrees. and steps from 70% to the polymerization can be carried out at temperatures down to -20.
Hope this helps. Bob Underwood Derm Research Center U of Washington
On Tue, 25 Mar 2003, shashi singh wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } Could some one give me tips on how to process plated } cells for immuno gold using LG gold or white. } and also } I have been processing some rabbit aortas with S Steel } stent in it for semi thick sections using } glycol-methacrylate. For some reason some blocks have } turned out be verysoft. Is it true that they cannot be } re processed and reembedded. Can I do some thing to } get sections of the region containing the stent } without disturbing the structure. } Shashi Singh } CCMB Hyderabad } INDIA } } __________________________________________________ } Do you Yahoo!? } Yahoo! Platinum - Watch CBS' NCAA March Madness, live on your desktop! } http://platinum.yahoo.com } }
Well, if you're gonna go this route, I cannot help but waste more bandwidth. "Traditional" microscopy probably refers to simple compound light microscopy. Putting a camera on microscope would be the first transition to "modern". "Modern" would refer to developments in the mid-20th C to the present, or 15-20 years ago (includes EM, mass produced infinity corrected optics, etc.). "Contemporary" (the term "postmodern" is loaded and banned and should only be used in an ironic fashion under erasure) microscopy would include the use of computers for image creation via mathematical convolutions of data collected by modern methods.
Practically, the person making the original question should be referred to an intro to microscopy course or Slayter's book, as already recommended.
I find the contrast between "modern" and "traditional" somewhat amusing.
Who determines when something changes from "modern" to "traditional".
For example, a brand new Olympus, Nikon or other "high end" optical microscope with all the trimmings and a digital camera could certainly be called a "modern" microscope, but I suspect it would be classified as "traditional" (in some cases "obsolete"?).
Sorry, I couldn't resist spending the bandwidth to share my amusement.
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Tuesday, March 25, 2003 10:19 AM To: Microscopy-at-sparc5.microscopy.com Cc: R.M. Carr
I'm not sure if it is possible draw the line between modern and traditional approaches to microscopy, but it seems that the advent of relatively cheap, user accessable computing power is one of the main differences between microscopic imaging today and what was involved a few years ago. Traditional photographic techniques and darkroom time are giving way to digital image capture because of considerations including cost, speed and ability to extract data from multidimensional data sets. We see this as a trend for retrofitting of existing "traditional microscopes" and for the development of new microscopies which are dependant on computing technology. Another factor that may be taken to separate modern microscopies from traditional methods are the advent of new illumination sources and sources of contrast. Lasers have revolutionized approaches towards microscopy, so have electron beams, so have light bulbs. Modern microscopes use modern physics. A third thing that comes to mind is the type of information that modern microscopes provide. Traditional microscopies may be thought of as providing structural information whereas modern methods also probe chemical properties, electro-magnetic properties, etc. at the microscale. -Karl G.
R.M. Carr wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
If the film is transparent interferometry can be used for the thickness measurements. Another promising approach is Rautherford Backscattering (RBS).
Mark
-----Original Message----- } From: wang-at-verrillon.com [mailto:wang-at-verrillon.com] Sent: Wednesday, March 26, 2003 7:39 PM To: MicroscopyListserver
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wang-at-verrillon.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 26, 2003 at 14:54:52 ---------------------------------------------------------------------------
Email: wang-at-verrillon.com Name: Chih-Hao Wang
Organization: Verrillon
Education: Graduate College
Location: Grafton, MA, 01721
Question: I want to measure the thickness for a very thin layer (20nm) of coating on the glass surface. I tried SEM but the resolution is not good enough for this.
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Emory Neurology Microscopy Core Laboratory is hosting a week-long workshop on Cryo-technique and immunogold labeling from May 4, through May 9, 2003 in Atlanta, Georgia, USA. The workshop curriculum will include the latest advances on cryo-fixation and substitution of biological samples, cryo-ultramicrotomy, and pre- and post-embedding immunogold labeling. Internationally well known experts in the fields, Dr. Kent McDonald (EM Laboratory at the University of California, Berkeley), Dr. Jan Luenissen (Aurion Immunogold Reagent & Accessories) and Mr. Helmut Gnaegi (Diatome) will be the instructors for the workshop. The workshop will include lectures, hands-on training, round table discussions, and presentations on applications. Also, participants of the workshop will be able to work on their own samples during the workshop. The industrial sponsors for the workshop are Leica Microsystems Inc., Aurion, EMS, and Diatome U.S.
The workshop will limit the number of participants to 15. Registration deadline is April 15. If you are interested in attending or need more information about the workshop, please contact the workshop technical coordinator Hong Yi by phone (404-712-8491) or email (hyi-at-emory.edu), or log on www.em-preparation.com.
We have some resin embedded skeletal muscle samples, and we are looking for end plates. However, end plates are usually aggregated along muscle fibers. Without knowing which blocks have end plates, or where in a block, we have to search on semi-thin sections first before cutting thin sections for EM. Can any one recommend a dye that stains end plates on epon semi-thin sections? It is very inefficient to search this way. Can anyone recommend a better approach? The samples have been treated, so somewhat precious.
Dear all, Our JEOL3010 alignment has got into a bad state, and I can't seem to get it back with the normal alignments described in the handbook. Probably a user has accidently altered a gun or condensor alignment. Does anybody have a set of instructions for a complete gun and column alignment, and not just the everyday procedures in the standard blue ring-binder?
(I can't get a beam on the phosphor screen at all in SA, in LM I see the beam but get weird caustic shapes and the best is with beam shifts way over on one side). There are no instructions in the manual about what to do with Gun deflectors or condensor deflectors, although playing with the Gun deflector did improve things slightly.
Thanks for any help you can give.
-- Ian MacLaren Technische Universität Darmstadt Material-und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany
Good morning all in the listserver, I had a question - how does on go about doing quality control on EM - both TEM and SEM. In other words, other than giving techs unknown samples and checking the end results - or just looking over their shoulders checking thicks and thin quality - is there another way? Thanks in advance, Connie Cummings
I was wondering if there exists a divided coverglass chamber where the chambers are close enough so that two chambers can be observed simultaneously with a 20 x objective. Has anyone seen such a thing?
Mike --
Michael J. Herron, U of MN, Entomology herro001-at-umn.edu 612-624-3688 Office, 624-3212 Lab, 625-5299 FAX
Here's a request for micrographs that's worth your attention; please respond directly to Alisia.
Wanted: images for NIH booklet We are looking for a handful of TEM, SEM, and laser scanning confocal micrographs to illustrate a science education booklet on cell biology. The booklet, a revised version of "Inside the Cell," is part of a series on basic biomedical topics published by the National Institute of General Medical Sciences (see http://www.nigms.nih.gov/news/science_ed/ for a list and online versions). The award-winning booklets are extremely popular with science teachers and are used in classrooms across the country.
We are primarily interested in images of cells, cell components, microbes, and other samples that relate to biomedicine. In addition, we would appreciate images of a single sample at a range of magnifications and/or as viewed through different types of scopes (light, TEM, SEM).
We will provide photo credits next to the images. Although we usually obtain images free of charge from scientists we fund, we are also willing to pay modestly for some images. As for all NIGMS publications, this booklet will be non-copyrighted and distributed free of charge. If you would like to submit images that have been previously published, please include complete publication reference information. We will give preference to unpublished images.
Please e-mail candidate images as low-resolution jpegs to alisa.machalek-at-nih.gov. For printing, we will need images of 300 dpi at a printed size of about 5"X5" (the size will vary depending on the image). We hope to obtain all images by April 17. For additional questions, feel free to contact Alisa Zapp Machalek at alisa.machalek-at-nih.gov or (301) 496-7301.
The Advanced Materials Research Institute (AMRI) at the University of New Orleansis looking for a post-doctoral scholar to work on a project about the characterization of transition metal oxides used as electrode material in rechargeable batteries. The successful candidate should have a background in crystallography and TEM. The position is to be filled immediately.
contact : Heike Gabrisch Assistant Professor of Chemistry and Material Sciences University of New Orleans College of Sciences Department of Chemistry/AMRI New Orleans, LA 70148
If the objective lens has gone down you may have a similar problem. As the instrument is aligned round a normally working objective lens the problems you describe could be attributed to that!
Look at you objective lens current - does it change when you change the focus controls or is it static? If it is static switch off and take a look at the objective lens output power transistors on the cooled board.
Alternatively your illumination alignment TILT is out of shape which would move the final image alignment well off centre. What worries me is the caustic that you are seeing on LM1. I do not know if JEOL use the objective just a little under these conditions, I would guess not, so we have to start suspecting the high voltage or the lens/high voltage standard levels are wrong it could be the EPROM?
You do not have a simple problem, maybe others have seen this before and know the route that the engineers took?
Come back with more detailed information and I would be pleased to help.
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "Ian MacLaren" {maclaren-at-tu-darmstadt.de} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Cc: "Ralf Theissmann" {ralf-at-steno.st.mw.tu-darmstadt.de} ; "Gerhard Miehe" {miehe-at-eddy.st.mw.tu-darmstadt.de} Sent: Monday, March 31, 2003 12:37 PM
The following is a response from one of the researchers working on a MD model in mice, who does EM of end plates . -----------------
The only way I can tell where the end plates are is to trim the muscle at the synaptic zone before I process it, because you can see the nerves with a dissecting scope. Then when I embed the muscle, I orient the square-trimmed end that I know has synapes to the top (or tip) of the resin block. Once the tissue has gone through Osmium, I think it is impossible to discern the synaptic zone. Thomas Proctor proctoth-at-ohsu.edu -------------------
Bob Kayton, PhD Histo/EM Core 503-494-2504-Lab 503-703-3938-Cell
As part of our restructuring, we will not renew the service contract on our trusty old Philips410LS. Can anyone recommend third party service in Northern California? I understand Ron Veil is no longer in business.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
A student would like to bring back phage and bacterial samples from Mexico to the UK.
With increased security at airports, he was considering bringing the pellets back in a sucrose/water solution rather than sucrose/cacodylate. The samples would be in this medium for approx. 15hours, and would be fixed in 2.5% Gluteraldehyde in 0.1M Sodium Cacodylate buffer. I am a bit concerned about leaving the samples in the sucrose/water medium, but what is the alternative?
Where he is going has no EM facilities, although they can obtain reagents.
Any advice would be greatly appreciated.
Linton Brown Institute of Aquaculture University of Stirling Stirling FK9 4LA Scotland UK -- The University of Stirling is a university established in Scotland by charter at Stirling, FK9 4LA. Privileged/Confidential Information may be contained in this message. If you are not the addressee indicated in this message (or responsible for delivery of the message to such person), you may not disclose, copy or deliver this message to anyone and any action taken or omitted to be taken in reliance on it, is prohibited and may be unlawful. In such case, you should destroy this message and kindly notify the sender by reply email. Please advise immediately if you or your employer do not consent to Internet email for messages of this kind. Opinions, conclusions and other information in this message that do not relate to the official business of the University of Stirling shall be understood as neither given nor endorsed by it.
Dear all, Thank you to all who replied and made suggestions about how we could fix things with the misaligned JEOL3010. The suggestion to use the "N" button and recover the engineers settings was a good one. I still didn't have much beam afterwards, but it was then close enough to adjust normally. The worst adjusted bits were the gun deflectors and condensor deflectors. The adjustment was made harder since the LAB6 crystal is a bit tilted to one side, but I have a good workable alignment now (even if at some point we have to look at the filament itself if it continues to tilt off).
So, thanks once again to all.
Best wishes
-- Ian MacLaren Technische Universität Darmstadt Material-und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany
This is an issue with many answers, depending on the type of work preformed.
Are you a quality control lab that is spot checking production type samples? If so, than a known defective product is given to the employee and asked to find any problems.
If you are a diagnostic TEM lab, than, are they responsible for actually making the diagnosis? Probably not, thus checking the quality of thicks and thins is the only objective way to due quality control. Look for useable thicks with minimal cutting and staining artifacts. When checking the thins, look for section quality, staining artifact and final micrograph quality. One can objectively review these items.
On the SEM, are you doing ED or WD x-ray analysis? Here a know sample can be used as a test material to determine if the proper x-ray spectrum is being determined. If bio. samples, look for CPD artifacts, such as ruptured membranes.
One way to help insure quality (if a bio. lab) is too have the techs become certified by MSA, thru their certification program.
This is not an easy answer, but if you can objectively critique the work on a daily basis, quality control should not be an issue.
Best of Luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} 03/31/03 07:42AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good morning all in the listserver, I had a question - how does on go about doing quality control on EM - both TEM and SEM. In other words, other than giving techs unknown samples and checking the end results - or just looking over their shoulders checking thicks and thin quality - is there another way? Thanks in advance, Connie Cummings
Why look for another service company, Philips is willing to work on a pay for service rate. Check with them, that way you still can maintain the current service person.
Best of Luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } Rick Harris {raharris-at-ucdavis.edu} 03/31/03 05:35PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As part of our restructuring, we will not renew the service contract on our trusty old Philips410LS. Can anyone recommend third party service in Northern California? I understand Ron Veil is no longer in business.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
} -------- } From: Linton Brown } Sent: 01 April 2003 12:09 } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Transporting samples } } A student would like to bring back phage and bacterial samples from Mexico } to the UK. } } With increased security at airports, he was considering bringing the } pellets back in a sucrose/water solution rather than sucrose/cacodylate. } The samples would be in this medium for approx. 15hours, and would be } fixed in 2.5% Gluteraldehyde in 0.1M Sodium Cacodylate buffer. I am a bit } concerned about leaving the samples in the sucrose/water medium, but what } is the alternative? } } Where he is going has no EM facilities, although they can obtain reagents. } } Any advice would be greatly appreciated. } } Linton Brown } Institute of Aquaculture } University of Stirling } Stirling } FK9 4LA } Scotland } UK } } -- The University of Stirling is a university established in Scotland by charter at Stirling, FK9 4LA. Privileged/Confidential Information may be contained in this message. If you are not the addressee indicated in this message (or responsible for delivery of the message to such person), you may not disclose, copy or deliver this message to anyone and any action taken or omitted to be taken in reliance on it, is prohibited and may be unlawful. In such case, you should destroy this message and kindly notify the sender by reply email. Please advise immediately if you or your employer do not consent to Internet email for messages of this kind. Opinions, conclusions and other information in this message that do not relate to the official business of the University of Stirling shall be understood as neither given nor endorsed by it.
Hi Does anyone know when the word "EDAX" was registered as a trademark? The reason I ask is that I have come across procedural documentation that calls for "...analysis by EDAX (Energy Dispersive Analysis of X-Rays)." One lab is now telling us that they cannot comply as the don't have EDAX - theirs is a PGT system !
Ady
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EDAX is the name of a company which provides EDXA (or EDS or EDX or...) equipment. They were founded in 1962 as Nuclear Diodes Inc. and have operated under the name EDAX since 1972. (see www.EDAX.com) EDAX is a tradename just as "Kleenex" is for facial tissue or "Xerox" is for photocopies.
A number of companies manufacture EDX equipment including: EDAX, Gresham, IXRF, Noran, Oxford, PGT, Rontec, ... (I may have missed some.)
There have been a number of discussions about the "proper" terminology for the Energy Dispersive method of X-Ray Analysis. There are even quibbles about the term "energy-dispersive".
The writer of your procedures was not careful enough about his terminology.
Cheers, Henk
At 06:37 AM 4/1/2003 -0800, ady jenkinson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
I have co-written a paper on "Quality in Electron Microscopy" which I would be pleased to send through to you if you are interested?
It deals with test procedures, instrument and staff assessment, quality control in an EM unit etc etc.
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, March 31, 2003 1:42 PM
Announcing an Additional Sunday Short Course Scheduled for Microscopy and Microanalysis 2003 San Antonio TX August 3, 2003
Application Pathways Solutions: From High Pressure Freezing to the Electron Microscope
Instructors: Kent McDonald Jan Slot
Location: San Antonio Convention Center, Room 008B Time: 9:00am to 5:00pm Attendance: Maximum 20 people Attendance Fee: same as other Sunday short courses. Payable with meeting registration or by contacting Meeting Management at:
Workshop outline (Combination of didactic and practical sessions): - Introduction of High Pressure Freezing, Why HPF? - Advantages over conventional chemical and or microwave fixation techniques. - Techniques and methods for Sample preparation after HPF including; - Freeze fracture - Frozen hydrated cryosectioning - Cryosectioning and immunolabeling - Cryoplaning - Freeze substitution. - Additional presentations
For registration information see the meeting site at: http://www.msa.microscopy.com/MSAMeetings/MM03/MMHomePage.html
Best- Jay ________________________________ AKA. W. Gray Jerome, Ph.D., FAHA Pathology Vanderbilt University Medical Center B-2101 MCN 1161 21st Ave, South Nashville, TN 37232-2561 615-322-5530 jay.jerome-at-vanderbilt.edu
Hello Ian, On the keyboard draw of the JEOL 3010 is a neutralize button (marked "N" under a clear plastic cover near the inset "reset" button). When this button is pressed the computer resets the presently selected alignment parameter to the last value set by the JEOL service technician when they last aligned your TEM in service mode.
If a user has put your alignment is some bizarre "wopper-jawed" state, pressing the neutralize button will return the setting to the last JEOL alignment. To accomplish this, press an alignment select button in the keyboard drawer then hit the neutralize button. Do this for all the alignment select buttons, e.g. Deflectors (Gun, Spot, Cond, Image, Proj), the Stimators, and Con Def Adj.
This should put your TEM in a useable state if gross misalignment is your only problem. You will need to fine tune the alignment after you neutralize the settings since the operational state of the instrument may have drifted since the last technician stored settings.
I will email you a PDF file of our detailed alignment procedures, which you may find of value.
Good luck and I hope this information helps.
Sincerely, Tyrone Daulton
Ian MacLaren wrote:
} } Dear all, } Our JEOL3010 alignment has got into a bad state, and I can't seem to get it } back with the normal alignments described in the handbook. Probably a user } has accidently altered a gun or condensor alignment. Does anybody have a } set of instructions for a complete gun and column alignment, and not just } the everyday procedures in the standard blue ring-binder? } } } -- } Ian MacLaren } Technische Universität Darmstadt } Material-und Geowissenschaften } Petersenstr. 23 } 64287 Darmstadt } Germany
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Facility Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
I have researcher gearing up to do a phytolith database (modern and ancient). I have never isolated these myself from fresh tissue and wondered if anyone had any pointers and good references I could look up. Thanks
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
EDAX is a tradename. It is not a trademark. It may have been at one time but not now. You can check for trademarks at
http://www.uspto.gov and do a search of the trademarks database. EDAX returns zero.
Have the procedure revised to kick the A out. EDX. That might work. Or, perhaps EDS?
gary g.
At 06:37 AM 4/1/2003, you wrote:
} Hi } Does anyone know when the word "EDAX" was registered } as a trademark? } The reason I ask is that I have come across procedural } documentation that calls for "...analysis by EDAX } (Energy Dispersive Analysis of X-Rays)." } One lab is now telling us that they cannot comply as } the don't have EDAX - theirs is a PGT system ! } } Ady
Below you will find the information on the EDAX trademark. I think it will answer your questions.
Best regards-
David
Word Mark EDAX Goods and Services IC 009. US 021 026 038. G & S: SCIENTIFIC MEASURING INSTRUMENTS-NAMELY, AN ENERGY DISPERSIVE X-RAY ANALYZER; ANALYTICAL X-RAY APPARATUS, NAMELY X-RAY DETECTORS, X-RAY SPECTROMETERS AND PARTS THEREFOR, ELEMENTAL ANALYZERS, X-RAY GENERATORS, VIDEO DISPLAY CONSOLES FOR X-RAY ANALYSIS, SAMPLE CHAMBERS, REMOTE CONTROLS AND COMPUTER PROGRAMS FOR X-RAY ANALYSIS; COMPUTER-BASED AIDED ANALYTICAL UNITS USING ENERGY DISPERSIVE ANALYSIS. FIRST USE: 19781229. FIRST USE IN COMMERCE: 19781229 Mark Drawing Code (3) DESIGN PLUS WORDS, LETTERS, AND/OR NUMBERS Design Search Code 170725 200308 261701 261704 Serial Number 73453331 Filing Date November 17, 1983 Published for Opposition March 24, 1987 Registration Number 1460660 Registration Date October 13, 1987 Owner (REGISTRANT) EDAX INTERNATIONAL, INC. CORPORATION DELAWARE 103 SHELTER ROAD P.O. BOX 135 PRAIRIE VIEW ILLINOIS 60069 (LAST LISTED OWNER) PHILIPS ELECTRONICS NORTH AMERICA CORPORATION CORPORATION BY CHANGE OF NAME FROM DELAWARE 100 EAST 42ND STREET NEW YORK NEW YORK 10017
Assignment Recorded ASSIGNMENT RECORDED Attorney of Record PAUL R. MILLER Prior Registrations 0925096 Disclaimer NO CLAIM IS MADE TO THE EXCLUSIVE RIGHT TO USE A DESIGN REPRESENTATION OF A GRAPH APART FROM THE MARK AS SHOWN Type of Mark TRADEMARK Register PRINCIPAL Affidavit Text SECT 15. SECT 8 (6-YR). Live/Dead Indicator LIVE
Word Mark EDAX Goods and Services IC 009. US 026. G & S: SCIENTIFIC MEASURING INSTRUMENTS-NAMELY, AN ENERGY DISPERSIVE X-RAY ANALYZER. FIRST USE: 19701000. FIRST USE IN COMMERCE: 19701000 Mark Drawing Code (1) TYPED DRAWING Serial Number 72391254 Filing Date May 6, 1971 Registration Number 0925096 Registration Date December 7, 1971 Owner (REGISTRANT) NUCLEAR DIODES, INC. CORPORATION ILLINOIS 103 SHELTER ROAD PRAIRIE VIEW ILLINOIS 60069 (LAST LISTED OWNER) EDAX, INC. CORPORATION BY CHANGE OF NAME, BY ASSIGNMENT, BY MERGER, BY CHANGE OF NAME, BY MERGER, BY CHANGE OF NAME, BY MERGER, BY ASSIGNMENT DELAWARE 91 MCKEE DRIVE MAHWAH NEW JERSEY 07430
Assignment Recorded ASSIGNMENT RECORDED Attorney of Record CHRISTOPHER R. LEWIS Type of Mark TRADEMARK Register PRINCIPAL Affidavit Text SECT 15. SECT 8 (6-YR). SECTION 8(10-YR) 20020516. Renewal 2ND RENEWAL 20020516 Live/Dead Indicator LIVE
ady jenkinson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 } Does anyone know when the word "EDAX" was registered } as a trademark? } The reason I ask is that I have come across procedural } documentation that calls for "...analysis by EDAX } (Energy Dispersive Analysis of X-Rays)." } One lab is now telling us that they cannot comply as } the don't have EDAX - theirs is a PGT system ! } } Ady } } __________________________________________________ } Do you Yahoo!? } Yahoo! Tax Center - File online, calculators, forms, and more } http://platinum.yahoo.com
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The problem for EDAX, the company, is that they have not defended their alleged trademark. None of their web pages show that EDAX is a registered trademark. Thus, they are in a minority position. Seemingly, a defensive position. But they may not even know this or be aware of this.
If one has a trademark, one must defend it....or risk losing it. The ® makes a big difference.
gary g.
At 06:39 PM 4/1/2003, you wrote: } -- [ From: Garber, Charles A. * EMC.Ver #3.1a ] -- } Attachment: earl.doc Code: 0KTIJ8M \ Created: 04-01-03, 09:36 PM } [124 Kb] } } I think you are incorrect, the term EDAX is a registered trade name with the } US Office of Patents and Trademarks. } Chuck } } } EDAX is a tradename. It is not a trademark. } } It may have been at one time but not now. } } You can check for trademarks at } } } } http://www.uspto.gov and do a search of } } the trademarks database. EDAX returns zero. } } } } Have the procedure revised to kick the A } } out. EDX. That might work. Or, perhaps EDS? } } } } gary g. } } } } } } } } At 06:37 AM 4/1/2003, you wrote: } } } } } Hi } } } Does anyone know when the word "EDAX" was registered } } } as a trademark? } } } The reason I ask is that I have come across procedural } } } documentation that calls for "...analysis by EDAX } } } (Energy Dispersive Analysis of X-Rays)." } } } One lab is now telling us that they cannot comply as } } } the don't have EDAX - theirs is a PGT system ! } } } } } } Ady } } } } } } -------- REPLY, End of original message -------- }
Reading the postings re JEOL 3010 alignment prompts me to ask whether any kind soul has a procedure for 840 alignment which is easier to follow than the method in the JEOL manual.
I will be grateful for a copy of same.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
John Russ helped start the confusion in terms. He popularized the "EDAX" term with the book "Energy Dispersive Analysis of X-rays" edited by John Russ and published by ASTM in 1971. John then worked for EDAX Inc. in the 1970s before going to North Caroline State.
Do you have any more comments on this history, John?
Ron Vane XEI Scientific
----- Original Message ----- } From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} To: "ady jenkinson" {ajenkinson-at-yahoo.com} ; {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, April 01, 2003 7:18 AM
Gary Whether EDAX is a trademark or not, it is the name of the trading company, and I think you'll find they will be defending it with the necessary, if not overwhelming, force. Interestingly, EDAX do not refer to EDX but use the acronyms EDS, WDS and XMS.
Chris
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Garber, Charles A." {cgarber-at-2spi.com} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 02, 2003 4:25 AM
This is an interesting fact. Thanks.
The problem for EDAX, the company, is that they have not defended their alleged trademark. None of their web pages show that EDAX is a registered trademark. Thus, they are in a minority position. Seemingly, a defensive position. But they may not even know this or be aware of this.
If one has a trademark, one must defend it....or risk losing it. The ® makes a big difference.
gary g.
At 06:39 PM 4/1/2003, you wrote: } -- [ From: Garber, Charles A. * EMC.Ver #3.1a ] -- } Attachment: earl.doc Code: 0KTIJ8M \ Created: 04-01-03, 09:36 PM } [124 Kb] } } I think you are incorrect, the term EDAX is a registered trade name with the } US Office of Patents and Trademarks. } Chuck } } } EDAX is a tradename. It is not a trademark. } } It may have been at one time but not now. } } You can check for trademarks at } } } } http://www.uspto.gov and do a search of } } the trademarks database. EDAX returns zero. } } } } Have the procedure revised to kick the A } } out. EDX. That might work. Or, perhaps EDS? } } } } gary g. } } } } } } } } At 06:37 AM 4/1/2003, you wrote: } } } } } Hi } } } Does anyone know when the word "EDAX" was registered } } } as a trademark? } } } The reason I ask is that I have come across procedural } } } documentation that calls for "...analysis by EDAX } } } (Energy Dispersive Analysis of X-Rays)." } } } One lab is now telling us that they cannot comply as } } } the don't have EDAX - theirs is a PGT system ! } } } } } } Ady } } } } } } -------- REPLY, End of original message -------- }
-----Message d'origine----- De : Edward Calomeni [mailto:calomeni-1-at-medctr.osu.edu] Envoyé : mardi 1 avril 2003 15:32 À : Microscopy-at-sparc5.microscopy.com Objet : Re: Third party microscope repairs and service
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Morning Rick,
Why look for another service company, Philips is willing to work on a pay for service rate. Check with them, that way you still can maintain the current service person.
Best of Luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } Rick Harris {raharris-at-ucdavis.edu} 03/31/03 05:35PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As part of our restructuring, we will not renew the service contract on our trusty old Philips410LS. Can anyone recommend third party service in Northern California? I understand Ron Veil is no longer in business.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
In a message dated 4/2/03 1:40:57 AM, RVane-at-Evactron.com writes:
} John Russ helped start the confusion in terms. He popularized the } "EDAX" term with the book "Energy Dispersive Analysis of X-rays" } edited by John Russ and published by ASTM in 1971. John then worked } for EDAX Inc. in the 1970s before going to North Caroline State. } } Do you have any more comments on this history, John?
Not really. I've been following the exchange with interest. EDAX Labs was started as a "division" of Nuclear Diodes in 1969, when I left JEOL to head it up. The name was actually proposed by Adrian Moggre, our European salesman. The name started to catch on, and we were a major, even dominant market player (along with PGT and Kevex), so in '71 we changed the name of company. The fact that people confused the name of the company with the name of the technique was mostly a welcome thing as it helped market penetration. EDS and EDX weren't pronouncable, but Edax was. Edax was sold to Philips in'74, and I left in '78 to come here to N. C. State U. (retired in '95). As far as I know there are only two people (Alan Sandborg, who preceded me, and Bob Shen, whom I hired) still at Edax from the "old days." Probably Alan could comment on whether any conscious decision was ever made to not register or defend the name as a trademark.
Dr. Guo Feng Zhang Division of Bioenginerring & Physical Science 301-451-3856 ZhangGuo-at-mail.nih.gov
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Sunday, March 30, 2003 8:18 PM To: NewSub-at-sparc5.microscopy.com
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I am interested in getting a second hand cryotransfer system for SEM. For example: Fisons LT 7400 Oxford CT 1500
Thank you for suggestions/offers.
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At 10:19 AM 4/2/03 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A colleague of mine is planning 3D reconstruction of mouse retina based on EM micrographs. My limited experience was from the manual tracing of scanned micrographs. I wonder any of you had done EM reconstruction lately? What's the recommended internal alignment marker? Any good imaging software to help with the image reconstruction? Any suggestions would be highly appreciated.
I am using Scion Image and a graphics tablet to measure areas of tissues under the microscope. I must use the line tool rather than the automatic function to find the areas, and thus I can get only one area at a time. There are 20 or more areas, sometimes overlapping, to measure per field, which makes the proces difficult if not impossible. Is there a workaround in Scion Image, or perhaps other software compitable with Scion cards, that can measure multiple areas per field? I would appreciate any advice.
If you are absolutely sure that manual tracing is the only way to get the measurements you need, why not put the image into a program like Photoshop (or PAint Shop Pro, etc.) and use it to draw outlines and fill the regions you are interested in with a unique color. then just have Image measure the colored regions.
John Russ =====
In a message dated 4/2/03 6:20:59 PM, gbarclay-at-trinidad.net writes:
} I am using Scion Image and a graphics tablet to measure areas of tissues } under the microscope. I must } use the line tool rather than the automatic function to find the areas, } and thus I can get only one area } at a time. There are 20 or more areas, sometimes overlapping, to measure } per field, which makes the } proces difficult if not impossible. Is there a workaround in Scion Image, } or perhaps other software } compitable with Scion cards, that can measure multiple areas per field? } I would appreciate any advice.
Are you making your paper universally available? If so I would like a copy too.
Cheers,
Jodi Reynolds. ---------- From: Steve Chapman [SMTP:protrain-at-emcourses.com] Sent: Wednesday, 2 April 2003 01:25 To: MSA Cc: ROSSCAC Subject: Quality in Electron Microscopy
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Hi
I have co-written a paper on "Quality in Electron Microscopy" which I would be pleased to send through to you if you are interested?
It deals with test procedures, instrument and staff assessment, quality control in an EM unit etc etc.
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, March 31, 2003 1:42 PM Subject: EM Quality Control
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good morning all in the listserver, } I had a question - how does on go about doing quality control on EM - both } TEM and SEM. In other words, other than giving techs unknown samples and } checking the end results - or just looking over their shoulders checking } thicks and thin quality - is there another way? } Thanks in advance, } Connie Cummings } }
Are there are any plant people out there who have used Yariv reagents to disrupt AGPs? I want to know how to prepare the reagents (purcahsed from Biosupplies Australia) so I can put them in a solid agar medium for culturing. Are there are any good references on the technical aspects of using the reagents?
Thanks for your help (I know, not strictly a microscope question!).
Are there are any plant people out there who have used Yariv reagents to disrupt AGPs? I want to know how to prepare the reagents (purcahsed from Biosupplies Australia) so I can put them in a solid agar medium for culturing. Are there are any good references on the technical aspects of using the reagents?
Thanks for your help (I know, not strictly a microscope question!).
Well if I had a $ for every email reply I have had for my offer I would now be planning a l o n g holiday??*!!
One MSA listserver star pointed out to me that to place the paper (now upgraded) on the web would save a good deal of work. So if you need a copy you are welcome to take it from www.emcourses.com.
Thank you all for your interest.
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
I would also appreciate a copy if available. Thank you
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com ----- Forwarded by Robert Fowler/TCU/TDK-US on 04/03/2003 07:15 AM -----
"Reynolds, Jodi JI" To: {microscopy-at-sparc5.microscopy.com} {ReynoldsJ-at-one cc: steel.com} Subject: RE: Quality in Electron Microscopy
04/02/2003 08:44 PM
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Hi Steve,
Are you making your paper universally available? If so I would like a copy too.
Cheers,
Jodi Reynolds. ---------- From: Steve Chapman [SMTP:protrain-at-emcourses.com] Sent: Wednesday, 2 April 2003 01:25 To: MSA Cc: ROSSCAC Subject: Quality in Electron Microscopy
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Hi
I have co-written a paper on "Quality in Electron Microscopy" which I would be pleased to send through to you if you are interested?
It deals with test procedures, instrument and staff assessment, quality control in an EM unit etc etc.
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, March 31, 2003 1:42 PM Subject: EM Quality Control
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good morning all in the listserver, } I had a question - how does on go about doing quality control on EM - both } TEM and SEM. In other words, other than giving techs unknown samples and } checking the end results - or just looking over their shoulders checking } thicks and thin quality - is there another way? } Thanks in advance, } Connie Cummings } }
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
When I have been asked in the past to comment on this topic, as a speaker, I have first asked the audience what their definition of quality was and it has always been an amazement to me how passionately individuals can become about "their" definition of quality.
I have always thought that quality meant "meeting or exceeding the customer's expectations". At least that has been our overall guiding philosophy in the management of both our own in-house electron microscopy services laboratory as well as the manufacturing and distribution of our SPI Supplies products.
Speaking specifically of the management of the EM laboratory part of our business, this means that in a sense, each customer could have their own perception of what quality means to them, and we have to understand that and we have to give them what they want in terms of expectations.
Some customers want the fastest possible turnaround time, yet others might want the highest possible resolution in an image. Others are more interested in contrast than necessarily resolution. Others are more interested in the detail and explanation in our final report. Some want the technician actually running the microscope to be readily available by telephone. Yet others, for example, someone measuring the size of latex particles, the correct magnification calibration of the microscopes is the single most important expectation.
And not to be forgotten is that customer who has a problem to solve and has the expectation that we will have the staff that has the ability to technically understand the problem and will take the time to sort things out to the degree that the right control and exemplar samples will submitted and the work will be done in a way that results will be generated to solve the problem.
The list becomes endless.
In order to enhance the chances that they will be operating at the expectation of their customers, there is needed a quality "process", a method by which there can be measured the degree to which the expectations of the customers are indeed being met. Some form of a total quality management system is needed. We ourselves have found that when a job had to be done over again, since the "cost of rework" in an EM lab is so high, it is always helpful to understand what went wrong and why it had to be done over again so that the management can learn from that experience and take steps to keep that kind of problem from happening in the future and thereby lowering the costs of rework and the overall costs of the laboratory's operation overall.
This kind of quality process is also an integral part of any credible laboratory accreditation program and the need to comply is what we needed to be the "stick" that brought about the needed mind set change. So we ourselves happen to see that whatever money we pay for the inspection to be accredited to the standard of ISO 17025, we get back in interest, so to speak, because of the way it has helped us to reduce the number of times a job has to be done over again and therefore also our cost of "rework".
So in the end, for an EM lab to operate with "high quality", it is a matter of whether it is being operated at a level that is either at or greater than the expectations of their customers and for that to happen, there has to be a constant process of feedback in terms of how close that laboratory is coming to meeting the expectations.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
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Steve,
I echo Jodi's response, I too would like a copy of your paper!
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
} -----Original Message----- } From: Reynolds, Jodi JI [SMTP:ReynoldsJ-at-onesteel.com] } Sent: Wednesday, April 02, 2003 8:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: RE: Quality in Electron Microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Steve, } } Are you making your paper universally available? If so I would like a } copy too. } } Cheers, } } Jodi Reynolds. } ---------- } From: Steve Chapman [SMTP:protrain-at-emcourses.com] } Sent: Wednesday, 2 April 2003 01:25 } To: MSA } Cc: ROSSCAC } Subject: Quality in Electron Microscopy } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } Hi } } I have co-written a paper on "Quality in Electron Microscopy" which } I would } be pleased to send through to you if you are interested? } } It deals with test procedures, instrument and staff assessment, } quality } control in an EM unit etc etc. } } Steve Chapman } Senior Consultant Protrain } Electron Microscopy Training and Consultancy World Wide } Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 } www.emcourses.com } } ----- Original Message ----- } } From: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Monday, March 31, 2003 1:42 PM } Subject: EM Quality Control } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Good morning all in the listserver, } } I had a question - how does on go about doing quality control on } EM - both } } TEM and SEM. In other words, other than giving techs unknown } samples and } } checking the end results - or just looking over their shoulders } checking } } thicks and thin quality - is there another way? } } Thanks in advance, } } Connie Cummings } } } } } } } }
I would also like to a copy if available, thanks a lot
Jinguo Wang
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, (814) 863-0637 email: jqw11-at-psu.edu At 07:16 AM 4/3/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The Z slice depth in the Zeiss LSM 510 software is limited to the working distance of the objective you are using. The objective you describe I believe has a working distance of only 90 ums. That means if you are asking it to image deeper into a sample than that it will not do it because it is impossible to image beyond its working distance Check its specs.
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University 609-258-5432
Visit us at http://www.molbio.princeton.edu/facility/confocal/index.html
All Many thanks for all the useful information. I think the best think to do is to remove the offending "A" from "EDAX" as an 'Editorial Change'in our procedure. That should keep everyone happy :-)
Regards Ady
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The Z slice depth in the Zeiss LSM 510 software is limited to the working distance of the objective you are using. The objective you describe I believe has a working distance of only 90 ums. That means if you are asking it to image deeper into a sample than that it will not do it because it is impossible to image beyond its working distance Check its specs.
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University 609-258-5432
Visit us at http://www.molbio.princeton.edu/facility/confocal/index.html
Marc Helvey VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 FAX: (408) 428-9555 E-Mail: marc.helvey-at-vlsistd.com {mailto:marc.helvey-at-vlsistd.com} Internet: http://www.vlsistandards.com {http://www.vlsistandards.com/}
-----Original Message----- } From: Sherwood, Margaret [mailto:MSHERWOOD-at-PARTNERS.ORG] Sent: Thursday, April 03, 2003 6:06 AM To: 'protrain-at-emcourses.com' Cc: 'microscopy-at-sparc5.microscopy.com'
Steve,
I echo Jodi's response, I too would like a copy of your paper!
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
} -----Original Message----- } From: Reynolds, Jodi JI [SMTP:ReynoldsJ-at-onesteel.com] } Sent: Wednesday, April 02, 2003 8:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: RE: Quality in Electron Microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Steve, } } Are you making your paper universally available? If so I would like a } copy too. } } Cheers, } } Jodi Reynolds. } ---------- } From: Steve Chapman [SMTP:protrain-at-emcourses.com] } Sent: Wednesday, 2 April 2003 01:25 } To: MSA } Cc: ROSSCAC } Subject: Quality in Electron Microscopy } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } Hi } } I have co-written a paper on "Quality in Electron Microscopy" which } I would } be pleased to send through to you if you are interested? } } It deals with test procedures, instrument and staff assessment, } quality } control in an EM unit etc etc. } } Steve Chapman } Senior Consultant Protrain } Electron Microscopy Training and Consultancy World Wide } Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 } www.emcourses.com } } ----- Original Message ----- } } From: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Monday, March 31, 2003 1:42 PM } Subject: EM Quality Control } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Good morning all in the listserver, } } I had a question - how does on go about doing quality control on } EM - both } } TEM and SEM. In other words, other than giving techs unknown } samples and } } checking the end results - or just looking over their shoulders } checking } } thicks and thin quality - is there another way? } } Thanks in advance, } } Connie Cummings } } } } } } } }
Quality is what meets the requirements of the job. Anything less has to be done over and anything more is a waste of time and money. Of course you have to aim a little higher than just good enough to do the job to eliminate the ones that fall short. But far too much money is spent on quality that isn't needed for the results desired.
How many times have you seen results to three decimal places based on measurement taken with instruments that are accurate to one decimal place. Computers have only made this worse. When we used slide rules we at least realized that the results were low resolution.
Realistically some one doing work for the public has to do higher quality than the customer would be satisfied with if he was doing it himself but when setting in house quality standards make them only as good as they need to be plus a reasonable safety factor.
Making every job a master piece looks great but it is terrible waste of time and money.
Gordon Couger Stillwater, OK www.couger.com/gcouger
----- Original Message ----- } From: "Garber, Charles A." {cgarber-at-2spi.com} : When I have been asked in the past to comment on this topic, as a speaker, I : have first asked the audience what their definition of quality was and it : has always been an amazement to me how passionately individuals can become : about "their" definition of quality. : : I have always thought that quality meant "meeting or exceeding the : customer's expectations". At least that has been our overall guiding : philosophy in the management of both our own in-house electron microscopy : services laboratory as well as the manufacturing and distribution of our SPI : Supplies products. : : Speaking specifically of the management of the EM laboratory part of our : business, this means that in a sense, each customer could have their own : perception of what quality means to them, and we have to understand that and : we have to give them what they want in terms of expectations. : : Some customers want the fastest possible turnaround time, yet others might : want the highest possible resolution in an image. Others are more interested : in contrast than necessarily resolution. Others are more interested in the : detail and explanation in our final report. Some want the technician : actually running the microscope to be readily available by telephone. Yet : others, for example, someone measuring the size of latex particles, the : correct magnification calibration of the microscopes is the single most : important expectation. : : And not to be forgotten is that customer who has a problem to solve and has : the expectation that we will have the staff that has the ability to : technically understand the problem and will take the time to sort things out : to the degree that the right control and exemplar samples will submitted and : the work will be done in a way that results will be generated to solve the : problem. : : The list becomes endless. : : In order to enhance the chances that they will be operating at the : expectation of their customers, there is needed a quality "process", a : method by which there can be measured the degree to which the expectations : of the customers are indeed being met. Some form of a total quality : management system is needed. We ourselves have found that when a job had to : be done over again, since the "cost of rework" in an EM lab is so high, it : is always helpful to understand what went wrong and why it had to be done : over again so that the management can learn from that experience and take : steps to keep that kind of problem from happening in the future and thereby : lowering the costs of rework and the overall costs of the laboratory's : operation overall. : : This kind of quality process is also an integral part of any credible : laboratory accreditation program and the need to comply is what we needed to : be the "stick" that brought about the needed mind set change. So we : ourselves happen to see that whatever money we pay for the inspection to be : accredited to the standard of ISO 17025, we get back in interest, so to : speak, because of the way it has helped us to reduce the number of times a : job has to be done over again and therefore also our cost of "rework". : : So in the end, for an EM lab to operate with "high quality", it is a matter : of whether it is being operated at a level that is either at or greater than : the expectations of their customers and for that to happen, there has to be : a constant process of feedback in terms of how close that laboratory is : coming to meeting the expectations. : : Chuck : : PS: Remember that we are striving to be 100% paperless, therefore there : are no paper copies kept of this correspondence. Please be sure to always : reply by way of "reply" on your software so that the entire string of : correspondence can be kept in one place. : ============================================ : : Charles A. Garber, Ph. D. Ph: 1-610-436-5400 : President 1-800-2424-SPI : SPI SUPPLIES FAX: 1-610-436-5755 : PO BOX 656 e-mail:cgarber-at-2spi.com : West Chester, PA 19381-0656 USA : Cust.Service: spi2spi-at-2spi.com : : Look for us! : ######################## : WWW: http://www.2spi.com : ######################## : ============================================ : :
I would also appreciate a copy if available. Thank you
dr. Pogány Lajos Senior Research Fellow, Metals Research Department, Research Institute for Solid State Physics and Optics, Hungarian Academy of Sciences Office Address: H-1121 Budapest, Konkoly-Thege ut 29-33 Letters: H-1525 Budapest, P.O.B. 49, Hungary Phone: (00)-36-1-392-2222/17-25; Fax: (00)-36-1-392-2215 e-mail:pogany-at-szfki.hu homepage: http://www.szfki.hu/~pogany/
The University of Texas Health Science Center at San Antonio will host a Symposium and Workshop sponsored by Hamamatsu Photonics KK on
Fluorescence Lifetime Imaging and Spectral Imaging
Take advantage of a unique opportunity to get Hands-On experience with commercially available Spectral Imaigng and Fluorescence Lifetime Imaging Microscopy systems.
Workshop Application Deadline: April 7, 2003 June 8-10, 2003 UT Health Science Center San Antonio, TX Tuition: $700 (includes room and board) 20 student limit/4 scholarships available
Prior to the Workshop a Symposium on Spectral Imaging and Fluorescence Lifetime Imaging Microscopy will be held at the Gunter Hotel. Separate regisistration for the symposium is required.
Symposium Registration Deadline: May 1, 2003 June 6-7, 2003 The Sheraton Gunter Hotel 205 E. Houston St. San Antonio, TX Student: $200 ($250 after May 1) Professional: $250 ($300 after May 1)
Academic Participants: Philippe Bastiaens (GER)*Robert Clegg (USA)*Mary Dickinson (USA)*Paul French (UK) Hans Gerritsen (Neth)*Brian Herman (USA)*Ammasi Periasamy (USA) Herbert Schneckenburger (GER)*Ton Visser (Neth)*Timo Zimmermann (GER)
-----Message d'origine----- De : Helvey, Marc [mailto:Marc.Helvey-at-vlsistd.com] Envoyé : vendredi 4 avril 2003 02:09 À : 'Sherwood, Margaret '; 'protrain-at-emcourses.com' Cc : 'microscopy-at-sparc5.microscopy.com' Objet : RE: Quality in Electron Microscopy
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In case someone hasn't already posted this:
http://www.emcourses.com/quality.htm
Marc Helvey VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 FAX: (408) 428-9555 E-Mail: marc.helvey-at-vlsistd.com {mailto:marc.helvey-at-vlsistd.com} Internet: http://www.vlsistandards.com {http://www.vlsistandards.com/}
-----Original Message----- } From: Sherwood, Margaret [mailto:MSHERWOOD-at-PARTNERS.ORG] Sent: Thursday, April 03, 2003 6:06 AM To: 'protrain-at-emcourses.com' Cc: 'microscopy-at-sparc5.microscopy.com'
Steve,
I echo Jodi's response, I too would like a copy of your paper!
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
} -----Original Message----- } From: Reynolds, Jodi JI [SMTP:ReynoldsJ-at-onesteel.com] } Sent: Wednesday, April 02, 2003 8:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: RE: Quality in Electron Microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Steve, } } Are you making your paper universally available? If so I would like a } copy too. } } Cheers, } } Jodi Reynolds. } ---------- } From: Steve Chapman [SMTP:protrain-at-emcourses.com] } Sent: Wednesday, 2 April 2003 01:25 } To: MSA } Cc: ROSSCAC } Subject: Quality in Electron Microscopy } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } Hi } } I have co-written a paper on "Quality in Electron Microscopy" which } I would } be pleased to send through to you if you are interested? } } It deals with test procedures, instrument and staff assessment, } quality } control in an EM unit etc etc. } } Steve Chapman } Senior Consultant Protrain } Electron Microscopy Training and Consultancy World Wide } Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 } www.emcourses.com } } ----- Original Message ----- } } From: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Monday, March 31, 2003 1:42 PM } Subject: EM Quality Control } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Good morning all in the listserver, } } I had a question - how does on go about doing quality control on } EM - both } } TEM and SEM. In other words, other than giving techs unknown } samples and } } checking the end results - or just looking over their shoulders } checking } } thicks and thin quality - is there another way? } } Thanks in advance, } } Connie Cummings } } } } } } } }
Eastman Chemical Company has a position in its corporate research and development labs for a microscopist. Extensive experience with both optical and electron microscopy is required and experience in AFM, computer programming, and/or particle size analysis would be a strong plus. Knowledge of polymer morphology and/or the morphology of coatings and inks is also highly desirable. A PhD is preferred. The laboratory is well equipped with several optical microscopes, an SEM, a TEM, an AFM, and a particle size analyzer. The successful candidate will be working with this equipment and other scientists and engineers at Eastman to develop new/improved polymers and chemicals and to help solve manufacturing problems. Candidates must be highly motivated, have good communication skills and be able to work with teams.
Eastman Chemical Company, a Fortune 500 company, is a major manufacturer of plastics, fibers, and specialty organic chemicals. This position is at the Kingsport, TN site. Kingsport is located in northeast Tennessee in the foothills of the Smoky Mountains. For great information on the Tri-Cities area check www.tricities.net {http://www.tricities.net} and www.johnsoncitytn.com {http://www.johnsoncitytn.com}
Eastman is an equal opportunity/affirmative action employer.
To submit your CV/Resume: www.eastman.com {http://www.eastman.com} , go to Employment, Positions Available and then click Submit Your Resume Online. Reference code ECO/1064HS.
Louis T. Germinario (Lou) Physical Chemistry Research Laboratory Eastman Chemical Company Lincoln Street, B-150B P.O. Box 1972 Kingsport, TN 37662-5150 (423) 229-4047 (423) 229-4558 (Fax) {mailto:germ-at-eastman.com}
The New England Society for Microscopy (NESM) and the Connecticut Microscopy Society (CMS) are pleased to announce the 20th. Annual Woods Hole Spring Symposium on Microscopy, to be held at the Marine Biological Laboratory, Woods Hole, Massachusetts, May 2-3, 2003.
Three scientific sessions are planned, together with a banquet and after-dinner talk, and an exhibition featuring the sposoring societies' sustaining members. A poster session and photomicrograph display will also be featured. Full details, including program information and registration details, are available in the NESM April newsletter, available online at:
All individuals interested in microscopy are invited to participate. Special registration rates are available for student members of the sponsoring societies. Corporate entities interested in sustaining membership are cordially invited to enquire for details as indicated in the Web pages.
The Electron Microscope Facility at the University of New Hampshire is in the process of buying a 120kV TEM. We are a core facility for the University. We are currently looking at Leo's 912AB Omega EFTEM, JEOL's JEM 1230 (HC) and FEI's Tecnai G2 12 Twin.
Do any list-server members have experience with any of these microscopes? Our applications tend to be predominantly biological although we do have some polymer and materials users/researchers.
Any responses would be helpful and can be e-mailed or telephoned directly to me.
Thank you very much.
Nancy Cherim Electron Microscope Facility University of New Hampshire Kendall Hall Room 6 Durham, NH 03824
The next meeting of the Metropolitan Microscopy Society will be held on April 9th, 2003 at the EDAX facility in Mahwah, NJ. EDAX has graciously agreed to host our meeting and to provide complimentary coffee and lunch to all the attendees.
The forthcoming meeting comprises five presentations offering a blend of microscopic, spectroscopic and image processing applications. Mike Marko will describe the cutting edge methods for tomographic reconstruction of biological organelles. Paul Kotula and Tom Hancewicz will discuss methods to extract maximum value from the data by analyzing spectral images. Matt Libera will illustrate ways to form novel surface patterns on polymeric materials, using electrons, for controlling biological activity and Del Redfern will highlight the advances in high-resolution imaging in projection x-ray microscopy.
We invite all members to attend and to inform their colleagues and fellow workers who may also wish to attend the meeting. This will not only offer an opportunity to listen to some of the experts in their respective fields but also a venue to meet colleagues and exchange ideas as well as to discuss some of the new items that are in the pipeline for this year.
It is important that members should pre-register as it will help us plan for lunches. The pre-registration deadline is April 4 and can be accomplished electronically. Please respond via email or fax to Evan Slow directly. For all attendees, the meeting fee, which includes lunch, will be $20.00.
For any additional information about the meeting, please contact any of the officers. We hope to see you on April 9th at the meeting in Mahwah!
Place: EDAX Inc, 91 McKee Drive, Mahwah, NJ 07430-2120. (201) 529-4880 Mapquest: http://www.mapquest.com/maps/map.adp?country=US&addtohistory=&address=18+mckee+drive&city=mahwah&state=nj&zipcode=&homesubmit=Get+Map · 9:30 - 9:55 Registration and Coffee· 9:55 - 10:00 Introduction Manoj Misra, Unilever Research and Development,Edgewater, NJ· 10:00 - 10:45 Electron tomography: techniques and applications Mike Marko, Wadsworth Research Center, Albany,NY· 10:45 - 11:30 Current Methods in Multivariate Spectroscopic ImageAnalysis Tom Hancewicz, Unilever Research andDevelopment, Edgewater, NJ· 11:30 - 12:15 Electron-beam surface-patterned polymers Matt Libera, Stevens Institute of Technology,Hoboken, NJ· 12:15 - 1:00 Lunch (included with registration ? pleasepre-register!)· 1:00 - 1:30 EDAX Tour· 1:30 - 2:15 Spectral Imaging: The Next Step in Microanalysis Paul Kotula, MAS Sponsored Tour Lecturer,Sandia National Lab, Albuquerque, NM · 2:15- 3:00 Projection X-ray microscopy in the SEM Del Redfern, EDAX, Mahwah, NJ
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id QAA03513 for dist-Microscopy; Mon, 7 Apr 2003 16:05:01 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id QAA03506 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Mon, 7 Apr 2003 16:04:30 -0500 (CDT) Received: from postmark.nist.gov (pushme.nist.gov [129.6.16.92]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id QAA03499 for {Microscopy-at-sparc5.Microscopy.Com} ; Mon, 7 Apr 2003 16:04:18 -0500 (CDT) Received: from SCOTT-OPTI.nist.gov (mag216.nist.gov [129.6.98.216]) by postmark.nist.gov (8.12.5/8.12.5) with ESMTP id h37KwKbe023464 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 7 Apr 2003 16:58:20 -0400 (EDT) Message-Id: {5.1.1.5.2.20030407163719.03b66118-at-postmark.nist.gov} X-Sender: js0b-at-postmark.nist.gov X-Mailer: QUALCOMM Windows Eudora Version 5.1.1
Meeting Announcement
REGISTRATION DEADLINE: Friday April 11, 2003
A 4-day MAS Special Topics Workshop on spectrum-imaging and multispectral & hyperspectral data analysis will be held at NIST in Gaithersburg, Maryland from April 28th through May 1st, 2003.
The event will cover forms of hyperspectral data relevant to microbeam analysis, including STEM-based EELS and XEDS, energy-filtered TEM imaging, XEDS in the SEM, spectrum imaging in mass spectrometry, XPS, and optical spectroscopies such as IR and cathodoluminescence.
While the original registration deadline has passed and the technical program is full, a limited number of attendance slots are available. There will also be room for additional posters if you wish to contribute or present scientific material or discuss your ideas on hyperspectral analysis.
If you would like to register for the workshop, please send an email to {mailto:johnhenry.scott-at-nist.gov} including:
Name Affiliation Telephone Number Country of Citizenship which days you will likely attend
I will reply as soon as possible to confirm your registration if slots are still available.
Details about the meeting can be found in the
Workshop Announcement PDF http://www.microprobe.org/masjh/TCs/2003-Hyperspectral/2003-Workshop-Announcement.pdf
or at the Workshop Announcement web page http://www.microprobe.org/masjh/TCs/2003-Hyperspectral/2003-Workshop-Announcement.html
Some Confirmed Invited Speakers:
· Ian Anderson, Oak Ridge National Laboratory · Jim Bentley, Oak Ridge National Laboratory · John Friel, Princeton Gamma-Tech, Inc. · Gerald Kothleitner, FELMI TU-Graz · Paul Kotula, Sandia National Laboratories · Richard Leapman, National Institutes of Health · Paul Mainwaring, Gatan, Inc. · Robert Martin, University of Strathclyde · Chris Michaels, NIST · Larry Nittler, Carnegie Institution of Washington · Michaeleen Pacholski, Rohm and Haas Company · Diane Peebles, Sandia National Laboratories · Peter Statham, Oxford Instruments · Masashi Watanabe, Lehigh University · Norman Wright, Digilab, LLC
Thanks to the generosity of NIST, MAS, and our corporate sponsors attending the workshop, there is no registration fee.
Sponsors
4Pi Analysis, Inc. Advanced Microbeam, Inc. EDAX International Eigenvector Research, Inc. EmiSpec Systems, Inc. FEI Company Gatan, Inc. JEOL USA, Inc. Oxford Instruments Princeton Gamma-Tech, Inc. Research Systems, Inc. Thermo NORAN Microbeam Analysis Society
I hope to see you in Gaithersburg at the Workshop,
-- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
I hope you will plan to join us this Thursday and Friday at the "Art of the Science Image" conference at the University of Tulsa. We're looking
forward to an exciting two days of lectures, workshops, and discussions about scientific imaging with speakers of great expertise and national reputation. For more detail about the conference, accomodations, and travel directions see the Oklahoma Microscopy Society (OMS) web page below.
Well explore the connection between human vision and imaging, delve into spectral imaging and x-ray mapping, tackle depth of field issues, and have the enviable opportunity of a half-day workshop on image analysis with renowned expert Dr. John Russ.
Our not-to-be-missed dinner event will feature a keynote address by David Scharf and a showing of the movie for which his unique blend of science and art received an Emmy award. Plan to enjoy continued conversation with the speakers and your fellow microscopists even after the event by staying at the conference hotel.
Well celebrate aesthetics with a workshop on how to create stunning polarized light images, a lecture on the basics of image composition, and a fascinating talk from author and art historian Lynn Gamwell on connections past and present between microscopy and art. An exhibit of scientific photography from the Rochester Institute of Technology will be on display along with works by David Scharf.
Corporate exhibitors and underwriters with whom you'll the the opportunity to meet and discuss equipment are: Gatan, MicroStar, IXRF, Olympus, Princeton Gamma Tech, Thermo NORAN, Meyer Instruments (Leica Microsytems
and Image-Pro), JEOL, Nikon, Atomic Spectroscopy Instruments, BioRad, and Hitachi.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu (405)325-4391 Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
I'm making up a .28 M stock solution for veronal acetate buffer. 1.47g barbituric acid, 0.97g sodium acetate trihydrate, carbon dioxide free water to 50 ml. I can't get this to dissolve. Does anyone know how to get this to go into solution? Jeannie Selker {jselker-at-molbio.uoregon.edu}
} I'm making up a .28 M stock solution for veronal acetate buffer. } 1.47g barbituric acid, 0.97g sodium acetate trihydrate, carbon } dioxide free water to 50 ml. I can't get this to dissolve. Does } anyone know how to get this to go into solution? Jeannie Selker } {jselker-at-molbio.uoregon.edu}
Are you sure you want barbituric acid as it is notoriously hard to dissolve? Most people use sodium barbitol (or veronal as it is sometimes called). Also it sounds too concentrated.
The formula that I use is: 2.96 g sodium barbitol, 1.94 g sodium acetate (hydrated) made up to 100 ml. You need to stir this vigorously (and even you can heat it to 70-80 deg. C with stirring). So, use a heated, magnetic stirrer that is vigorously stirring the solution. Be patient as it may take a long time to dissolve (sometimes over an hour).
The buffer is then completed by mixing: 12.5 ml of your fixative (2% osmium for example, in ddwater), 5.0 ml of the above veronal solution, 2.5 ml of ddwater, and 0.1N HCl to adjust to the final pH (around 5 ml for a pH of 7.4).
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 ##############################################################
We have the fei Technai 12 TEM. The scope is really nice. We have a twin lens and not the biotwin which suits us fine since we are suppose to serve all disciplines from one scope. The scope is great with relative easy alignment for biological science. "Auto stitch" is a very useful function (only free if negotiated during purchase!) stitching more than one pic eg. 3X3 together to increase your resolution. It works nice for biological sciences but the maximum magnification at which it works nice is a bit low for other sciences. The stigmators are the difficult part in the alignment procedure (I might be the only user struggling with it!). The problems we had is with the third party peripherals. Which ever scope you buy just make sure that it all is working properly and that the scope will do what you like in the future. Service and the service provider to me is part of the scope specs since it is horrible to be left without good or slow support. These are my comments. Hope it is of some help.
-----Original Message----- } From: Nancy Cherim [mailto:nac-at-cisunix.unh.edu] Sent: Monday, April 07, 2003 8:46 PM To: Microscopy-at-sparc5.microscopy.com
The Electron Microscope Facility at the University of New Hampshire is in the process of buying a 120kV TEM. We are a core facility for the University. We are currently looking at Leo's 912AB Omega EFTEM, JEOL's JEM 1230 (HC) and FEI's Tecnai G2 12 Twin.
Do any list-server members have experience with any of these microscopes? Our applications tend to be predominantly biological although we do have some polymer and materials users/researchers.
Any responses would be helpful and can be e-mailed or telephoned directly to me.
Thank you very much.
Nancy Cherim Electron Microscope Facility University of New Hampshire Kendall Hall Room 6 Durham, NH 03824
Further, with regard to barbituric acid, I just checked the Merck Index and they say "Difficultly sol in cold water; freely sol in hot water, in dil acids."
Regarding sodium barbitol: "One gram dissolves in 5 ml water, 2.5 ml boiling water, 400 ml alc. Aq soln is alkaline to litmus and phenophthalein. Ph of 0.1 molar aq soln, 9.4."
So, hot water is the way to go.
############################################################## 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/~image/ ##############################################################
Hi, I was wondering if anybody can tell me where to buy a centrifugation adapter. I have dilute virus samples that I would like to pellet directly onto a grid.
I'm not a member of this group. Therefore, I would appreciate it if you could send your response directly to my e-mail address in addition to posting it.
Thank a lot.
Thomas
thomas.weber-at-mssm.edu -- Dr. Thomas Weber Assistant Professor The Carl C. Icahn Center for Gene Therapy and Molecular Medicine Box 1496 Mount Sinai School of Medicine 1425 Madison Avenue New York, NY 10029-6574 United States of America
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I have a researcher trying to get decent images of mouse egg microvilli. So far no luck. Early SEM examples had an obvious layer of gunk (PVA?) on the the egg. Recent trials show collapsed or adherent microvilli. I have indicated below the articles they were referencing for their prep. Currently, they are fixing, removing zona, dehydrating in EtOH, then CPD. Any suggestions would be appreciated.
For SEM:
-Phillips, D.M. and Shalgi, R. Surface Architecture of the Mouse and Hamster Zona Pellucida and Oocyte. 1980; J. of Ultrastructure Research 72, 1-12
-Shalgi, R. and Phillips, D.M. Mechanics of in vitro Fertilization in the Hamster Biology of Reprod. 1980; 23, 433-444
-Fulka Jr, J., Flechon, B. and Flechon, J.E. Fusion of Mammalian oocytes: SEM observations of surface changes. Reprod. Nutr. Dev. 1989; 29, 551-558
For TEM:
1: Albertini DF, Combelles CM, Benecchi E, Carabatsos MJ. Cellular basis for paracrine regulation of ovarian follicle development. Reproduction. 2001 May;121(5):647-53. Review. PMID: 11427152 [PubMed - indexed for MEDLINE]
2: Albertini DF, Rider V. Patterns of intercellular connectivity in the mammalian cumulus-oocyte complex. Microsc Res Tech. 1994 Feb 1;27(2):125-33. PMID: 8123905 [PubMed - indexed for MEDLINE]
3: Ducibella T, Anderson E, Albertini DF, Aalberg J, Rangarajan S. Quantitative studies of changes in cortical granule number and distribution in the mouse oocyte during meiotic maturation. Dev Biol. 1988 Nov;130(1):184-97. PMID: 3141231 [PubMed - indexed for MEDLINE]
I have an Oxford QX2000 free to a good home, donee to pay packing costs and freight.
It was in working order when it was taken offline a few years ago, apart from a seriously drifty XP2 pulse-processor board.
Unit is complete with monitor and keyboard, and has a few options, the exact nature of which I can't remember, but which included the ability to show a BSE image and position the beam from it.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
first, i have no commercial interest, but i was involved in the early work developing the use of the beckman Airfuge EM-90 rotor. i have used it extensively for quantification of virus, determining species of subviral components produced and their proportions, immunoEM, and about everything else i can think of doing with suspensions of viruses and bacteria. two papers bracketing the work and covering your specific questions are:
Hammond, GW, Hazelton, PR, Chuang, I, and Klisko, B. 1981. Improved Detection of viruses by electron microscopy after direct ultracentrifuge preparation of specimens. J. Clin. Micro., 14:210-221.
Hazelton, PR and Coombs, KM. 1999. The Reovirus Mutant tsA279 L2 Temperature- Sensitive Lesion Is Associated with Production of a Core Particle Deficient in the lambda 2 Core Spike Protein and Minor Core Protein sigma 2. Journal of Virology, 73:2298-2308.
also, i would refer you to the recent review article by hans gelderblom and i, where we discuss a number of concentration techniques.
Hazelton, PR and Gelderblom, HR. 2003. The use of the electron microscope for rapid diagnosis of viral agents in emergent situations. Emerging Infectious Diseases 9:294-303.
if you have any questions, or need any help, do not hesitate to contact me.
paul r. hazelton, PhD electron microscopist, Department of Medical Microbiology University of Manitoba 531 Basic Medical Sciences 730 William Avenue Winnipeg, MB, Canada, R3E 0W3 Clinical Virology consultant, Cadham Provincial Laboratory Winnipeg, Manitoba, Canada telephone 204-789-3313 pager: 204-931-9354 fax: 204-789-3926 e-mail: paul_hazelton-at-umanitiba.ca
We have the remains of two Oxford Instruments AN10000s - somewhat knocked about but still a valuable source of spares - and an entire LEMAS stage/spectrometer control system. The LEMAS was configured for a 4-WD spectrometer microprobe and in good working order when it was taken out last year - again a very useful source of spares for anybody still keeping one of those going.
Cheers,
Stu ---------------------------- Stuart Kearns Department of Earth Sciences University of Bristol Bristol, UK BS8 1RJ Tel: +44 117 954 5435 Fax: +44 117 925 3385 Stuart.Kearns-at-bristol.ac.uk ----------------------------
Is there freeware out there or a plug in to ifranview for batch printing of images 4 images per page of all selected images?
I really love this list. I just want to thank you all for your helpful comments on questions posted here. Thank you Nestor for keeping it up and running.
Stephan H Coetzee Department of Physics EMU Private Bag 0704 Gaborone, Botswana
I am not sure about the plugins, but if you use windows XP, it has "Windows picture and fax viewer" which does the batch printing.
Pavel
----- Original Message ----- } From: "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} To: "Listserver Microscopy (E-mail)" {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 09, 2003 5:54 AM
Hi everyone,
I have a Hitachi H600 TEM (20 years old). I think the HV cable is dying. My evidence for this is:
1. Unstable voltage readouts. When we set the voltage to, say, 50 Kv, it does not read 50 Kv but reads 75Kv. 2. There is a 'hissing' sound when we try to change from 75Kv to 100Kv. The hissing appears to be coming from behind the gun. 3. Our filaments, when they break, are showing signs of over-heating as evidenced by a round blob of tungsten on each of the broken ends. 4. When I open the gun, it smells.
We need to keep it alive for another 12-18 months by which time we anticipate replacing it.
Are my conclusions re the HV cable correct? Does anyone out there know if it is possible to repair the HV cable and if so, what is involved?
cheers
Sarah Ellis
Manager, Microscopy Imaging and Research Core Facility Peter MacCallum Cancer Centre Locked Bag #1 A'Beckett Street East Melbourne 8006
Sorry to be a bother, but I have a couple of queries.
I have looked in the archives and hope that I haven't missed any relevant info.
Firstly, I'm looking for the name/product no. of a Dow Corning product that can be used for moulding to make replicas for microscopy. In the past I have used simple latex, but heard Dow did something better. I have looked at the D-C web site but couldn't work out exactly what product to buy. I want to use this stuff for cleaning purposes (AFM standards) and also for making actual replicas, so the name of the right casting product would also be very much appreciated.
Secondly, I'm interested in pattern recognition (for AFM, SEM, TEM etc.). Am I right in thinking this should be a relatively simple branch of image analysis? I know that one can use image analysis to do things like count cells on agar plates via a live camera, or from an image. Can it be done for things like recurring patterns in say crystals? Is there anything available out there that will do this and is free or quite cheap (is there any add to Scion Image that will do it without the need for too many macros etc.?)? Finally, is there any web-based resource/text on image analysis that would cover from beginnings up to a reasonable sophistication, or what is/are the best available books at present?
Thank you all for your patience and thanks in advance to anyone who is able to help.
Are there any references or experienced folks out there that are doing immunoEM for TEM and antigen retreival for archived material and stuff soaking in 2.5% glut?
I'm checking into the method and have come up with nothing so far.
Thanks!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
There are several options, none that I am aware of that are free or shareware.
The one I us is Thumbs Plus from Cerious Software. It costs about $30. Another program is Image AXS, about the same price. These allow variable number of thumbs per page, depending on the size of the thumbs and page size.
If you make thumbnails as a contact page, you can output them to Acrobat Distiller and save as PDF. They are small file sized and high fidelity output. Current Acrobat is 5.0.
gary g.
At 02:54 AM 4/9/2003, you wrote:
} Dear All } } Is there freeware out there or a plug in to ifranview for batch printing of } images 4 images per page of all selected images? } } I really love this list. I just want to thank you all for your helpful } comments on questions posted here. Thank you Nestor for keeping it up and } running. } } Stephan H Coetzee } Department of Physics } EMU } Private Bag 0704 } Gaborone, } Botswana } } Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} } } Telephone: (+267) 355 2462 } Fax: (+267) 3185 097 } Telephone: (+267) 355 0000 Switchboard
Hi listers My compressor on the cooling tower for my JEOL T-220A has failed. Can anyone refer me to a local (stateside) resource for acquiring this compressor. The system I have now is R-12. I would prefer to change over to R-134. Thank you
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com
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If you are not the intended recipient, be advised that any use, dissemination, distribution or duplication of this transmission is strictly prohibited. If you received this transmission in error, please notify the sender immediately by electronic reply to this transmission or by phone (847-803-6100). Thank you.
Stephan, You should have a look at XnView ( www.xnview.com ). It doesn't do batch printing, but it will create batch contact sheets that you can then batch print. Its a great all round program for working with piles of images. ACDSee does a good job with batch printing, but it's not free (CAD$ 70).
Cheers
Glenn
Glenn Poirier
Microbeam Specialist Mineralogy and Metallurgical Processing Mining and Mineral Sciences Laboratory, CANMET 555 Booth St Ottawa, ON K1A OG1
Spécialiste en microfaisceau Minéralogie et procédés métallurgiques Laboratoires des mines et des sciences minérales de CANMET 555, rue Booth Ottawa, Ontario K1A 0G1
tel: (613) 947-9833 FAX: (613) 996-9673 -----Original Message----- } From: Coetzee, Mr S. H Physics Science [mailto:COETZEES-at-mopipi.ub.bw] Sent: 09 April, 2003 05:54 AM To: Listserver Microscopy (E-mail)
Dear All
Is there freeware out there or a plug in to ifranview for batch printing of images 4 images per page of all selected images?
I really love this list. I just want to thank you all for your helpful comments on questions posted here. Thank you Nestor for keeping it up and running.
Stephan H Coetzee Department of Physics EMU Private Bag 0704 Gaborone, Botswana
Photoshop 7 (and 5 too, but with fewer options) has a File--} Automate--} Contact Sheet II feature for making contact sheets of variable # of images, names, PPI, etc. which can the be printed. Basically, you chhose the directory of images and the format you want and it automatically makes the sheets. It does not, however, batch save or print them.
At 07:31 AM 4/9/2003 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
I have been using Qimage Pro 2003 that does a great job and has quite a few capabilities. It allows you to print different numbers of images per page, put the file name underneath the image, draw a line around the image, etc, etc. You can check it out and get a trial copy at
www.ddisoftware.com/qimage
it costs about $35
Usual disclaimer, Damian Neuberger Senior Research Scientist Baxter Healthcare Corp.
----- Original Message ----- } From: "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} To: "Listserver Microscopy (E-mail)" {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 09, 2003 4:54 AM
Rick,
Aside from the fixation and sample prep, what type of SEM are you using. I did some extensive work with mouse peritoneal microvilli using approximately the same general prep methods. I took some very nice images of the microvilli at fairly high magnification using our JEOL Field Emission SEM at a couple of keV and low beam current. Unfortunately, near the end of the study, I got the call for my lung transplant and so couldn't complete the SEM work on the last samples. They sent them to an outside lab and the photos showed the same kind of damage to the microvilli you describe. Seems that the other lab used a standard tungsten filament SEM at 20keV and unknown beam currrent. It looked like beam damage to me. Could this be an issue for mouse egg samples?
Damian Neuberger Senior Research Scientist Baxter Healthcare Corp.
----- Original Message ----- } From: "Rick Harris" {raharris-at-ucdavis.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, April 08, 2003 3:48 PM
Dear Stephan,
You wrote:
} Is there freeware out there or a plug in to ifranview for batch printing } of images 4 images per page of all selected images? } For some time we have been using a program called PrintStation which will allow you to have from one to however many images you want per page. It is very easy to set up and all our users love it. At US$19.95 it is also very cheap. You can find it at : http://www.picmeta.com/printstation.htm
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-cam.ac.uk
I have tried to maintain a good database of third-party service providers in the past, but recent expansions in this field have probably outdated the resources I currently list. I would appreciate it if anyone out there who provides services for SEM, TEM and x-ray microanalysers would send their particulars (company name, contact name, email, web site, phone number, fax number, address, coverage area, instruments serviced, services offered).
I receive many requests for sales, service, de-installation, installation and training that are outside the area I cover (the great fly-over country of the American Midwest) - from as far away as Pakistan. I always like to provide competitive referrals whenever possible.
I'd be happy to provide compilations to those who express an interest, and frankly, will soon update my website to provide such referrals in a simple format.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
I can't say for certain whether it is your cable that's going. But I did have an old Siemens IA cable go on me back in the 1970s. It was quite spectacular because first there was a loud crack and for a little while there seemed to be a bit of a plasma-like glow at the breakdown point.
Cables may differ but I can tell you that ours went quickly giving no warning and the only recourse we had was to look for a second hand or re-conditioned cable, which we managed to source. I think the only form of repair possible then would have been to shorten the cable but the cost of re-connecting to the gun would be as much as a second hand cable and would not be provided by the manufacturer so there was a risk that it might not be reliable. Try asking a friendly e.m. engineer or Hitachi - they can be quite friendly sometimes.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: "Ellis, Sarah" {s.ellis-at-pmci.unimelb.edu.au} (by way of MicroscopyListserver)
Hi Sarah,
We had a HV cable problem with our Hitachi 2500 SEM and we heard a hissing noise whenever we tried to increase magnification and it would go worse and worse each time.We removed the suspect HV cable and it was cracked inside which caused arching when we turned it on.We were not able to repair it but since our SEM has 2 detectors and we only use one of them we just switched the cables and observation detector while we were waiting for a new cable.
Hope this helps
Virginia Soares INESC Microsistemas e Nanotecnologias R.Alves Redol,9 1000-029 Lisboa PORTUGAL Tel:+351 21 3100300 ext 2504 Fax: +351 21 314 58 43 URL: www.inesc-mn.pt
Well it looks as if you may be correct but there are two steps that you need to take.
1. Follow a strict gun cleaning procedure - take out the cable (care heavy) and if you know how remove the gun chamber (also heavy). If not we can still go ahead. Clean the chamber (if it is in place WITH GREAT CARE NOT TO GET THE MEDI NEAR ANY JOINTS as it will be hard to remove) with a metal polish soluble in ammonia solution. Clean this away with 10:1 water plus ammonia solution on a fluff free but absorbent tissue. Do not let the tissue get too wet, the solution must not run off the tissue when you wipe the chamber. Clean the glazed gun ceramic and all its components with metal polish or where possible in an ultrasonic cleaner in 1:3 water plus ammonia solution (strong). Clean each piece in the US cleaner on its own to prevent scratching. Heat the gun chamber and the metal pieces to drive off residual media and check the gun pieces with a 10X hand lens before re assembly. Check all "o" ring surfaces and "o" rings as you re assemble. This may be enough?
2. The "final" test. Switch off the high voltage and clean round the connection which goes into the high voltage tank. Have some clean paper towel ready, the type that will not leave fluff on a surface. Release the bolts and carefully remove the high voltage cable connection from the tank. Cover the hole in the tank with the clean paper after covering the high voltage connection. Switch on the microscope. If the high voltage levels and standing current are back to normal YES you do have a cable problem.
For greater detail have a look at our CD "Monitoring and Maintaining EM".
If the cable is the problem you are able to have it repaired in your own country. Any organisation that works with high voltage (e.g. hospital x-ray sets etc) will be able to attach a new cable to your old cable ends.
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "Ellis, Sarah (by way of MicroscopyListserver)" {s.ellis-at-pmci.unimelb.edu.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 09, 2003 1:46 PM
Does anyone know of a current phone number/address for the ETP/SEMRA company, who manufactured the Robinson detectors? My last address for them was Livermore, CA; however, that phone number is not valid anymore. This company may have merged with someone else. Any help would be greatly appreciated. Thank you in advance. Cathy Kelloes
we are trying to make a in-situ heating stage for a vg501 STEM and are unable to get factory drawings of the stage or sample cartridge. we are concerned with the tapered fit that postions the cartridge. any help would be welcome.
Michael T. Marshall Research Engineer University of Illinois Frederick Seitz Materials Research Laboratory 104 South Goodwin Avenue Urbana, IL 61801 Voice: 217/265-5380 Fax: 217/244-2278 marshall-at-mrl.uiuc.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jimekstrom-at-attbi.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, April 10, 2003 at 08:09:24 ---------------------------------------------------------------------------
Email: jimekstrom-at-attbi.com Name: James Ekstrom
Organization: Phillips Exeter Academy
Education: 9-12th Grade High School
Location: Exeter, NH USA
Question: I am looking for a copy of the manual for a Nikon Inverted - Phase Contrast microscope. (If you have such a manual I can make a copy and send the original back to you.) I do not have the model number, but I can send along a small JPEG image of the scope if you think you might have a manual for this older microscope. Any assistance would be appreciated.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It sounds to me like you may have a problem with your vacuum system. I'm not really familiar with Hitachi microscopes but on the JEOL microscopes a smell in the gun usually indicated backstreaming oil from the diffusion pump. Poor vacuum combined with oil in the gun chamber would cause discharging in the gun at higher accelerating voltages. That could also explain the hissing (discharging) sound and the problems with filaments.
Bill
Hi everyone,
I have a Hitachi H600 TEM (20 years old). I think the HV cable is dying. My evidence for this is:
1. Unstable voltage readouts. When we set the voltage to, say, 50 Kv, it does not read 50 Kv but reads 75Kv. 2. There is a 'hissing' sound when we try to change from 75Kv to 100Kv. The hissing appears to be coming from behind the gun. 3. Our filaments, when they break, are showing signs of over-heating as evidenced by a round blob of tungsten on each of the broken ends. 4. When I open the gun, it smells.
We need to keep it alive for another 12-18 months by which time we anticipate replacing it.
Are my conclusions re the HV cable correct? Does anyone out there know if it is possible to repair the HV cable and if so, what is involved?
cheers
Sarah Ellis
Manager, Microscopy Imaging and Research Core Facility Peter MacCallum Cancer Centre Locked Bag #1 A'Beckett Street East Melbourne 8006
You can get quite a few varieties of impression materials from any dental supplier. I used to use them, especially the silicone ones, to make replicas. The casting material is epoxy, such as an Epon substitute or one of the resins made by Epotek. Hope this helps.
Lesley Weston.
on 09/04/2003 5:45 AM, Chris Peppiatt (by way of MicroscopyListserver) at chris.peppiatt-at-nuigalway.ie wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } Sorry to be a bother, but I have a couple of queries. } } I have looked in the archives and hope that I haven't missed any relevant } info. } } Firstly, I'm looking for the name/product no. of a Dow Corning } product that can be used for moulding to make replicas for } microscopy. In the past I have used simple latex, but heard Dow did } something better. I have looked at the D-C web site but couldn't work } out exactly what product to buy. I want to use this stuff for } cleaning purposes (AFM standards) and also for making actual } replicas, so the name of the right casting product would also be very } much appreciated. } } Secondly, I'm interested in pattern recognition (for AFM, SEM, TEM } etc.). Am I right in thinking this should be a relatively simple } branch of image analysis? I know that one can use image analysis to } do things like count cells on agar plates via a live camera, or from } an image. Can it be done for things like recurring patterns in say } crystals? Is there anything available out there that will do this and } is free or quite cheap (is there any add to Scion Image that will do } it without the need for too many macros etc.?)? Finally, is there any } web-based resource/text on image analysis that would cover from } beginnings up to a reasonable sophistication, or what is/are the best } available books at present? } } Thank you all for your patience and thanks in advance to anyone who } is able to help. } } Chris Peppiatt }
} ---------------------------------------------------------- } -------------- The Microscopy ListServer -- Sponsor: The } Microscopy Society of America To Subscribe/Unsubscribe -- } Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm } l } ---------------------------------------------------------- } -------------. } } Does anyone know of a current phone number/address for the } ETP/SEMRA company, who manufactured the Robinson } detectors? My last address for them was Livermore, CA; } however, that phone number is not valid anymore. This } company may have merged with someone else. Any help would } be greatly appreciated. Thank you in advance. Cathy } Kelloes
Here's a followup on my not-so-recent query of The List:
} ... bought a suitable video adapter with C mount for my company's } stereomicroscope and then bought a Kodak MDS 100 camera, both on } eBay,
Took two tries. The first firm set impossible conditions and so my money order was returned unclaimed. The second fellow was quite responsive and the camera arrived factory sealed and installed like a dream, thanks to Kodak's software provider and a second CD hastily stuffed inside the box at "the factory."
} ... in my price range (less than $500 in it so far).
.. snippage ...
One question remains:
} Are there any wavelength issues, such as poor image quality due } to infrared seeping through all that glass ?
Several Listers warned me about this, and sure enough, the image is too red (correctable) and rather fuzzy. There is clearly the need for a filter transparent to visible light but opaque to the long- wavelength infrared light that's causing what I presume to be lots of chromatic abberation. What's such a thing called ?
Best regards, George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
Hello, I must purchase indoor oil mist exhaust filters for six rotary pumps that are attached to EMs, sputter and carbon coaters and a freeze dryer. Can someone please suggest the most practical, easy to maintain economical units from their experience? Thank you. Regards, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
Here are the contact details of Dr Viv Robinson, and the company details.
Dr. Vivian Robinson ETP Semra Pty Ltd
244 Canterbury Rd Canterbury, NSW 2193
Ph: +61 (2) 9718 1444 Fax: +61 (2) 9718 8222
E-mail: viv-at-etpsemra.com.au
Regards George
----Original Message----- } From: Kelloes, Cathy L [mailto:KELLOECL-at-bp.com] Sent: Thursday, 10 April 2003 9:24 PM To: MSA (E-mail)
Does anyone know of a current phone number/address for the ETP/SEMRA company, who manufactured the Robinson detectors? My last address for them was Livermore, CA; however, that phone number is not valid anymore. This company may have merged with someone else. Any help would be greatly appreciated. Thank you in advance. Cathy Kelloes
Marc Helvey Strategic Accounts Manager VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 E-Mail: marc.helvey-at-vlsistd.com {mailto:marc.helvey-at-vlsistd.com} Internet: http://www.vlsistandards.com {http://www.vlsistandards.com/}
-----Original Message----- } From: Kelloes, Cathy L [mailto:KELLOECL-at-bp.com] Sent: Thursday, April 10, 2003 4:24 AM To: MSA (E-mail)
Does anyone know of a current phone number/address for the ETP/SEMRA company, who manufactured the Robinson detectors? My last address for them was Livermore, CA; however, that phone number is not valid anymore. This company may have merged with someone else. Any help would be greatly appreciated. Thank you in advance. Cathy Kelloes
A daylight LBD filter will be the first one needed followed by an IR filter. Each goes in different places, depending on your particular scope. The IR filter is usually at the lamp house. The LBD is at the transport lens or in the vertical illuminator.
gary g.
At 02:09 PM 4/10/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Assuming no toxic exhaust is present, and pumps are not huge, I would use the following:
Some of the best filters are made by Edwards Vacuum (now BOC Edwards). Less expensive OK filters made by Alcatel. Both have easily replaceable cartridges.
See both at these sites, or look up direct links (google will do) http://www.kurtlesker.com/ http://www.duniway.com/ Both these sites have variety of filters.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
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----- Original Message ----- } From: "Jim Romanow" {bsgphy3-at-uconnvm.uconn.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, April 10, 2003 6:43 PM
Thanks to all of you who responded in reference to my request on contacts for the address of ETP-USA.
Jim, Also check out Balston Filters (part of the Parker Corporation). The link is : http://www.parker.com/parkersql/default.asp?type=2&id=344
Gary M. Easton Scanners Corporation 410-857-7633
----- Original Message ----- } From: "Vitaly Feingold" {vitalylazar-at-worldnet.att.net} To: {microscopy-at-sparc5.microscopy.com} ; "Jim Romanow" {bsgphy3-at-uconnvm.uconn.edu} Sent: Thursday, April 10, 2003 9:14 PM
Hi, Does anyone know where I could get a pure polystyrene latex for to create a performance specimen as described in "Quality in Electron Microscopy" by Tony Bruton, Steve Chapman, and Paul Harding.
Hi Jim, I frankly don't know what you can do without spending a real bundle. I agree with Vitaly that Edwards BOC is the best bet but you probably have a mixture of pumps. The caveat with respect to the new Edwards oil mist filters is that they fill up with oil, at least on the new 5's and 8's, and if you don't want to be in constant maintenance, you have to purchase the Edwards recirculator packs. Finally, even their refills aren't inexpensive.
For Welch pumps, try: http://www.welchvacuum.com/
For Edwards pumps, try: http://www.bocedwards.com/vacuum/rotary_vane_pump/overview.html
Kurt J. Lesker Vacuum handles about everything you could want and might be the one to give the best prices.
http://www.lesker.com/
Rietschle Pumps can be found here:
http://www.rietschlepumps.com/home.htm
First thing I would recommend is to get a list of Model and serial numbers then you know how much leverage you will have.
Hope this helps, and I do not envy you this task. If it isn't done right, the effort will become a true waste of money as the filter media become saturated and non-functional. Welsh has drains available for their oil mist filters as well. Even when you have the mist depressors, a really good installation will take the air output from the filter to an easily refillable air filter to get the rest of the oil out.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: Jim Romanow [mailto:bsgphy3-at-uconnvm.uconn.edu] Sent: Thursday, April 10, 2003 6:43 PM To: microscopy-at-sparc5.microscopy.com
Hello, I must purchase indoor oil mist exhaust filters for six rotary pumps that are attached to EMs, sputter and carbon coaters and a freeze dryer. Can someone please suggest the most practical, easy to maintain economical units from their experience? Thank you. Regards, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
We are trying to measure the proliferation of EPC cells after they have been injected into a host. Prior to injection, we label the cells with CMTMR, a live cell tracker, for identification and subsequently want to do BrdU labelling as a cell proliferation assay.
I fear that the harsh conditions necessary to denature the dsDNA in the BrdU staining protocol will quench the CMTMR signal. First we will test the system in cell culture but even if that works, such conditions may not be as severe as the ones which include paraffin embedding and possible antigen retrieval (cryo sections are probably better) of tissue.
Does anyone have experience with these protocols or are there any suggestions for a more compatible pair of cell marker/proliferation assay products?
Thank you - all suggestions will be highly appreciated.
Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Queen 30 Bond St. Toronto, ON M5B 1W8 Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
The Solid Latex test specimen requires a large quantity (high density) of latex, much more than you can purchase from most suppliers for other applications.
Chuck Garber of Spi has been working on the preparation of a latex test specimen so you could try Spi, or your local university chemistry department. Many of these departments make latex in vast quantities as part of one of their student experiments.
Regards
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "SBC ATC SEM" {atcsem-at-sbcglobal.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, April 11, 2003 3:54 PM
Torino 14 April 2003 (Italy) Hi, I'm an amateur naturalist and I like to improve my optical microscope. So I'm going to change the traditional illumination with tungsten lamp by a LED light source with a 12v and 25 w alimentation (max. 20 mA current at the diode) I noticed that the light is quit strong. I wonder if it wuould be dangerous for my eies, although I use the cyan filter during the observation. Also I can lower the light intensity by a variable resistance. Thank you, With my Best Regards,
I am a PhD student in the Netherlands. I work with the fluorescent tracer Fluoro-Gold. I am trying to do plastic embedding of peripheral nerve and to observe the tracing. However the polymerisation step is performed at 60 deg. C, which is quite detrimental to my signal. Can somebody advise me what kind of plastic should I use. I do not want to heat up my specimens more than 40-45 deg. C.
regards
} Dr. Dimiter Prodanov } Neuroregulation Group, } Department of Neurosurgery, } Leiden University Medical Center, } P.O. Box 9604, 2300 RC Leiden, } The Netherlands } } Tel : +31 71 527 -6760, -6749 } Fax : +31 71 527 6782 }
Greetings, Plastic embedding with UV polymerization allows embedding at 4 C (or even colder). We use a mixture of butyl and methyl methacrylate that is easy and gives good preservation. Its main advantage is that it is extractable with acetone after sectioning so antibodies get great access to their antigens, boosting the signal. The disadvantage is that the resin is a bit soft (no crosslinker) so it is not perfect for TEM use. If you would like a protocol for using butylmethyl methacrylate, contact me off line. Tobias
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Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
Another question: is anyone experiencing what we call "concrete water" in the diamond knife boat??? The symptoms of "concrete water" are that the initial section cuts ok, but subsequent sections pile up becuase they will not move out across the water surface. Even when you manage to separate one section, it is almost impossible to move it about on the surface of the water with a one-hair brush...it just will not move...the water is like "concrete" (Note: this is NOT like the problem of an unclean knife edge when sections pile up for that reason. It is something entirely different.)
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello all,
I am a PhD student in the Netherlands. I work with the fluorescent tracer Fluoro-Gold. I am trying to do plastic embedding of peripheral nerve and to observe the tracing. However the polymerisation step is performed at 60 deg. C, which is quite detrimental to my signal. Can somebody advise me what kind of plastic should I use. I do not want to heat up my specimens more than 40-45 deg. C.
regards
} Dr. Dimiter Prodanov } Neuroregulation Group, } Department of Neurosurgery, } Leiden University Medical Center, } P.O. Box 9604, 2300 RC Leiden, } The Netherlands } } Tel : +31 71 527 -6760, -6749 } Fax : +31 71 527 6782 }
} Another question: is anyone experiencing what we call "concrete water" } in the diamond knife boat??? The symptoms of "concrete water" are that } the initial section cuts ok, but subsequent sections pile up becuase } they will not move out across the water surface. Even when you manage } to separate one section, it is almost impossible to move it about on the } surface of the water with a one-hair brush...it just will not } move...the water is like "concrete" (Note: this is NOT like the problem } of an unclean knife edge when sections pile up for that reason. It is } something entirely different.) } } Any ideas???
Yes, I know exactly what you mean: the water behaves like molasses so that the sections are nearly impossible to move. In our experience, we found the cause was a thin film of either protein or oil on the water surface since cleaning the water surface with a lintless lens paper (Rosmarin brand, for example) got rid of the phenomenon (at least temporarily).
You need to eliminate the source of the contamination. This could be traced to either: a dirty knife trough (clean with a gentle detergent), a dirty eyelash probe (caused by cleaning the eyelash by pinching between fingers), dirty glassware that contains the distilled water, contaminated micropore filtration apparatus (change micropore filter and rinse entire apparatus, especially the new micropore filter, with sterile, clean distilled water). Finally, water purification systems (like reverse osmosis or deionization) that become overloaded (or have been recently "recharged") are more likely to cause this than traditionally prepared distilled water.
So, clean everything mentioned. If the problem persists, borrow or purchase some distilled water from a different source and check it out.
############################################################## 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/~image/ ##############################################################
Does anybody have experience with mito-tracker dye which can distinguish between inactive (dead) mitochondria and active mitochondria. It is probably mostly used with animal or human tissue; however, we would like to use it with corn anthers. The dye seems to have difficulties penetrating into tissue deep enough. If anybody can give me some information or tip, I would be delighted. Thank you in advance. Marianne B. Smith, Schnable-Lab, Agronomy, Iowa State University, Ames, IA 50011
} From: "Massimo" {max_gra-at-libero.it} : : Torino 14 April 2003 : (Italy) : Hi, : I'm an amateur naturalist and I like to improve my optical microscope. : So I'm going to change the traditional illumination with tungsten lamp by a : LED light source with a 12v and 25 w alimentation (max. 20 mA current at the : diode) : I noticed that the light is quit strong. I wonder if it wuould be dangerous : for my eies, although I use the cyan filter during the observation. Also I : can lower the light intensity by a variable resistance.
There are two ways to lower the intensity of a LED. The simplest is to put a variable resistor in series with on leg of the LED and the more complicated is to very the time it turned on and off at frequency of 500 Hz or greater.
Fortunately a LED unlike a conventional bulb does not change color when you limit the current going to it.
Get a data sheet on the LED if you can and see what spectra it puts out. The cyan filter may be redundant. White LEDs are not continuous spectrum devices.
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
I have received tons of mails from you, which made my mail flow fall into a congested state. Please please delete my e-address fro your list. I do not want to receive e-mail from you. Thank you.
in our lab we have attached to a TEM a Noran Ge-EDX detector with a possibly broken or damaged diamond window. It seems that either the window is broken or that the window is not anymore attached correctly to the counterpart.
Is there anything that we could repair by ourself (either "glueing" or exchang the window)? Does anyone know of any kind of repair kit for such broken windows?
Any advice would be greatly appreciated.
Johannes Bernardi Vienna University of Technology / E052
Back in the days when 4489 was new I too had problems with patchy negatives.
Fortunately Ilford and Kodak were clients so I went along and independently asked them what I was doing wrong. In short almost everything!
They both came up with this procedure
1. Develop sheet film using the dunk and tilt method. 2. Place the film rack in the developer which must be within 2 degs of the ideal 20 deg C if not warm or cool as appropriate. Do not use the time variation to compensate if the temperature is outside this range. 3. After 15 secs lift the film out of the developer and tilt over at 90 degrees for 3 seconds then return to the developer. 4. Repeat the procedure for the desired time, tilting in opposite directions each time. 5. At the development time place the rack of film in and out of water at the same temperature as the developer for 30 seconds. 6. Place the rack of film in the fixer and repeat the dunk and tilt method for the first minute and a half. The said patching could even come from the fixation process; that was new to me.
Problems!
1. The film fogs! Why, because no one replaces safe lights (do they?) and after five years they will have faded and are no longer efficient.
Results!
So good were the negatives that I was able to use a point source enlarger and they are famous for picking out every single defect in a negative! I was running the Hitachi European demo lab at the time, with such cracking pictures we wiped up the London biology market!
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "bonnie sheppard" {bls4u-at-cstone.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, April 14, 2003 8:35 PM
Dear Dimiter, } The alternative to heat polymerisation of epoxy resins (e.g. Epon, Spurr) } is to use UV light. Resins used are "London resins" or "Lowicryls", e.g. } "HM20". } Typically this is done after cryofixation and a process called "Freeze } Substition" (FS). During FS the specimen is brought from approx. minus90 } degrees C } to approx. minus30 degrees C ("freeze") while the water in the specimen is } exchanged against solvent ("substitution"). } Once the substitution is complete, infiltration in Lowicryl and } polymerisation under UVlight also happen at approx. -30°C. } The hardened blocks can be sectioned with an ultramicrotome semithin for LM } and ultrathin for TEM. Both types of sections can be immunolabelled. } } There will be a course on "Cryosectioning and Immunolabelling" in Utrecht } this July that will also include a part on cryofixation and FS. } If you need more info, please have a look on the Leica website } "www.em-preparation.com" under "events" or get in contact with Jan Slot and } George Posthuma } in Utrecht: Dr. George Posthuma, Department of Cell Biology } University Medical Center Utrecht, AZU room G.02.525 } Heidelberglaan 100, 3584 CX Utrecht, The Netherlands } Tel. : + 31 30 2506548 } Fax. : + 31 30 2541797 } Email: g.posthuma-at-lab.azu.nl } } } best regards from Vienna, } 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 } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } } I am a PhD student in the Netherlands. I work with the fluorescent tracer } Fluoro-Gold. I am trying to do plastic embedding of peripheral nerve and to } observe the tracing. } However the polymerisation step is performed at 60 deg. C, which is quite } detrimental to my signal. Can somebody advise me what kind of plastic } should } I use. I do not want to heat up my specimens more than 40-45 deg. C. } } regards } } } Dr. Dimiter Prodanov } } Neuroregulation Group, } } Department of Neurosurgery, } } Leiden University Medical Center, } } P.O. Box 9604, 2300 RC Leiden, } } The Netherlands } } } } Tel : +31 71 527 -6760, -6749 } } Fax : +31 71 527 6782
Bonnie, I have not yet actually used the new 4489 film, but spoke with someone at Kodak before purchasing it. Seeing all the negative reports on it, I wanted to "get the scoop" before trying it out. They told me that the best things to try are additional agitation in both the D-19 and Fixer. If the negatives still appear uneven or muddy try using a 1:1 dilution of D-19 instead of the usual 1:2. Good luck to you and anyone else having to use the new film! I hope I won't have too much trouble. Have a good one!
Donna R. Clarkson
Northrop Grumman Information Technology for U S Army Medical Research Detachment at Brooks City-Base Phone (210) 536-1416 FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil
-----Original Message----- } From: bonnie sheppard [mailto:bls4u-at-cstone.net] Sent: Monday, April 14, 2003 2:35 PM To: Microscopy-at-sparc5.microscopy.com
Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
Yesterday I posted a message asking if anyone is having trouble with streaking, blotchiness, etc. on kodak 4489 TEM film. I have received no answers, and maybe it is because I did not put my email address in the message.
I have never heard of such a thing. Perhaps one exists, but I doubt it.
The volume behind the window is normally evacuated so special procedures would be needed to replace it. There is also a risk of contamination of the detector crystal if the window is broken. We recently had to send a detector in for repair of a small leak in the window and it was deemed necessary to replace the detector crystal as well. On another occasion years before, just the window was replaced.
In short, I doubt that this project is one that can be done in one's own lab.
I presume most EDS vendors would offer detector repair services. There are also multiple third party vendors available. We have gone both routes. One unit was repaired by Kevex and the other by Gresham. Both were satisfactory.
Warren
At 12:47 PM 4/15/2003 +0200, you wrote:
} Dear all, } } in our lab we have attached to a TEM a Noran Ge-EDX detector with a } possibly broken or damaged diamond window. } It seems that either the window is broken or that the window is not } anymore attached correctly to the counterpart. } } Is there anything that we could repair by ourself (either "glueing" or } exchang the window)? } Does anyone know of any kind of repair kit for such broken windows? } } Any advice would be greatly appreciated. } } Johannes Bernardi } Vienna University of Technology / E052
A vacancy for a technical assistant in the Structural Virology Group of Purdue University is anticipated. This position is ideal for an individual interested in cryo-electron microscopy studies of viruses. The position will involve sample preparation, initial sample assessment, sample freezing, data collection, image analysis and interpretation. A BS with a major in biochemistry or a related area of biology and experience with transmission electron microscopy is considered a minimal requirement. Additional backgrounds in physics and computing along with a willingness to learn and the ability to balance multiple projects would be highly desirable. Employment will entail comprehensive training during the first year and extensive daily interactions with a team of graduate students, post-doctoral scholars and faculty. Our facilities include state of the art 200 and 300kv TEMs. Applications or requests for further information should be sent to Michael Rossmann or Richard Kuhn, Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA. Tel: 765-494-4911; FAX 765-496-1189; e-mail: mgr-at-indiana.bio.purdue.edu or rjkuhn-at-bragg.bio.purdue.edu.
Paul Chipman Research Electron Microscopist Purdue University Structural Virology
Moxtek definitely sells EDS windows and may sell such a kit. I'm not sure, ask them. www.moxtek.com
Be aware that even if you are successful in gluing in a new window you'll have to re-evacuate (pump out) the cryostat again. That will require a diffusion or turbo pumping station and special adapter to attach it to the detector. A leak detector is also useful.
If you've never undertaken this before it may be better for you in the overall to have an expert do this for you.
Owen
At 12:47 PM 4/15/2003 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Sounds like: 1. some gunk on the block is building up on the knife edge causing the sections to stick. or 2. some contamination in the boat water that is floating on the surface so the sections can't leave the knife edge. 3. Both 1 and 2.
Try cleaning the face and sides of the block, and the knife edge and the boat. Then get some fresh distilled water and a syringe filter to fill the boat.
Geoff
bonnie sheppard wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Hi Bonnie, This problem was dealt with recently at length. You should search the archives.
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: bonnie sheppard [mailto:bls4u-at-cstone.net] Sent: Monday, April 14, 2003 3:35 PM To: Microscopy-at-sparc5.microscopy.com
Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
We were also taken by surprise by a box of the "New Formulation" 4489 that we didn't even know we had, until we started using it. This seems to be the film from hell. We solved the density problem by increasing our nitrogen burst agitation to 2 seconds every 10 seconds, but we still are getting strange agitation marks.
I started out years ago using the method described by Steve Chapman, which is the Kodak recommendation, and only started using nitrogen burst when I moved to my present job in 1999. Absolutely no problems until the new film showed up, then we had very thin negatives with horrible streaks. As I said, the density problem has been solved, but we're still fighting the streaks.
What we are doing is a series of exposure trials using "blank" exposed negatives, i.e., exposing the films in the scope with no specimen in place to give an even field of detail-less illumination. This is guaranteed to show any agitation irregularities. We will then vary agitation procedures and times until we get acceptable results. Preliminary results are showing that the plastic film holders with narrow sides, rather then the ones that reach nearly to the bottom of the films, cause fewer agitation problems. Other first results are showing that a combination of "lift and tilt" with nitrogen burst evens things out. Finally, we were getting a "thumb-print" on the bottom of the negatives that turned out to be caused by an agitation swirl around the plastic support rod that holds the films in place. We solved that one by lifting the films up slighty so they don't touch the rod, but that only works on holders that are narrow enough to bend the films when they're inserted.
When we complete our tests, I hope to post images and procedures on our website and post a link to the listserver. In the meantime, I'm following this thread with great interest.
My question is: why would Kodak substitute this for a perfectly good film without any warning? The package insert from a box of the old film was identical to that in the new film, but it seems as if processing procedures need to be radically altered. This has cost us some unhappy clients and no end of headaches, not to mention one embarassing incident where an employee was blamed for something that wasn't his fault. Unless I missed some major press release, this seems like a highly irresponsible action. Does anyone have any insight into this?
Agitatedly, Randy
Randy Tindall EM Core Facility University of Missouri, Columbia
-----Original Message----- } From: Clarkson Donna R Contr USAMRD [mailto:donna.clarkson-at-brooks.af.mil] Sent: Tue 4/15/2003 8:09 AM To: bonnie sheppard; Microscopy-at-sparc5.microscopy.com Cc:
Bonnie, I have not yet actually used the new 4489 film, but spoke with someone at Kodak before purchasing it. Seeing all the negative reports on it, I wanted to "get the scoop" before trying it out. They told me that the best things to try are additional agitation in both the D-19 and Fixer. If the negatives still appear uneven or muddy try using a 1:1 dilution of D-19 instead of the usual 1:2. Good luck to you and anyone else having to use the new film! I hope I won't have too much trouble. Have a good one!
Donna R. Clarkson
Northrop Grumman Information Technology for U S Army Medical Research Detachment at Brooks City-Base Phone (210) 536-1416 FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil
-----Original Message----- } From: bonnie sheppard [mailto:bls4u-at-cstone.net] Sent: Monday, April 14, 2003 2:35 PM To: Microscopy-at-sparc5.microscopy.com
Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
I have to make a recommendation whether to buy one or the other cryostat. Do any of you have experience with one or the other or both? Microm is relatively new, gets a lot of recommendations, while Leica, being a long time around seems to receive complaints about service and parts, but we know that Leica is a good microtome. So I would very much like some feedback. Please!! Thank you all in advance.
We are using the new formulation 4489 film, and use nitrogen bursts during developing and fixing (1 second every 10 seconds). We haven't had any problem with fogging, etc. It might be a worthwhile investment to consider...
Marc
On Tuesday, April 15, 2003, at 05:55 AM, Steve Chapman wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Hi } } Back in the days when 4489 was new I too had problems with patchy } negatives. } } Fortunately Ilford and Kodak were clients so I went along and } independently } asked them what I was doing wrong. In short almost everything! } } They both came up with this procedure } } 1. Develop sheet film using the dunk and tilt method. } 2. Place the film rack in the developer which must be within 2 degs } of } the ideal 20 deg C if not warm or cool as appropriate. Do not use the } time } variation to compensate if the temperature is outside this range. } 3. After 15 secs lift the film out of the developer and tilt over } at 90 } degrees for 3 seconds then return to the developer. } 4. Repeat the procedure for the desired time, tilting in opposite } directions each time. } 5. At the development time place the rack of film in and out of } water at } the same temperature as the developer for 30 seconds. } 6. Place the rack of film in the fixer and repeat the dunk and tilt } method for the first minute and a half. The said patching could even } come } from the fixation process; that was new to me. } } Problems! } } 1. The film fogs! Why, because no one replaces safe lights (do } they?) } and after five years they will have faded and are no longer efficient. } } Results! } } So good were the negatives that I was able to use a point source } enlarger } and they are famous for picking out every single defect in a negative! } I was running the Hitachi European demo lab at the time, with such } cracking } pictures we wiped up the London biology market! } } Steve Chapman } Senior Consultant Protrain } Electron Microscopy Training and Consultancy World Wide } Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 } www.emcourses.com } } ----- Original Message ----- } } From: "bonnie sheppard" {bls4u-at-cstone.net} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Monday, April 14, 2003 8:35 PM } Subject: Kodak 4489 film } } } } } } ---------------------------------------------------------------------- } } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } -. } } } } } } Is anyone else out there having trouble with "shadows and unevenness" } } on } } 4489 film? We have gone to extensive lengths to be sure our } } developing } } is consistent, and still we are having annoying problems with uneven } } (blotchy) negatives. This is not a problem when there is ample tissue } } detail in the negative...but it is horrible and a real printiong } } problem } } for folks looking at DNA spreads, where there is little tissue to } } negative. } } } } Help! } } } } } } } } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
We are not looking forward to trying the new formulation. Perhaps we will go digital. Can anyone tell me the rough cost of a nitrogen burst system?
Dave
On Tue, 15 Apr 2003 18:53:50 -0400 Marc Pypaert {marc.pypaert-at-yale.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are using the new formulation 4489 film, and use nitrogen bursts } during developing and fixing (1 second every 10 seconds). } We haven't had any problem with fogging, etc. } It might be a worthwhile investment to consider... } } Marc } } On Tuesday, April 15, 2003, at 05:55 AM, Steve Chapman wrote: } } } ----------------------------------------------------------------------- } } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } } . } } } } } } Hi } } } } Back in the days when 4489 was new I too had problems with patchy } } negatives. } } } } Fortunately Ilford and Kodak were clients so I went along and } } independently } } asked them what I was doing wrong. In short almost everything! } } } } They both came up with this procedure } } } } 1. Develop sheet film using the dunk and tilt method. } } 2. Place the film rack in the developer which must be within 2 degs } } of } } the ideal 20 deg C if not warm or cool as appropriate. Do not use the } } time } } variation to compensate if the temperature is outside this range. } } 3. After 15 secs lift the film out of the developer and tilt over } } at 90 } } degrees for 3 seconds then return to the developer. } } 4. Repeat the procedure for the desired time, tilting in opposite } } directions each time. } } 5. At the development time place the rack of film in and out of } } water at } } the same temperature as the developer for 30 seconds. } } 6. Place the rack of film in the fixer and repeat the dunk and tilt } } method for the first minute and a half. The said patching could even } } come } } from the fixation process; that was new to me. } } } } Problems! } } } } 1. The film fogs! Why, because no one replaces safe lights (do } } they?) } } and after five years they will have faded and are no longer efficient. } } } } Results! } } } } So good were the negatives that I was able to use a point source } } enlarger } } and they are famous for picking out every single defect in a negative! } } I was running the Hitachi European demo lab at the time, with such } } cracking } } pictures we wiped up the London biology market! } } } } Steve Chapman } } Senior Consultant Protrain } } Electron Microscopy Training and Consultancy World Wide } } Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 } } www.emcourses.com } } } } ----- Original Message ----- } } } From: "bonnie sheppard" {bls4u-at-cstone.net} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Monday, April 14, 2003 8:35 PM } } Subject: Kodak 4489 film } } } } } } } } } } ---------------------------------------------------------------------- } } } -- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } } } -. } } } } } } } } } Is anyone else out there having trouble with "shadows and unevenness" } } } on } } } 4489 film? We have gone to extensive lengths to be sure our } } } developing } } } is consistent, and still we are having annoying problems with uneven } } } (blotchy) negatives. This is not a problem when there is ample tissue } } } detail in the negative...but it is horrible and a real printiong } } } problem } } } for folks looking at DNA spreads, where there is little tissue to } } } negative. } } } } } } Help! } } } } } } } } } } } } } } } } } } } } } -- } Marc Pypaert } Department of Cell Biology } Center for Cell and Molecular Imaging } Ludwig Institute for Cancer Research } Yale University School of Medicine } 333 Cedar Street, PO Box 208002 } New Haven, CT 06520-8002 } TEL 203-785 3681 } FAX 203-785 7446 } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Dear SIR, I am Mr.Bliss Marayi and my sister is miss Maureen Marayi we are the children of late Chief John Marayi from Sierra Leone.I am writing you in absolute confidence primarily to seek your assistance to transfer our cash of 18 Million Dollars($18,000.000.00) now in the custody of a private Security trust firm in Europe, the money is in two trunk boxes deposited and declared as family valuables by my late father as a matter of fact the company does not know the content as money, although my father made them to understand that the boxes belongs to his foreign partner. Source of the money: My late father Chief John Marayi , a native of Mende District in the Northerh province of Sierra Leone, was the General Manager of Sierra Leone Mining co-operation (S.L.M.C.) Freetown . According to my father,this money was the income accrued from Mining Co-operation's over draft and minor sales before the peak of the civil war between the rebels forces of Major Paul Koroma and the combined forces of ECOMOG peace keeping operation that almost destroyed my country, following the forceful removal from power of the civilian elected President of Ahmed Tejan Kabbah by the rebels. My father had already made arrangement for his family thats talking about my mother, my little sister and myself to be evacuated to Abidjan Cote d' Ivoire with the CERTIFICATE OF DEPOSIT he made with the security firm in Europe through the aid of U.N evacuation team. During the war in my country, and following the indiscriminate looting of Public and Government properties by the rebel forces, the Sierra Leone mining coop Was one of the target looted and it was destroyed. My father including other top Government functionaries were attacked and killed by the rebels in November 2000 because of his relationship with the civilian Government of Ahmed Tejan Kabbah. As a result of my father's death , and with the news of my uncle's involvement in the air crash in January it dashed our hope of survival.The untimely deaths caused my mother's heart failure and other related complications of which she later died in the hospital after we must have spent a lot of money on her early this year . Now my 18 years old sister and myself are alone in this strange country suffering without any care or help. Without any relation, we are now like refugees and orphans. Our only hope now is in you and the boxes deposited in the Security Firm.To this effect, I humbly solicit your assistance in the followings ways. 1. to assist me claim this boxes from the security Firm as the beneficiary. 2. to transfer this money (USD$18M) in your name to your country. 3. to make a good arrangement for a joint business investment on our behalf in your country and you, our Adviser/ Manager. For your assistance, I have agreed with my younger sister that 20% of the total amount will be for your effort and another 10 % to cover all expenses that may be incured during the business transaction, Lastly, I urge you to keep this transaction strictly confidential as no one knows our where about. Please as you show your willingness, Forward to us your full name,address, Tel and Fax numbers to me via my private email address as indicated bellow, this is for security reasons as i will only be accessing my private email earnestly awaiting your response. Thanks. May God bless you as you assist us.
Dear SIR, I am Mr.Bliss Marayi and my sister is miss Maureen Marayi we are the children of late Chief John Marayi from Sierra Leone.I am writing you in absolute confidence primarily to seek your assistance to transfer our cash of 18 Million Dollars($18,000.000.00) now in the custody of a private Security trust firm in Europe, the money is in two trunk boxes deposited and declared as family valuables by my late father as a matter of fact the company does not know the content as money, although my father made them to understand that the boxes belongs to his foreign partner. Source of the money: My late father Chief John Marayi , a native of Mende District in the Northerh province of Sierra Leone, was the General Manager of Sierra Leone Mining co-operation (S.L.M.C.) Freetown . According to my father,this money was the income accrued from Mining Co-operation's over draft and minor sales before the peak of the civil war between the rebels forces of Major Paul Koroma and the combined forces of ECOMOG peace keeping operation that almost destroyed my country, following the forceful removal from power of the civilian elected President of Ahmed Tejan Kabbah by the rebels. My father had already made arrangement for his family thats talking about my mother, my little sister and myself to be evacuated to Abidjan Cote d' Ivoire with the CERTIFICATE OF DEPOSIT he made with the security firm in Europe through the aid of U.N evacuation team. During the war in my country, and following the indiscriminate looting of Public and Government properties by the rebel forces, the Sierra Leone mining coop Was one of the target looted and it was destroyed. My father including other top Government functionaries were attacked and killed by the rebels in November 2000 because of his relationship with the civilian Government of Ahmed Tejan Kabbah. As a result of my father's death , and with the news of my uncle's involvement in the air crash in January it dashed our hope of survival.The untimely deaths caused my mother's heart failure and other related complications of which she later died in the hospital after we must have spent a lot of money on her early this year . Now my 18 years old sister and myself are alone in this strange country suffering without any care or help. Without any relation, we are now like refugees and orphans. Our only hope now is in you and the boxes deposited in the Security Firm.To this effect, I humbly solicit your assistance in the followings ways. 1. to assist me claim this boxes from the security Firm as the beneficiary. 2. to transfer this money (USD$18M) in your name to your country. 3. to make a good arrangement for a joint business investment on our behalf in your country and you, our Adviser/ Manager. For your assistance, I have agreed with my younger sister that 20% of the total amount will be for your effort and another 10 % to cover all expenses that may be incured during the business transaction, Lastly, I urge you to keep this transaction strictly confidential as no one knows our where about. Please as you show your willingness, Forward to us your full name,address, Tel and Fax numbers to me via my private email address as indicated bellow, this is for security reasons as i will only be accessing my private email earnestly awaiting your response. Thanks. May God bless you as you assist us.
I am having an odd problem staining cell monolayers with a directly labled FITC antibody to an intracellular antigen. If I try to stain the cells in either a labtek slide chamber (glass) or in a 96 well plate I get no staining. If I remove the cells with trypsin and make a cytospin the cells stain perfectly.
I really need to stain the intact monolayers for routine work, but nothing I have tried seems to work.
Some of the permutations I have tried include:
Fixing with PFA then permeabilizing with either Triton or MeOH then stain
Remove the media and let the monolayer AIR DRY, then permeabilizing with either Triton or MeOH then stain
Fix/permeabilize the wet monolayer with MeOH then stain
Suggestions? --
Michael J. Herron, U of MN, Entomology herro001-at-umn.edu 612-624-3688 Office, 624-3212 Lab, 625-5299 FAX
Hi Marianne, I have been using a Leica cryostat, motorized. It is really good and I am happy with it. For the parts, I can't say anything because it is new and therefore didn't need any service. In my area (Montreal, Quebec) the tech rep is really good but very busy, but so far I he always gave a good service (for other equipments). I guess it depends on who is in your area. Hope this helps! Emmanuelle
Randy, I'm sure it's got something to do with Kodak saving a fraction of a penny per film box. Remember Coca-Cola's new formulation years ago? Seems like Kodak should've taken a lesson. There's a Murphy's Law corollary which states, " If it ain't broke, don't fix it!"
WWW ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor Cannon Electron Microscopy Lab Carolinas Medical Center P.O. Box 32861; Charlotte, NC 28232-2861 Ship to: 1000 Blythe Blvd; Charlotte, NC 28203 Ofc:704-355-1267 ; Lab:704-355-7220 ; Fax 704-355-0589 E-mail: {mailto:WWiggins-at-Carolinas.org} Winston.Wiggins-at-CarolinasHealthCare.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
-----Original Message----- } From: Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] Sent: Tuesday, April 15, 2003 1:33 PM To: Clarkson Donna R Contr USAMRD Cc: microscopy-at-sparc5.microscopy.com
Hi,
We were also taken by surprise by a box of the "New Formulation" 4489 that we didn't even know we had, until we started using it. This seems to be the film from hell. We solved the density problem by increasing our nitrogen burst agitation to 2 seconds every 10 seconds, but we still are getting strange agitation marks.
I started out years ago using the method described by Steve Chapman, which is the Kodak recommendation, and only started using nitrogen burst when I moved to my present job in 1999. Absolutely no problems until the new film showed up, then we had very thin negatives with horrible streaks. As I said, the density problem has been solved, but we're still fighting the streaks.
What we are doing is a series of exposure trials using "blank" exposed negatives, i.e., exposing the films in the scope with no specimen in place to give an even field of detail-less illumination. This is guaranteed to show any agitation irregularities. We will then vary agitation procedures and times until we get acceptable results. Preliminary results are showing that the plastic film holders with narrow sides, rather then the ones that reach nearly to the bottom of the films, cause fewer agitation problems. Other first results are showing that a combination of "lift and tilt" with nitrogen burst evens things out. Finally, we were getting a "thumb-print" on the bottom of the negatives that turned out to be caused by an agitation swirl around the plastic support rod that holds the films in place. We solved that one by lifting the films up slighty so they don't touch the rod, but that only works on holders that are narrow enough to bend the films when they're inserted.
When we complete our tests, I hope to post images and procedures on our website and post a link to the listserver. In the meantime, I'm following this thread with great interest.
My question is: why would Kodak substitute this for a perfectly good film without any warning? The package insert from a box of the old film was identical to that in the new film, but it seems as if processing procedures need to be radically altered. This has cost us some unhappy clients and no end of headaches, not to mention one embarassing incident where an employee was blamed for something that wasn't his fault. Unless I missed some major press release, this seems like a highly irresponsible action. Does anyone have any insight into this?
Agitatedly, Randy
Randy Tindall EM Core Facility University of Missouri, Columbia
-----Original Message----- } From: Clarkson Donna R Contr USAMRD } [ {mailto:donna.clarkson-at-brooks.af.mil} mailto:donna.clarkson-at-brooks.af.mil] Sent: Tue 4/15/2003 8:09 AM To: bonnie sheppard; Microscopy-at-sparc5.microscopy.com Cc:
Bonnie, I have not yet actually used the new 4489 film, but spoke with someone at Kodak before purchasing it. Seeing all the negative reports on it, I wanted to "get the scoop" before trying it out. They told me that the best things to try are additional agitation in both the D-19 and Fixer. If the negatives still appear uneven or muddy try using a 1:1 dilution of D-19 instead of the usual 1:2. Good luck to you and anyone else having to use the new film! I hope I won't have too much trouble. Have a good one!
Donna R. Clarkson
Northrop Grumman Information Technology for U S Army Medical Research Detachment at Brooks City-Base Phone (210) 536-1416 FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil
-----Original Message----- } From: bonnie sheppard [ {mailto:bls4u-at-cstone.net} mailto:bls4u-at-cstone.net] Sent: Monday, April 14, 2003 2:35 PM To: Microscopy-at-sparc5.microscopy.com
Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
Help!
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We, at the high voltae microscope had used 4489 for years. A few years ago we noticed streaking, particulary from top to bottom. Attributing it to the way the films are held verical in the developing tank and that for the most part the nitrogen burst bubbles followed thesame paths repeatedly; we stopped the bursting for most of the developing time. Instead we would move the racks up and down and sideways so that the flow of developer was for the most part parallel to the film. This seemed to help but it took a lot more care and time.
As far as digital goes, I would suggest that film, particularly 4489, can give much more information and micrograph quality than almost any digital systems.
What you gain with digital is time efficiency ( no developing), better quantitation (CCDs are very linear with exposure), and perhapse more sensativity. The results are not subject to variation and whims of the development process. But what you loose with digital is the large uniform field of information ( 4489 would exceed 5000x5000 resolution easily), Digital tends to have more grainieness due to the scintillator and fiber optic transfer plates. If you are using 4489 instead of SO163 you are probably trying to get a very fine grained smooth image - going digital is going the other way. The dynamic range of 4489 film is greater than all but the most spectacular digital systems (probably unaffordable). This effects how well you can resolve micrographs with high brightness relief - with bright areas of detail and dark areas on the same frame.
If you have the time film is good.
Dave -- David Barnard Wadsworth Ctr NYS Dept Health Albany NY 12201-0509 barnard-at-wadsworth.org 518 473-5299 voice 518 474-7992 fax
I would like to say "Thank you very much" to all the people who took the time to respond to my request for information about and experience with the various 120kV TEMs.
This is Lifeng Dong from Department of Physics, Portland State University, USA. I am planning to attend M&M 2003, and reserved a double room (two beds) from Hampton Inn Downtown San Antonio. The price including tax is $121.42 per night, which is a little expensive for a graduate student. If you or some of your male colleagues will attend this conference and are looking for a hotel, I would like to share it with you or him.
If you have any question, contact me at lifengd-at-pdx.edu.
Hi, I want to thanks all answered my question about illumination by LED and at the same time I want to assure about my project that I am not going to use a Laser LED but a simple led lamp just like you are get used to see on some device with colors red, green. The difference is that my light is white. In spite of that I think would be better to insert an infrared filter or similar as you have recommended me. The device that I'm making is like that you can find at this address: http://www.microscopy-uk.org.uk/mag/artjul99/bdoumic.html Thank you. With my Best Regards,
Cost depends on what you already have. A gas burst timer is approximately $400.00. A setup with 3-1gal tanks in a water jacket with floating lids, a cover, and plenums in two tanks would be approximately $1100. This does not include the timer or water temperature control. A similar setup with 3-3.5gal tanks would be approximately $1400. These are estimates..
George
George Laing National Graphic Supply 800.223.7130 x3109 scisales-at-ngscorp.com
} } We are not looking forward to trying the new formulation. } Perhaps we will go digital. Can anyone tell me the rough } cost of a nitrogen burst system? } } Dave
I have the option to buy either a 5 or 3 megapixel camera. What I want to know is whether a 5 megapixel camera is better than a 3 megapixel camera, if used exclusively on a microscope, or whether the extra pixels (and cost) are wasted. Assume bit depth is the same.
I'd like someone to check my math and my reasoning.
I understand that microscope resolution is limited by diffraction and a good rule of thumb for displaying images at normal reading distance is a final image mag of no more than 1000x the numerical aperture of the objective. On my microscope this means about 100x for images taken with the 4x lens and 1250x for images with the 100x oil lens. Assuming I use a 1x relay eyepiece to connect the CCD camera, the image at the detector could be magnified 25 times for the 4x objective, and 12.5 times for the 100x objective, before getting into empty mag.
On the camera side, I understand that the more pixels the better the resolution. The 3 megapixel camera has 2080 horizontal pixels and the 5mp has 2580. Let's ignore the vertical pixels for the moment.
One more assumption is that for printing, it seems that the highest practical output resolution is 266-300 dpi. This means that the 3 mp camera produces an image that optimally prints an image 2080/266 = 7.8 inches or 198 mm horizontally, or less, and the 5 mp can print an image about 243 mm horizontally or less. Enlarging above these dimensions is possible but the print image will need interpolation and might be degraded.
Going back to the microscope, the maximum horizontal field of view with the 4x objective captured by the camera is 1.6 mm. It could be printed up to 100 x 1.6 = 160 mm before empty mag sets in. But I have already calculated that the 3 mp can print up to 198 mm and the 5 mp up to 243 mm before the print image degrades.
If these calculations are correct (please tell me if it they are not) then both the 3 mp and 5 mp have more than enough resolution to capture what the microscope makes available at 4x, and the situation is even better for the 100x lens because the lens does most of the heavy lifting, magnification-wise.
One final condition that might matter is that both are RBG cameras with Bayer filters, so that the real resolution is approximately 1/3 of the nominal pixel resolution. I'm not sure how to figure that into the calculations.
So, at approximately a dollar-per-pixel, do I need the extra 2000 pixels of a 5 mp camera?
Gary P. Radice gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond 804-289-8233 (FAX) Richmond VA 23173 http://www.richmond.edu/~gradice
When teaching an EM course, how many additional non-EM courses might others teach per semester? I am presently teaching 2 courses, one of which is on SEM. I also teach microtechnique and a TEM course.
Obviously, the answer depends on EM course formats and campus size.
My courses are intensive, usually involving 5-6 students, sometimes more. An SEM course may take 15-25 hours per week, including all formally scheduled lab and lecture times, weekly tutorials, solutions preparation etc.(there is no assistant). Our campus is small, perhaps 14,000 students.
Since I teach so few students, my Chair is wondering about the fairness of my workload relative to others in my department who teach many more students in straight lecture courses (the larger courses have assistants).
Your answers will be most helpful in clarifying a "normal" academic work load that includes EM instruction at other similar sized universities. Please send your replies to me at the e-mail address below. I will summarize the results and post them on the listserver.
Thank you in advance!
Jerry
--------------------- Jerry D. Kudenov Dept. Biological Sciences University of Alaska Anchorage 3211 Providence Drive Anchorage, Alaska 99508
} My question is: why would Kodak substitute this for a perfectly good film } without any warning? The package insert from a box of the old film was } identical to that in the new film, but it seems as if processing } procedures need to be radically altered. This has cost us some unhappy } clients and no end of headaches, not to mention one embarrassing incident } where an employee was blamed for something that wasn't his fault. Unless I } missed some major press release, this seems like a highly irresponsible } action. Does anyone have any insight into this? } } Agitatedly, } Randy
Normally I don't like litigation but the pain and suffering and the lack of warning seems to point to a class action suit, and at least an official explanation accompanying the new film and properly labeled new film. I'm maybe just surprised to not have seen that suggestion in the previous discussion on 4489 formula change.
I know this has been gone over in length before. We do not have any nitrogen capabilities in our darkroom, and I am dreading the next batch of 4489 film. Maybe Kodak is using this like Coca-cola did when they made "new" coke, and then switched back to "classic" but that change let them go from using expensive beet sugar to corn sugar as the primary sweetener saving millions of dollars. Who knows - but from the great discussion here I at least have a plan for when we get the new formula (not that I will be able to implement it).
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
Hello Gary Your calculations sounds reasonable. The things you always have to keep in mind going "digital" are that most problems appeared when you need to crop (and possible magnify after all) your image in order to show some small details etc. It happening frequently when image prepared for publication. There you will face the real problem with amount of pixels you have. Sergey
At 11:30 AM 4/16/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
On Wednesday, April 16, 2003, at 03:16 AM, Patton, David wrote:
} We are not looking forward to trying the new formulation. } Perhaps we will go digital. Can anyone tell me the rough } cost of a nitrogen burst system? } Dear David, We just got one from Pella for ~$1150. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
In respond on my message on the "first wave" of the 4489 new formulation discussion I got respond from Kodak that most customers are very happy with new formulation and ONLY a few were unhappy. I am too lazy to find that message in my archive. I wrote back asking why they did not change the product name if altering development process on "new formulation" - I never had respond back. Sergey
At 02:12 PM 4/16/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ramg21-at-yahoo.co.in) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, April 16, 2003 at 11:36:56 ---------------------------------------------------------------------------
Email: ramg21-at-yahoo.co.in Name: ram varma
Organization: univ of louisiana lafayette
Education: Graduate College
Location: lafayette,LA, USA
Question: could you tell me where i can find comprehensive information about
1] near field optical microscopy 2] reflectometry 3] ellipsometry
I would like to know all the details viz. equipment specification,opration,theory of operation etc.
Sounds like the paint crew I hired once to paint my house- all other customers were always happy, no further response.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
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----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 16, 2003 10:05 PM
} From: "Massimo" {max_gra-at-libero.it} : : Torino 16 April 2003 (ITALY) : : Hi, : I want to thanks all answered my question about illumination by LED and at : the same time I want to assure about my project that I am not going to use a : Laser LED but a simple led lamp just like you are get used to see on some : device with colors red, green. The difference is that my light is white. : In spite of that I think would be better to insert an infrared filter or : similar as you have recommended me. : The device that I'm making is like that you can find at this address: : http://www.microscopy-uk.org.uk/mag/artjul99/bdoumic.html
That will work fine. The LED you are using will have a rating in milliamps or 1/1000 amps. The resistor R1 is what limits the current to keep from burning the LED out. The value of R1 is found R1=Voltage divided by the maximum current that the LED can carry.
e.g. If you have 20 ma LED and are using 5 volts R1=5/.02 = 250 ohms. The variable resistor should be a 1,000 ohms. If you are using a ultra bright LED you need to consider the power that the resistor carries that is represented by the voltage times the current and is represented in watts in this case it is 250 *.02 = 0.1 watt so a .25 watt resistor will do. An ultra bright LED will need a half watt or possibly a 1 watt resistor in both R1 and R2.
Any voltage over 3 volts will work. With a 1.5 volt battery you don't need R1 in most cases and with less than 3 volts you may not get full brightness from the LED.
None of the LED materials that emit in the visible range emit infra red see: http://www.theledlight.com/technical3.html
Here is a lot more information on LED construction. http://www.theledlight.com/technical.html
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
Having read of the problems experienced by many users of Kodak 4489 film, after the formulation or something else in the production process appears to have changed, we faced using our new stock with much fear and trepidation. Therefore, in order to try to see what effect this would have in our lab, and to see what we could do to minimize the problems faced by other users, I decided to fill a film cassette half with "old" 4489 film and half with the "new" film and then instructed our darkroom technician to process the film exactly as he had always done, but to increase manual agitation of the film (we do not have nitrogen burst agitation) to 10 seconds at a time at 10 second intervals throughout development, i.e. 3 x 10 second agitations per minute.
After development the difference between the "old" and the "new" was immediately obvious, but only in that the "new" negatives were significantly denser (darker) than the old. There does not appear to be any steaking or inconsistency in the development that others have reported. Although the "new" negatives are not so dark as to be difficult to print, I have since adjusted the TEM's film sensitivity setting to allow us to obtain negatives that are similar in density to those that we obtained from the previous 4489 film.
Perhaps the fact that we do not experience the same problems as reported by others has something to do with the development conditions that we use? We use Ilford PQ Universal developer diluted 1 + 9 for 5 minutes at 20C.
I hope this does not further confuse the issue!
Regards
Rob
=====================================
Rob Cross Director : EM Unit, Rhodes University tel: (046) 603 8168/9
We also use Ilford PQ Universal with Kodak film, 4489 and SO-163, because it is much more convenient. We only use D-19 very very occaisionally for a forced development protocol.
Sally Stowe
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University, Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 http://www.anu.edu.au/EMU
} } } "R. Cross" {r.cross-at-ru.ac.za} 04/17/03 05:10PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear 4489 users
Having read of the problems experienced by many users of Kodak 4489 film, after the formulation or something else in the production process appears to have changed, we faced using our new stock with much fear and trepidation. Therefore, in order to try to see what effect this would have in our lab, and to see what we could do to minimize the problems faced by other users, I decided to fill a film cassette half with "old" 4489 film and half with the "new" film and then instructed our darkroom technician to process the film exactly as he had always done, but to increase manual agitation of the film (we do not have nitrogen burst agitation) to 10 seconds at a time at 10 second intervals throughout development, i.e. 3 x 10 second agitations per minute.
After development the difference between the "old" and the "new" was immediately obvious, but only in that the "new" negatives were significantly denser (darker) than the old. There does not appear to be any steaking or inconsistency in the development that others have reported. Although the "new" negatives are not so dark as to be difficult to print, I have since adjusted the TEM's film sensitivity setting to allow us to obtain negatives that are similar in density to those that we obtained from the previous 4489 film.
Perhaps the fact that we do not experience the same problems as reported by others has something to do with the development conditions that we use? We use Ilford PQ Universal developer diluted 1 + 9 for 5 minutes at 20C.
I hope this does not further confuse the issue!
Regards
Rob
=====================================
Rob Cross Director : EM Unit, Rhodes University tel: (046) 603 8168/9
you are right. From the point of microscopy there are only two reasons to have more pixels:
#1 To print of large images (e.g. for posters or for other representations) without loss of details. #2 For digital processing of your images (e.g. to 'improve' resolution of instrument or to extract details).
Or are there other reasons?
} From a major interest should be the data depth (possible grey steps in each pixel) for a computer processing of brightness and contrast. If you have high dynamics, even wrong lighted images are able to convert into better results or you are able to improve contrast of low differences in an image region, which is uniform in colour and brightness with first views.
Frank Eggert
http://www.microanalyst.net
Gary Radice schrieb:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have the option to buy either a 5 or 3 megapixel camera. What I want } to know is whether a 5 megapixel camera is better than a 3 megapixel } camera, if used exclusively on a microscope, or whether the extra } pixels (and cost) are wasted. Assume bit depth is the same. } } I'd like someone to check my math and my reasoning. } } I understand that microscope resolution is limited by diffraction and a } good rule of thumb for displaying images at normal reading distance is } a final image mag of no more than 1000x the numerical aperture of the } objective. On my microscope this means about 100x for images taken with } the 4x lens and 1250x for images with the 100x oil lens. Assuming I use } a 1x relay eyepiece to connect the CCD camera, the image at the } detector could be magnified 25 times for the 4x objective, and 12.5 } times for the 100x objective, before getting into empty mag. } } On the camera side, I understand that the more pixels the better the } resolution. The 3 megapixel camera has 2080 horizontal pixels and the } 5mp has 2580. Let's ignore the vertical pixels for the moment. } } One more assumption is that for printing, it seems that the highest } practical output resolution is 266-300 dpi. This means that the 3 mp } camera produces an image that optimally prints an image 2080/266 = 7.8 } inches or 198 mm horizontally, or less, and the 5 mp can print an image } about 243 mm horizontally or less. Enlarging above these dimensions is } possible but the print image will need interpolation and might be } degraded. } } Going back to the microscope, the maximum horizontal field of view with } the 4x objective captured by the camera is 1.6 mm. It could be printed } up to 100 x 1.6 = 160 mm before empty mag sets in. But I have already } calculated that the 3 mp can print up to 198 mm and the 5 mp up to 243 } mm before the print image degrades. } } If these calculations are correct (please tell me if it they are not) } then both the 3 mp and 5 mp have more than enough resolution to capture } what the microscope makes available at 4x, and the situation is even } better for the 100x lens because the lens does most of the heavy } lifting, magnification-wise. } } One final condition that might matter is that both are RBG cameras with } Bayer filters, so that the real resolution is approximately 1/3 of the } nominal pixel resolution. I'm not sure how to figure that into the } calculations. } } So, at approximately a dollar-per-pixel, do I need the extra 2000 } pixels of a 5 mp camera? } } Gary P. Radice gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond 804-289-8233 (FAX) } Richmond VA 23173 http://www.richmond.edu/~gradice
At least with Coke there are other soft drinks. As far as I can tell 4489 is the only film available for routine biological tissue work. I have formed the impression from this listserver that S0163 (?) is for more specialised use. Therefore I think we are over a barrel re voting with our feet (mixed metaphors anyone?).
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Randy, } I'm sure it's got something to do with Kodak saving a fraction of a } penny per film box. Remember Coca-Cola's new formulation years ago? } Seems like Kodak should've taken a lesson. There's a Murphy's Law } corollary which states, " If it ain't broke, don't fix it!" } } WWW } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Winston W. Wiggins, Supervisor } Cannon Electron Microscopy Lab } Carolinas Medical Center } P.O. Box 32861; Charlotte, NC 28232-2861 } Ship to: 1000 Blythe Blvd; Charlotte, NC 28203 } Ofc:704-355-1267 ; Lab:704-355-7220 ; Fax 704-355-0589 } E-mail: {mailto:WWiggins-at-Carolinas.org} Winston.Wiggins-at-CarolinasHealthCare.org } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } -----Original Message----- } } From: Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] } Sent: Tuesday, April 15, 2003 1:33 PM } To: Clarkson Donna R Contr USAMRD } Cc: microscopy-at-sparc5.microscopy.com } Subject: RE: Kodak 4489 film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } We were also taken by surprise by a box of the "New Formulation" 4489 } that we didn't even know we had, until we started using it. This } seems to be the film from hell. We solved the density problem by } increasing our nitrogen burst agitation to 2 seconds every 10 } seconds, but we still are getting strange agitation marks. } } I started out years ago using the method described by Steve Chapman, } which is the Kodak recommendation, and only started using nitrogen } burst when I moved to my present job in 1999. Absolutely no problems } until the new film showed up, then we had very thin negatives with } horrible streaks. As I said, the density problem has been solved, } but we're still fighting the streaks. } } What we are doing is a series of exposure trials using "blank" } exposed negatives, i.e., exposing the films in the scope with no } specimen in place to give an even field of detail-less illumination. } This is guaranteed to show any agitation irregularities. We will } then vary agitation procedures and times until we get acceptable } results. Preliminary results are showing that the plastic film } holders with narrow sides, rather then the ones that reach nearly to } the bottom of the films, cause fewer agitation problems. Other first } results are showing that a combination of "lift and tilt" with } nitrogen burst evens things out. Finally, we were getting a } "thumb-print" on the bottom of the negatives that turned out to be } caused by an agitation swirl around the plastic support rod that } holds the films in place. We solved that one by lifting the films up } slighty so they don't touch the rod, but that only works on holders } that are narrow enough to bend the films when they're inserted. } } When we complete our tests, I hope to post images and procedures on } our website and post a link to the listserver. In the meantime, I'm } following this thread with great interest. } } My question is: why would Kodak substitute this for a perfectly good } film without any warning? The package insert from a box of the old } film was identical to that in the new film, but it seems as if } processing procedures need to be radically altered. This has cost us } some unhappy clients and no end of headaches, not to mention one } embarassing incident where an employee was blamed for something that } wasn't his fault. Unless I missed some major press release, this } seems like a highly irresponsible action. Does anyone have any } insight into this? } } Agitatedly, } Randy } } Randy Tindall } EM Core Facility } University of Missouri, Columbia } } } } } } -----Original Message----- } } From: Clarkson Donna R Contr USAMRD } } [ {mailto:donna.clarkson-at-brooks.af.mil} mailto:donna.clarkson-at-brooks.af.mil] } Sent: Tue 4/15/2003 8:09 AM } To: bonnie sheppard; Microscopy-at-sparc5.microscopy.com } Cc: } Subject: RE: Kodak 4489 film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Bonnie, } I have not yet actually used the new 4489 film, but spoke with someone } at Kodak before purchasing it. Seeing all the negative reports on it, I } wanted to "get the scoop" before trying it out. They told me that the best } things to try are additional agitation in both the D-19 and Fixer. If the } negatives still appear uneven or muddy try using a 1:1 dilution of D-19 } instead of the usual 1:2. } Good luck to you and anyone else having to use the new film! I hope I } won't have too much trouble. } Have a good one! } } Donna R. Clarkson } } Northrop Grumman Information Technology } for U S Army Medical Research Detachment } at Brooks City-Base } Phone (210) 536-1416 } FAX (210) 536-1449 } e-mail donna.clarkson-at-brooks.af.mil } } } -----Original Message----- } } From: bonnie sheppard [ {mailto:bls4u-at-cstone.net} mailto:bls4u-at-cstone.net] } Sent: Monday, April 14, 2003 2:35 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Kodak 4489 film } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is anyone else out there having trouble with "shadows and unevenness" on } 4489 film? We have gone to extensive lengths to be sure our developing } is consistent, and still we are having annoying problems with uneven } (blotchy) negatives. This is not a problem when there is ample tissue } detail in the negative...but it is horrible and a real printiong problem } for folks looking at DNA spreads, where there is little tissue to } negative. } } Help! } } } } } } } } } *********************************************************************** } This electronic message may contain information that is confidential } and/or legally privileged. It is intended only for the use of the } individual(s) and entity named as recipients in the message. If you } are not an intended recipient of this message, please notify the } sender immediately and delete the material from any computer. Do not } deliver, distribute or copy this message, and do not disclose its } contents or take any action in reliance on the information it } contains. Thank you. }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
The first place to start is a Google search on each of the terms. A similar search could be done under PubMed (National Library of Medicine). I have found that the Google searches find good on-line references from educational institutions that range from the basic to in-depth.
Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals Ridgefield, CT -- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (ramg21-at-yahoo.co.in) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } April 16, 2003 at 11:36:56 } --------------------------------------------------------------------------- } } Email: ramg21-at-yahoo.co.in } Name: ram varma } } Organization: univ of louisiana lafayette } } Education: Graduate College } } Location: lafayette,LA, USA } } Question: could you tell me where i can find comprehensive information about } } 1] near field optical microscopy } 2] reflectometry } 3] ellipsometry } } I would like to know all the details viz. equipment } specification,opration,theory of operation etc. } } --------------------------------------------------------------------------- }
Capturing the spatial resolution of your microscope is a matter of both inner and outer resolution. The inner resolution is the capability of you camera to resolve the spatial information provided by your microscope. This is determined by the spatial resolution of the microscope, total magnification and the pixel size of the CCD-grid on your camera. The Nyquist sampling theorem and the Whittaker-Shannon Sampling theorem deals with this issue:
The larger the individual pixels of the camera, the more you will need to magnify the image, given the same spatial resolution. This is important for quantification, but can be more relaxed for visualisation or publications.
The outer resolution is determined by the fraction of the field of view the camera can capture on its CCD. The larger the field covered by the camera in one image the less you will need to move the microscope stage to view more of the sample.
So in a sense the digital image is defined by its inner and outer resolution. This is only a short description of this issue and there are of course more and more precise thing to say about it.
===================================================== } } I have the option to buy either a 5 or 3 megapixel camera. What I want to } } know is whether a 5 megapixel camera is better than a 3 megapixel } } camera, if used exclusively on a microscope, or whether the extra pixels } } (and cost) are wasted. Assume bit depth is the same. } } } } I'd like someone to check my math and my reasoning. } } } } I understand that microscope resolution is limited by diffraction and a } } good rule of thumb for displaying images at normal reading distance is a } } final image mag of no more than 1000x the numerical aperture of the } } objective. On my microscope this means about 100x for images taken with } } the 4x lens and 1250x for images with the 100x oil lens. Assuming I use a } } 1x relay eyepiece to connect the CCD camera, the image at the detector } } could be magnified 25 times for the 4x objective, and 12.5 times for the } } 100x objective, before getting into empty mag. } } } } On the camera side, I understand that the more pixels the better the } } resolution. The 3 megapixel camera has 2080 horizontal pixels and the 5mp } } has 2580. Let's ignore the vertical pixels for the moment. } } } } One more assumption is that for printing, it seems that the highest } } practical output resolution is 266-300 dpi. This means that the 3 mp } } camera produces an image that optimally prints an image 2080/266 = 7.8 } } inches or 198 mm horizontally, or less, and the 5 mp can print an image } } about 243 mm horizontally or less. Enlarging above these dimensions is } } possible but the print image will need interpolation and might be degraded. } } } } Going back to the microscope, the maximum horizontal field of view with } } the 4x objective captured by the camera is 1.6 mm. It could be printed up } } to 100 x 1.6 = 160 mm before empty mag sets in. But I have already } } calculated that the 3 mp can print up to 198 mm and the 5 mp up to 243 mm } } before the print image degrades. } } } } If these calculations are correct (please tell me if it they are not) then } } both the 3 mp and 5 mp have more than enough resolution to capture what } } the microscope makes available at 4x, and the situation is even better for } } the 100x lens because the lens does most of the heavy lifting, } } magnification-wise. } } } } One final condition that might matter is that both are RBG cameras with } } Bayer filters, so that the real resolution is approximately 1/3 of the } } nominal pixel resolution. I'm not sure how to figure that into the } } calculations. } } } } So, at approximately a dollar-per-pixel, do I need the extra 2000 pixels } } of a 5 mp camera? } } } } } } Gary P. Radice gradice-at-richmond.edu } } Department of Biology 804-289-8107 (voice) } } University of Richmond 804-289-8233 (FAX) } } Richmond VA 23173 http://www.richmond.edu/~gradice } }
I suggest that everyone unhappy with the new formulation of 4489 should contact Eric Ambrose, and respond to his post of Jan 24th.
Jim
PS: Our users are some of the "few" unhappy.
} } Eric Ambrose } } eric.ambrose-at-kodak.com } } Silver Halide Product Manager } } Scientific Imaging Systems } } Eastman Kodak Company
Jim
Sergey wrote: } } In respond on my message on the "first wave" of the 4489 new formulation } discussion I got respond from Kodak that most customers are very happy with } new formulation and ONLY a few were unhappy. I am too lazy to find that } message in my archive. I wrote back asking why they did not change the } product name if altering development process on "new formulation" - I never } had respond back. Sergey } } }
} } From MicroscopyL-request-at-sparc5.microscopy.com Fri Jan 24 05:38:36 2003 } } Date: Thu, 23 Jan 2003 18:25:06 -0600 } } To: Microscopy-at-sparc5.microscopy.com } } From: eric.ambrose-at-kodak.com (by way of MicroscopyListserver) } } Subject: TEM Imaging: Kodak 4489 EM Film Reformulation } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } } } Subject: TEM Imaging: Kodak 4489 EM Film Reformulation } } } } Recently, there have been a number of messages posted on the MSA ListServer } } regarding issues with the reformulated KODAK Electron Microscope Film 4489. } } As the Kodak product manager for the films for electron micrography, I } } would like to offer the following information in response to the recent } } inquiries on Kodak EM films. } } } } Scientific Imaging Systems, a division of Eastman Kodak Company, remains } } committed to supplying the highest quality scientific imaging products } } required to meet the critical needs of today's research scientists. Kodak } } manufactures and sells two different films for use in TEM imaging, KODAK } } Electron Microscope Film 4489 and KODAK Electron Image Film SO-163. } } } } Over the years, for many different reasons, Kodak has made changes to the } } formulations of these films. Some changes have been minor while others have } } been more significant. The decision to make a formulation change is one of } } the most contentious and difficult decisions that we address. Most of } } these changes are forced upon us in response to manufacturing equipment } } requirements, market demand, and/or issues with supply of raw material. } } Whatever the reason, Kodak makes these required changes not only to ensure } } the continued availability of electron micrography products to scientific } } customers, but also to ensure the products perform to the highest quality } } standards. Every change to a formulation is accompanied by extensive } } in-house and customer testing to ensure the product meets its } } specifications. Various techniques performed in the research community can } } result in some instances where modifications to some protocols are required } } to achieve comparable results with a reformulated film compared to its } } predecessor. } } } } Scientific Imaging Systems (SIS) has a dedicated team of technical support } } staff ready to assist anyone with questions or concerns about the } } performance of the Kodak EM films, as well as any other products sold by } } SIS. People should not hesitate to contact the SIS Technical Support Group } } with questions, concerns, and suggestions. We value our customers and } } will make every effort to assist you. Please email us at: } } sis-support-at-kodak.com or call 1-877-SIS-HELP or 203-786-5657. } } } } In regards to some of the comments and issues recently expressed on the MSA } } Microscopy ListServer, I have personally contacted, or have attempted to } } contact, each person listed with a concern. As in many instances, we need } } to see the films themselves to make a determination on the issue and how to } } best assist the customer going forth. Most customers that have used the } } reformulated 4489 film have found that it supplies the same high quality } } results they have always received from Kodak EM films. However, as seen on } } the Microscopy ListServer, we do have a small number of customers that } } appear to have an issue of some kind. As is often the case, these issues } } may not be related to the reformulation and/or each other. We see every } } one of our customers as very important; we are committed to understanding } } the nature and cause of the difficulty, and finding a way to resolve it. } } Should anyone have a concern or question, please feel free to contact our } } technical support group. } } } } Thank you for your interest in Eastman Kodak Company products. } } } } Respectfully, } } } } Eric Ambrose } } Silver Halide Product Manager } } Scientific Imaging Systems } } Eastman Kodak Company } }
To our knowledge also the Kodak SO163 changed. Does anybody have more information about this? We tried to get information from Kodak about the changed emulsion, but were referred to the sales people ... who did not have the information. The film seems to have gotten less sensitive. I am concerned that this may pose a serious problem for low-dose electron microscopy.
Regards, Michael
__________________________________________ Michael Radermacher University of Vermont College of Medicine Dept. Mol. Physiol. & Biophysics Burlington, VT 05405
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This is Lifeng Dong from Department of Physics, Portland State University, USA. I am planning to attend M&M 2003, and reserved a double room (two beds) from Hampton Inn Downtown San Antonio. The price including tax is $121.42 per night, which is a little expensive for a graduate student. If you or some of your male colleagues will attend this conference and are looking for a hotel, I would like to share it with you or him.
If you have any question, contact me at lifengd-at-pdx.edu.
Post for a friend. -shuyou. ------------------------------------------------------------- Dear All,
I have some troubles on my results. Any comments would be appreciated.
It is about characterization of epitaxial beta-FeSi2. The substrate is Si (111). The thickness of epitaxial Fe film is 10 angstroms. Form beta-FeSi2 islands after annealing at 600C. Bulk beta-FeSi2 belongs to based centered orthorhombic lattice with a=0.9863nm, b=0.7791nm and c=0.7833nm. Epitaxial relationship is (110)beta//(111)Si with [001]beta//[1-10]Si. The fact is that beta-FeSi2 can be viewed as a layered structure based on (110)beta plane, and the atoms on each layer always distribute in the form of distorted hexagon similar to that of Si (111) plane.
Now my question is whether the superimposed diffraction pattern along [111]Si zone axis are same (similar) as the FFT result obtained from the HRTEM images having [111]Si orientation or not.
In my opinion, they are different. The reason is that in real space (110)beta // (111)Si according to the epitaxial relationship. It is clear that [111]Si is perpendicular to (111)Si. But [110]beta is not perpendicular to (110)beta because beta is orthorhombic. On the other hand, the crystal directions should always be perpendicular to the reciprocal planes with the same indices as crystal directions. As a result, in reciprocal space two reciprocal plane (111)*Si and (110)*beta are not parallel each other. The reciprocal plane (111)*Si is perpendicular to electron beam when the orientation is exact [111]Si zone axis, whereas the reciprocal (110)*beta plane is not perpendicular to beam. Another thing is the reciprocal lattice points on (110)*beta plane should distribute in a form of RECTANGLE. So we can see magnified projection of thess rectangles in the superimposed pattern.
We can observe nice plan-view HRTEM images when the orientation is [111]Si. As for FFT of HRTEM image taken from [111]Si orientation, the dots resulting from overlaping beta and Si each other distribute in the form of distorted hexagon. The FFT results show there are two sets of spots, near the Si reflection spot there is always another adjacent spot. Both of them display HEXAGONAL shape in FFT results. I think this is because the atoms distribute in the forms of hexagon in Si and distorted hexagon in beta. In this case both (110)beta and (111)Si planes are perpendicular to beam, but no beta zone axis with simple indices are parallel to beam. The reason why we can see HRTEM images is that there indeed exist some atomic (atom group) planes perpendicular to (110)beta plane although these atomic planes do not have simple indices.
My conclusion is that with same orientation the electron diffraction pattern and FFT of HRTEM image are not always same (similar) if one of them is non-cubic crystal. But somebody tell me this conclusion is stupid wrong. I can not find which step is wrong. Also he did not let me know where is my mistake. Till now I believe it is true. I beg your kind comment for my above conclusion. I am anxious to know any comments. Thank you very much in advance.
Best Wishes,
M. HAN HAN.ming-at-nims.go.jp
_____________________________ Shu-You Li, Ph.D. Electron Microscopist, EPIC NUANCE Northwestern University http://www.nuance.northwestern.edu/EPIC
Having followed several messages on this site, I wanted to know if I already had the "new formulation" in my stock so I asked a few questions and the result of two messages resulted in the following Quotes and the directions that have already been posted about nitrogen bursts/extra agitation with tilting. I also requested that Kodak go back to the old formulation.
"All of the new formulation film is marked on the outside of the container "New Formulation". We started producing the "new formulation" approximately last June. The films expiration date is typically 2 years from the date manufactured. Therefore, you if the film that you have does not specify "new formulation", you may still have the original formulation film."
"The reason for reformulating KODAK Electron Microscope Film 4489 was the direct result of the unavailability of some raw materials necessary in the manufacture of the prior version of this product. Every change to a formulation is accompanied by in-house and customer testing to ensure the product meets its specification. Both in-house and customer testing confirmed that reformulated KODAK Electron Microscope Film 4489 does meet the product specification. Most customers that have used the reformulated KODAK Electron Microscope Film 4489 have found it supplies the same high quality results they have always received from Kodak EM films. However, the various processing techniques performed in the research community can result in some instances where modifications to some protocols are required to achieve comparable results with a reformulated film compared to its predecessor."
This answered my questions and I hope that it will do the same for you.
Remember when Kodak discontinued SO-163 a few decades ago? That was when we switched to 4489 after an extended trial and error period. Or is my mind playing tricks on me?
Pat Connelly Dept. of Biology University of Pennsylvania Philadelphia, PA 19104-6018
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } My question is: why would Kodak substitute this for a perfectly good } film without any warning? The package insert from a box of the old } film was identical to that in the new film, but it seems as if } processing procedures need to be radically altered. This has cost } us some unhappy clients and no end of headaches, not to mention } one embarrassing incident where an employee was blamed for something } that wasn't his fault. } Unless I missed some major press release, this seems like a highly } irresponsible action. Does anyone have any insight into this? } Agitatedly, } Randy } === } Normally I don't like litigation but the pain and suffering and the lack } of warning seems to point to a class action suit, and at least an } official explanation accompanying the new film and properly labeled new } film. I'm maybe just surprised to not have seen that suggestion in the } previous discussion on 4489 formula change. } } I know this has been gone over in length before. We do not have any } nitrogen capabilities in our darkroom, and I am dreading the next batch } of 4489 film. Maybe Kodak is using this like Coca-cola did when they } made "new" coke, and then switched back to "classic" but that change let } them go from using expensive beet sugar to corn sugar as the primary } sweetener saving millions of dollars. Who knows - but from the great } discussion here I at least have a plan for when we get the new formula } (not that I will be able to implement it). } } Geoff Williams } Microscopy Facility Supervisor } CMU Biology Department Microscopy Facility web page. } http://www.cst.cmich.edu/centers/microscopy/
Greetings, In the early and mid 1990's I was the proud owner of a Panasonic Optical Drive, model LF 7010, until it stopped working a few years ago. I now find myself in the embarassing position of wanting to get some data off a disk written by that drive (I could have sworn we backed the stuff up on CD but if so the back up is lost). The Panasonic drive was operated by a Mac, as a scsi device. The disk in question is a 1.5 GB Panasonic Optical Disk (LM-R1500A). I know this is a long shot, but is there anyone out there who might be able to read this disk?? (Panasonic tech support only has PC's nowadays, I checked with their Midwest branch).
It's been my experience that this sort of comment comes from a complete lack of understanding of EM techniques. I have always maintained that it is much, much easier to teach a standard lecture course to 50-100+ students, or a standard lecture & lab course to 20-30 students, than it is to teach an intensive EM course to 8-16 students. The students need much more intensive instruction in lab techniques and equipment operation than in a standard lab course; they usually work at their own convenience (since we don't have an EM or microtome for each student), so they need help throughout the day and week; they usually work on a course project to hone skills, and that requires lots of mentoring and personal problem solving; and let's not forget about the time it takes to keep the equipment running when novices are hard on it. Try counting the time you spend on the course (not limited to in-class time), and compare it to 3 hours of lecture per week. And good luck. --------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Division of Natural Sciences Purchase College State University of New York Purchase, NY 10577 --------------------------------------- Ofc. Tel: 914-251-6659 Ofc. Fax: 914-251-6635 E-mail: jfactor-at-purvid.ns.purchase.edu ---------------------------------------
JERRY KUDENOV wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Colleagues, } } When teaching an EM course, how many additional non-EM courses might } others teach per semester? I am presently teaching 2 courses, one } of which is on SEM. I also teach microtechnique and a TEM course. } } Obviously, the answer depends on EM course formats and campus size. } } My courses are intensive, usually involving 5-6 students, sometimes } more. An SEM course may take 15-25 hours per week, including all formally } scheduled lab and lecture times, weekly tutorials, solutions preparation } etc.(there is no assistant). Our campus is small, perhaps 14,000 students. } } Since I teach so few students, my Chair is wondering about the fairness } of my workload relative to others in my department who teach many more } students in straight lecture courses (the larger courses have assistants). } } Your answers will be most helpful in clarifying a "normal" academic work } load that includes EM instruction at other similar sized universities. } Please send your replies to me at the e-mail address below. I will } summarize the results and post them on the listserver. } } Thank you in advance! } } Jerry } } --------------------- } Jerry D. Kudenov } Dept. Biological Sciences } University of Alaska Anchorage } 3211 Providence Drive } Anchorage, Alaska 99508 } } afjdk-at-uaa.alaska.edu } Office: 1-907.786.1769 } Fax: 1-907.786.4607 } ----------------------
The annual meeting of the Pacific Northwest Microscopy Society will be held this year at the Pacific Northwest National Laboratory (PNNL) in Richland, WA on May 29 and 30, 2003.
We are soliciting abstracts for contributed and poster presentations in all aspects of light and electron microscopy, as well as in technology advances using microscopy as the analytical technique.
Two parallel sessions will run separately in biological and material science. Biological sciences will focus on applications in fluorescent microscopy including confocal and multi-photon imaging, cathodoluminescence, flow cytometry and immunocytochemistry, and 3-d reconstruction by electron tomography. Contributions to the "technologist's forum" are encouraged. The material science division will have a great Focused Ion Beam (FIB) session; our invited MSA sponsored speaker Dr.Lucille Gianuzzi (University of Central Florida) will give a presentation on this topic.
Abstracts - for both posters and platform presentations should be sent to the program chair, Jim Young (jim.young-at-pnl.gov) in an electronic version (either Word or pdf file), with text and figures not to exceed one page. Deadline for abstract submission is April 30, 2003. Please specify platform or poster session. All accepted papers will be printed (as submitted) in meeting proceedings. We encourage both undergraduate and graduate students to participate in a poster session. Cash prizes of $300, $200 and $100 will be awarded to the best student posters in both biological and material science categories. Vendors - of companies manufacturing or selling products and instrumentation related to microscopy will participate in the exhibit during the meeting. Social event - an evening social including a complimentary catered dinner and wine tasting will be held at the Gordon Brothers winery in Pasco on Thursday evening for all registered participants and spouses. Accommodation - the new PNNL user-housing facility, conveniently located just a short walk from the conference rooms, will be available to the meeting participants at a discounted rate. Rooms with a queen bed and a private bathroom at $50/night includes in room continental breakfast. We have reserved 30 of these rooms at a group rate. You can make a reservation on-line at uhf-at-pnl.gov or call 509-372-6736. Additional accommodation available in Richland hotels. Registration - for the meeting will be free of charge to all current PNMS members (contact Jim Young for membership info). Non-members $10 symposia only, $20 including the Thursday night reception. Badging - for those unfamiliar with the PNNL policies, a visitor badge is required for getting in all PNNL labs. Therefore, each participant wishing to tour the lab facility needs to have a visitor's badge. Conference rooms and the auditorium doesn't require a badge, however we strongly recommend to apply for one in order to participate in in-lab demos and tours. The following information is required: 1. Full name, including middle name 2. Social Security Number 3. Citizenship 4. Date of birth 5. Employer and address 6. Telephone number Additional information is required for foreign nationals, and they should contact Jim Young as soon as possible since the security clearance takes 4-6 weeks.
We are looking forward to see you in Richland!
Alice Dohnalkova PNMS president 2003 PNNL, Richland WA (509)372-0692
Jim Young WA program chair jim.young-at-pnl.gov PNNL, Richland WA (509) 376-2797
Alice Dohnalkova Environmental Microbiology Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 tel. (509) 372-0692 office (509) 376-3654 TEM lab fax (509) 376-1321
At 06:26 AM 4/17/2003, 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
SO 163 actually has an advantage over 4489 for biological work because it is faster (less problem with moving specimens. which in our lab is THE main problem for biologists). So unless you have a lot of users who routinely enlarge the negative 8 or 10 times....why not try it?
cheers
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University, Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 http://www.anu.edu.au/EMU
} } } "Patton, David" {David.Patton-at-uwe.ac.uk} 04/17/03 08:45PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
At least with Coke there are other soft drinks. As far as I can tell 4489 is the only film available for routine biological tissue work. I have formed the impression from this listserver that S0163 (?) is for more specialised use. Therefore I think we are over a barrel re voting with our feet (mixed metaphors anyone?).
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} -----------------------------------------------------------------------. } } } Randy, } I'm sure it's got something to do with Kodak saving a fraction of a } penny per film box. Remember Coca-Cola's new formulation years ago? } Seems like Kodak should've taken a lesson. There's a Murphy's Law } corollary which states, " If it ain't broke, don't fix it!" } } WWW } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Winston W. Wiggins, Supervisor } Cannon Electron Microscopy Lab } Carolinas Medical Center } P.O. Box 32861; Charlotte, NC 28232-2861 } Ship to: 1000 Blythe Blvd; Charlotte, NC 28203 } Ofc:704-355-1267 ; Lab:704-355-7220 ; Fax 704-355-0589 } E-mail: {mailto:WWiggins-at-Carolinas.org} Winston.Wiggins-at-CarolinasHealthCare.org
} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } -----Original Message----- } } From: Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] } Sent: Tuesday, April 15, 2003 1:33 PM } To: Clarkson Donna R Contr USAMRD } Cc: microscopy-at-sparc5.microscopy.com } Subject: RE: Kodak 4489 film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver
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} -----------------------------------------------------------------------. } } } Hi, } } We were also taken by surprise by a box of the "New Formulation" 4489 } that we didn't even know we had, until we started using it. This } seems to be the film from hell. We solved the density problem by } increasing our nitrogen burst agitation to 2 seconds every 10 } seconds, but we still are getting strange agitation marks. } } I started out years ago using the method described by Steve Chapman, } which is the Kodak recommendation, and only started using nitrogen } burst when I moved to my present job in 1999. Absolutely no problems } until the new film showed up, then we had very thin negatives with } horrible streaks. As I said, the density problem has been solved, } but we're still fighting the streaks. } } What we are doing is a series of exposure trials using "blank" } exposed negatives, i.e., exposing the films in the scope with no } specimen in place to give an even field of detail-less illumination. } This is guaranteed to show any agitation irregularities. We will } then vary agitation procedures and times until we get acceptable } results. Preliminary results are showing that the plastic film } holders with narrow sides, rather then the ones that reach nearly to } the bottom of the films, cause fewer agitation problems. Other first } results are showing that a combination of "lift and tilt" with } nitrogen burst evens things out. Finally, we were getting a } "thumb-print" on the bottom of the negatives that turned out to be } caused by an agitation swirl around the plastic support rod that } holds the films in place. We solved that one by lifting the films up } slighty so they don't touch the rod, but that only works on holders } that are narrow enough to bend the films when they're inserted. } } When we complete our tests, I hope to post images and procedures on } our website and post a link to the listserver. In the meantime, I'm } following this thread with great interest. } } My question is: why would Kodak substitute this for a perfectly good } film without any warning? The package insert from a box of the old } film was identical to that in the new film, but it seems as if } processing procedures need to be radically altered. This has cost us } some unhappy clients and no end of headaches, not to mention one } embarassing incident where an employee was blamed for something that } wasn't his fault. Unless I missed some major press release, this } seems like a highly irresponsible action. Does anyone have any } insight into this? } } Agitatedly, } Randy } } Randy Tindall } EM Core Facility } University of Missouri, Columbia } } } } } } -----Original Message----- } } From: Clarkson Donna R Contr USAMRD } } [ {mailto:donna.clarkson-at-brooks.af.mil} mailto:donna.clarkson-at-brooks.af.mil]
} Sent: Tue 4/15/2003 8:09 AM } To: bonnie sheppard; Microscopy-at-sparc5.microscopy.com } Cc: } Subject: RE: Kodak 4489 film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver
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} -----------------------------------------------------------------------. } } } Bonnie, } I have not yet actually used the new 4489 film, but spoke with someone } at Kodak before purchasing it. Seeing all the negative reports on it, I } wanted to "get the scoop" before trying it out. They told me that the best } things to try are additional agitation in both the D-19 and Fixer. If the } negatives still appear uneven or muddy try using a 1:1 dilution of D-19 } instead of the usual 1:2. } Good luck to you and anyone else having to use the new film! I hope I } won't have too much trouble. } Have a good one! } } Donna R. Clarkson } } Northrop Grumman Information Technology } for U S Army Medical Research Detachment } at Brooks City-Base } Phone (210) 536-1416 } FAX (210) 536-1449 } e-mail donna.clarkson-at-brooks.af.mil } } } -----Original Message----- } } From: bonnie sheppard [ {mailto:bls4u-at-cstone.net} mailto:bls4u-at-cstone.net]
} Sent: Monday, April 14, 2003 2:35 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Kodak 4489 film } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver
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} -----------------------------------------------------------------------. } } } Is anyone else out there having trouble with "shadows and unevenness" on } 4489 film? We have gone to extensive lengths to be sure our developing } is consistent, and still we are having annoying problems with uneven } (blotchy) negatives. This is not a problem when there is ample tissue } detail in the negative...but it is horrible and a real printiong problem } for folks looking at DNA spreads, where there is little tissue to } negative. } } Help! } } } } } } } } } *********************************************************************** } This electronic message may contain information that is confidential } and/or legally privileged. It is intended only for the use of the } individual(s) and entity named as recipients in the message. If you } are not an intended recipient of this message, please notify the } sender immediately and delete the material from any computer. Do not } deliver, distribute or copy this message, and do not disclose its } contents or take any action in reliance on the information it } contains. Thank you. }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
On Thursday, April 17, 2003, at 03:45 AM, Patton, David wrote:
} I have formed the impression from } this listserver that S0163 (?) is for more specialised use.
Dear David, SO163 is more sensitive than 4489, and it can be push-processed to double its sensitivity. The grain size is small enough that the negatives can be scanned with a small pixel size, say 5 micrometers, and get acceptable quantitation. These features make it better for low-dose EM than 4489. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
I have some troubles on my results. Any comments would be appreciated.
It is about characterization of epitaxial beta-FeSi2. The substrate is Si (111). The thickness of deposited Fe film is 10 angstroms. Form beta-FeSi2 islands after annealing. Bulk beta-FeSi2 belongs to based centered orthorhombic lattice with a=0.9863nm, b=0.7791nm and c=0.7833nm. Epitaxial relationship is (110)beta//(111)Si with [001]beta//[1-10]Si. The fact is that beta-FeSi2 can be viewed as a layered structure based on (110)beta plane, and the atoms on each layer always distribute in the form of distorted hexagons similar to that of Si (111) plane.
Now my question is whether the superimposed diffraction pattern along [111]Si zone axis are same (similar) as the FFT result obtained from the HRTEM images having [111]Si orientation or not.
In my opinion, they are different. The reason is that in real space (110)beta // (111)Si according to the epitaxial relationship. It is clear that [111]Si is perpendicular to (111)Si. But [110]beta is not perpendicular to (110)beta because beta is orthorhombic. On the other hand, the crystal directions should always be perpendicular to the reciprocal planes with the same indices as crystal directions. As a result, in reciprocal space two reciprocal plane (111)*Si and (110)*beta are not parallel each other. The reciprocal plane (111)*Si is perpendicular to electron beam when the orientation is exact [111]Si zone axis, whereas the reciprocal (110)*beta plane is not perpendicular to beam. Another thing is the reciprocal lattice points on (110)*beta plane should distribute in a form of RECTANGLE. So we can see the magnified projection of thess rectangles in the superimposed pattern.
We can also observe nice plan-view HRTEM images when the orientation is [111]Si. As for FFT of HRTEM image taken from [111]Si orientation, the dots resulting from overlaping beta and Si each other distribute in the form of distorted hexagon. The FFT results show there are two sets of spots, near the Si reflection spot there is always another adjacent spot. Both of them display HEXAGONAL shape in FFT results. I think this is because the atoms distribute in the forms of hexagon in Si and distorted hexagon in beta. In this case both (110)beta and (111)Si planes in real space are perpendicular to beam, but no beta zone axis with simple indices are parallel to beam. The reason why we can see HRTEM images in this case is that there indeed exist some atomic (atom group) planes perpendicular to (110)beta plane although these atomic planes do not have simple indices.
My conclusion is that with same orientation the electron diffraction pattern and FFT of HRTEM image are not always same (similar) if one of them is non-cubic crystal. But somebody tell me this conclusion is stupid wrong. I can not find which step is wrong. Also he did not let me know where is my mistake. Till now I believe it is true. I beg your kind comment for my above conclusion. I am anxious to know any comments. Thank you very much in advance.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cperalta-at-wisc.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, April 18, 2003 at 08:18:09 ---------------------------------------------------------------------------
Email: cperalta-at-wisc.edu Name: Carlos Peralta
Organization: University of Wisconsin
Education: Graduate College
Location: Madison, Wisconsin, USA
Question: Hello: I am getting ready to mount conodonts on glass slides. I'll be using gum tragacanth. Can you tell me what compound should I add to prevent fungal growth. Any answer or suggested sources will be greatly appreciated.
I already asked my colleagues but I didn't get a good answer.
Thank you to everyone who responded to my question regarding the best oil mist filtration system for venting my vacuum pumps indoors.
I received several responses that boiled down to about three basic ideas.
1) Using a favorite brand name (their were several) filter on each pump.
2) Connecting one very large brand name filter to all of the pumps.
3) Venting to the outside air with or without some kind of filter.
Someone also suggested using modified generic automobile engine oil filters.
Regards, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
Pat, I do not know where you got your information but Kodak has been making SO-163 for years and is still doing so. We have used it since the late 80's and have not had any problem with developing (or new formulations so far!). It gives excellent images. Those of you having problems with 4489 might want to try this film. Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 4/17/03 2:02 PM, "Pat Connelly" {psconnel-at-sas.upenn.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Having followed several messages on this site, I wanted to know if I } already had the "new formulation" in my stock so I asked a few } questions and the result of two messages resulted in the following } Quotes and the directions that have already been posted about } nitrogen bursts/extra agitation with tilting. I also requested that } Kodak go back to the old formulation. } } "All of the new formulation film is marked on the outside of the } container "New Formulation". We started producing the "new formulation" } approximately last June. The films expiration date is typically 2 } years from the date manufactured. Therefore, you if the film that you } have does not specify "new formulation", you may still have the original } formulation film." } } "The reason for reformulating KODAK Electron Microscope Film 4489 was the } direct result of the unavailability of some raw materials necessary in } the manufacture of the prior version of this product. Every change to a } formulation is accompanied by in-house and customer testing to ensure } the product meets its specification. Both in-house and customer testing } confirmed that reformulated KODAK Electron Microscope Film 4489 does } meet the product specification. Most customers that have used the } reformulated KODAK Electron Microscope Film 4489 have found it supplies } the same high quality results they have always received from Kodak EM } films. However, the various processing techniques performed in the } research community can result in some instances where modifications to } some protocols are required to achieve comparable results with a } reformulated film compared to its predecessor." } } This answered my questions and I hope that it will do the same for you. } } Remember when Kodak discontinued SO-163 a few decades ago? That was } when we switched to 4489 after an extended trial and error period. Or } is my mind playing tricks on me? } } Pat Connelly } Dept. of Biology } University of Pennsylvania } Philadelphia, PA 19104-6018 } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } My question is: why would Kodak substitute this for a perfectly good } } film without any warning? The package insert from a box of the old } } film was identical to that in the new film, but it seems as if } } processing procedures need to be radically altered. This has cost } } us some unhappy clients and no end of headaches, not to mention } } one embarrassing incident where an employee was blamed for something } } that wasn't his fault. } } Unless I missed some major press release, this seems like a highly } } irresponsible action. Does anyone have any insight into this? } } Agitatedly, } } Randy } } === } } Normally I don't like litigation but the pain and suffering and the lack } } of warning seems to point to a class action suit, and at least an } } official explanation accompanying the new film and properly labeled new } } film. I'm maybe just surprised to not have seen that suggestion in the } } previous discussion on 4489 formula change. } } } } I know this has been gone over in length before. We do not have any } } nitrogen capabilities in our darkroom, and I am dreading the next batch } } of 4489 film. Maybe Kodak is using this like Coca-cola did when they } } made "new" coke, and then switched back to "classic" but that change let } } them go from using expensive beet sugar to corn sugar as the primary } } sweetener saving millions of dollars. Who knows - but from the great } } discussion here I at least have a plan for when we get the new formula } } (not that I will be able to implement it). } } } } Geoff Williams } } Microscopy Facility Supervisor } } CMU Biology Department Microscopy Facility web page. } } http://www.cst.cmich.edu/centers/microscopy/ } } }
Does anyone out there have an Amray SEM not being used that has the External Beam Interface plate? I would appreciate getting one of these plates. Will pay packing and shipping.
It is a small aluminum plate inside the rear of the main console. There are two BNC connectors ( X Y ) and a screw terminal strip. coming out of the plate is a cable with a 6-pin AMP connector that says P9 on it.
I think this needs a bit clarification. More is not always better.
The theoretical requirements for optimum sample frequency were calculated by Shannon and Nyquist some time ago. They basically arrive at the same results: To measure or transmit a frequency X without artifacts, you need to sample the signal with frequency 2X. Staying below this frequency will introduce artifacts, going above it will not yield much improvement.
In other words, if your microscope is capable of resolving 1 micron, you need to use a camera that can resolve 0.5 microns. To find out the matching magnification on the microscope (b/w camera), you solve PS/mag=0.5, where PS is the Pixel size of the camera, and mag is the total magnification between object and camera chip (note that there may be other lenses in the beam path).
For color cameras the situation is more complex, as the resolution is actually color dependend. A Bayer-type color chip has twice as many green pixels as red and blue. For green object, therefore, the resolution is half that of the chip, for red and blue one quarter. If you have a grey object, you'd get full resolution as all pixels are sensitive to some of the grey spectrum.
As for the printing mentioned below: If you are limited by the instrument resolution, and you are using a camera with "too many" pixels, all you get is empty resolution, which is the same as using a camera with a lower resolution (fewer pixels) and artificially increasing the number of pixels and interpolate. The same is true for posters or large prints: Whether you "grossly oversample" the image by using a camera with way too many pixels or artificially increase the image size with interpolation doesn't matter. All you get is empty resolution.
Finally, the "resolution improvement". Image processing is no magic bullet. I have seen those scenes on TV or in Movies, where somebody takes a blurry picture of a crowd, and someone takes a single face, which now looks like a grey blob, then "sharpens" the image to a point where you can see the stubbles of the man's five 'o clock shadow. That's fiction. There are some ways of improving resolution by using deconvolution techniques, but they usually require additional information about the measurement setup (modulation transfer functions, etc). Most other image processing functions make certain information in the image more visible (or less visible), but don't actually improve the resolution. For example a "sharpen" filter does not add information to the image. It does make contrast transitions more visible and thus creates the impression of a "sharper" image, but it also increases noise.
So much for my 2 cents worth...
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Frank Eggert [mailto:Eggert-at-mikroanalytik.de] Sent: Thursday, April 17, 2003 1:41 AM To: Gary Radice Cc: Microscopy-at-sparc5.microscopy.com
Gary,
you are right. From the point of microscopy there are only two reasons to have more pixels:
#1 To print of large images (e.g. for posters or for other representations) without loss of details. #2 For digital processing of your images (e.g. to 'improve' resolution of instrument or to extract details).
Or are there other reasons?
} From a major interest should be the data depth (possible grey steps in each pixel) for a computer processing of brightness and contrast. If you have high dynamics, even wrong lighted images are able to convert into better results or you are able to improve contrast of low differences in an image region, which is uniform in colour and brightness with first views.
Frank Eggert
http://www.microanalyst.net
Gary Radice schrieb:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have the option to buy either a 5 or 3 megapixel camera. What I want } to know is whether a 5 megapixel camera is better than a 3 megapixel } camera, if used exclusively on a microscope, or whether the extra } pixels (and cost) are wasted. Assume bit depth is the same. } } I'd like someone to check my math and my reasoning. } } I understand that microscope resolution is limited by diffraction and a } good rule of thumb for displaying images at normal reading distance is } a final image mag of no more than 1000x the numerical aperture of the } objective. On my microscope this means about 100x for images taken with } the 4x lens and 1250x for images with the 100x oil lens. Assuming I use } a 1x relay eyepiece to connect the CCD camera, the image at the } detector could be magnified 25 times for the 4x objective, and 12.5 } times for the 100x objective, before getting into empty mag. } } On the camera side, I understand that the more pixels the better the } resolution. The 3 megapixel camera has 2080 horizontal pixels and the } 5mp has 2580. Let's ignore the vertical pixels for the moment. } } One more assumption is that for printing, it seems that the highest } practical output resolution is 266-300 dpi. This means that the 3 mp } camera produces an image that optimally prints an image 2080/266 = 7.8 } inches or 198 mm horizontally, or less, and the 5 mp can print an image } about 243 mm horizontally or less. Enlarging above these dimensions is } possible but the print image will need interpolation and might be } degraded. } } Going back to the microscope, the maximum horizontal field of view with } the 4x objective captured by the camera is 1.6 mm. It could be printed } up to 100 x 1.6 = 160 mm before empty mag sets in. But I have already } calculated that the 3 mp can print up to 198 mm and the 5 mp up to 243 } mm before the print image degrades. } } If these calculations are correct (please tell me if it they are not) } then both the 3 mp and 5 mp have more than enough resolution to capture } what the microscope makes available at 4x, and the situation is even } better for the 100x lens because the lens does most of the heavy } lifting, magnification-wise. } } One final condition that might matter is that both are RBG cameras with } Bayer filters, so that the real resolution is approximately 1/3 of the } nominal pixel resolution. I'm not sure how to figure that into the } calculations. } } So, at approximately a dollar-per-pixel, do I need the extra 2000 } pixels of a 5 mp camera? } } Gary P. Radice gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond 804-289-8233 (FAX) } Richmond VA 23173 http://www.richmond.edu/~gradice
Debby, When I started at U of P in 1976, the film in use was the SO-163 and it was in use until either the late 70's or early 80's. I took off three years, had two children and when I returned, the film had been switched to Kodak 4489. I believe that so many researchers complained, that Kodak re-issued the film but we decided not to switch back from the 4489 that we were, at that time, using as the replacement. I am considering the switch back to the SO-163 now, in that by using either it or the "new formulation 4489" I will have to adjust my protocol and scope (Philips 200) settings. (There are no nitrogen burst tanks here.)
Can anyone tell me when using the new formulation 4489, if the negatives are darker or lighter than the old formulation before making adjustments to exposure time or sensitivity or is the total problem just the proper agitation of the developer?
I am blessed to have ordered, by chance, a quantity of film while the old supply still was available.
Thanks, Pat Connelly
} Pat, } I do not know where you got your information but Kodak has been making } SO-163 for years and is still doing so. We have used it since the late } 80's and have not had any problem with developing (or new formulations so } far!). } It gives excellent images. Those of you having problems with 4489 might } want to try this film. } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } } On 4/17/03 2:02 PM, "Pat Connelly" {psconnel-at-sas.upenn.edu} wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } -----------------------------------------------------------------------. } } Remember when Kodak discontinued SO-163 a few decades ago? That was } } when we switched to 4489 after an extended trial and error period. Or } } is my mind playing tricks on me? } } Pat Connelly } } Dept. of Biology } } University of Pennsylvania } } Philadelphia, PA 19104-6018
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
This is exactly the test I performed (we alternated old/new sheets) with exactly the same density results, when photographing a "typical" biological sample. I used freshly made Kodak D-19 developer diluted 1:2 (as recommended by Kodak) and Kodak Rapid Fixer; the temperature was within 1/2 degree of the recommended 20 degrees C, using constant agitation, occasionally lifting the negatives completely out of the developer, tilting first to one side then the next to get the developer in the grooves of the Lucite holder. All of the negatives were evenly developed, but the new formulation was significantly darker than the old formulation. I have not yet adjusted the exposure (sensitivity) of the microscope to the film since we still have a little of the old film left, but I expect that this should be much easier (and cheaper!) than trying to implement a nitrogen bust system, which we've never had nor needed despite Kodak's instructions.
I've been following this discussion since early February when I first posted a question myself about the film, and was rather surprised to see that my test result was quite different from everything I had read. Glad to see that someone's results finally corroborate mine!
Valerie
At 09:10 AM 4/17/2003 +0200, R. Cross wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Valerie M. Knowlton Research Assistant/Teaching Technician Center for Electron Microscopy 1219 Gardner Hall, Box 7615 North Carolina State University Raleigh, NC 27695
I have a user who wants to make TEM carbon platinum replicas of mammalian cells growing on glass slides with attached tissue tek culture chambers. He figures to fix the cells, dehydrate, critical point dry, then coat the slides (minus the growth chambers) and float the replicas off the slide the same way he floats formvar off a slide when making coated grids. In his first attempt, he is unable to float the metal coating off the slide. A query to this listserver quite a while back mentioned using hydrofluoric acid to remove the replica. does anyone have details of this procedure, or preferably an alternative procedure that will enable my user to make his replicas?
Thanks for any insights in this regard
Steve -- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
A friend from our Soil Science department wants to compare roots from live plants grown in good soil and dead plants grown in poisoned soil. So far we have been looking at them under dark-field optical microscopy. It would be good if we could look at them under SEM, but we don't have an ESEM, and we don't want the root shape to collapse under vacuum dessication.
When I have searched for root fixation I come across things like glutaraldehyde and cacodylate buffer, and we'd like to avoid using these if possible. Does anyone know simpler ways of fixing the external shape of roots?
It would be nice if we could do the same thing for moulds, to show visiting students "this is what you get if you leave bread lying around in your accommodation!".
Any ideas, please?
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +----------------------------------------+ Phone:+44 (0) 118 9318572 Fax: +44 (0) 118 9750203 University internal extension 7867 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +----------------------------------------+
Dear Steve If it's Pt or Pt/C coating to remove replica from the glass you may use some HF solution. I don't remember exact concentration, but I would think, 10% (v/v) should work. If not, you may try higher concentration. Please, keep in mind: HF is extremely aggressive chemical: it's strong irritant and will corrode everything on it's way including SS EM tweezers. It should be stored in the plastic (NO GLASS) and used with precautions. Basically, you use the same technique as for floating Formvar film except everything should be plastic and HF solution instead of water. When replica is detached from the glass, you may move it with Pt loop to the water to wash it a few times and then mount on the grids. Good luck. Sergey.
At 12:55 PM 4/21/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Dear Ming, This problem is in principle quite simple. Any diffraction pattern can be seen as a Fourier transform of the diffracting object. You can find this in any undergraduate optics textbook. Normally in TEM you will then have to convulute the periodicity of the periodicity of the object with a top- hat function describing its very limited thickness. This leads to the well-known reciprocal lattice rods for thin samples.
So, an SAD pattern is a Fourier transform including specimen periodicity and specimen thickness.
An HRTEM image can also be Fourier transformed. As you should know, however, the HRTEM image is strongly influenced by the contrast transfer function (i.e. highly defocus dependent). That means that the intensities in a Fourier-transform of a HRTEM image are strongly affected by image defocus. You see this effect particularly strongly for amorphous material.
So, an SAD pattern and a Fourier-transformed HRTEM image have the same geometry, but may have rather different intensities in the different spots since the latter is so focus-dependent.
That said, in your case the main point is that the geometry of the pattern is identical for SAD or for Fourier-transformed HRTEM.
Now, if your orientation relationship is correct, then when you orient along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are matching the (111) Si planes. I reckon on a quick calculation that the angle between the (110)beta plane normal and [110]beta is 13.4 °, much too large to see the [110]beta diffraction pattern. But the axis [230]beta lies only 1.86° from the (110)beta plane normal. Are you seeing this diffraction pattern? At such a small angle off axis, you will still see the diffraction pattern and an HRTEM image (even if itâs not very good or symmetric).
Best wishes
Ian MacLaren
------- Forwarded message ------- } From: HAN.Ming-at-momokusa.nims.go.jp To: Microscopy-at-sparc5.microscopy.com
ATTN:
} From the Desk of MR MIKE OBI AFRI Bank of Nigeria, Lagos-Nigeria.
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Greetings, Fixation will distort the organ, no matter what. But you should still be able to compare your samples. Aldehydes give the best fixation, as a rule, but there is no reason to use cacodylate for your purpose (actually, some would say no reason ever, but that is another story). Plant roots can be fixed in FAA, dehydrated to absolute ethanol, critically point dried, coated with metal, and viewed. As an alternative to FAA, you could use 4% paraformaldehyde in say PBS (or the dilute buffer of your choice, near pH7), rinse in the buffer, dehydrate etc as above. I expect that 2 h in 4% paraformaldehyde fix would be fine. You can check for shrinkage and other untoward affects by looking at your fixed or dehydrated roots under darkfield.
Hope this helps, Tobias } } } } A friend from our Soil Science department wants to compare roots from live } plants grown in good soil and dead plants grown in poisoned soil. So far } we have been looking at them under dark-field optical microscopy. It } would be good if we could look at them under SEM, but we don't have an } ESEM, and we don't want the root shape to collapse under vacuum } dessication. } } When I have searched for root fixation I come across things like } glutaraldehyde and cacodylate buffer, and we'd like to avoid using these } if possible. Does anyone know simpler ways of fixing the external shape } of roots? } } It would be nice if we could do the same thing for moulds, to show } visiting students "this is what you get if you leave bread lying around in } your accommodation!". } } Any ideas, please? } } +-----------------------------------------+ } Robert H.Olley } J.J.Thomson Physical Laboratory } University of Reading } Whiteknights } Reading RG6 6AF } England } +----------------------------------------+ } Phone:+44 (0) 118 9318572 } Fax: +44 (0) 118 9750203 } University internal extension 7867 } Email: R.H.Olley-at-reading.ac.uk } URL: http://www.reading.ac.uk/~spsolley } +----------------------------------------+
IŽm interested in labelling bacterial surfaces with colloidal gold for negative stain and TEM. So far negative stain works OK, also indirect immunofluorescence with a primary mouse monoclonal. However, secondary antibodies conjugated with 10-15 nm gold doesnŽt work at all. We have adhered fimbriated E Coli to Formvar-coated gold grids with or without fixation in various concentrations of formaldehyde, labelled them and fixed with glutaraldehyde. Any tips and tricks?? Regards, Margaretha
--
Margaretha Lindroth, Ph D Phone: 4613-222616 Dept of Medical Microbiology Fax: 4613-224789 Faculty of Health Sciences e-mail: Margaretha.Lindroth-at-hul.liu.se Linköpings Universitet SE-581 85 Linköping
I would greatly appreciate your reponse. I believe you spent much time answering my stupid question.
In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this sphere is not a plane. Sometimes it brings some troubles to us. Let me show an example. If the specimen normal is NOT parallel to incident beam, or if the orientation is NOT exact, the intersections between relrod and sphere are not symmetric. In SAD pattern, not only intensity but also the distance is different, in other words, SAD pattern is NOT symmetric. On the other hand, FFT always symetric even though zone axis is not exact. This let me believe FFT and SAD is not so identical.
Thanks for your kind calculation. Near [111]Si orientation, we can find SAD pattern. As you said, it is not an exact zone axis. I took three patterns with different tilting angles (x=-9, +2, +8) and spent two weeks understanding these patterns. After calibrating the tilting angles by Kikuchi pattens, my indexing results let me believe they come from an identical reciprocal plane (130)*. I believe your calculation is right. I guess the reasons why it is [130] rather than [230] maybe come from two points. 1. beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. (640) spot is too far from transmitted spot. 2. The epitaxial phase is under distorted state. The real angle maybe different from those calculated based on bulk data.
Till now I still think FFT is not completely identical to SAD. I believe the discussion itself is significant. In my opinion, we can view Ewald sphere as a plane and based on this approximation we can think FFT=SAD. But for epitaxy, we can not ignore this approximation if we want to discuss some slight mismatch information. Maybe I made some mistakes in my analysis. I hope to hear different sounds and look forward to hearing from you again.
Thanks again,
Best wishes,
Han
} Dear Ming, } This problem is in principle quite simple. Any diffraction pattern can be } seen as a Fourier transform of the diffracting object. You can find this } in any undergraduate optics textbook. Normally in TEM you will then have } to convulute the periodicity of the periodicity of the object with a top- } hat function describing its very limited thickness. This leads to the } well-known reciprocal lattice rods for thin samples. } } So, an SAD pattern is a Fourier transform including specimen periodicity } and specimen thickness. } } An HRTEM image can also be Fourier transformed. As you should know, } however, the HRTEM image is strongly influenced by the contrast transfer } function (i.e. highly defocus dependent). That means that the intensities } in a Fourier-transform of a HRTEM image are strongly affected by image } defocus. You see this effect particularly strongly for amorphous material. } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } geometry, but may have rather different intensities in the different spots } since the latter is so focus-dependent. } } That said, in your case the main point is that the geometry of the pattern } is identical for SAD or for Fourier-transformed HRTEM. } } Now, if your orientation relationship is correct, then when you orient } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } matching the (111) Si planes. I reckon on a quick calculation that the } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much too } large to see the [110]beta diffraction pattern. But the axis [230]beta } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } diffraction pattern? At such a small angle off axis, you will still see } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } symmetric). } } Best wishes } } Ian MacLaren }
The Pt-C replicas can be detached from the glass substrate by submerging the specimen at a shallow angle (} 45º) into a small petri dish containing 6 to 10% HF. If the concentration is too high, the replica may disintegrate into small pieces. To clean the replicas of any biological material adhering to the replica undersurface, you may have to transfer to 70% sulphuric acid. Another choice would be 1- 2 M NaOH. After a few hours, the replicas should be transferred to distilled water and then to 400 mesh TEM grids. Any remaining water should be removed with filter paper. A good technical reference is Zeile, U. (2000) Fundamentals of cryo preparation and replica technique. 7th Asia-Pacific Electron Microscopy Conference. Reprints are available from Bal-Tac. However, other methods to avoid the creation of artefacts in biological specimens is to use an Environmetal SEM (LV-SEM, VP-SEM) or the cryo method for rapid freezing (plunge freezing, jet-freezing, or high pressure freezing) and the use a freeze-fracture system for Pt-C replicas.
-- Kelly
} } Dear Steve } If it's Pt or Pt/C coating to remove replica from the glass you may } use some HF solution. I don't remember exact concentration, but I } would think, 10% (v/v) should work. If not, you may try higher } concentration. Please, keep in mind: HF is extremely aggressive } chemical: it's strong irritant and will corrode everything on it's way } including SS EM tweezers. It should be stored in the plastic (NO } GLASS) and used with precautions. Basically, you use the same } technique as for floating Formvar film except everything should be } plastic and HF solution instead of water. When replica is detached } from the glass, you may move it with Pt loop to the water to wash it a } few times and then mount on the grids. Good luck. Sergey. } } At 12:55 PM 4/21/2003, you wrote: } } } } } } } Dear all } } } } I have a user who wants to make TEM carbon platinum replicas of } } mammalian cells growing on glass slides with attached tissue tek } } culture chambers. He figures to fix the cells, dehydrate, critical } } point dry, then coat the slides (minus the growth chambers) and float } } the replicas off the slide the same way he floats formvar off a slide } } when making coated grids. In his first attempt, he is unable to } } float the metal coating off the slide. A query to this listserver } } quite a while back mentioned using hydrofluoric acid to remove the } } replica. does anyone have details of this procedure, or preferably } } an alternative procedure that will enable my user to make his replicas? } } } } Thanks for any insights in this regard }
Mike; Yes, Nyquist was an optimist. The 44 hz frequency rate for CDs is for stereo, and the 22hz sampling rate per channel has been abandoned in the newer SACD standard which oversamples 8 times (that's 176 kz).
John Mardinly
John Mardinly Intel
-----Original Message----- } From: Mike Bode [mailto:mb-at-Soft-Imaging.com] Sent: Monday, April 21, 2003 1:11 PM To: 'Microscopy-at-MSA.Microscopy.Com'
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
} } FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD.
I'm afraid not. In fact, the FFT (of the image) is NEVER the same as the SAD. This is a point that I have always tried to stress when teaching at the NCEM Summer Schools.
The FFT (or image diffractogram) is always complex hermitian because it is the FT of a real function (the image intensity) -- that means that the intensities of the FFT (or image diffractogram) are always symmetric (as you point out) for ANY image at ANY specimen tilt or ANY misorientation.
For a crystalline specimen, producing strong Bragg spots in the diffraction pattern, the spots in the FFT (or image diffractogram) are well-defined combinations of interferences of the Bragg spots in the diffraction pattern. These interferences come from the auto-correlation (a kind of convolution) of the (complex) Bragg spot amplitudes (after the Bragg spot amplitudes have been modulated by the phase changes imposed by the objective lens). For a brief discussion of how the spots in the FFT (or image diffractogram, or image intensity spectrum) are formed from the Bragg spots in the diffraction pattern, see "Resolution-damping functions in non-linear images", M.A. O'Keefe, in 37th Ann. Proc. EMSA, San Antonio, Texas (1979) 556-557.
Of course, none of the above helps you with your orientation problem -- sorry.
Mike O'Keefe
$B4Z(J $BL-at-(J wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Ian MacLaren, } } I would greatly appreciate your reponse. I believe you spent much time answering my } stupid question. } } In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is true if Ewald } sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this sphere is not } a plane. Sometimes it brings some troubles to us. Let me show an example. If the specimen } normal is NOT parallel to incident beam, or if the orientation is NOT exact, the } intersections between relrod and sphere are not symmetric. In SAD pattern, not only } intensity but also the distance is different, in other words, SAD pattern is NOT } symmetric. On the other hand, FFT always symetric even though zone axis is not exact. } This let me believe FFT and SAD is not so identical. } } Thanks for your kind calculation. Near [111]Si orientation, we can find SAD pattern. As } you said, it is not an exact zone axis. I took three patterns with different tilting } angles (x=-9, +2, +8) and spent two weeks understanding these patterns. After } calibrating the tilting angles by Kikuchi pattens, my indexing results let me believe } they come from an identical reciprocal plane (130)*. I believe your calculation is right. } I guess the reasons why it is [130] rather than [230] maybe come from two points. 1. } beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. (640) spot is } too far from transmitted spot. 2. The epitaxial phase is under distorted state. The real } angle maybe different from those calculated based on bulk data. } } Till now I still think FFT is not completely identical to SAD. I believe the discussion } itself is significant. In my opinion, we can view Ewald sphere as a plane and based on } this approximation we can think FFT=SAD. But for epitaxy, we can not ignore this } approximation if we want to discuss some slight mismatch information. Maybe I made some } mistakes in my analysis. I hope to hear different sounds and look forward to hearing } from you again. } } Thanks again, } } Best wishes, } } Han } } } Dear Ming, } } This problem is in principle quite simple. Any diffraction pattern can be } } seen as a Fourier transform of the diffracting object. You can find this } } in any undergraduate optics textbook. Normally in TEM you will then have } } to convulute the periodicity of the periodicity of the object with a top- } } hat function describing its very limited thickness. This leads to the } } well-known reciprocal lattice rods for thin samples. } } } } So, an SAD pattern is a Fourier transform including specimen periodicity } } and specimen thickness. } } } } An HRTEM image can also be Fourier transformed. As you should know, } } however, the HRTEM image is strongly influenced by the contrast transfer } } function (i.e. highly defocus dependent). That means that the intensities } } in a Fourier-transform of a HRTEM image are strongly affected by image } } defocus. You see this effect particularly strongly for amorphous material. } } } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } } geometry, but may have rather different intensities in the different spots } } since the latter is so focus-dependent. } } } } That said, in your case the main point is that the geometry of the pattern } } is identical for SAD or for Fourier-transformed HRTEM. } } } } Now, if your orientation relationship is correct, then when you orient } } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } } matching the (111) Si planes. I reckon on a quick calculation that the } } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much too } } large to see the [110]beta diffraction pattern. But the axis [230]beta } } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } } diffraction pattern? At such a small angle off axis, you will still see } } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } } symmetric). } } } } Best wishes } } } } Ian MacLaren } }
The three-day intensive hands-on workshop on Image Processing and Measurement presented by John Russ (author of âThe Image Processing Handbookâ and âPractical Stereologyâ) through the North Carolina State University Department of Continuing and Professional Education is now in its 21st year. The course dates for 2002 are May 21 - 23 in Raleigh, NC. This course has generated highly favorable reviews from the thousands of previous students. The primary focus is on images from various types of microscopy, with practical guidance in correcting imaging defects, enhancing the images for presentation and measurement, and performing stereological meaningful measurements on them. Textbooks and computer software are provided to attendees. Lab sessions with an opportunity to bring your own images makes this course immediately useful and highly productive.
For full information on the course, including outlines, faculty information, a downloadable brochure, and on-line registration, go to http://members.AOL.com/IPCourse/ or call Cindy Allen at 919 515 8171. Act now, because only a few vacancies are still available.
SACD was launched in 1999 and is based on a different technology for the CDs (you won't be able to play the SAcDs on your regular CD player). I looked this up because, frankly, I had not even heard of SACD. They also claim they can reproduce sound up to 100 kHz (on the Sony SACD site). That's even too high for my dog, perhaps bats can hear that. And what speaker can reproduce that? But again: 176 kHz sampling rate for a 100 kHz signal is again roughly a factor of 2.
What they are saying is, that the reproduction of higher frequencies can produce better sound. That may well be. In order to sample the higher frequencies, they have to go to higher sampling rates, and they end up again at roughly Nyquist.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, April 22, 2003 10:42 AM To: Mike Bode; Microscopy-at-MSA.Microscopy.Com
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
I can't help Tobias, but if anybody out there has a need for a Panasonic LF-5090 (sold through Corel) I can let them have it for the cost of shipping.
This unit writes to LM-D501W disks (940 MB double sided - 470 MB per side). I don't have any spare media, though. I know it works as I lashed it together over the Christmas holidays to transfer all of my files from it to CD. I've never been able to get it to work under Windows, but it works under DOS on my old, original Compaq Portable just fine (my age is showing). I've got the ISA SCSI card that it seems to like, all the documentation, and several (I think) versions of the software (available at www.driverguide.com as well). It's just taking up space here, so if anybody needs it to keep an old piece of equipment viable, let me know.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
} Greetings, } In the early and mid 1990's I was the proud owner of a } Panasonic Optical Drive, model LF 7010, until it stopped working a } few years ago. I now find myself in the embarassing position of } wanting to get some data off a disk written by that drive (I could } have sworn we backed the stuff up on CD but if so the back up is } lost). The Panasonic drive was operated by a Mac, as a scsi device. } The disk in question is a 1.5 GB Panasonic Optical Disk (LM-R1500A). } I know this is a long shot, but is there anyone out there who might } be able to read this disk?? (Panasonic tech support only has PC's } nowadays, I checked with their Midwest branch). } } Thanks, } Tobias
Mike, you are missing the point: as pointed out by Professor Marks, just Nyquist just doesn't quite cut it.
John Mardinly Intel
-----Original Message----- } From: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Tuesday, April 22, 2003 10:41 AM To: 'Microscopy-at-MSA.Microscopy.Com' Cc: Mardinly, John
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
Several weeks ago, I asked on references of dose measurement in microscope. I am still looking for that. This time, I want to more specify my question. I would appreciate any comments you have.
Here, I simply consider current density instead of dose. There is a relation between current densities on specimen (Is) and Faraday cup (If).
Is = If x M x M
Where M is a magnification on the Faraday cup.
The relation between magnifications on Faraday cup (M) and film (m) are given by m/M = l/L where L indicates the distance between Projection Lens and Faraday cup, and l indicates the distance between Projection Lens and film. We know magnification on film (m) and can measure the current density on Faraday cup, so we can calculate the current density on the specimen.
I am looking for a textbook or paper that described the method I wrote above. Please advise.
Thank you,
Hiromi Konishi, Ph.D Dept. of Earth and Planetary Sciences The University of New Mexico
i am sure you will get dozens of responses, all with good advice. the only thing i would mention is some early work we did with chlamydial elementary bodies. the problem was that the monoclonal antibody worked very well with the western blot and, supposedly with immunofluorescent microscopy. i tried making my own gold and labelling it directly with the antibody, using protein A as an interface with the gold and antibody, and indirect, with purchased gold anti-mouse. none of them worked. my only explaination is, to this day, that the problem with monoclonal antibodies is accessability and conformation of their target site. if it is inside the cell, or buried in a hydrophobic fold or globular structure, the antibody will not work. it sounds as if one answer is that your antigenic target is not accessable. for what it is worth, i have had good success with each of the gold labelling methods, direct Ab to gold, using protein A, or using store bought stuff. and the store bought stuff is soooooo much easier because you do not have to make it yourself.
i would suggest that you try repeating the work with polyclonal antisera. i have always had much better success with them. the chances of an antibody which will recognize an accessable site is much greater.
paul hazelton, PhD EM Unit Medical Microbiology and Infectious Diseases University of Manitoba Winnipeg, Manitoba, Canada phone 204-789-3313 fax: 204-789-3926 pager: 204-931-9354
Ok, John, maybe I AM not getting the point, but here is as I see it:
1) Both Nyquist and Shannon calculated the THEORETICAL sampling frequency to measure a frequency without introducing artifacts. They both arrive at 2X. In a practical setting (camera with noise), the frequency may well be higher. How much a few percent noise will add to the sampling frequency I don't know, but my guess is, it's not a lot.
2) You mentioned the new SACDs. They use a higher sampling frequency to be able to reproduce higher acoustic frequencies because the claim is, that the music sounds better then (which I can't make any statement about). However, their own web site claims, that they can reproduce up to 100 kHz. With a 176 kHz sampling frequency, they are staying BELOW Nyquist and claim they can reproduce 100 kHz sound. I am not saying that they can or can't or that it sounds better or worse, just an observation that the guys at Sony and Philips use numbers similar to Nyquist.
Maybe we should move this discussion off-line before we start boring people.
I'd be interested to hear if you have any calculations or papers that show the effects of noise and/or pixel shape on the resolution.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, April 22, 2003 2:27 PM To: Mike Bode; Microscopy-at-MSA.Microscopy.Com
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
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Hello all,
IŽm interested in labelling bacterial surfaces with colloidal gold for negative stain and TEM. So far negative stain works OK, also indirect immunofluorescence with a primary mouse monoclonal. However, secondary antibodies conjugated with 10-15 nm gold doesnŽt work at all. We have adhered fimbriated E Coli to Formvar-coated gold grids with or without fixation in various concentrations of formaldehyde, labelled them and fixed with glutaraldehyde. Any tips and tricks?? Regards, Margaretha
--
Margaretha Lindroth, Ph D Phone: 4613-222616 Dept of Medical Microbiology Fax: 4613-224789 Faculty of Health Sciences e-mail: Margaretha.Lindroth-at-hul.liu.se Linköpings Universitet SE-581 85 Linköping
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Dear Ian MacLaren,
I would greatly appreciate your reponse. I believe you spent much time answering my stupid question.
In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this sphere is not a plane. Sometimes it brings some troubles to us. Let me show an example. If the specimen normal is NOT parallel to incident beam, or if the orientation is NOT exact, the intersections between relrod and sphere are not symmetric. In SAD pattern, not only intensity but also the distance is different, in other words, SAD pattern is NOT symmetric. On the other hand, FFT always symetric even though zone axis is not exact. This let me believe FFT and SAD is not so identical.
Thanks for your kind calculation. Near [111]Si orientation, we can find SAD pattern. As you said, it is not an exact zone axis. I took three patterns with different tilting angles (x=-9, +2, +8) and spent two weeks understanding these patterns. After calibrating the tilting angles by Kikuchi pattens, my indexing results let me believe they come from an identical reciprocal plane (130)*. I believe your calculation is right. I guess the reasons why it is [130] rather than [230] maybe come from two points. 1. beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. (640) spot is too far from transmitted spot. 2. The epitaxial phase is under distorted state. The real angle maybe different from those calculated based on bulk data.
Till now I still think FFT is not completely identical to SAD. I believe the discussion itself is significant. In my opinion, we can view Ewald sphere as a plane and based on this approximation we can think FFT=SAD. But for epitaxy, we can not ignore this approximation if we want to discuss some slight mismatch information. Maybe I made some mistakes in my analysis. I hope to hear different sounds and look forward to hearing from you again.
Thanks again,
Best wishes,
Han
} Dear Ming, } This problem is in principle quite simple. Any diffraction pattern can be } seen as a Fourier transform of the diffracting object. You can find this } in any undergraduate optics textbook. Normally in TEM you will then have } to convulute the periodicity of the periodicity of the object with a top- } hat function describing its very limited thickness. This leads to the } well-known reciprocal lattice rods for thin samples. } } So, an SAD pattern is a Fourier transform including specimen periodicity } and specimen thickness. } } An HRTEM image can also be Fourier transformed. As you should know, } however, the HRTEM image is strongly influenced by the contrast transfer } function (i.e. highly defocus dependent). That means that the intensities } in a Fourier-transform of a HRTEM image are strongly affected by image } defocus. You see this effect particularly strongly for amorphous material. } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } geometry, but may have rather different intensities in the different spots } since the latter is so focus-dependent. } } That said, in your case the main point is that the geometry of the pattern } is identical for SAD or for Fourier-transformed HRTEM. } } Now, if your orientation relationship is correct, then when you orient } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } matching the (111) Si planes. I reckon on a quick calculation that the } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much too } large to see the [110]beta diffraction pattern. But the axis [230]beta } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } diffraction pattern? At such a small angle off axis, you will still see } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } symmetric). } } Best wishes } } Ian MacLaren }
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The Pt-C replicas can be detached from the glass substrate by submerging the specimen at a shallow angle (} 45º) into a small petri dish containing 6 to 10% HF. If the concentration is too high, the replica may disintegrate into small pieces. To clean the replicas of any biological material adhering to the replica undersurface, you may have to transfer to 70% sulphuric acid. Another choice would be 1- 2 M NaOH. After a few hours, the replicas should be transferred to distilled water and then to 400 mesh TEM grids. Any remaining water should be removed with filter paper. A good technical reference is Zeile, U. (2000) Fundamentals of cryo preparation and replica technique. 7th Asia-Pacific Electron Microscopy Conference. Reprints are available from Bal-Tac. However, other methods to avoid the creation of artefacts in biological specimens is to use an Environmetal SEM (LV-SEM, VP-SEM) or the cryo method for rapid freezing (plunge freezing, jet-freezing, or high pressure freezing) and the use a freeze-fracture system for Pt-C replicas.
-- Kelly
} } Dear Steve } If it's Pt or Pt/C coating to remove replica from the glass you may } use some HF solution. I don't remember exact concentration, but I } would think, 10% (v/v) should work. If not, you may try higher } concentration. Please, keep in mind: HF is extremely aggressive } chemical: it's strong irritant and will corrode everything on it's way } including SS EM tweezers. It should be stored in the plastic (NO } GLASS) and used with precautions. Basically, you use the same } technique as for floating Formvar film except everything should be } plastic and HF solution instead of water. When replica is detached } from the glass, you may move it with Pt loop to the water to wash it a } few times and then mount on the grids. Good luck. Sergey. } } At 12:55 PM 4/21/2003, you wrote: } } } } } } } Dear all } } } } I have a user who wants to make TEM carbon platinum replicas of } } mammalian cells growing on glass slides with attached tissue tek } } culture chambers. He figures to fix the cells, dehydrate, critical } } point dry, then coat the slides (minus the growth chambers) and float } } the replicas off the slide the same way he floats formvar off a slide } } when making coated grids. In his first attempt, he is unable to } } float the metal coating off the slide. A query to this listserver } } quite a while back mentioned using hydrofluoric acid to remove the } } replica. does anyone have details of this procedure, or preferably } } an alternative procedure that will enable my user to make his replicas? } } } } Thanks for any insights in this regard }
Mike, You are correct in your statement about Nyquist & Shannon, but this is not in fact how a camera works. If you take an image I(r), and measure it ONLY at a set of points (and nowhere else) what you said is completely true. (This is the case that they considered.) However, what a camera actually does is measure the integrated intensity over a pixel. This integration is equivalent to a convolution in the image plane, i.e. it introduces an envelope that modifies what you have in reciprocal (Fourier) space. If it is a square pixel this is a sinc function along the x & y directions; if you had a round pixel it would be a Bessel-type function (J1(ar)/ar if I remember right). When you say 2% noise, you have also to be a bit careful. If the measurement noise for a single pixel is 2%, you will get 2% noise at the Nyquist limit. If you take a coarser frequency, say twice this (in the image) the average noise is 2%/sqrt(2) - less. (Similarly for other frequencies). This gives the classic 1/f noise.
On Tue, 22 Apr 2003, Mike Bode wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ok, John, maybe I AM not getting the point, but here is as I see it: } } 1) Both Nyquist and Shannon calculated the THEORETICAL sampling frequency to } measure a frequency without introducing artifacts. They both arrive at 2X. } In a practical setting (camera with noise), the frequency may well be } higher. How much a few percent noise will add to the sampling frequency I } don't know, but my guess is, it's not a lot. } } 2) You mentioned the new SACDs. They use a higher sampling frequency to be } able to reproduce higher acoustic frequencies because the claim is, that the } music sounds better then (which I can't make any statement about). However, } their own web site claims, that they can reproduce up to 100 kHz. With a 176 } kHz sampling frequency, they are staying BELOW Nyquist and claim they can } reproduce 100 kHz sound. I am not saying that they can or can't or that it } sounds better or worse, just an observation that the guys at Sony and } Philips use numbers similar to Nyquist. } } Maybe we should move this discussion off-line before we start boring people. } } I'd be interested to hear if you have any calculations or papers that show } the effects of noise and/or pixel shape on the resolution. } } mike } }
SUMMER I 2003 COURSE ANNOUNCEMENT - Scanning Electron Microscopy (BIO. 222-Section BA)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I, 2003 semester, course in Biological Scanning Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered four days per week (Monday through Thursday) between the hours of 8:00 am and NOON. Classes will begin on May 27 and end on June 26, 2003.
This is a "hands-on" course emphasizing biological specimen preparation, student operation of the SEM (Hitachi S-2400) and the production of electron micrographs through the process of black & white photography and digital image capture, along with electron micrograph analysis. Students will work on a number of biological samples with the goal of producing a portfolio of high quality SEM photomicrographs.
The course is widely transferrable and the cost per credit is reasonable at $106 per credit (for Nassau County residents or New York State residents with a certificate of residency).
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and student gallery of EM photomicrographs is available at our web site. The URL is {http://www.ncc.edu/users/becks} .
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students.
If you have further questions, you should e-mail me directly at the address below.
For information about mail or telephone registration (Dial-a-Course) point your browser to http://www.sunynassau.edu/courses/sum03/pdf/dialacourse.pdf . The phone registration option is available until 5/1/03 by calling 516-572-7131 or 7372 or 7425.
P.S. A Fall 2003 TEM course is also being offered (BIO 221 - Section E2) on Thursday evenings beginning at 5:30 PM.
Stephen J. Beck Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
I have included the formulas I used to calculate the sampling frequencies for digital microscopy on my webpage.
The calculation is based on the Rayleigh formula and the Nyquist sampling rate is twice the resolution given by the Rayleigh formula, the Rayleigh formula gives a better approximation for high N.A. lenses than the Abbe formula. For the wavelength I have used green light at a wavelength (lambda) of 0.520 micrometer (um). The image pixel-unit is calculated from the CCD pixel size and the total magnification.
Abbe formula for resolving power of a microscope: d = lambda / 2 * N.A. Rayleigh formula for resolving power of a microscope: d = 0.61 * lambda / N.A.
I use the Rayleigh formula in another form for my own clarity: Rayleigh (um) = 1.220 * lambda(um) / ( 2.0 * Numerical Aperture ) Nyquist samplingrate = 0.5 * Rayleigh / image pixel-unit
} From this formula you can understand that turning down the light of a microscope by lowering the voltage of the lightsource will reduce the resolving power. The light will become "reddish" and the wavelength will shift from 520 to +/- 800. Using more "blueish" ligh will improve the resolving power for the microscope. You should use neutral density filters to reduce the amount of light instead of lowerin the voltage of your lightsource.
Using cameras with smaller pixelsizes will allow a lower magnification, which will increase the amount of light falling on the CCD chip in fluorescence microscopy where the intensity (I) is given by:
I = N.A.^4 / Mag.^2
However larger CCD pixel sizes give you a bigger "bucket", which allows to capture more photons.
As usual there is a lot more to say, but I stop here.
P.S.: in the discussion of the appropriate sampling rate, people tend to forget that for sound one also has to sample the harmonics. Musical instruments do not create a "pure" sound, but create complex sounds, composed by interacting harmonics.
I think we definitely shouldn't move the discussion offline when it's getting interesting.
I don't understand Prof. Marks' point about noise and the need for oversampling.
The sampling theorem states that any scene with a maximum frequency can be represented without loss by samples taken at twice that frequency. (That means two samples per shortest wavelength in the original scene.) This requirement that the scene is bandlimited also applies to whatever noise is present in the original scene or is added somewhere in the recording process before the actual sampling takes place. The effect of the finite size of the detector pixels is simply to multiply the Fourier transform of the image with a sinc function (for square pixels). Since this sinc function doesn't go to zero below the Nyqist frequency no information is lost and the correct amplitudes of all Fourier components can be easily recovered. (I'm assuming that the physical pixel size is not bigger than the distance between samples, so no overlap.) Assuming this correction is done the sampling theorem seems to state that the original signal including whatever noise there is can be perfectly reconstructed from the mentioned samples. I don't see why noise would require oversampling as long as it is also bandlimited. Actually I don't see how you distinguish between noise and signal from just one image. The sampling theorem will reconstruct whatever it samples, no matter whether that happens to be signal or noise.
I've heard about the need to oversample in a different context, namely when images need to be interpoated in order to align and average them. I have equally big problems with that. I wonder if anybody can point me to a publication?
Philip
----- Original Message ----- } From: "L. D. Marks" {ldm-at-risc4.numis.nwu.edu} To: "Mike Bode" {mb-at-Soft-Imaging.com} Cc: "'Mardinly, John'" {john.mardinly-at-intel.com} ; "'Microscopy-at-MSA.Microscopy.Com'" } } } Mike, } You are correct in your statement about Nyquist & Shannon, but } this is not in fact how a camera works. If you take an image I(r), and } measure it ONLY at a set of points (and nowhere else) what you said } is completely true. (This is the case that they considered.) However, } what a camera actually does is measure the integrated intensity over a } pixel. This integration is equivalent to a convolution in the image plane, } i.e. it introduces an envelope that modifies what you have in reciprocal } (Fourier) space. If it is a square pixel this is a sinc function along the } x & y directions; if you had a round pixel it would be a Bessel-type } function (J1(ar)/ar if I remember right). } } } } Ok, John, maybe I AM not getting the point, but here is as I see it: } } } } 1) Both Nyquist and Shannon calculated the THEORETICAL sampling frequency to } } measure a frequency without introducing artifacts. They both arrive at 2X. } } In a practical setting (camera with noise), the frequency may well be } } higher. How much a few percent noise will add to the sampling frequency I } } don't know, but my guess is, it's not a lot.
as you are looking at the surface of the root, you can just fix the root in 95% Ethanol, then to several times 100% ethanol. Then either you can go CPD or HMDS before coating. Aldehyde fixation can preserve well the organells, but I dont think it matters so much for your goals. I know a SEM expert who used this method to study the morphology of tree buds.
Good luck
Haixin Xu (Ph.D) Biological Sciences University of Maryland Baltimore County On Tue, 22 Apr 2003, Tobias Baskin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings, } Fixation will distort the organ, no matter what. But you } should still be able to compare your samples. Aldehydes give the } best fixation, as a rule, but there is no reason to use cacodylate } for your purpose (actually, some would say no reason ever, but that } is another story). Plant roots can be fixed in FAA, dehydrated to } absolute ethanol, critically point dried, coated with metal, and } viewed. As an alternative to FAA, you could use 4% paraformaldehyde } in say PBS (or the dilute buffer of your choice, near pH7), rinse in } the buffer, dehydrate etc as above. I expect that 2 h in 4% } paraformaldehyde fix would be fine. You can check for shrinkage and } other untoward affects by looking at your fixed or dehydrated roots } under darkfield. } } Hope this helps, } Tobias } } } } } } } } A friend from our Soil Science department wants to compare roots from live } } plants grown in good soil and dead plants grown in poisoned soil. So far } } we have been looking at them under dark-field optical microscopy. It } } would be good if we could look at them under SEM, but we don't have an } } ESEM, and we don't want the root shape to collapse under vacuum } } dessication. } } } } When I have searched for root fixation I come across things like } } glutaraldehyde and cacodylate buffer, and we'd like to avoid using these } } if possible. Does anyone know simpler ways of fixing the external shape } } of roots? } } } } It would be nice if we could do the same thing for moulds, to show } } visiting students "this is what you get if you leave bread lying around in } } your accommodation!". } } } } Any ideas, please? } } } } +-----------------------------------------+ } } Robert H.Olley } } J.J.Thomson Physical Laboratory } } University of Reading } } Whiteknights } } Reading RG6 6AF } } England } } +----------------------------------------+ } } Phone:+44 (0) 118 9318572 } } Fax: +44 (0) 118 9750203 } } University internal extension 7867 } } Email: R.H.Olley-at-reading.ac.uk } } URL: http://www.reading.ac.uk/~spsolley } } +----------------------------------------+ } } } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 Tucker Hall } / / / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ University of Missouri } / / / \ \ \ Columbia, MO USA } / / / \ \ \ 65211-7400 } / / ___ / \ \__/ \ ____ voice: 573-882-0173 } fax: 573-882-0123 } http://www.biosci.missouri.edu/Baskin/baskin_lab__home_page.htm }
If I only compare 2D primitive cell shapes between FFT and SAD, I think the statement I said last time is right, i.e. FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD.
Is it right? Thank you very much. I also look forward to hearing your kind comment.
Best wishes,
Han
} } } FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as } SAD. } } I'm afraid not. In fact, the FFT (of the image) is NEVER the same as the SAD. This is a } point that I have always tried to stress when teaching at the NCEM Summer Schools. } } The FFT (or image diffractogram) is always complex hermitian because it is the FT of a real } function (the image intensity) -- that means that the intensities of the FFT (or image } diffractogram) are always symmetric (as you point out) for ANY image at ANY specimen tilt or } ANY misorientation. } } For a crystalline specimen, producing strong Bragg spots in the diffraction pattern, the } spots in the FFT (or image diffractogram) are well-defined combinations of interferences of } the Bragg spots in the diffraction pattern. These interferences come from the } auto-correlation (a kind of convolution) of the (complex) Bragg spot amplitudes (after the } Bragg spot amplitudes have been modulated by the phase changes imposed by the objective } lens). For a brief discussion of how the spots in the FFT (or image diffractogram, or image } intensity spectrum) are formed from the Bragg spots in the diffraction pattern, see } "Resolution-damping functions in non-linear images", M.A. O'Keefe, in 37th Ann. Proc. EMSA, } San Antonio, Texas (1979) 556-557. } } Of course, none of the above helps you with your orientation problem -- sorry. } } Mike O'Keefe } } } $B4Z(J $BL-at-(J wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear Ian MacLaren, } } } } I would greatly appreciate your reponse. I believe you spent much time answering my } } stupid question. } } } } In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is true if Ewald } } sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this sphere is not } } a plane. Sometimes it brings some troubles to us. Let me show an example. If the specimen } } normal is NOT parallel to incident beam, or if the orientation is NOT exact, the } } intersections between relrod and sphere are not symmetric. In SAD pattern, not only } } intensity but also the distance is different, in other words, SAD pattern is NOT } } symmetric. On the other hand, FFT always symetric even though zone axis is not exact. } } This let me believe FFT and SAD is not so identical. } } } } Thanks for your kind calculation. Near [111]Si orientation, we can find SAD pattern. As } } you said, it is not an exact zone axis. I took three patterns with different tilting } } angles (x=-9, +2, +8) and spent two weeks understanding these patterns. After } } calibrating the tilting angles by Kikuchi pattens, my indexing results let me believe } } they come from an identical reciprocal plane (130)*. I believe your calculation is right. } } I guess the reasons why it is [130] rather than [230] maybe come from two points. 1. } } beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. (640) spot is } } too far from transmitted spot. 2. The epitaxial phase is under distorted state. The real } } angle maybe different from those calculated based on bulk data. } } } } Till now I still think FFT is not completely identical to SAD. I believe the discussion } } itself is significant. In my opinion, we can view Ewald sphere as a plane and based on } } this approximation we can think FFT=SAD. But for epitaxy, we can not ignore this } } approximation if we want to discuss some slight mismatch information. Maybe I made some } } mistakes in my analysis. I hope to hear different sounds and look forward to hearing } } from you again. } } } } Thanks again, } } } } Best wishes, } } } } Han } } } } } Dear Ming, } } } This problem is in principle quite simple. Any diffraction pattern can be } } } seen as a Fourier transform of the diffracting object. You can find this } } } in any undergraduate optics textbook. Normally in TEM you will then have } } } to convulute the periodicity of the periodicity of the object with a top- } } } hat function describing its very limited thickness. This leads to the } } } well-known reciprocal lattice rods for thin samples. } } } } } } So, an SAD pattern is a Fourier transform including specimen periodicity } } } and specimen thickness. } } } } } } An HRTEM image can also be Fourier transformed. As you should know, } } } however, the HRTEM image is strongly influenced by the contrast transfer } } } function (i.e. highly defocus dependent). That means that the intensities } } } in a Fourier-transform of a HRTEM image are strongly affected by image } } } defocus. You see this effect particularly strongly for amorphous material. } } } } } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } } } geometry, but may have rather different intensities in the different spots } } } since the latter is so focus-dependent. } } } } } } That said, in your case the main point is that the geometry of the pattern } } } is identical for SAD or for Fourier-transformed HRTEM. } } } } } } Now, if your orientation relationship is correct, then when you orient } } } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } } } matching the (111) Si planes. I reckon on a quick calculation that the } } } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much too } } } large to see the [110]beta diffraction pattern. But the axis [230]beta } } } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } } } diffraction pattern? At such a small angle off axis, you will still see } } } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } } } symmetric). } } } } } } Best wishes } } } } } } Ian MacLaren } } } }
*********************************** Dr. Ming HAN Nanomaterials Laboratory National Institute for Materials Science 3-13, Sakura Tsukuba 305-0003, Ibaraki Japan Tel: +81-29-863-5548 Fax: +81-29-863-5616 E-mail: HAN.Ming-at-nims.go.jp http://www.nims.go.jp/nano_char/index.html
-- I have been asked to under take a project looking at junctional complexes in the lens of mouse eye. I understand, from personal conversations, that lens is very difficult to fix and embed for TEM. A combination fixative, including Picric acid, is recommended in the literature (Sakai et al 1991). Do any of you listers have experience of handling and processing of mouse lens for ultrastructural TEM studies?
Thanks for your kind comments. It is the reason that I propose this question. Actually I can not tilt the specimen to get both exact zone axes for epitaxial phase and substrate.
Best wishes,
Han
} Han, } another point to consider is that in cases of mismatched epitaxy } such as this, there will inevitably be some tilt between the different } crystals, giving a small deviation from the nominal orientation relationship } (typically {5 degrees). This is a combination of misfit strain and the } dislocations which are present at interfacial steps, which produce a } low-angle grain boundary type misorientation. } } Richard } } _______________________________ } Richard Beanland } Analytical Services } Bookham Technology plc } Caswell, } Towcester, } Northamptonshire NN12 8EQ } UK } Tel: +44 (0) 1327 356362 } Fax: +44 (0) 1327 356775 } http://www.bookham.com } } } } -----Original Message----- } From: HAN.Ming-at-momokusa.nims.go.jp [mailto:HAN.Ming-at-momokusa.nims.go.jp] } Sent: 22 April 2003 16:51 } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: EDP and FFT } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Ian MacLaren, } } I would greatly appreciate your reponse. I believe you spent much time } answering my } stupid question. } } In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is } true if Ewald } sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this } sphere is not } a plane. Sometimes it brings some troubles to us. Let me show an example. If } the specimen } normal is NOT parallel to incident beam, or if the orientation is NOT exact, } the } intersections between relrod and sphere are not symmetric. In SAD pattern, } not only } intensity but also the distance is different, in other words, SAD pattern is } NOT } symmetric. On the other hand, FFT always symetric even though zone axis is } not exact. } This let me believe FFT and SAD is not so identical. } } Thanks for your kind calculation. Near [111]Si orientation, we can find SAD } pattern. As } you said, it is not an exact zone axis. I took three patterns with different } tilting } angles (x=-9, +2, +8) and spent two weeks understanding these patterns. } After } calibrating the tilting angles by Kikuchi pattens, my indexing results let } me believe } they come from an identical reciprocal plane (130)*. I believe your } calculation is right. } I guess the reasons why it is [130] rather than [230] maybe come from two } points. 1. } beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. } (640) spot is } too far from transmitted spot. 2. The epitaxial phase is under distorted } state. The real } angle maybe different from those calculated based on bulk data. } } Till now I still think FFT is not completely identical to SAD. I believe the } discussion } itself is significant. In my opinion, we can view Ewald sphere as a plane } and based on } this approximation we can think FFT=SAD. But for epitaxy, we can not ignore } this } approximation if we want to discuss some slight mismatch information. Maybe } I made some } mistakes in my analysis. I hope to hear different sounds and look forward to } hearing } from you again. } } Thanks again, } } Best wishes, } } Han } } } } Dear Ming, } } This problem is in principle quite simple. Any diffraction pattern can be } } } seen as a Fourier transform of the diffracting object. You can find this } } in any undergraduate optics textbook. Normally in TEM you will then have } } to convulute the periodicity of the periodicity of the object with a top- } } hat function describing its very limited thickness. This leads to the } } well-known reciprocal lattice rods for thin samples. } } } } So, an SAD pattern is a Fourier transform including specimen periodicity } } and specimen thickness. } } } } An HRTEM image can also be Fourier transformed. As you should know, } } however, the HRTEM image is strongly influenced by the contrast transfer } } function (i.e. highly defocus dependent). That means that the intensities } } } in a Fourier-transform of a HRTEM image are strongly affected by image } } defocus. You see this effect particularly strongly for amorphous } material. } } } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } } geometry, but may have rather different intensities in the different spots } } } since the latter is so focus-dependent. } } } } That said, in your case the main point is that the geometry of the pattern } } } is identical for SAD or for Fourier-transformed HRTEM. } } } } Now, if your orientation relationship is correct, then when you orient } } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } } } matching the (111) Si planes. I reckon on a quick calculation that the } } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much } too } } large to see the [110]beta diffraction pattern. But the axis [230]beta } } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } } diffraction pattern? At such a small angle off axis, you will still see } } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } } symmetric). } } } } Best wishes } } } } Ian MacLaren } } } } } } ======================================================================= } This e-mail is intended for the person it is addressed to only. The } information contained in it may be confidential and/or protected by } law. If you are not the intended recipient of this message, you must } not make any use of this information, or copy or show it to any } person. Please contact us immediately to tell us that you have } received this e-mail, and return the original to us. Any use, } forwarding, printing or copying of this message is strictly prohibited. } No part of this message can be considered a request for goods or } services. } ======================================================================= } Any questions about Bookham's E-Mail service should be directed to postmaster-at-bookham.com. }
Film Users: About a year ago we noticed a slight change in density of low dose micrographs on SO163 film which had been developed in full strength D19 for 12 minutes, the recommended protocol by Kodak. Specific information on the SO163 film change has been difficult to obtain but through a supplier we heard that there was a change in late 2001 (something to do with manufacturing process and raw material availability). It was suggested to us that a 10-15% change in developing time may be sufficient to compensate. To identify the film lots, if the first three digits of the eight digit emulsion number on the front of each multipak of film is 205 or higher, then that pack is "new" film. Additionally, we stumpled upon a couple of multipaks of SO163 film with an unusual emulsion number: the first four digits were 9801. This turned out to be a lot of the "old" or original film which was manufactured in Canada. Don
Donald Gantz Dept. Physiology & Biophysics Center Advanced Biomedical Research Boston Univ. School of Medicine Boston, MA 02118 Email: gantz-at-biophysics.bumc.bu.edu Phone: 617-638-4017 (voice mail)
The Facility for Electron Microscopy Research at McGill University is looking to purchase new web based calendar software for the reservation of instruments and equipment. Some of the needs that we have identified are:
- Pre-defined time-slots - Time slots that can be customized - Can create many reservation pages for all the instruments/equipment - Can customize time slots per instrument - Can assign individual user names and passwords for the booking of each instrument - Keep historical reservation information
Using the criteria listed above, I would appreciate any recommendations.
Thanks,
Kelly
--
S. Kelly Sears, Ph.D., B.F.A. Manager, Electron Microscopy Centre McGill University, 3640 University Street, Montreal, QC H3A 2B2 (T) 514.398.6334; (C) 514.576.1926; (F) 514.398.5047; (E) sksears-at-eps.mcgill.ca http://www.medicine.mcgill.ca/emcentre/s__kelly_sears.htm
Could someone please tell me what cellular components/substances that Toluidine Blue stains? (We use 1% Toluidine Blue O and 1% Sodium Tetraborate in 30% Ethanol on 1 micron-thick Durcupan sections.)
We are investigating histopathological changes in the mouse cochlea.
Thank you,
J.M. Lett
Harold W. Siebens Hearing Research Center Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
John, you are great to advertise himself. Is clear advertisement permitted on ListServer? Or opposite: responds on advertisement are not permitted? It sounds the case: I had troubles when respond on Cryo-EM course (Vancouver) advertisement. Have a good day. Sergey.
At 01:35 PM 4/22/03 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think, this discussion is very interesting and useful. Moreover: I think such discussions are most important things on our ListServer. Many thanks to all participants. Sergey.
At 03:00 PM 4/23/03 +0200, 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
120 film is a lot bigger than the little rectangles of x-ray film my dentist uses. Will the film even fit in the processor? I suggest you call Kodak on the phone and talk to them. If you want a machine to process roll film try a company called Jobo.
Geoff
Teresa Flores wrote:
} We have borrowed a Dent-X developer and have been trying to go online to } KODAK to see if previous users have imputed times for developing } Technical } Pan-120 Kodak film using a Dent-X automatic developer (Used in Dentist } office to developer x-rays). } There are three choices to choose from in the Dent-X auto-developer. } 4 & 1/2; 6 and Endo. } Any help would be appreciated, even from KODAK to let us know where we } could look up the correct time to use for the Technical Pan - 120 film } (used to take TEM photos in a Zeiss 109). } Any imput is greatly appreciated and will save rolls of film. } Teresa Flores } LSUHSC } EM Lab } Path Dept } New Orleans,LA } } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
We have borrowed a Dent-X developer and have been trying to go online to KODAK to see if previous users have imputed times for developing Technical Pan-120 Kodak film using a Dent-X automatic developer (Used in Dentist office to developer x-rays). There are three choices to choose from in the Dent-X auto-developer. 4 & 1/2; 6 and Endo. Any help would be appreciated, even from KODAK to let us know where we could look up the correct time to use for the Technical Pan - 120 film (used to take TEM photos in a Zeiss 109). Any imput is greatly appreciated and will save rolls of film. Teresa Flores LSUHSC EM Lab Path Dept New Orleans,LA
Having looked at several packages myself, I would appreciate any leads you are given that do not appear to the list. Here is a partial list of what I have examined so far that may match your criteria, though you did not specify what platform to run it on. In addition to your criteria we are looking for a package that does not require installation of a client, requires little administrative overhead does not cost a fortune, and requires only a few clicks to use. That said I have not found anything suitable, though the closest contender is perlcal. One other drawback to them all is in tracking sessions added in series. Suppose you sign-up for every M,W,F 3-4. This booking goes in as one "unit" If you later do not show up on day or need to adjust the time slot, all slots are affected. I have not found a way to individually edit a serial booking in any of the packages I have tried. Another nice feature is e-mail reminders. Anyway, there are a bunch out there. Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } "S. Kelly Sears" {sksears-at-eps.mcgill.ca} 04/23/03 03:50PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The Facility for Electron Microscopy Research at McGill University is looking to purchase new web based calendar software for the reservation of instruments and equipment. Some of the needs that we have identified are:
- Pre-defined time-slots - Time slots that can be customized - Can create many reservation pages for all the instruments/equipment - Can customize time slots per instrument - Can assign individual user names and passwords for the booking of each instrument - Keep historical reservation information
Using the criteria listed above, I would appreciate any recommendations.
Thanks,
Kelly
--
S. Kelly Sears, Ph.D., B.F.A. Manager, Electron Microscopy Centre McGill University, 3640 University Street, Montreal, QC H3A 2B2 (T) 514.398.6334; (C) 514.576.1926; (F) 514.398.5047; (E) sksears-at-eps.mcgill.ca http://www.medicine.mcgill.ca/emcentre/s__kelly_sears.htm
Over the years I have had to gradually revised the policy on announcements of all courses.
Originally only "non-profit" courses were allowed. Non-profit meant (to me ) you may charge costs but don't make any money, but there were loop holes which were constantly cropping up. You can envision any number if you think about how to declare you don't make a profit on a short course.
To be honest it was beginning to be alot of hassel with individuals as well as organizations saying that they were just covering all costs. I'm neither an auditor nor a lawyer so , in the long run and to try to be fair to everyone I changed the rules to treat all short course/workshop announcements equally.
Thus, it is currently the policy to allow a "single" short posting of an announcement for a short courses of interest to the Listserver community. (See theListserver FAQ)
The benefits to the general community of allowing a single posting of a course/workshop in my opinion are sufficient to warrant this change.
I will use my judgement and if I perceive any individual or organization abusing this single short announcement policy then I will take appropriate steps.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } Does anyone have a good contact for buying M-Bond 610 in Germany? } } Our previous contact seems to have gone dead. We ring the phone but } get no answer. This was Vishay measurements group. } } Any other ideas? } } Best wishes }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
We are considering the purchase of an Alto system and are interested in better understanding the capabilities and ease of use of these systems. Applications will involve, at least in part, polymers.
Feel free to contact me off-line.
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Ian MacLaren wrote: ============================================================== Does anyone have a good contact for buying M-Bond 610 in Germany?
Our previous contact seems to have gone dead. We ring the phone but get no answer. This was Vishay measurements group.
Any other ideas? ============================================================== M-Bond 610 has been available in Germany for many years via or long time distributor:
Mr.Adi Hassel Ing.-Büro fur Prozeßtechnik und Instrumentelle Analytik Connollystraße 29 D-80809 München 40
I am sure he would be more than happy to help you with your requirement for M-Bond 610! Details on M-Bond 610 are given on URL http://www.2spi.com/catalog/spec_prep/glue.shtml
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
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Au contraire my friend. We, at the Lab have Meeting Maker labwide. It is used for the reservation of meeting rooms and I think it would be great for the reservation of scope time. Didn't think of it until your posting. And there it was in the list of applications. I am checking into it today!
I have made a series of appointments, on my MM calendar, regarding a certain meeting I attend on Tuesday mornings. And I have gone in later to change some parameter. A box pops up asking me if this will affect this particular date or all future dates of this activity. Also, if the microscopist would like to have the investigator/scientist on hand while studying the sample, all one has to do is send a proposal to that person through Meeting Maker.
Annie --
+++++++++++++++++++++++++++++
R. Ann Bliss Technician, Chemistry and Materials Science Materials Science and Technology Division Lawrence Livermore National Laboratory
I have found a Roll-Film Apron (long plastic strip with wavy edges) for the 120 film that was made for the Kodak Roll-Film Tanks that are no longer available. It is in very good condition and will hold approx. 36 inches of film. If anyone, like Teresa Flores, would like to have it for the price of shipping or to TRADE for an Apron for 35mm film (mine are getting brittle) I will be willing to part with it.
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
I have found a Roll-Film Apron (long plastic strip with wavy edges) for the 120 film that was made for the Kodak Roll-Film Tanks that are no longer available. It is in very good condition and will hold approx. 36 inches of film. If anyone, like Teresa Flores, would like to have it for the price of shipping or to TRADE for an Apron for 35mm film (mine are getting brittle) I will be willing to part with it.
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
I have found a Roll-Film Apron (long plastic strip with wavy edges) for the 120 film that was made for the Kodak Roll-Film Tanks that are no longer available. It is in very good condition and will hold approx. 36 inches of film. If anyone, like Teresa Flores, would like to have it for the price of shipping or to TRADE for an Apron for 35mm film (mine are getting brittle) I will be willing to part with it.
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
} On Tuesday, April 22, 2003, at 08:51 AM, Han wrote: } } In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD } is true if Ewald } sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But } this sphere is not } a plane. Sometimes it brings some troubles to us. Let me show an } example. If the specimen } normal is NOT parallel to incident beam, or if the orientation is NOT } exact, the } intersections between relrod and sphere are not symmetric. In SAD } pattern, not only } intensity but also the distance is different, in other words, SAD } pattern is NOT } symmetric. On the other hand, FFT always symetric even though zone } axis is not exact. } This let me believe FFT and SAD is not so identical. } Dear Han, In addition, there are effects due to dynamical and secondary scattering that change the diffraction amplitudes (both magnitude and phase) so that they are not identical to the FFT of the scattering potential (the diffracting object). Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
I've been following the discussion of pixel number and empty mag with interest, although I haven't been able to pick out the take-home message (are 3 megapixels enough or do I need 5?).
Coincidently, during this discussion I was contacted by a large company that makes LCD computer monitors. At first they were merely interested in using one of my MicroAngela pictures to show off their new 9.2 megapixel LCD display. However, this evolved into a discussion of the usefulness of large, high-res displays for scientific use, such as microscopy.
I realized that I have a 1kx1k cooled CCD on my EFTEM, a couple of 3.2 megapixel cameras on light microscopes, my confocal users typically use 512x512 pixels, but we can up that, and I have completely forgotten the number of lines my A-to-D converter picks up from my FESEM, final size 3.2MB.
The first question is, are there cameras out there that people are using, such as on newer types of instruments, that approach anywhere near 9 megapixels?
Would there by any reason to have a 9.2 megapixel monitor (3840 x 2400 pixels), 20.2 inches, for viewing micrographs? Other than the fact that having a big, bright, expensive high-res monitor on your desk would make you smile, of course...!
Trying to apply some of Gary Radice's math, if a 3mp camera will give you a 7.8 inch print at 300 dpi, will that image displayed on your monitor at 72 dpi require at least a 32.5 inch monitor to see all the information?
Comments?
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Toluidine blue at highly alkaline pH is pretty non-specific. Most things are varing shades of blue. In dilute aqueous solutions tol blue stains a variety of intercellular secretory granules and extracellular components, depending on the (acidic) pH.
Geoff
Jaclynn Lett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
We are considering the purchase of an Alto system and are interested in better understanding the capabilities and ease of use of these systems. Applications will involve, at least in part, polymers.
Feel free to contact me off-line.
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
I did as David suggested and discovered that the Hopkins calendar appears to be a development that is at least introduced here:
http://www.macromedia.com/devnet/mx/dreamweaver/articles/aspnet_calendar_02. html Hopkins might even do better than charge heavily for the code, but even if they did, it would be worth a try to see how their IT folks implemented such a nice method.
In response to Scott's attempts to find a program that permits action on elements of a series, I go back to Outlook and its ability to calendar via mail/Exchange with calendars in public folders in which resources may be scheduled. Wherever that is used, however, would be private rather than web-based.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: David Elliott Ph.D. [mailto:David.Elliott-at-yale.edu] Sent: Thursday, April 24, 2003 9:08 AM To: Microscopy-at-sparc5.microscopy.com
I would suggest going to http://www.hopkinsmedicine.org/micfac/reservations.cfm check out there page. It may work for you. David
} } } }
} } } } } } } } } } ------------------------------------------------------------------- -- -- } } } } } - } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } } } America } } } } } To Subscribe/Unsubscribe -- } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } On-Line Help } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } } ------------------------------------------------------------------- -- -- } } } } } . } } } } } } } } } } } } } } } The Facility for Electron Microscopy Research at McGill University is } } } } } looking to purchase new web based calendar software for the } } } } } reservation of instruments and equipment. Some of the needs that we } } } } } have identified are: } } } } } } } } } } - Pre-defined time-slots } } } } } - Time slots that can be customized } } } } } - Can create many reservation pages for all the instruments/equipment } } } } } - Can customize time slots per instrument } } } } } - Can assign individual user names and passwords for the booking of } } } } } each instrument } } } } } - Keep historical reservation information } } } } } } } } } } Using the criteria listed above, I would appreciate any } } } } } recommendations. } } } } } } } } } } Thanks, } } } } } } } } } } Kelly } } } } } } } } } } } } } } } -- } } } } } S. Kelly Sears, Ph.D., B.F.A. } } } } } Manager, Electron Microscopy Centre } } } } } McGill University, 3640 University Street, Montreal, QC H3A 2B2 } } } } } (T) 514.398.6334; (C) 514.576.1926; (F) 514.398.5047; (E) } } } } } sksears-at-eps.mcgill.ca } } } } } http://www.medicine.mcgill.ca/emcentre/s__kelly_sears.htm
On Wednesday, April 23, 2003, at 04:08 PM, Robert Kayton wrote:
} A faculty member asked me if knew anything about an ultra-small gold } particle, for immunogold labeling in E.M. } } Any help is appreciated. } Dear Bob, EM supply vendors sell colloidal gold in various sizes, and this can be conjugated to antibodies for immunogold labeling. Depending on your requirements, you may be able to purchase secondary antibody already conjugated. (I assume that your antigen is not common enough that you could get primary antibody commercially.) Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Another group on campus that is handling scheduling for a research boat and is using this software "csCalendar". http://www.cgiscript.net/cgi-script/csNews/csNews.cgi?database=cgi.db&command=viewone&id=35. I haven't tried this yet but we are going to. I'll let you know how it works. Price is sure right.
Owen
At 08:10 AM 4/24/2003 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Oops - a 3840 x 2400 pixel, 20.2 inch display has more than 72 dpi. The 20.2 inch designation is diagonal, of course, and I didn't take the time to do the math, but the display has closer to 190 dpi.
Sorry!
Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
On Tuesday, April 22, 2003, at 03:39 PM, Hiromi Konishi wrote:
} Several weeks ago, I asked on references of dose measurement } in microscope. I am still looking for that. This time, I want to more } specify my question. I would appreciate any comments you have. } } Here, I simply consider current density instead of dose. } There is a relation between current densities on specimen (Is) } and Faraday cup (If). } } Is = If x M x M } } Where M is a magnification on the Faraday cup. } } The relation between magnifications on Faraday cup (M) and film } (m) are given by m/M = l/L where L indicates the distance between } Projection Lens and Faraday cup, and l indicates the distance } between Projection Lens and film. We know magnification on film } (m) and can measure the current density on Faraday cup, so we } can calculate the current density on the specimen. } } I am looking for a textbook or paper that described the method I } wrote above. } Please advise. } Dear Hiromi, I performed such a measurement a few years ago, but did not publish the results. Our shop fabricated a high-precision Faraday cup that could be mounted in the same plane as the film, so the mag calibrations taken on film would also apply to the FC. Since M is squared, it is important to use the calibrated mag, not just the nominal mag. It is also important to have a FC that has the correct aspect ratio so that backscattered electrons do not escape--a paper by J.N. Turner (and perhaps other authors) discusses this. I had a piece of low-Z material inserted into the bottom of the FC to reduce backscattering further; either carbon or polyethylene will do. I was able accurately to measure the area of the hole through which the electrons could enter the FC, which is, of course, important. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Using Tech Pan film there are a very wide range of results you can get depending on the developer, agitation and time.
Your best bet is to call Kodak support explain to them what you are doing and how you are processing the film. They can get you started with the right developer, time etc. With Tech Pan these you can spend a lot of time trying to get what you want by experimentation. You need to be very consistent with all your development technique.
USA Support phone numbers: http://www.kodak.com/global/en/service/contactKodak/kodakPhoneNumbers.jhtml# 0012
International contact phone numbers: http://www.kodak.com/include/international.shtml ----- Original Message ----- } From: "Teresa Flores" {tflore-at-lsuhsc.edu} To: "Histotechnologist communicat" {histonet-at-pathology.swmed.edu} Cc: {ListServer-at-MSA.Microscopy.Com} Sent: Wednesday, April 23, 2003 8:47 AM
Add one more to your list to check out... www-sched at http://wilfred.berkeley.edu/~gordon/www-sched-download
You can vew a real sample of it at: http://wilfred.berkeley.edu/~gordon/www-sched/sched.cgi?fac=test2 I won't give out the passwords to this one yet, as we are doing some work on it this weekend.
The new version allows multiple schedules for different instruments/rooms and has a really nice web based interface for administration. You can manage all the schedules, including customizing their appearance, schedulable hours, how many people can sign up on one spot... etc.
Let me know what you think, as we are doing final customizations to it this weekend. The older www-sched is stand alone, and is meant to work with one schedule, unless you are adept at Perl. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Thu, 24 Apr 2003, Scott Whittaker wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Having looked at several packages myself, I would appreciate any leads you } are given that do not appear to the list. Here is a partial list of what I } have examined so far that may match your criteria, though you did not } specify what platform to run it on. In addition to your criteria we are } looking for a package that does not require installation of a client, } requires little administrative overhead does not cost a fortune, and } requires only a few clicks to use. That said I have not found anything } suitable, though the closest contender is perlcal. One other drawback to } them all is in tracking sessions added in series. Suppose you sign-up for } every M,W,F 3-4. This booking goes in as one "unit" If you later do not show } up on day or need to adjust the time slot, all slots are affected. I have } not found a way to individually edit a serial booking in any of the packages } I have tried. Another nice feature is e-mail reminders. Anyway, there are a } bunch out there. Good luck } } Ical-http://www.brownbearsw.com/ical/icalpage.html unix/pc } Webcal-http://bulldog.tzo.org/webcal/webcal.html unix } perlcal-http://www.perlcal.com/ unix/pc } Meeting maker- http://www.meetingmaker.com/home.cfm pc } webevent-http://www.webevent.com/demo/index.html } } Scott Whittaker } Laboratories of Analytical Biology } Smithsonian Institution } National Museum of Natural History } PO Box 37012 MRC104 } Washington DC 20013-7012 } 202-357-1651 } } } } } } "S. Kelly Sears" {sksears-at-eps.mcgill.ca} 04/23/03 03:50PM } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } The Facility for Electron Microscopy Research at McGill University is } looking to purchase new web based calendar software for the reservation } of instruments and equipment. Some of the needs that we have identified } are: } } - Pre-defined time-slots } - Time slots that can be customized } - Can create many reservation pages for all the instruments/equipment } - Can customize time slots per instrument } - Can assign individual user names and passwords for the booking of each } instrument } - Keep historical reservation information } } Using the criteria listed above, I would appreciate any recommendations. } } Thanks, } } Kelly } } } -- } } S. Kelly Sears, Ph.D., B.F.A. } Manager, Electron Microscopy Centre } McGill University, 3640 University Street, Montreal, QC H3A 2B2 } (T) 514.398.6334; (C) 514.576.1926; (F) 514.398.5047; (E) } sksears-at-eps.mcgill.ca } http://www.medicine.mcgill.ca/emcentre/s__kelly_sears.htm } } } } }
Ian; I would have though that the electrons forming SAD spots also would have gone through the objective lens, and have similar CTF convolution, but the part of the specimen sampled would be different between FTF and SAD.
John Mardinly Intel
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: Tuesday, April 22, 2003 2:07 AM To: HAN.Ming-at-momokusa.nims.go.jp Cc: Microscopy Listserver
Dear Ming, This problem is in principle quite simple. Any diffraction pattern can be seen as a Fourier transform of the diffracting object. You can find this in any undergraduate optics textbook. Normally in TEM you will then have to convulute the periodicity of the periodicity of the object with a top- hat function describing its very limited thickness. This leads to the well-known reciprocal lattice rods for thin samples.
So, an SAD pattern is a Fourier transform including specimen periodicity and specimen thickness.
An HRTEM image can also be Fourier transformed. As you should know, however, the HRTEM image is strongly influenced by the contrast transfer function (i.e. highly defocus dependent). That means that the intensities in a Fourier-transform of a HRTEM image are strongly affected by image defocus. You see this effect particularly strongly for amorphous material.
So, an SAD pattern and a Fourier-transformed HRTEM image have the same geometry, but may have rather different intensities in the different spots since the latter is so focus-dependent.
That said, in your case the main point is that the geometry of the pattern is identical for SAD or for Fourier-transformed HRTEM.
Now, if your orientation relationship is correct, then when you orient along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are matching the (111) Si planes. I reckon on a quick calculation that the angle between the (110)beta plane normal and [110]beta is 13.4 °, much too large to see the [110]beta diffraction pattern. But the axis [230]beta lies only 1.86° from the (110)beta plane normal. Are you seeing this diffraction pattern? At such a small angle off axis, you will still see the diffraction pattern and an HRTEM image (even if itâs not very good or symmetric).
Best wishes
Ian MacLaren
------- Forwarded message ------- } From: HAN.Ming-at-momokusa.nims.go.jp To: Microscopy-at-sparc5.microscopy.com
The gold label in question is likely the 1.4nm Nanogold particle, available from Nanoprobes, Inc. and probably from other distributors. Nanoprobes Web address is http://www.nanoprobes.com
-- _______________________________________________________________________ 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-onid.orst.edu
The smallest gold particles known to me are "undecaGold" 0.8 nm diameter and "nanoGold", 1.4 nm diameter. Both do have reactive group to bind with different compounds (antibodies etc). I do believe both compounds are still manufactured by NanoProbe: http://www.nanoprobes.com Sergey
At 02:17 PM 4/24/2003, 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
Hello Nestor Does these changes mean that any institution including commercial may post a single advertisement on any course/workshop? It would be nice to establish some rules how to post such things. I would suggest to use some template, like: Course's title, date, place, short description and reference to particular WEB-site or contact person and positively NO self-advertising or words how successful that course was last year. "Short" should be short, right? Thanks. Sergey
At 05:47 AM 4/24/2003, 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
Not to beat a dead horse (no, not the Amray SEM--it is quite alive), does anyone have any experience with EDS detectors on the 1810/1910 flat lens Amray systems? I am trying to add EDS to this 1910FE system but am frustrated by WD, take-off angle and physical position.
I need to be able to do spot analysis at about 6mm, ROI analysis at 6-10mm and mapping at between 6-15mm. Is this possible or just a dream?
The flat lens has distinct advantages over conical lenses. But it also has disadvantages. Such is life...and SEMs.
All inputs, public and private are appreciated.
gary g. Microtechnics, Inc. http://www.microtechnics.com Granite Bay, CA 95746 916.791.8191
Hello everyone It's not absolutely necessary to spent money on expensive huge LCDs. I do find that even on my 19" LCD with 1280x1024 the letters sometime too small to read comfortably. In most cases we need more space, not resolution (we are talking about monitors). If you are running Win2K you may use two monitors simultaneously. They act altogether as one big monitor. It mean that you could place your menu bars in one place (monitor) and use another for fine work. If it's a case, people usually use expensive, good monitor as a working place and another cheaper (and usually smaller) to store icons, menu-bars etc... Mouse and other controls act as it's single monitor. Such set-up is very popular between graphic designers. Sergey
At 12:46 PM 4/24/2003, 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
Have you tried changing your font size to larger than it is?
gary g.
At 06:43 PM 4/24/2003, you wrote:
} Hello everyone } It's not absolutely necessary to spent money on expensive huge LCDs. I do } find that even on my 19" LCD with 1280x1024 the letters sometime too small } to read comfortably. In most cases we need more space, not resolution (we } are talking about monitors). If you are running Win2K you may use two } monitors simultaneously. They act altogether as one big monitor. It mean } that you could place your menu bars in one place (monitor) and use another } for fine work. If it's a case, people usually use expensive, good monitor } as a working place and another cheaper (and usually smaller) to store } icons, menu-bars etc... Mouse and other controls act as it's single } monitor. Such set-up is very popular between graphic designers. Sergey } } At 12:46 PM 4/24/2003, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It's actually quite easy to make colloidal gold with diameters between 3.5 and 15 nm using a method such as Slot, Geuze 1985, Europ. J. Cell Biology 38, 87-93.
If you want even smaller labels you need clusters such as "nanogold", which is about 1.5 nm in diameter (Hainfeld, Furuya 1992, J. Histochemistry and Cytochemistry 40, 177-184)
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em
----- Original Message ----- } From: "Bill Tivol" {tivol-at-caltech.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, April 24, 2003 11:17 PM
Yes, I could, but, for instance on some WEB-pages you could meet very small fonts and nothing to do with that. Personally, I am really happy with my monitor. My point was, that sometime we need more space, not necessary so many pixels when we are talking about monitors. Two monitors also is very economical solution. Sergey.
At 09:48 PM 4/24/2003, you wrote: } Have you tried changing your font size to } larger than it is? } } gary g. } } } } At 06:43 PM 4/24/2003, you wrote: } } } Hello everyone } } It's not absolutely necessary to spent money on expensive huge LCDs. I do } } find that even on my 19" LCD with 1280x1024 the letters sometime too } } small to read comfortably. In most cases we need more space, not } } resolution (we are talking about monitors). If you are running Win2K you } } may use two monitors simultaneously. They act altogether as one big } } monitor. It mean that you could place your menu bars in one place } } (monitor) and use another for fine work. If it's a case, people usually } } use expensive, good monitor as a working place and another cheaper (and } } usually smaller) to store icons, menu-bars etc... Mouse and other } } controls act as it's single monitor. Such set-up is very popular between } } graphic designers. Sergey } } } } At 12:46 PM 4/24/2003, 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
Hello: Well, our SEM with LaB6 started to behave very strange. Turning on high voltage creates an emission current even with cold filament. About 12 kV the curren is small so we can eventually work. Higher high voltage, lets say about 20 kV, causes so high emission current (with no filament heating, Filamet=0) that there is emergency turn off. Any hint or suggestion would be greatly appreciated. Maybe I should add that the filament works "only" two months.
I have several large monitors on my desk. Not so that I can look at images on the whole screen, but so that when I am looking at a high res image, I can still have all of the editing tools on the screen. You need more (sometimes, much more) space on your monitor that is needed for the image. David
On Thursday, April 24, 2003, at 03:46 PM, Tina Carvalho wrote:
} Would there by any reason to have a 9.2 megapixel monitor (3840 x 2400 } pixels), 20.2 inches, for viewing micrographs? Other than the fact that } having a big, bright, expensive high-res monitor on your desk would } make } you smile, of course...!
Yes, you're right. The diffraction pattern is in fact distorted by the objective lens spherical aberration, i.e. the phase distortion implied by the CTF is also a geometrical distortion in the pattern (wave optics and ray optics agree on this point).
But the amount of distortion in the pattern is typically tiny. It gets bigger for very high-order reflections, but is essentially not measurable under normal circumstances. What is important for imaging is that when the beams interfere to form the image, this tiny directional shift translates into very signifacant phase shifts of the Fourier components of the image.
For the story as relates to SADP, the details are covered well in Hirsch, section 1.8 'Accuracy of selected area diffraction'.
Regards,
Wharton **************************************************************** Wharton Sinkler, PhD. UOP LLC 25 E. Algonquin Rd. Des Plaines, IL 60017-5017 tel. 847-391-3878 fax. 847-391-3719 mailto Wharton.Sinkler-at-uop.com
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Thursday, April 24, 2003 7:13 PM To: ian.maclaren-at-physics.org; HAN.Ming-at-momokusa.nims.go.jp Cc: Microscopy Listserver
Dear Dr. Kayton:
Ultrasmall colloidal gold particles was first introduce by Dr. Jan Leunissen (The founder of Aurion) as a marker for immunogold labeling. Antibodies conjugated to +/- one nanometer gold particles became commercially available in late 80's. Currently there are several companies carrying this type of products with different trade names. The following is a list. Even though "Ultra small gold" was first and still is used by Aurion as the trade name for their products, the term has become popular and often been used casually to refer to the whole category of small gold conjugates.
Ultra Small Gold: Aurion Immunogold Reagents and Accessories (distributed in the US by Electron Microscopy Sciences, www.emsdiasum.com) AuroProbe One: Amersham Biosciences, (www.amersham.com) One nanometer gold: British BioCell International (distributed in the US by Ted Pella, www.tedpella.com) Nanogold, Nanoprobes (www.nanoprobes.com)
Hong
==================== Hong Yi Emory Neurology Microscopy Core Emory University School of Medicine 6215 Woodruff Memorial Research Building 1639 Pierce Drive Atlanta, GA 30322
List, To add to the discussion, ultrasmall gold has advantages over larger gold conjugates. The size of US-gold is on the order of the size of the antibody allowing much better penetration, especially in tissue such as vibratome sections of brain. It can also result in increased labeling density. The US-gold is then silver enhanced to increase its visibility in the TEM. Tom
Thomas Moninger (thomas-moninger-at-uiowa.edu) University of Iowa Central Microscopy Research Facility (www.uiowa.edu/~cemrf) View expressed are my own.
In one sentence, ultrasmall gold probe has much higher sensitivity (meaning that you can dilute your primary Ab an order of magnitude further down), and yet you can grow it by silver enhancement as big as you wish, even to be visible under LM. The developing part is now much easier than it used to be.
I thought this was unfair that noone mentioned Aurion (http://www.aurion.nl/). They make a line of ultrasmall (0.8 nm) conjugates which may actually be a better fit for a histo-related application. I of course have no material interest in Aurion or its US distributor, but their Goat Anti-Rabbit F(ab')2-Ultrasmall has really done wonders for two of my applications. You are welcome to contact me with more specific questions, Bob.
Vlad -- ------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
} } From: darryl krueger {dkruege-at-clemson.edu} } Subject: TEM single crystal sample } } I have a friend that is doing Diffraction, who is looking for a source for } single crystals on TEM grids. If there is source someone knows of could } you please let us know. TIA } } Darryl
Tina; That monitor has a 205 dpi pixel density, so the pixels are .0049 inches apart. The resolution of the human eye is approximately 1/60 of a degree but that is only at the center of the field of view. The density of rods and cones on the retina is less off axis, At 10 inches viewing distance (nearsighted people can get closer and farsighted and presbyopic people would be farther away, without glasses, but that is another optical element...). This works out to approximately .0029 inches, so you should be able to easily see the pixels when you are ten inches away from the monitor. However, I have difficulty holding my eyes 10 inches away from a monitor while operating a microscope... Another plus would be the ability to display six 1 megapixel images without any overlap, if you need that.
John Mardinly Intel
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Thursday, April 24, 2003 12:46 PM To: Microscopy Listserver
Hi, All-
I've been following the discussion of pixel number and empty mag with interest, although I haven't been able to pick out the take-home message (are 3 megapixels enough or do I need 5?).
Coincidently, during this discussion I was contacted by a large company that makes LCD computer monitors. At first they were merely interested in using one of my MicroAngela pictures to show off their new 9.2 megapixel LCD display. However, this evolved into a discussion of the usefulness of large, high-res displays for scientific use, such as microscopy.
I realized that I have a 1kx1k cooled CCD on my EFTEM, a couple of 3.2 megapixel cameras on light microscopes, my confocal users typically use 512x512 pixels, but we can up that, and I have completely forgotten the number of lines my A-to-D converter picks up from my FESEM, final size 3.2MB.
The first question is, are there cameras out there that people are using, such as on newer types of instruments, that approach anywhere near 9 megapixels?
Would there by any reason to have a 9.2 megapixel monitor (3840 x 2400 pixels), 20.2 inches, for viewing micrographs? Other than the fact that having a big, bright, expensive high-res monitor on your desk would make you smile, of course...!
Trying to apply some of Gary Radice's math, if a 3mp camera will give you a 7.8 inch print at 300 dpi, will that image displayed on your monitor at 72 dpi require at least a 32.5 inch monitor to see all the information?
Comments?
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Greetings, Does anyone have a protocol for immunolabeling intracellular constituents in small A. thaliana seedlings? I would like to label microtubules in a whole mount for confocal microscopy, but I anticipate that those nasty cell walls will pose a problem. Is there an effective way to permeablize the little weeds or is this a silly approach? Thanks in advance for any advice. Regards, Karl G.
On Thursday, April 24, 2003, at 12:46 PM, Tina Carvalho wrote:
Dear Tina,
} I've been following the discussion of pixel number and empty mag with } interest, although I haven't been able to pick out the take-home } message } (are 3 megapixels enough or do I need 5?). } } Coincidently, during this discussion I was contacted by a large company } that makes LCD computer monitors. At first they were merely interested } in } using one of my MicroAngela pictures to show off their new 9.2 } megapixel } LCD display. However, this evolved into a discussion of the usefulness } of } large, high-res displays for scientific use, such as microscopy. } } I realized that I have a 1kx1k cooled CCD on my EFTEM, a couple of 3.2 } megapixel cameras on light microscopes, my confocal users typically use } 512x512 pixels, but we can up that, and I have completely forgotten the } number of lines my A-to-D converter picks up from my FESEM, final size } 3.2MB. } } The first question is, are there cameras out there that people are } using, } such as on newer types of instruments, that approach anywhere near 9 } megapixels?
Yes. We are getting a 4k x 4k CCD. } } Would there by any reason to have a 9.2 megapixel monitor (3840 x 2400 } pixels), 20.2 inches, for viewing micrographs? Other than the fact that } having a big, bright, expensive high-res monitor on your desk would } make } you smile, of course...!
Yes, again--assuming that you wanted to look at several pics at the same time. } } Trying to apply some of Gary Radice's math, if a 3mp camera will give } you } a 7.8 inch print at 300 dpi, will that image displayed on your monitor } at } 72 dpi require at least a 32.5 inch monitor to see all the information?
Yes. However, if the 3mp was in the usual 3:2 ratio, the camera would be 1414 x 2121 pixels, so the image could be displayed at full resolution with a large border. Either the monitor is, indeed, as large as you calculate, or the display is at } 72 dpi. You might want to zoom in on a detail that wouldn't have apparent pixillation on the monitor, so the extra size could be warranted for that. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
On Thursday, April 24, 2003, at 05:13 PM, Mardinly, John wrote:
} I would have though that the electrons forming SAD spots also would } have gone through the objective lens, and have similar CTF } convolution, but the part of the specimen sampled would be different } between FTF and SAD. } Dear John, The electrons do go through the objective lens, and the lens is at precisely the same current for both SAD and SA imaging modes, but the projector lenses image the back focal plane in SAD, and there is no CTF convolution. This can easily be seen by taking an ED pattern of something like Au, where the intensities of the spots do not have too much structure--the unit cell is pretty simple--and taking the FFT of the corresponding image. Even if the objective lens is a little out of focus, you will not see the diffraction spots disappear at the zeros of the CTF, but you will see it in the FFT. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
} The smallest gold particles known to me are "undecaGold" 0.8 nm } diameter and "nanoGold", 1.4 nm diameter. } There is the problem, however, of seeing the gold in your specimen, and undecagold is not visible (unless it can be decorated). Furthermore, if the specimen is stained, the gold must be distinguished from the other heavy metals, so it must be large enough to have an identifiably distinct shape. If it just looks like the clusters of stain, it won't be very useful. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Karl, Oh this is not silly, you can do it. Of course, chewing up the cell wall takes its toll, but you can get some nice images. Several groups have published on this. You can look at Sugimoto et al 2000 Plant Physiology 124: 1493-1506 for a protocol that has been used fairly widely.
A couple of points.
If you grow arabidopsis on agar plates, keep the plates vertical to get straight roots, on the surface of the plate, and when their time has come pour the fix directly on the plates (put them horizontal first!). This is the gentlest way, the plants die happy, and they stay submerged. No need to mess about with vacuums. When you are ready to add antibody, a nice idea is to put a droplet, 50 microL say, on a piece of parafilm covering the bottom of a Petri dish. Then you can put 5 or so root tips in the same drop and lay the rest of the root and the cotyledons out across the parafilm. The cotyledons are handy for picking up the seedlings and moving them from solution to solution (ie for rinsing) so don't cut them off until you are ready to mount.
One change that I recommend from the Sugimoto protocol is to replace the EGTA in the fix buffer with a little bit of CaCl2 (say 0.5 mM). EGTA is toxic and rips calcium out of the membranes and generally wreaks havoc on the cell. Not a good idea.
Hope this helps. Please feel free to contact me if you have further questions.
As ever, Tobias
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi all, The discussion on camera pixel numbers has been interesting but I haven't the expertise to contribute anything but have learned something. Thanks to all contributors.
However, I did make the jump to dual monitors as Sergey describes and find it very helpful in several ways: 1) When using Photoshop I keep the tools and maybe another app windows on one monitor, and the image on the Photoshop desktop on the primary monitor. This is great also when you are documenting in another app every step in manipulating the image; 2) when working on more that one document; and 3) when answering email like this one.
I did run into one issue with using dual monitors. Depending on which type of video board you get, you could end up with the desktop spread over two monitors making mousing across a wide field an annoyance but allowing you to open two images to a fairly large size (depending on screen width) with no overlap. Other boards make one monitor the primary and the other a secondary so that the desktop is on one monitor and the other just a blank work area where you can open a second app or a duplicate app. An example of this is Matrox G450 versus Matrox G550, the former puts the desktop across both screens, the latter puts the desktop on the primary screen (of your choice). I still perfer the CRT for my primary Photoshop screen as I think it still gives slightly better colors than an LCD but the gap is closing fast.
Damian Neuberger
} } Hello everyone } It's not absolutely necessary to spent money on expensive huge LCDs. I do } find that even on my 19" LCD with 1280x1024 the letters sometime too small } to read comfortably. In most cases we need more space, not resolution (we } are talking about monitors). If you are running Win2K you may use two } monitors simultaneously. They act altogether as one big monitor. It mean } that you could place your menu bars in one place (monitor) and use another } for fine work. If it's a case, people usually use expensive, good monitor } as a working place and another cheaper (and usually smaller) to store } icons, menu-bars etc... Mouse and other controls act as it's single } monitor. Such set-up is very popular between graphic designers. Sergey
I have a friend that is doing Difraction, who is looking for a source for single crystals on TEM grids. If there is source someone knows of could you please let us know. TIA
Darryl Krueger wrote: ===================================================== I have a friend that is doing Difraction, who is looking for a source for single crystals on TEM grids. If there is source someone knows of could you please let us know. TIA ===================================================== Actually there are number of such "samples" depending on the spacing you are wanting to calibrate and measure. These are all available from SPI Supplies as well as some of our major competitors:
The single crystal catalase specimen is probably the most commonly requested , see URL http://www.2spi.com/catalog/standards/othertem11.shtml
Another popular such test sample is the molybdenum trioxide specimen, see URL http://www.2spi.com/catalog/standards/othertem19.shtml
We also have the chloro-Cu-phthalocyanine test specimen favored by some, see URL http://www.2spi.com/catalog/standards/othertem9.shtml
And finally, there is the gold crystal specimen, see URL http://www.2spi.com/catalog/standards/othertem4.shtml
Other single crystal specimens can be found on URL http://www.2spi.com/catalog/standards/othertem.html
I am sure that there are others but these seem to be the most popular of the various possibilities, perhaps because they are low cost and are reasonably robust and seem to last the typical user quite some time.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
In general a digital microsocope is calibrated by using grids and measuring the distance between two points on the grid. The sampling error made when calibrating the microscope and the time spent, made us think of another way of calibrating a microscope by using the physical proersties of the CCD grid of the camera and the properties of the digitizer/framegrabber.
I want to know if this procedure is reliable and which problems may arise ? The result of the calculation will be the "size" of one pixel and as such can be used to calibrate the system. This example is based on a traditional PAL video camera (NTSC differs) and a framegrabber, modern digital cameras will need a different approach of course.
First we need to determine the chip area used on the CCD for acquireing, we have to take care only to use the active chip area in our calculations and not the entire chip. The placement of the CCD chip in the lightbeam of the microsocope will have an influence on the final magnification also, and there is no "standard" position for various cameras and microscopes. In addition when a camera dosn't have rectangular pixels, we will have to take this into account.
e.g. A (B/W) camera with the following specifications and square pixels:
A 1/2 inch CCD and a CCD diameter of 18 mm with a 3/4 aspect ratio, an array size 800 x 581 pixels (of which 756 active horizontaly) will result in:
The effective area is 756 x 581 pixels and not 800 x 581, thus the aspect ratio is not 4/3 ( 768/576 ), so in the end this will give us a (square) pixel size of :
Which instrument do you use? Some manufacturers have a "overnight" heating system which runs the filament at half power when you have switched OFF the high voltage. In this way they keep the filament out gassed when it is not in operation, which is a very good idea.
Could this system have failed to switch over to normal filament operation when you switched the high voltage on?
Please tell us more?
Regards
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "Witold Zielinski" {WIZIEL-at-INMAT.PW.EDU.PL} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, April 25, 2003 11:28 AM
I am looking for a RMC MT 6000-XL specimen holder (the whole thing with arc segment mount etc., second hand or new). Anybody an idea where I could purchase one?
I guess I did not read you mail as well as I thought. I would suggest you have a microscope (Philips or FEI) that always has a degree of filament heating even if the "filament" supply is turned off!
With that in mind I think you need to clean the cathode. I believe you have a whisker shorting out the bias circuit. If you have a whisker of material between the filament and the cathode cap (Whenelt, or whatever you call it) the bias circuit is no longer in operation and the current is massive. Add to that the stand-by heating and you will easily obtain an image. Push the high voltage up too high and the safety circuit will trip in as you will pull too much current.
Sorry I did not spot this when I first read your mail.
Regards
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "Witold Zielinski" {WIZIEL-at-INMAT.PW.EDU.PL} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, April 25, 2003 11:28 AM
} From: Coetzee, Mr S. H Physics Science } Sent: Monday, April 28, 2003 12:24 PM } To: Listserver Microscopy (E-mail) } Subject: Needs assessment - EM project for kids } } } Dear All } I would love to call this e-mail: Needs assessment - video or DVD on EM } and LM under subject but due to juncmail filters I had to change } the subject to } get it to the list. } } Life is always interesting and full of fun when you are doing EM. } We are doing a needs assessment on the following: } "Introduction video/DVD and LM for kids" } We want to target 13 - 18 year old kids and hopefully be able to reach 1st } and second year students as well. } The idea is to improve the number of students in Science with the focus on } EM. Input will be greatly appreciated. } We need this input for fund application. } } } Stephan H Coetzee } Department of Physics } EMU } Private Bag 0704 } Gaborone, } Botswana } } Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} } } Telephone: (+267) 355 2462 } Fax: (+267) 3185 097 } Telephone: (+267) 355 0000 Switchboard } }
When you make an image, the electrons that are scattered by the sample in different directions have to be brought back together by the lens. The phase relation between the electrons that leave the sample in different directions is important. If it is wrong, the image will be wrong. The contrast transfer function tells you how the electrons in different directions are shifted in phase. It controls the quality of the image.
In diffraction, whether selected area diffraction or convergent beam diffraction (which would be a better choice nearly always), the electrons that leave the sample in different directions are never brought back together. They go to different places in the diffraction pattern. Therefore the phase shift has no effect on the pattern. The contrast transfer function plays no role. This is why a diffraction pattern is still good if the microscope is misaligned, but an image is greatly degraded if the microscope is misaligned.
Alwyn Eades -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
This is a commercial reply relative to the request from: Gary S. Laevsky, Ph.D. Research Associate The Scripps Research Institute " I'm looking for the "best" source for an objective warmer."
Below is a clip and paste from our brochure on objective heating. But first a word of caution to all on this subject.
Bioptechs developed the first patented commercial objective heating system over ten years ago. Now there are several companies making similar devices. There are several very important factors that should not be overlooked when selecting an objective warming device.
a. When heat is applied to an objective it must be applied efficiently to the central core (where the lenses are). This means that all of the heat must go into the objective. There can be no excess radiation. Reason, it will require X amount of energy to warm the objective above ambient plus a continuous amount of additional energy to overcome the heat-sink effect of the nosepiece. Therefore, if the heat is not applied efficiently, the heat that is not transferred to the objective will radiate out and upward toward your specimen. This excess heat can "cook" your specimen.
b. Not all objectives have the same thermal profile. Therefore, the control system must be able to compensate for these variations.
c. Not all objectives can be thermally regulated. When lenses are designed there is little, probably no, thought given by the manufacturers to the prospect of temperature control for live-cell observation. I think this is an oversight on their part but that is another story for another time. Further detailed information is available on the Bioptechs web site. http:.//www.bioptechs.com.
Snip The Problem: When high numeric aperture objectives are used to observe temperature sensitive specimens, heat from the specimen is transferred through the optical coupling medium (oil, glycerine, or water) to the colder objective. Therefore when live-cell microscopy requires the use of high numeric aperture lenses, it is necessary to control the temperature of the objective. The thermal mass of a fluid coupled objective is overwhelming when compared to the thermal mass of the cells. Unfortunately, microscope manufacturers do not offer temperature controlled objectives, nor do they consider the need for thermal regulation in the design of their objectives.
The Solution: To eliminate this thermal gradient, Bioptechs offers a patented Objective Heater System, which includes a heater/sensor and an electronic controller. The heater/sensor is comprised of an adjustable thin film heating band which surrounds 3/4 of the diameter of the upper region of the central retracting tube of the objective. A surface probe thermal sensor positioned in the gap formed between the ends of the heating band measures the temperature of the objective. This heater/sensor assembly is supported on an adjustable metal mounting to fit objectives ranging from 16 to 34mm in diameter. The heater loop requires a minimum of 5mm longitudinal, physical contact with the cylindrical objective surface. In some cases there may be a decorative collar on the objective which must be removed in order to permit adequate surface contact.
The Controller is specifically designed to slowly heat the objective over a fifteen-minute warm-up period, then hold the objective at the setpoint value within 0.2°C. The Controller operates from ambient to 50 °C and has special safety circuitry which utilizes a 0.9°C error window to shut down the Controller and sound an alarm if, for any reason, the temperature of the objective deviates after it has reached setpoint. A user calibration test is also built in to the Controller. End snip
Dan
-- Dan Focht Bioptechs 3560 Beck Road Butler, PA 16002 dan-at-bioptechs.com www.bioptechs.com V 724-282-7145 F 724-282-0745
I am not sure what you are getting at. Calibrating a microscope means, that you try to get rid of the errors and non-linearities of the optical system (lenses, etc.) and other influences that affect the true magnification of the system, so that you can do true measurements on an image. I have the impression, your calculation starts from known numbers about the chip size and ends up with a number for the pixel size, which you can also get from the vendor. If that is your goal, then yes, you can do it that way. However, you have now "calibrated" the chip or camera, but not the microscope.
If you think, that the errors introduced by the measurement of a standard are larger than those introduced by the lenses and other elements, you only need to take the total magnification between the sample and the camera chip and divide the pixel size by this number, and you arrive at the pixel calibration for the image.
Calibrating a microscope takes about 5 minutes, and you really don't need to redo it unless you change anything on the microscope (objectives, camera position, etc.).
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Peter Van Osta [mailto:pvosta-at-unionbio-eu.com] Sent: Monday, April 28, 2003 1:48 AM To: MSA
Hi,
In general a digital microsocope is calibrated by using grids and measuring the distance between two points on the grid. The sampling error made when calibrating the microscope and the time spent, made us think of another way of calibrating a microscope by using the physical proersties of the CCD grid of the camera and the properties of the digitizer/framegrabber.
I want to know if this procedure is reliable and which problems may arise ? The result of the calculation will be the "size" of one pixel and as such can be used to calibrate the system. This example is based on a traditional PAL video camera (NTSC differs) and a framegrabber, modern digital cameras will need a different approach of course.
First we need to determine the chip area used on the CCD for acquireing, we have to take care only to use the active chip area in our calculations and not the entire chip. The placement of the CCD chip in the lightbeam of the microsocope will have an influence on the final magnification also, and there is no "standard" position for various cameras and microscopes. In addition when a camera dosn't have rectangular pixels, we will have to take this into account.
e.g. A (B/W) camera with the following specifications and square pixels:
A 1/2 inch CCD and a CCD diameter of 18 mm with a 3/4 aspect ratio, an array size 800 x 581 pixels (of which 756 active horizontaly) will result in:
The effective area is 756 x 581 pixels and not 800 x 581, thus the aspect ratio is not 4/3 ( 768/576 ), so in the end this will give us a (square) pixel size of :
Does anyone know which company sell (make) a motorized Z-drive for light microscope? I am only interested in the simple add-on device to collect z-stacks of light images, I don't need a motorized stage. The microscope used is Olympus BX51.
Any shared information will be highly appreciated.
Xinran Liu
Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center at Dallas Phone: 214-648-1830 Fax: 214-648-1801 E-mail: xinran.liu-at-utsouthwestern.edu
Hi Stephen, TEM is hard work, but while SEM is even harder to understand, with ESEM you can make movies - the perfect solution for we biologists! Here's a recipe that I just worked out.
You need to find some dehydrated, over the counter, crosslinked polyacrylamide that is used in diapers and in some crazy toys, like alligators that grow in the bathtub. I got mine in a bag of crystals sold for the purpose of helping me keep our Christmas tree sanguine for as long as possible. These crystals imbibe water and swell to at least 50x their original size. Thus, if you find the same raw material as was in my little bag, you will have to do as I did and crush a couple crystals with mortar and pestle until you have one VERY tiny crystal that you can place (with #5 tweezers) at the bottom of the smallest depression in your cold stage insert set. Settle the insert in the cold stage and bring the temperature to 5oC and the pressure to 840 Pascal. You should home the stage (WITHOUT ROTATION, of course), and you should connect focus to z ASAP, and finally raise the stage to as close to 10 mm as possible. You should be able to image the crystal and it should be on the order of 100-200 um in size. Set up the movie to capture 1 image every 30 seconds and start recording. Then raise the pressure to 860, 880, 900, and finally 920 Pascal at which point you should begin to see puddles (dark blotches in the image) begin to form. NOTE: you will have to continuously tweak brightness and contrast to keep the image balanced properly. Once the water begins to condense on the stage, you should soon see changes in the crystal. The crystal will grow to its maximum size (1-2mm) and then you can start reducing pressure from 920 back to 840 by increments of 20. Again keep tweaking the brightness and contrast to keep the image right, until you are back to your crystal. NOTE: If the crystal is strange enough, it will probably resemble a cartoon character or two during its ascent or descent from totally bilious.
Finally, the routine is not perfect yet, but I am working on it before I bring elementary kids in on the joke. On the first one I did, the crystal went through a growing flower stage on the way up and a dying chicken stage on the way down.
My next trick will not be so silly, though I do plan to try rice and unhusked wheat. I plan to reproduce something I have not yet seen but was done somewhere in Mississippi with an old Electroscan. A movie of opening and closing stomata on a leaf.
Finally, during my very shallow learning curve, I managed to figure out a way to pull the ribbon cable out of the stage, thus disconnecting it from it's brain. It has been repaired, and has worked well since, as long as I am on my knees when I close the stage door and home the stage before doing that.
Hope this works for you, if you can try it, and I would like your results too.
I am copying this to the Q200-600 users list as well.
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: Coetzee, Mr S. H Physics Science [mailto:COETZEES-at-mopipi.ub.bw] Sent: Monday, April 28, 2003 8:24 AM To: Microscopy-at-sparc5.microscopy.com
} From: Coetzee, Mr S. H Physics Science } Sent: Monday, April 28, 2003 12:24 PM } To: Listserver Microscopy (E-mail) } Subject: Needs assessment - EM project for kids } } } Dear All } I would love to call this e-mail: Needs assessment - video or DVD on EM } and LM under subject but due to juncmail filters I had to change } the subject to } get it to the list. } } Life is always interesting and full of fun when you are doing EM. } We are doing a needs assessment on the following: } "Introduction video/DVD and LM for kids" } We want to target 13 - 18 year old kids and hopefully be able to reach 1st } and second year students as well. } The idea is to improve the number of students in Science with the focus on } EM. Input will be greatly appreciated. } We need this input for fund application. } } } Stephan H Coetzee } Department of Physics } EMU } Private Bag 0704 } Gaborone, } Botswana } } Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} } } Telephone: (+267) 355 2462 } Fax: (+267) 3185 097 } Telephone: (+267) 355 0000 Switchboard } }
Bill Usually the "silver enhancer" used to enlarge particle size to made it visible in EM or even in LM. This technique is quite popular when you use pre-embedding "staining" with antibodies. In this case the probe size is critical, so people tends to use the smallest possible probe. Another good things about "undecaGold" and "NanoGold" (I am not familiar with other brands) that it has only one specific reactive group, maleimide for instance. So, you may attach these particles very precisely: such probe is more "direct". Sergey
At 04:46 PM 4/25/03 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Anyone know where I could get someting coated with a thin layer of silicon dioxide?
A researcher here wants to coat a gold microelectrode with a 100 nm layer of silicon dioxide to use in a quartz crystal microbalance. He asked me because I have done other coating jobs for him, but I can't do silicon dioxide.
If you can do it or know where to go to get it done, let me know and I will pass the info on.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Damian If I understand correctly, you are using "dual" monitor video-cards. Nevertheless is it true or not, you may use two independent video-cards. Personally, I like Nvidia Gforce line (ATI is good choice as well). I am using one AGP-video for primary monitor, and another PCI-video card - for secondary. Personally, I like to have Win toolbar on the single monitor, so I am using "smaller" monitor as a primary. I was suspicious about LCD monitors for years, but things are changed now. Samsung monitors are very good (no interest in company) and deliver very good image quality. My favorite setup is 17" (1280x1024) + 15" in portrait mode (780x1024). It comes with "Colorific (?)" software for color calibration. It's first monitor in my life, which delivers exact matching with color printer. Another good thing about LCD - it's not flickering. My eyes tend to get tired after a few hours of working with high-quality CRT monitor, with LCD I could work comfortably much longer. Have a good day. Sergey
At 11:51 PM 4/25/03 -0500, you wrote: } Hi all, } The discussion on camera pixel numbers has been interesting but I haven't } the expertise to contribute anything but have learned something. Thanks to } all contributors. } } However, I did make the jump to dual monitors as Sergey describes and find } it very helpful in several ways: 1) When using Photoshop I keep the tools } and maybe another app windows on one monitor, and the image on the Photoshop } desktop on the primary monitor. This is great also when you are documenting } in another app every step in manipulating the image; 2) when working on more } that one document; and 3) when answering email like this one. } } I did run into one issue with using dual monitors. Depending on which type } of video board you get, you could end up with the desktop spread over two } monitors making mousing across a wide field an annoyance but allowing you to } open two images to a fairly large size (depending on screen width) with no } overlap. Other boards make one monitor the primary and the other a } secondary so that the desktop is on one monitor and the other just a blank } work area where you can open a second app or a duplicate app. An example of } this is Matrox G450 versus Matrox G550, the former puts the desktop across } both screens, the latter puts the desktop on the primary screen (of your } choice). I still perfer the CRT for my primary Photoshop screen as I think } it still gives slightly better colors than an LCD but the gap is closing } fast. } } Damian Neuberger } } } } } } Hello everyone } } It's not absolutely necessary to spent money on expensive huge LCDs. I do } } find that even on my 19" LCD with 1280x1024 the letters sometime too small } } to read comfortably. In most cases we need more space, not resolution (we } } are talking about monitors). If you are running Win2K you may use two } } monitors simultaneously. They act altogether as one big monitor. It mean } } that you could place your menu bars in one place (monitor) and use another } } for fine work. If it's a case, people usually use expensive, good monitor } } as a working place and another cheaper (and usually smaller) to store } } icons, menu-bars etc... Mouse and other controls act as it's single } } monitor. Such set-up is very popular between graphic designers. Sergey
} I am looking for a RMC MT 6000-XL specimen holder (the whole thing with } arc segment mount etc., second hand or new). Anybody an idea where I } could purchase one? } Stefan -
Contact Dave Roberts of RMC (now a division of Boeckeler Instruments) at dave-at-boeckeler.com.
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 need to perform plunge freezing of solutions for CryoTEM. I was wondering if anyone had some advice on how to handle these explosive cryogens while using the Gatan cryotransfer system.
Any and all advice would be welcome as well as additional protocals for plunge freezing.
What kind of something? I might be able to get a TEOS wafer or two (SiO2 on Si).
gary g.
At 01:46 PM 4/28/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What kind of something? I might be able to get a TEOS wafer or two (SiO2 on Si).
gary g.
At 01:46 PM 4/28/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In a message dated 04/28/2003 10:05:23 AM US Mountain Standard Time, xinran.liu-at-utsouthwestern.edu writes:
{ { Does anyone know which company sell (make) a motorized Z-drive for light microscope? I am only interested in the simple add-on device to collect z-stacks of light images, I don't need a motorized stage. The microscope used is Olympus BX51.
Any shared information will be highly appreciated.
Xinran Liu } }
Xinran,
I would recommend either Ludl Electronic Products at {www.ludl.com} or Prior Scientific at {www.prior.com} .
On the Prior website, look under "focus controls." The product you're looking for is the "Laser Autofocus" system.
Best regards,
Bob
Bob Chiovetti GTI Microsystems Leica Regional Dealer Desert Southwest Region / USA
***Disclaimer: GTI Microsystems represents Ludl and Prior, along with several other manufacturers of peripheral microscope control systems, but not in your region. This information is offered only for your information. Please contact your local representing agency for more details.***
We have decided to install AC/DC field cancellations to improve the performance of our FE-TEM and FE-SEM. We also plan to put acoustic tiles on walls in both rooms. I was wondering if anyone in the community could share some experience with me. Off-line vendor responses are welcome.
**************************************** Chaoying Ni The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
Are we talking about calibrating the ability of the camera to measure distances (spatial resolution)?
What about using a graticule? Just use an appropriate graticule once with each lens and as long as you don't change the lenses or other system components it will 'never' need to be done again.
Graticules are expensive so for one-off jobs perhaps you could borrow?
There was a company in UK called Graticules Limited who have now been taken over by Pyser-SGI Limited (see http://www.vertmedia.co.uk/pyser/contact.htm) or write to graticules-at-pyser-sgi.com.
I am not connected with the company in any way and if anyone from the company is reading this - do you think you could make your website a little more user-friendly?
Also try searching on www.google.com using graticule(s) as a search term.
I hope this helps. Med vänliga hälsningar/With best regards
Gareth
Take a look at http://www.vardforbundet.se/ifbls2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
I have never made one for a microscope but I have made them for many other devices using 4 solid state relays, a power supply, stepper motor and two timing belt pulleys and a timing belt. The pulleys are select to give the resolution needed using a 200 step per revolution stepper motor.
The stepper motor is driven by a simple program from the printer port driving the realys.
The only unknown is how you trigger your camera. It really needs to be triggered by the computer as well after a few seconds wait for vibration to die down.
If you only need one building one of off the rack parts is probably the least expensive choice. If you need several replacing the solid state relays with discreet components will save money. I expect 10 or 15 would be the break even point.
I can help you with the design and software or I expect I can get them built for you and furnish the software.
If you are having them built or building them yourself consider what other features your would like in the future. A few dollars spent on a more complex controller at the start will save scrapping what you have an starting over.
Good Luck
Gordon Couger Stillwater, OK www.couger.com/gcouger
} From: "RCHIOVETTI-at-aol.com"-at-sparc5.microscopy.com : : In a message dated 04/28/2003 10:05:23 AM US Mountain Standard Time, : xinran.liu-at-utsouthwestern.edu writes: : : { { Does anyone know which company sell (make) a motorized Z-drive for light : microscope? I am only interested in the simple add-on device to collect : z-stacks of light images, I don't need a motorized stage. The microscope : used is Olympus BX51. : : Any shared information will be highly appreciated. : : Xinran Liu : } } : : Xinran, : : I would recommend either Ludl Electronic Products at {www.ludl.com} or Prior : Scientific at {www.prior.com} . : : On the Prior website, look under "focus controls." The product you're : looking for is the "Laser Autofocus" system. : : Best regards, : : Bob : : Bob Chiovetti : GTI Microsystems : Leica Regional Dealer : Desert Southwest Region / USA : : ***Disclaimer: GTI Microsystems represents Ludl and Prior, along with several : other manufacturers of peripheral microscope control systems, but not in your : region. This information is offered only for your information. Please : contact your local representing agency for more details.*** :
we sell a motorized Z-drive which only controls the focus. We also have controllers which allow to connect the Z-drive and a motorized X/Y- stage. You could use the Z-drive now and add the X/Y-stage later. Our system is based on a Märzhäuser Z-drive.
Please contact me if you would like to receive a quotation.
Listers, I have a person wanting to look at rhihzobium involvement in root nodules. What is the best way to prepare this sample? I'm most concerned
about how best to expose the fungal involvement with the least amount of
artifact. Any suggestions or references will be appreciated. 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 =============================================================
Hello, this is my first post to the list. I need to find the specification of a Philips both the CM10 and CM100 water chillers, so I can be sure that the replacements are up to the specification the microscopes require. If anyone could help, I would be very grateful,
You can buy stage micrometers for about $150 at the usual microscopy supply places. If I am not mistaken, they are even traceable. For an additional price you can also get certified micrometers.
I didn't get that far looking for graticule. "Stage micrometer" seems to be a better phrase to look for.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Gareth Morgan [mailto:Gareth.Morgan-at-labmed.ki.se] Sent: Tuesday, April 29, 2003 12:09 AM To: Microscopy-at-sparc5.microscopy.com
Hi
Are we talking about calibrating the ability of the camera to measure distances (spatial resolution)?
What about using a graticule? Just use an appropriate graticule once with each lens and as long as you don't change the lenses or other system components it will 'never' need to be done again.
Graticules are expensive so for one-off jobs perhaps you could borrow?
There was a company in UK called Graticules Limited who have now been taken over by Pyser-SGI Limited (see http://www.vertmedia.co.uk/pyser/contact.htm) or write to graticules-at-pyser-sgi.com.
I am not connected with the company in any way and if anyone from the company is reading this - do you think you could make your website a little more user-friendly?
Also try searching on www.google.com using graticule(s) as a search term.
I hope this helps. Med vänliga hälsningar/With best regards
Gareth
Take a look at http://www.vardforbundet.se/ifbls2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
I got 11,300 hits for graticule and 18,700 for stage micrometer. Why do we call the same thing such very different names? Med vänliga hälsningar/With best regards
Gareth
Take a look at http://www.vardforbundet.se/ifbls2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
} We are doing a needs assessment on the following: } "Introduction video/DVD and LM for kids" } We want to target 13 - 18 year old kids and hopefully be able to reach 1st } and second year students as well. } The idea is to improve the number of students in Science with the focus on } EM. Input will be greatly appreciated. } We need this input for fund application.
Mr. Coetzee -
I'd like to help, but it isn't clear what sort of input you need. If a reviewed list of what is currently available on video & CD-ROM (mostly at the middle & secondary level) will help, I can send the updated MICRO information directly to you as an attachment. It will be available at the MICRO website (URL below) as a searchable database within the month.
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
You didn't say if you wanted TEM or SEM, but I've prepared nodules for both, so here goes:
For TEM, I usually just cut the nodule into small pieces and do a standard TEM prep---primary fix in buffered gluteraldehyde or paraformaldehyde/glut mixture, buffer washes, osmium post-fix, water washes, uranyl acetate enbloc stain/fix, washes, dehydration, infiltration (recommend Spurr's or Spurr's/Epon for better infiltration), embedding, and ultramicrotomy. Vacuum and/or microwave-assisted fixation/infiltration often helps get the reagents in. Sometimes the best part of the nodule is in the center, if you're looking for bacteria (rhizobial strains), otherwise you may find a lot of empty cells.
For SEM, I have done standard fixation (again, vacuum and/or microwave-assist helps with infiltration of reagents), up to about 90% ethanol. At this point, you can drop the nodules into liquid nitrogen until they stop fizzing, then slice/fracture with a razor blade, complete the ethanol dehydration, critical point dry, mount and view. You should be able to see good internal cell structure in the nodule, with lots of rhizobial bacteria/bacteroids inside the cavaties.
About the fungal involvement, if you mean the rhizobia they are actually nitrogen-fixing species of bacteria, such as Rhizobia or Bradyrhizobia. Did you perhaps mean mycorrhizae (hope I spelled that right)? If the latter, I have no knowledge or experience at viewing them in nodules, or if they're even present in nodules.
Hope this helps. Let me know if you need more detail.
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Bill Chissoe [mailto:wchiss-at-ou.edu] Sent: Tuesday, April 29, 2003 8:52 AM To: MSA listserver
Listers, I have a person wanting to look at rhihzobium involvement in root nodules. What is the best way to prepare this sample? I'm most concerned
about how best to expose the fungal involvement with the least amount of
artifact. Any suggestions or references will be appreciated. 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 =============================================================
Thank you for your reply. I am aware that calibrating a microscope with a graticule is the classiacla way to do. We use the apporach I mentioned before for our automated microsopy based system. It has multiple cameras and of course different objectives and other lenses. We sue the calculation I mentioed before to avoid the calibration step when we exchange lenses.
An automated dual (or triple) camera setup with 6 objectives and 3 intermediate lenses plus an additional lens in the camera coupler of one of the cameras would take us some time to calibrate the entire system. By suing th approach I mentioned before, we back-calculate the size an iamg epixel has when going trhough the entire system; objective -} intermediate lens -} camera -} framegrabber -} memory.
The formula I mentioned, allows to take into account the effect of the camera-framegrabber combination on the "physical" dimensions of an image pixel.
Best regards,
Peter Van Osta
=========================================== Gareth Morgan wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi } } Are we talking about calibrating the ability of the camera to measure } distances (spatial resolution)? } } What about using a graticule? Just use an appropriate graticule once with } each lens and as long as you don't change the lenses or other system } components it will 'never' need to be done again. } } Graticules are expensive so for one-off jobs perhaps you could borrow? } } There was a company in UK called Graticules Limited who have now been taken } over by Pyser-SGI Limited } (see http://www.vertmedia.co.uk/pyser/contact.htm) or write to } graticules-at-pyser-sgi.com. } } I am not connected with the company in any way and if anyone from the } company is reading this - do you think you could make your website a little } more user-friendly? } } Also try searching on www.google.com using graticule(s) as a search term. } } I hope this helps. } Med vänliga hälsningar/With best regards } } Gareth } } Take a look at http://www.vardforbundet.se/ifbls2004 } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-labmed.ki.se } } Tel +46 8 5858 1038 } Fax +46 8 5858 7730 } } Gareth Morgan MPhil MSc FIBMS, } Department of Laboratory Medicine (Labmed), } Karolinska Institutet, } Huddinge Universitetssjukhus, F46 } SE 141 86 Stockholm } Sweden } } OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet } för klinisk patologi och cytologi. } } NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. } Clinical Histo- and Cytopathology Laboratory.
Response to the following comments from Dr Ian S. Harper. Mon, 28 Apr 2003
Reply:
I am happy to take this opportunity to clarify any misconceptions regarding the PeCon objective heating product and Bioptechs Products. Bioptechs began producing a patented commercial objective heater in 1992. Although it is a common sense solution to an existing problem, it has been a difficult concept to mainstream into live-cell time lapse applications. Back in 1992 every major microscope manufacturer was petitioned to either adopt our objective heater or at least make objectives in a manner that an optimal heat transfer surface would be exposed. None of the manufacturers were interested. They all said there was not enough of a market for temperature controlled objectives to be worthwhile.
The Bioptechs system is designed to meet the specific needs of high resolution live-cell time lapse imagers and takes into account both the safety of the objective and the cells under investigation. It is not simply a heating device that mounts on an objective. Now that time lapse imaging has become a more commonly used methodology, there are several companies producing devices that claim to have similar capabilities as the Bioptechs system.
Be it known that none of these companies are affiliated in any way with Bioptechs. We do not re-label, re-bag, or permit any other representation of our products. We have a distributor network but they are not permitted to re-label our products.
PeCon has a very similar product in appearance sold for the same application as ours but that is as far as it goes.
I again suggest referring to my Monday April, 28th posting relative to important factors with respect to selecting an objective warmer. Especially the part concerning excess radiated heat.
Further info on this subject is available on our web site http://www.bioptechs.com.
Dan Focht
Original message:
} For general information: Objective heaters (heating device plus } controller) are commercially available from: } 1. Bioptechs } (http://www.bioptechs.com/Instructions/Obj_Htr_i/obj_htr_i.html). } The manufacturers recommend the removal of the objective sleeve so allow } adequate contact between heating loop and objective barrel. A thermistor } against the objective barrel senses the temp and is part of feedback } control. Two heater loop sizes avail. } } 2. PeCon (http://www.pe-con.de/PeCon/PeCon-News.htm).. for specific } Zeiss and Leica lenses. Design look rather like the Bioptechs system - } does anyone know if this is a rebadged product ? } } 3. Our local uni instrument maker designed and built a system for us } (now available through a company Scientific concepts } (http://www.scientificconcepts.com.au/) - 'scuse the plug but people } might be interested in other live cell imaging accessories they have. (I } get no kickbacks, in case you're wondering, am merely a satisfied } owner). I suggest you make contact with S.C. via the website as the } heater does not yet feature on the website. The design is based on a } metal heating cylinder made to fit each lens (not an issue really, as, } for example, all our Olympus immersion lenses have more or less the same } diameter). Contact area between heater and barrel is large, and when } coupled with a heating chamber at same temperature, I have measured } {0.2ºC variation over the radius of a 25mm coverslip. (Compare this to a } 5ºC under similar conditions with non-heated immersion lens !!!).
} 4. 20/20 Technology } http://www.20-20tech.com/objective_heater.htm } To fit Leica, Nikon and Olympus lenses. } } 5. Zeiss } http://www.zeiss.de/C12567BE0045ACF1/allBySubject/2E19B0FD92393E6CC1256CB400326879 } } 6. Scientifica Life Sciences - a low cost thermal foil wrapping for } objective heating. } http://www.scientifica.uk.com/a.html } http://www.scientifica.uk.com/a_categorydetails.php?id=6
} Dr Ian S. Harper, Director } Monash Micro Imaging } The Microscopy & Imaging Research Facility } Building 13C, Monash University, Clayton, VIC 3800, Australia
-- Dan Focht Bioptechs 3560 Beck Road Butler, PA 16002 dan-at-bioptechs.com www.bioptechs.com V 724-282-7145 F 724-282-0745
University of Washington Employment Opportunities Research Coordinator (Imaging)
Department: Biology Date Available: May 1, 2003 Closing Date: Open until filled.
General Duties/Description: Under faculty overview, oversee the operation of a Bio-Rad MRC-600 laser scanning confocal microscope (LSCM), a Bio-Rad Radiance 2000 multi-photon LSCM microscope, a Philips CM-100 transmission electron microscope (TEM), a JEOL 840A scanning electron microscope (SEM), a sample preparation lab and photographic facilities. In addition to oversight, this position has a substantial "hands-on" component.
Training/consultation responsibilities: train facility users (faculty, postdocs, grad students and technicians) on equipment and techniques; instruct users in photographic and darkroom techniques; analyze and solve problems in methodology, software and hardware; consult with users on technical aspects of research projects related to microscopy; perform TEM, SEM, or confocal microscopy for potential users to make sure the techniques suit their needs before they go through training on the instruments (this may include sectioning samples or teaching sectioning); and collaborate with faculty and students to carry out long-term research projects. Maintenance responsibilities: determine service needs and schedule repairs; perform maintenance and necessary adjustments to equipment; recommend the upgrade of facilities by evaluating new equipment and software for purchase; and install new equipment and software (includes configuring the computer). Assist with computer programs designed for image analysis, graphics output and three-dimensional reconstruction: provide technical support in software use age; customize software via configuration or macros to fit users' needs; write and update support documentation; troubleshoot problems; and assist users with variable computer skills. General responsibilities: ensure imaging suite and darkrooms are safe and uncontaminated environments; properly dispose of waste chemicals for imaging and photographic suite; and order and oversee use of supplies, calculate recharges for instrument usage.
Requirements: A Bachelor's degree and a minimum of 2 years' experience with TEM (fixation, embedding, sectioning, staining of biological samples); prior experience with SEM, confocal or multiphoton microscopy would be useful but not essential (we will provide training for the successful applicant); computer literate on DOS and Macintosh (able to write macros, experience with Photoshop and NIH Image desirable). An equivalent combination of education and experience may substitute for stated requirements.
Salary: Salary and benefits are competitive. Salary is commensurate with qualifications and experience.
Send a resume and letter describing relevant experience and education to:
Sharon Larsson, Administrator, Biology, Box 351800, University of Washington Seattle, WA 98195.
slarsson-at-u.washington.edu 206 685-8241
The University of Washington is an equal opportunity, affirmative action employer.
Can someone point me in the direction of a public domain program which can take as input raw (peak minus background) wds element peak intensities of standards and samples and convert the sample intensities into wt % concentrations?
thanks
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
There have been several listers who seem to be calling graticules and stage micrometers the same thing. In my experience in UK light microscopy a stage micrometer is a specific accurate 'ruler' which may be mounted on a light microscope slide. It is an absolute size such as 1 or 10mm and is subdivided for accurate absolute measurements. Most graticules are eyepiece devices which may have up to 100 linear divisions or be divided up in some other way such as by angles. You can calibrate an eyepiece graticule using a stage micrometer for a particular objective lens configuration but the graticule is still an arbitrary measure.
I suppose you could argue that a stage micrometer was a specific absolute graticule but that does not make the terms interchangeable. If I am being pedantic, I apologise.
Thanks for your attention
Malcolm Haswell e.m. unit University of Sunderland UK
We observed similar behaviour in gold labeling E.coli outer membrane proteins quite some time back. We were not succesful in trying to label even unfixed intact cells, whereas labeling was not a problem at all on ultrathin cryosections of aldehyde fixed cells. A small percentage of the intact cells would label however, being about the same number as the number of mutant cells with shorter LPS molecules. That was the basis of our conclusion that long carbohydrate chains of LPS caused steric hindrance in intact cells, whereas an approach 'from the side' on sections was just working fine.
You mention using 10/15 nm particles, I am not sure whether ultra small particles would have the same limitations if LPS carbohydrates are causing this phenomenon.
IŽm interested in labelling bacterial surfaces with colloidal gold for negative stain and TEM. So far negative stain works OK, also indirect immunofluorescence with a primary mouse monoclonal. However, secondary antibodies conjugated with 10-15 nm gold doesnŽt work at all. We have adhered fimbriated E Coli to Formvar-coated gold grids with or without fixation in various concentrations of formaldehyde, labelled them and fixed with glutaraldehyde. Any tips and tricks??
I would suggest a detailed site survey of your field problems before going ahead and buying any field cancelling system. You should really find out if fields are adversely affecting your scope rooms (preferably without the scopes there, I know this may be difficult!) and if the fields are of a nature which a field cancelling systems can deal with. This is because our field cancelling system was actually unable to cancel our largest field problem as the frequency was too low to damp (we ended up digging up and re-installing old mains cables to solve the problem). It is also important to realise that a field cancellation system can typically only damp fields at one point (where the measurement probe is), which begs the question where do you put it? At the objective lens? Or close to the gun/HT? Anybody who has tested the position effect more closely, I would love to hear your input!
Acoustic tiles are however a simpler matter and I would definately recomend you install these.
Hope that helped!
Dr M Weyland, 1851 Research Fellow, Electron Microscopy group --------------------------------------------------------------------------- Department of Materials Science & Metallurgy University of Cambridge Pembroke St Cambridge CB2 3QZ
--On 28 April 2003 22:11 -0400 Chaoying Ni {cni-at-udel.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } } We have decided to install AC/DC field cancellations to improve the } performance of our FE-TEM and FE-SEM. We also plan to put acoustic tiles } on walls in both rooms. I was wondering if anyone in the community could } share some experience with me. Off-line vendor responses are welcome. } } **************************************** } Chaoying Ni } The W.M. Keck Electron Microscope Facility } Facility location: 022 Spencer Laboratory } Mailing address: } 201 duPont Hall } Dept of Matls Sci & Eng } University of Delaware } Newark, DE 19716 } (302) 831-8354 (O); -2318(L); -4545(Fax) } http://eml.masc.udel.edu } ***************************************** }
Hi: We are currently investigating various freeze sub and freeze plunge systems and would appreciate any pros or cons people have towards the different systems. Vendors feel free to email me with your products and please include the prices. Thanks Charlie Murphy murphyc-at-ba.ars.usda.gov
Charles Murphy USDA, ARS, Electron Microscopy Unit Bldg. 177B, BARC East Beltsville, MD, 20705 (301) 504-8046 (301) 504-8923 fax murphyc-at-ba.ars.usda.gov
Does anyone have any information about whether osmium tetroxide can be used in xylene?
In our clinical pathology EM lab I am frequently called upon to *rescue* a piece of tissue from a paraffin block and work it up for EM. The standard protocol is to deparaffinate the tissue in xylene, then hydrate the tissue to osmicate it then dehydrate it again to go on to infiltrating with resin and finally embedding/polymerizing.
I have a faint recollection of a discussion years ago that osmium could be suspended in xylene saving the whole hydration/osmication/dehydration step. Thus one would deparaffinate the tissue in xylene then osmicate in xylene and go on to infiltration from there.
Any comments?
Karen Bovard Creighton University Medical Center Department of Pathology Electron Microscopy Laboratory Omaha, Nebraska
Jon; If you have a Gatan duo mill, you can put a piece of quartz where the sample normally goes, and the sputtered SiO2 will deposit on your target substrate held near it.
John Mardinly Intel
-----Original Message----- } From: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Monday, April 28, 2003 1:46 PM To: Microscopy-at-sparc5.microscopy.com
Anyone know where I could get someting coated with a thin layer of silicon dioxide?
A researcher here wants to coat a gold microelectrode with a 100 nm layer of silicon dioxide to use in a quartz crystal microbalance. He asked me because I have done other coating jobs for him, but I can't do silicon dioxide.
If you can do it or know where to go to get it done, let me know and I will pass the info on.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I don't know about xylene but I do know it works well in 100% ethanol in many different freeze-substitution protocols. So even if xylene won't work, you might not have to do a full dehydration. But the strength of the osmication reaction in ethanol at room temperature is much stronger than in water so I would try low osmium concentrations.
At 11:23 AM 4/30/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Hello all, I am trying to gather information about saving digital images. Currently I import all images to Quartz PCI and save them as TIFF files on our network. I would like to continue to do this however, the IT department is threatening to cut me off if I continue to use so much space on the server. We do occasional image analysis, and since JPEG compression is lossy and introduces artifacts, I am leary of heading in that direction. I thought that if I could convince them that this is the best practice for saving images we could come to some sort of agreement. Any information of how you handle saving SEM images would be helpful in my quest for peace. Thank you
~Amanda Marchese
PQ Corporation Research & Development Center 280 Cedar Grove Road Conshohocken PA 19428 (610) 651-4668
On Monday, April 28, 2003, at 07:11 PM, Chaoying Ni wrote:
} We have decided to install AC/DC field cancellations to improve the } performance of our FE-TEM and FE-SEM. We also plan to put acoustic } tiles } on walls in both rooms. I was wondering if anyone in the community } could } share some experience with me. Off-line vendor responses are welcome.
Dear Chaoying, We are installing a FE-TEM, and we had some acoustics problems (our fields were very low). We eliminated the acoustical vibrations at the source, and we hung thick acoustical curtains, which solved all but one problem, which we are still working on. During the course of our investigations, we found that acoustic tiles are very good for reducing noise in the few-hundred Hertz range and above, but they were poor for reducing lower frequencies. Depending on the noise spectrum at your facility, tiles may either be useful or useless. Getting rid of, say, 60 Hz noise requires mass, so even anechoic tiles will be impractical--they must be at least 1/4 wavelength thick, which is impractical. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Offer to split the cost or buy your IT department a new hard drive for the storage of your images.
Don't sacrifice image information if you don't have to.
Mike
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ============================================================ Michael Dunlap office (530) 752-0284 University of California lab (530) 752-5489 Chemical Engineering & Material Science Fax (530) 752-9554 110A EU-II mrdunlap-at-ucdavis.edu One Shields Ave. http://www.chms.ucdavis.edu/ Davis CA, 95616 ============================================================
You can dissolve the osmium in xylene. Remove the paraffin first with xylene, then mix the osmium tetroxide in xylene directly before use. The osmium reduces over time, so the mixture does not store well. We buy 0.1gram ampules of crystalline osmium tetroxide and use it at a concentration of 1% in xylene. Time in osmium is dependent on the size of the sample: sections on slides-15 minutes, 1mm3 tissue blocks-45 min to 1 hour. Rinse afterwards with several changes of xylene, then on to propylene oxide and resin. To dispose of the osmium mixture, we combine it with corn oil (1 part Os waste: 2 parts corn oil).
Vicky
Victoria Madden Microscopy Services Laboratory Pathology and Laboratory Medicine University of North Carolina at Chapel Hill vmadden-at-med.unc.edu
Possible solutions with increasing costs would be to 1. Buy a larger hard drive to save the images on your computer and share it to the network (I've seen 120 GB drives for $99/50,000+ images, assuming 2 MB images), 2. Burn images to CD's, $2/300 images, 3. Buy a network storage device, like a SNAP server made by Quantum, Iomega, Dell, and others, 200 GB systems are about $1500-2000/ 100,000 images. Or tell your management you could go back to Polaroids at $2 to $3 an image versus about $.002/image on your hard drive. The network storage device is nice because the images are easily accessed by others without affecting the computer where the images are collected.
At 1:16 PM -0400 4/30/03, Amanda Marchese(RD) wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
Yes. Osmium dissloves in xylene. I have used a similar procedure. Biggest problem is measuring a tiny amount of osmium, since such a small amount is needed. Our Mettler is not in a hood, only an old swinging arm balance. I keep one vial of OsO4 crystals for such use. On the whole, over the years, such "rescued" tissue does not give good results and in most cases the results are uninterpretable and non-diagnostic. It is much more effective to put a snip of tisue for any case which might possibly need EM in EM fixative for potential use. The cost of processing one case for EM with poor results far outweighs the cost of a batch (or 2) of EM fixative and vials.
Becky Garrison Pathology Supervisor
-----Original Message----- } From: "kbovard-at-creighton.edu"-at-sparc5.microscopy.com [mailto:"kbovard-at-creighton.edu"-at-sparc5.microscopy.com] Sent: Wednesday, April 30, 2003 12:23 PM To: Microscopy-at-sparc5.microscopy.com
Does anyone have any information about whether osmium tetroxide can be used in xylene?
In our clinical pathology EM lab I am frequently called upon to *rescue* a piece of tissue from a paraffin block and work it up for EM. The standard protocol is to deparaffinate the tissue in xylene, then hydrate the tissue to osmicate it then dehydrate it again to go on to infiltrating with resin and finally embedding/polymerizing.
I have a faint recollection of a discussion years ago that osmium could be suspended in xylene saving the whole hydration/osmication/dehydration step. Thus one would deparaffinate the tissue in xylene then osmicate in xylene and go on to infiltration from there.
Any comments?
Karen Bovard Creighton University Medical Center Department of Pathology Electron Microscopy Laboratory Omaha, Nebraska
I'd be glad to explain how I handle this problem. It may not be extendable to your situation. But you can pick and choose if you find something useful.
My premise is that the digital images are priceless. Trying to re-create them is a horrible prospect. So redundancy is paramount. I have perhaps 10,000 image files, ranging from LM, macro and SEM plus film images that have been scanned. Some images are 5MB while others can be up to 100MB. They are all over the place, size-wise.
Once the images are captured on MO disk (Fujitsu magneto optical 3.5"), the images are transferred to RAID IDE drives on my main computer. These images are then processed and finalized. The original images are then sent to a set of Dell 715N NAS servers (1TB total) for fast access archiving. Then, I do periodic backups of the servers to Ultrium 1 LTO tape (redundant). Each media holds between 100-200MB, depending on compression ability. This is all done over 100BaseT LAN using Novastore NovaBack network software. Once a full backup is done, all the others are incremental ones. These tapes are stored off-site in a bonded vault (very low cost).
Final output is also stored on the NAS servers and LTO tapes but also on redundant DVD-R media (I used to use CDs).
So, depending on how much data you have that makes your IT people complain, consider getting a DVD-R burner, or CD burner as a minimum. Then, if necessary, get one NAS SNAP server. Dell, Maxtor, and others make these. They are about $1200 used, and vary in price based on total capacity (size of the four internal IDE drives). These run under Win2K but you can also store Mac data to them using DAVE and standard TCP/IP protocol.
Another option is to get two external Firewire or USB hard drives. Plug them in and store the data to both drives. These drives are pretty low cost. Furthermore, if you can accommodate them, removeable IDE drives are very handy. You install a tray in your system box and put a drive in a pull out drawer. They show up in the OS as a regular hard drive. I have about eight drives ranging from 30GB to 200GB. Again, the price of the lost data is $????
Hope that this gives you some ideas.
gary g. Microtechnics, Inc.
At 10:16 AM 4/30/2003, you wrote:
} Hello all, } I am trying to gather information about saving digital images. } Currently I import all images to Quartz PCI and save them as TIFF files on } our network. I would like to continue to do this however, the IT department } is threatening to cut me off if I continue to use so much space on the } server. We do occasional image analysis, and since JPEG compression is lossy } and introduces artifacts, I am leary of heading in that direction. } I thought that if I could convince them that this is the best practice for } saving images we could come to some sort of agreement. Any information of } how you handle saving SEM images would be helpful in my quest for peace. } Thank you } } ~Amanda Marchese } } } PQ Corporation } Research & Development Center } 280 Cedar Grove Road } Conshohocken PA 19428 } (610) 651-4668
if you want to or have to save the images on a server and can't afford to tolerate some changes or artifacts in the images, you only have the option for a loss-less compression. There are several options to do so, the most commonly known is probably the ZIP utilities, but there are others, for example JPEG2000. The last one actually offers both lossy and loss-less compression. I don't know if Quartz PCI supports these compressions or file formats, we have implemented in our software, specifically for this reason, a JPEG2000 compression within the TIF format, so you get the best of both worlds.
If you can use a different storage location, you can probably relax this a lot. For example, you can buy so-called "Network attached storage devices" (basically a rudimentary PC with huge hard disks, often with redundant storage systems) and simply hook them up to the network. Perhaps you can convince your IT department to let you buy them one of those and keep it with the servers? Because you are not running any applications on these boxes (except the administration interface), there is usually very little administration necessary.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Amanda Marchese(RD) [mailto:Amanda.Marchese-at-PQCorp.com] Sent: Wednesday, April 30, 2003 11:16 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Hello all, I am trying to gather information about saving digital images. Currently I import all images to Quartz PCI and save them as TIFF files on our network. I would like to continue to do this however, the IT department is threatening to cut me off if I continue to use so much space on the server. We do occasional image analysis, and since JPEG compression is lossy and introduces artifacts, I am leary of heading in that direction. I thought that if I could convince them that this is the best practice for saving images we could come to some sort of agreement. Any information of how you handle saving SEM images would be helpful in my quest for peace. Thank you
~Amanda Marchese
PQ Corporation Research & Development Center 280 Cedar Grove Road Conshohocken PA 19428 (610) 651-4668
This is sooo strange to hear. Disk drives are so cheap, it is like $80 for a 120 Gigabyte drive now. Just buy your IT department a new drive for your own data, and tell them you'll throw in lunch.
You can also do the storage internally in your own lab. Buy a server with a large hard disk and dump your data onto there. Servers more than adequate for this run as low as $400-$800. Add a DVD burner and you can back up all your images onto more permanent storage on a monthly basis.
I've implemented the above in our laboratory. If you need help with it, contact me offline.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Wed, 30 Apr 2003, Amanda Marchese(RD) wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } I am trying to gather information about saving digital images. } Currently I import all images to Quartz PCI and save them as TIFF files on } our network. I would like to continue to do this however, the IT department } is threatening to cut me off if I continue to use so much space on the } server. We do occasional image analysis, and since JPEG compression is lossy } and introduces artifacts, I am leary of heading in that direction. } I thought that if I could convince them that this is the best practice for } saving images we could come to some sort of agreement. Any information of } how you handle saving SEM images would be helpful in my quest for peace. } Thank you } } ~Amanda Marchese } } } PQ Corporation } Research & Development Center } 280 Cedar Grove Road } Conshohocken PA 19428 } (610) 651-4668 } }
Precise language with universally accepted definitions form the basis of rational scientific discussion. If this is pedantry, it seems to be warranted here.
Betty Abajian-Seaman Wyeth Pearl River, NY
} } } Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} 4/30/2003 4:32:28 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
There have been several listers who seem to be calling graticules and stage micrometers the same thing. In my experience in UK light microscopy a stage micrometer is a specific accurate 'ruler' which may be mounted on a light microscope slide. It is an absolute size such as 1 or 10mm and is subdivided for accurate absolute measurements. Most graticules are eyepiece devices which may have up to 100 linear divisions or be divided up in some other way such as by angles. You can calibrate an eyepiece graticule using a stage micrometer for a particular objective lens configuration but the graticule is still an arbitrary measure.
I suppose you could argue that a stage micrometer was a specific absolute graticule but that does not make the terms interchangeable. If I am being pedantic, I apologise.
Thanks for your attention
Malcolm Haswell e.m. unit University of Sunderland UK
Hi Karen, We have been using Xylene/Oso4 for many years.It works quickly and with good results if original fixation was buffered formalin. The reference is as follows: Kai Chien,R.L.Van deVeldeand R.C.Hauser, Fortieth Annual EMSA Meeting,1982,pp356-357 I don't know if it was ever published any where else. We use .01grm of OsO4 rather than using a whole grm,since we only do it a few times a year. Our procedure is as follows: two 20 minute changes in xylene one 20 minute change in xylene/OsO4 two 10 minute changes in Propylene oxide 15 minutes in 3:1 P.O./resin mixture 30 minutes 1:1 " " " 30 minutes 1:3 " " " 20 minutes in fresh resin-- embed
Almost easier than starting with fresh tissue! Cheers, Connie
-----Original Message----- } From: "kbovard-at-creighton.edu"-at-sparc5.microscopy.com [mailto:"kbovard-at-creighton.edu"-at-sparc5.microscopy.com] Sent: Wednesday, April 30, 2003 12:23 PM To: Microscopy-at-sparc5.microscopy.com
Does anyone have any information about whether osmium tetroxide can be used in xylene?
In our clinical pathology EM lab I am frequently called upon to *rescue* a piece of tissue from a paraffin block and work it up for EM. The standard protocol is to deparaffinate the tissue in xylene, then hydrate the tissue to osmicate it then dehydrate it again to go on to infiltrating with resin and finally embedding/polymerizing.
I have a faint recollection of a discussion years ago that osmium could be suspended in xylene saving the whole hydration/osmication/dehydration step. Thus one would deparaffinate the tissue in xylene then osmicate in xylene and go on to infiltration from there.
Any comments?
Karen Bovard Creighton University Medical Center Department of Pathology Electron Microscopy Laboratory Omaha, Nebraska
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (WOwsiany-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, April 30, 2003 at 11:59:57 ---------------------------------------------------------------------------
Email: WOwsiany-at-aol.com Name: Walter J. Owsiany
Organization: Vineyards Country Club
Education: Undergraduate College
Location: Naples, Florida 34119
Question: I am a golf course superintendent, what type of scope would I need to look at turfgrasses diseases in my office. Spending no more than $8oo to $1000.
Amanda, We convinced our IT dept. to install SNAP servers to accommodate the images generated by our Optical Microscopy Core Facility, and the rest of the department. It has worked out quite well. Lee
} -----------------------------------------------------------------------. } } } Hello all, } I am trying to gather information about saving digital images. } Currently I import all images to Quartz PCI and save them as TIFF files on } our network. I would like to continue to do this however, the IT department } is threatening to cut me off if I continue to use so much space on the } server. We do occasional image analysis, and since JPEG compression is lossy } and introduces artifacts, I am leary of heading in that direction. } I thought that if I could convince them that this is the best practice for } saving images we could come to some sort of agreement. Any information of } how you handle saving SEM images would be helpful in my quest for peace. } Thank you } } ~Amanda Marchese } } } PQ Corporation } Research & Development Center } 280 Cedar Grove Road } Conshohocken PA 19428 } (610) 651-4668
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I concur. We are a pathology lab for a research group, and that is standard procedure: fixing a piece of each for LM and EM. It's a lot of work to deparaffinize and re-fix a piece of tissue for such poor results! However, sometimes it's unavoidable, if the tissue is archival and now they need to look at ultrastructure.
I do so few retrieval samples, that I was interested to learn about mixing osmium in xylene. Thanks for the information!
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
} -----Original Message----- } From: Garrison, Becky [SMTP:becky.garrison-at-jax.ufl.edu] } Sent: Wednesday, April 30, 2003 4:18 PM } To: '"kbovard-at-creighton.edu"-at-sparc5.microscopy.com'; } Microscopy-at-sparc5.microscopy.com } Subject: RE: Osmium in xylene } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Yes. Osmium dissloves in xylene. I have used a similar procedure. Biggest } problem is measuring a tiny amount of osmium, since such a small amount is } needed. Our Mettler is not in a hood, only an old swinging arm balance. I } keep one vial of OsO4 crystals for such use. } On the whole, over the years, such "rescued" tissue does not give good } results and in most cases the results are uninterpretable and } non-diagnostic. } It is much more effective to put a snip of tisue for any case which might } possibly need EM in EM fixative for potential use. The cost of processing } one case for EM with poor results far outweighs the cost of a batch (or 2) } of EM fixative and vials. } } Becky Garrison } Pathology Supervisor } } } -----Original Message----- } } From: "kbovard-at-creighton.edu"-at-sparc5.microscopy.com } [mailto:"kbovard-at-creighton.edu"-at-sparc5.microscopy.com] } Sent: Wednesday, April 30, 2003 12:23 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Osmium in xylene } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have any information about whether osmium tetroxide can be } used in xylene? } } In our clinical pathology EM lab I am frequently called } upon to *rescue* a piece of tissue from a paraffin block and work it up } for EM. The standard protocol is to deparaffinate the tissue in xylene, } then hydrate the tissue to osmicate it then dehydrate it again to go on to } infiltrating with resin and finally embedding/polymerizing. } } I have a faint recollection of a discussion years ago that osmium could be } suspended in xylene saving the whole hydration/osmication/dehydration } step. Thus one would deparaffinate the tissue in xylene then osmicate in } xylene and go on to infiltration from there. } } Any comments? } } } Karen Bovard } Creighton University Medical Center } Department of Pathology } Electron Microscopy Laboratory } Omaha, Nebraska
We also use PCI and, as you are aware, the big advantage of a database system is the availability of images. I certainly believe you are using a sound strategy and suggest you try to keep your images on the server. If you are not currently performing daily back-ups I would also suggest that you consider implementing a routine in order to protect against data loss.
Isn't it amazing? The IT department is charged with the difficult task of supporting an ever changing network environment. In this silicon world of flux they lose sight of their mission....to support the users of the network!!! Amanda, they work for you, or at least that's how it should work! My suggestion is to convince the IT department that it is in everyone's best interests if the IT "experts" assist with a network solution to the problem. They need to provide you with network storage space and a timely backup routine in order to protect your valuable research, which is much more costly that a few gigabits of hard drive space. Finally, I would avoid setting up a local server since it places much more responsibility on you now and, more importantly, in the future. Things change.....and look how difficult it is for the experts to keep things running! :)
Good luck, jr
John A. Robson Boehringer Ingelheim Pharmaceuticals, Inc. PO Box 368 900 Ridgebury Rd Ridgefield, CT 06877
-----Original Message----- } From: Amanda Marchese(RD) [mailto:Amanda.Marchese-at-PQCorp.com] Sent: Wednesday, April 30, 2003 1:16 PM To: 'Microscopy-at-MSA.Microscopy.Com'
Hello all, I am trying to gather information about saving digital images. Currently I import all images to Quartz PCI and save them as TIFF files on our network. I would like to continue to do this however, the IT department is threatening to cut me off if I continue to use so much space on the server. We do occasional image analysis, and since JPEG compression is lossy and introduces artifacts, I am leary of heading in that direction. I thought that if I could convince them that this is the best practice for saving images we could come to some sort of agreement. Any information of how you handle saving SEM images would be helpful in my quest for peace. Thank you
~Amanda Marchese
PQ Corporation Research & Development Center 280 Cedar Grove Road Conshohocken PA 19428 (610) 651-4668
On Wednesday, April 30, 2003, at 07:40 AM, Charles Murphy wrote:
} Hi: We are currently investigating various freeze sub and freeze } plunge systems and would appreciate any pros or cons people have } towards the different systems. Vendors feel free to email me with } your products and please include the prices. } Dear Charlie, If your budget will cover it, I strongly recommend getting a Vitrobot for plunge freezing; the cost is ~$70k. It provides a consistent, controlled environment and gives excellent results. I have no financial interest in FEI, which sells the Vitrobot; I'm just a satisfied customer. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
I'm a researcher, currently unaffiliated, and I hope to purchase a compound scope and a dissecting scope to use in my mycological work. I'll be studying mycorrhizae and endophytic fungi in grasses, and I need scopes that will allow me to examine fungal hyphae and spores, as well as grass inflorescences. (I have a master's in botany and will begin a Ph.D. in the fall of 2004, when I'll have access to high-quality equipment.)
I am looking for scopes that are durable (I'll be traveling with them to field stations), low maintenance, inexpensive (under $1000 each, and preferably less), and of course, adequate to the task of viewing fungi. Which brands and models would you recommend? Is it reasonable to buy used ones, say from ebay?
has anyone swapped out the 486DX2 that was common on many mid-late 90's 982s with a newer processor board. i have a PII 233 SBC that i'd like to install, but there seems to be a "timing" issue between it and the microscope hardware. any help would be appreciated...
Dennis Bellotto Research Scientist Pulmonary Research Mail code:9034 UT Southwestern Medical Center 5323 Harry Hines Blvd. Dallas, TX 75390-9034 ph:(214)648-3597/fax:(214)648-8027 pager: (214)822-0541 dennis.bellotto-at-utsouthwestern.edu Come to my aerobics classes: Dynastep Tue/Thur 5:20pm and Mon/Fri 12:15 at the Bryan Williams Center
Malcolm, that's always the way I learnt it, but then I also larned dis-section rather than di-section.
Being pedantic saddles one with the "p" word which is a lot better than being encumbered with either a "d" or "b" word.
Cheers,
Fred Monson
P.S. Speaking of reticles, reticules(#), graticles, graticules and mIcromEters, does anyone know where to find a 'reasonably' priced finder/locater (master) slide (square array of letters and numbers)? I purchased one from Scientific Products here in the USA some years back for around $50.00 and now the only ones I can find are closer to a very much bigger number. Any help would be appreciated. Apparently these things were all a lot less expensive to make when manufactured by hand by skilled technicians, but that was before automated lasers began turning them out. I guess now you don't have to send the locater slide with the specimen slide when you want to show your colleague where to look. Beer is more expensive and so are graticules, but now there is also a big market for brilliant yellow HummVeeez! Progress! Must be close to Friday.
-----Original Message----- } From: Malcolm Haswell [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Wednesday, April 30, 2003 4:32 AM To: MSA microscopy listserver
There have been several listers who seem to be calling graticules and stage micrometers the same thing. In my experience in UK light microscopy a stage micrometer is a specific accurate 'ruler' which may be mounted on a light microscope slide. It is an absolute size such as 1 or 10mm and is subdivided for accurate absolute measurements. Most graticules are eyepiece devices which may have up to 100 linear divisions or be divided up in some other way such as by angles. You can calibrate an eyepiece graticule using a stage micrometer for a particular objective lens configuration but the graticule is still an arbitrary measure.
I suppose you could argue that a stage micrometer was a specific absolute graticule but that does not make the terms interchangeable. If I am being pedantic, I apologise.
Thanks for your attention
Malcolm Haswell e.m. unit University of Sunderland UK
We want to reduce/eliminate vibration associated with some water cooling lines on our TEM. One suggestion has been to mount the water coolers (Haskris) on a platform supported by truck tire inner tubes (to prevent ground transmission) and put some constrictions in the water line to reduce vibration transmission. The first one sounds good; I suspect that some sort of in-line filter would be better to remove vibrations in the water.
I would be interested in how others have approached this; I think it is a fairly general issue.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
I know it's not the same thing but we have a Hitachi S3000N scanning e.m. It is controlled by Windows NT 4.0 and we have been instructed that we shouldn't even install the latest service pack for the operating system because of potential conflicts especially with peripherals and the electron microscope controls.
What you are proposing sounds much more drastic and it might be worth getting advice from LEO or a service engineer. Have you got a particular reason for the upgrade because any possible gains might be far outweighed by the problems you may get? I suspect you would definitely be in the area of "invalidating the manufacturer's warranty".
I get the impression that many of the computerised facilities in modern instruments are tied down to particular hardware and/or software/firmware with little hope of upgrades to keep up with the pace of modern computing technologies. An obvious example might be if we need to upgrade to a new printer or removeable disc technology - my answer to date has been to network a modern computer to our system for archiving and driving newer technologies when we need them.
My concern is that it's not just the storage media (CD, DVD, magneto optical) on modern systems will be obsolete in a few years but the computing technologies (Operating systems, software and hardware) will be unsupported. We have used Win 95, 98 and NT 4.0 which I believe Microsoft is about to drop or already has dropped technical support for.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: brian mcintyre {mcintyre-at-optics.rochester.edu}
Dear Amanda,
One other note on image storage: the Quartz PCI database will keep track of the CDs or DVDs that you burn full of images, if you tell the database, and tell you to put in e.g. "CD #12345" when you want to access that image.
Regards, Mary Mager
Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "Amanda Marchese(RD)" {Amanda.Marchese-at-PQCorp.com} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 30, 2003 10:16 AM
Dear Ritchie, I used to use the DOS code from John Donovan at Berkeley called CITZAF. I see there is an up-dated version of it on the MSA web site. Look for "Reference and Education Activities" on the bottom of the first page, then "Microscopy and Microanalysis Software Libraries". Under "03-MASlib/" you will find "CITZAF2" and "CITZAF3". They should help, although I haven't tried them myself. Good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} ; "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, April 29, 2003 1:44 PM
I would like to thank those of you who have provided me with the information regarding add-on of the Z-motorized function on my Olympus BX51 microscope.
We are currently considering to go with Prior OptiScan ES101, which fits our purpose best.
Have a nice weekend.
Xinran Liu
Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center at Dallas Phone: 214-648-1830 Fax: 214-648-1801 E-mail: xinran.liu-at-utsouthwestern.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bop00rav-at-sheffield.ac.uk) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 4, 2003 at 10:41:32 ---------------------------------------------------------------------------
Email: bop00rav-at-sheffield.ac.uk Name: robert vasey
Organization: university of sheffield
Education: Graduate College
Location: sheffield, uk
Question: I am fixing and cutting sections of plant roots for light microscopy. I am embedding in LR White resin. My question relates to fixation. Is it better to use sodium phosphate buffer or sodium cacodylate buffer? If one is better than the other could you tell me why.
We have someone who would like to examine osteoclast ruffled border formation on adult mouse vertebrae through TEM. We compared two protocols that were used on osteoclast study. From those protocols, we are planning to perfuse the mouse with 2% Formaldehyde (EM grade), 2.5% glutaraldehyde in 0.1M cacodylate buffer, followed by immersion fixation for 24 hours at room temp with the same fixative as the perfusate fixative but with 0.5% Calcium Chloride. Each vertebra is approximately 4mm x 2mm x 3mm in dimension. Although the recommended size of the sample for EM must be less than 1 mm in dimension, is the fixative concentration and length of fixation sufficient enough to fix the sample? It wasn't mentioned whether the samples were fixed for 24 hours either at room temp or in the refrigerator? Is fixation at room temp for 24 hours reasonable? Are there other means of cutting each vertebra into smaller dimensions for EM during the primary fixation? Following immersion fixation, the sample will be decalcified with 14% EDTA for 2 days. The samples will then be further cut into smaller dimensions, washed with cacodylate buffer, post-fixed with 2% osmium tetroxide for 1 hour, followed by gradual dehydration with ethanol, intermediate dehydration with propylene oxide, and gradual embedding with Glauert (Med) EMBED. We appreciate your suggestions. Thank you for your time.
Sincerely,
Claire Haueter
Claire Haueter EM Technician Integrated Microscopy Core Baylor College of Medicine 713-798-4952
Im working with meosporous materials. I would like to get a computer simulation of the MCM41 structure with a multislice program. Can anyone give me some information about it?
Thanks
Emma.
Emma Rossinyol
Departament d'Electrònica Universitat de Barcelona Martí i Franqués 1 08028 Barcelona
Morning Walter, From what I can see, the great majority of turfgrass diseases can be observed by the naked eye or with a magnfiying glass, so for closer inspection, without any other need than identification or confirmation of identification, my suggestion would be for a dissecting microscope with either a ring light (fluorescent or fiber optic light) or directed light (separate or integrated) You must decide whether you need both incident and transmitted light, but it would appear that you should start with a scope such as the one on the web page below: http://www.wme-inc.com/WebPages/MetEqPgs/b&l4.htm others here: http://www.wme-inc.com/WebPages/MetEqPgs/Met01.htm#Microscopes
This used equipment is being sold by a company in Fort Walton Beach, but here is a company with reconditioned scopes and prices. http://www.midwestbioservicecompany.com/used.asp
B&L dissecting scopes are usually very good. I have had several for years.
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: WOwsiany-at-aol.com [mailto:WOwsiany-at-aol.com] Sent: Thursday, May 01, 2003 8:34 AM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (WOwsiany-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, April 30, 2003 at 11:59:57 ---------------------------------------------------------------------------
Email: WOwsiany-at-aol.com Name: Walter J. Owsiany
Organization: Vineyards Country Club
Education: Undergraduate College
Location: Naples, Florida 34119
Question: I am a golf course superintendent, what type of scope would I need to look at turfgrasses diseases in my office. Spending no more than $8oo to $1000.
This is to thank everybody who replied with a wealth of ideas concerning Roots and SEM. If haven't had any feedback yet from our soil science department, so I can't as yet report any progress on that front.
However, in regard to moulds, I inadvertently (honest!) let some bread go mouldy recently. So I've looked through all the suggestions. For fixing, one can't slowly go up through the progressions of increasingly concentrated ethanol, since the bread will go intractably soggy (does anyone remember the song "Mouldy Old Dough"?) Neither do I want to hit the mould with neat alcohol. So what I'm doing, as a first stage at least, is to put thin mouldy crusts of the bread over methanol in a vapour tank, and letting it stand over the long weekend (we British are having our May Day Holiday on Monday 5th!). If anything useful results, I'll keep you posted.
Bye for now,
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +----------------------------------------+ Phone:+44 (0) 118 9318572 Fax: +44 (0) 118 9750203 University internal extension 7867 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +----------------------------------------+
Greetings, There is probably no absolute 'better'. The choice of reagent typically depends on what questions you want to ask, and hence what protocols you plan to follow.
It is worth knowing that cacodylate contains arsenic and so is quite toxic and must be handled and disposed with care. Generally, cacodylate buffers are used to avoid precipitation between phosphate and heavy metal, esp osmium. If you are doing light microscopy, your most likely won't be osmitcating, etc, and so the simple phosphate buffer should work fine. There may exist some specific protocols for LM that feature cacodylate. If you are going to do immuno localization, most of protocols for that call for fixation in an organic buffer, pipes, but I don't know that anyone has ever tested rigorously whether that is really better than phosphate. Good control of pH probably is needed.
Hope this helps, TObias
} } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (bop00rav-at-sheffield.ac.uk) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May } 4, 2003 at 10:41:32 } --------------------------------------------------------------------------- } } Email: bop00rav-at-sheffield.ac.uk } Name: robert vasey } } Organization: university of sheffield } } Education: Graduate College } } Location: sheffield, uk } } Question: I am fixing and cutting sections of plant roots for light } microscopy. I am embedding in LR White resin. My question relates } to fixation. Is it better to use sodium phosphate buffer or sodium } cacodylate buffer? If one is better than the other could you tell } me why. } } Thanks. } } Bob. } } ---------------------------------------------------------------------------
Phosphate can lead to calcium precipitation which is noticeable in the TEM. Cacodylate doesn't have this problem but it is arsenic based and therefore toxic. As a hazardous waste, it needs proper disposal which can be expensive but more importantly, something that should be minimized unless absolutely necessary. I recommend HEPES or PIPES.
At 11:42 AM 5/4/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
The Materials Characterization Facility at the University of Pennsylvania is interested in selling a Phillips 420T TEM/STEM. The instrument has been maintained under service contract and is equipped with a Kevex 8204 Delta 1 EDS system and a Gatan serial electron energy loss spectrometer. The instrument comes with single-tilt and double-tilt, tilt-rotate, tensile, heating and cryogenic holders.
Additional information (specifications, picture, etc.) can be found at:
Please contact Doug Yates for further details regarding terms of sale at dmyates-at-lrsm.upenn.edu. The purchaser will be responsible for all relocation costs.
********************************************************* Douglas M. Yates, Ph.D. Technical Director Materials Characterization Facility 215-898-2013 dmyates-at-lrsm.upenn.edu Department of Materials Science & Engineering 3231 Walnut Street Philadelphia, PA 19104 *********************************************************
I used to do a lot of bones and teeth for EM, using a protocol similar to the one you describe but with some differences. I perfused rats with glutaraldehyde in phosphate buffer, then continued the fixation in Karnowski's fixative (paraformaldehyde and glutaraldehyde in cacodylate) for 24h at 4 degrees with continuous stirring. I was looking at pieces of rat skull much bigger than mouse vertebrae, and they seemed to be fixed properly. For the decalcification stage, you might want to check the end-point by x-ray, if possible, rather than giving all the samples a standard two days. Hope this helps.
Lesley Weston.
on 04/05/2003 8:10 PM, chaueter at chaueter-at-bcm.tmc.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } We have someone who would like to examine osteoclast ruffled border } formation on adult mouse vertebrae through TEM. We compared two protocols } that were used on osteoclast study. From those protocols, we are planning to } perfuse the mouse with 2% Formaldehyde (EM grade), 2.5% glutaraldehyde in 0.1M } cacodylate buffer, followed by immersion fixation for 24 hours at room temp } with the same fixative as the perfusate fixative but with 0.5% Calcium } Chloride. Each vertebra is approximately 4mm x 2mm x 3mm in dimension. } Although the recommended size of the sample for EM must be less than 1 mm in } dimension, is the fixative concentration and length of fixation sufficient } enough to fix the sample? It wasn't mentioned whether the samples were fixed } for 24 hours either at room temp or in the refrigerator? Is fixation at room } temp for 24 hours reasonable? Are there other means of cutting each vertebra } into smaller dimensions for EM during the primary fixation? } Following immersion fixation, the sample will be decalcified with 14% EDTA } for 2 days. The samples will then be further cut into smaller dimensions, } washed with cacodylate buffer, post-fixed with 2% osmium tetroxide for 1 hour, } followed by gradual dehydration with ethanol, intermediate dehydration with } propylene oxide, and gradual embedding with Glauert (Med) EMBED. } We appreciate your suggestions. Thank you for your time. } } Sincerely, } } Claire Haueter } } Claire Haueter } EM Technician } Integrated Microscopy Core } Baylor College of Medicine } 713-798-4952 } }
There is a straight-foward and easy approach to eliminating vibrations that emanate from the chillers: Add an extra 10-20 feet of tubing between chiller and microscope, wind up the extra tubing and place on the floor (preferably somewhere out of the way). The coil of extra tubing absorbs the chiller vibration, thus solving the problem. This is an old solution that I have successfully used on several microscopes. Of course, it always helps to locate the chiller as far from the instrument as possible. Contact Haskris for this maximum distance.
I doubt that mounting the chiller on an anti-vibration platform will not work because you are only isolating the vibration from the floor, not from the tubing leading to and from the microscope.
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"L. D. Marks" {ldm-at-risc4.numis.n To: Microscopy List wu.edu} {microscopy-at-sparc5.microscopy.com} cc: Subject: Antivibration measures in water lines (TEM) 05/02/03 06:24 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We want to reduce/eliminate vibration associated with some water cooling lines on our TEM. One suggestion has been to mount the water coolers (Haskris) on a platform supported by truck tire inner tubes (to prevent ground transmission) and put some constrictions in the water line to reduce vibration transmission. The first one sounds good; I suspect that some sort of in-line filter would be better to remove vibrations in the water.
I would be interested in how others have approached this; I think it is a fairly general issue.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
A holding tank, and gravity feed of water to the microscopes is in use at the JEOL factory. Seems to work quite well. Inner tubes for floor vibration sounds like a great idea, but are truck tire inner tubes perhaps a bit big? I would have thought motorcycle inner tubes to be more like the right size, especially for stability of the platform..
John Mardinly Intel
-----Original Message----- } From: L. D. Marks [mailto:ldm-at-risc4.numis.nwu.edu] Sent: Friday, May 02, 2003 4:24 AM To: Microscopy List
We want to reduce/eliminate vibration associated with some water cooling lines on our TEM. One suggestion has been to mount the water coolers (Haskris) on a platform supported by truck tire inner tubes (to prevent ground transmission) and put some constrictions in the water line to reduce vibration transmission. The first one sounds good; I suspect that some sort of in-line filter would be better to remove vibrations in the water.
I would be interested in how others have approached this; I think it is a fairly general issue.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
We want to examine negative-stained liposomes that contain a protein inserted in the lipid membrane, using the TEM.
The technique we use for plant viruses is:
Place a drop of the suspension on parafilm Place a couple of formvar coated grids on the suspension for 5 minutes Blot Place grids on UA for 1-5 minutes Blot
Can we use the same technique for the liposomes? Are there tricks involved to maintain structural integrity and to avoid loss of specimens from the grid surface?
Thanks,
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
On Friday, May 2, 2003, at 04:24 AM, L. D. Marks wrote:
} We want to reduce/eliminate vibration associated with some } water cooling lines on our TEM. One suggestion has been to } mount the water coolers (Haskris) on a platform supported } by truck tire inner tubes (to prevent ground transmission) } and put some constrictions in the water line to reduce } vibration transmission. The first one sounds good; I suspect } that some sort of in-line filter would be better to remove } vibrations in the water. } } I would be interested in how others have approached this; } I think it is a fairly general issue. } Dear Laurence, In Albany, we added vibration dampers, which were cylinders with inflatable bladders inside. We maintained the bladders at about 2/3 the pressure of the water--which varied in the several locations and between the inflow and outflow lines. These dampers were recommended for a low-frequency range of vibrations, tens to a hundred or so Hz. They were mounted on T's off the water lines in locations near the Haskris, the lines to the lenses, the electronics racks and the camera DP. We also had in-line filters to keep the cooling water clean. Both types of installation were equipped with bypass valves, so we could measure the bladder pressure with and without water flow to the damper or the flow rate with and without the filter. This way we could maintain the system, which we did at regular intervals. I hope there is still someone at Albany who could give you more detailed info, such as the manufacturers, costs, etc. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
A vacancy for a technical assistant in the Structural Virology Group of Purdue University is now available. This position is ideal for an individual interested in cryo-electron microscopy studies of viruses. The position will involve sample preparation, initial sample assessment, sample freezing, data collection, image analysis and interpretation. A BS with a major in biochemistry or a related area of biology and experience with transmission electron microscopy is considered a minimal requirement. Additional backgrounds in physics and computing along with a willingness to learn and the ability to balance multiple projects would be highly desirable. Employment will entail comprehensive training during the first year and extensive daily interactions with a team of graduate students, post-doctoral scholars and faculty. Our facilities include state of the art 200 and 300kv TEMs. Applicants can find the official posting on Purdue University's website (http://www.adpc.purdue.edu/Personnel/currjobs/lablist.htm). Requests for further information should be sent to Michael Rossmann or Richard Kuhn, Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA. Tel: 765-494-4911; FAX 765-496-1189; e-mail: mgr-at-indiana.bio.purdue.edu or rjkuhn-at-bragg.bio.purdue.edu.
Paul Chipman Research Electron Microscopist Purdue University Structural Virology
I just recei ved an inquiry from a safety engineer asking about the nature of the oil that was used in the high voltage tank of the RCA EML and EMU-3 TEMs. It seem that their university has one of these instruments that has been standing unused for a long time, and which now is to be disposed of, and which he has the task of doing the disposing. His problem is that he doesn't know what the character of the several gallons of oil in the high voltage tank might be, and what the proper disposal procedures are for it.
As I recall the high voltage tanks on some TEMs of that vintage were filled with polychlorinated biphenyl fluids. Then, after these fluids fell into disfavor highly refined hydrocarbon oils were used. However, I don't know what RCA was using at the time (circa 1955).
Any reliable info anyone can offer will be much appreciated. Incidentalyy, if anyone would like to have the instrument for a museum piece (it is now nearly 50 years old and therefore officially an antique) let me know before the disposal process goes too far.
Best regards, Wil Bigelow -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Dear Wil I would suspect, the oil in the old EM is pretty the same as in the X-ray generators. Probably, regulation for X-ray Roentgen stations in the hospital should be OK for the vintage EMs. Your safe engineer, probably, don't need to know the exact formulation for disposal purpose. If s/he is so curious, s/he may perform mass-spectrum analysis for exact formulation. Another idea comes to me: high voltage transformers in power grill. Sergey
At 02:09 PM 5/5/2003, 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
Rick Perhaps, you need some more sophisticated technique to see protein incorporated into lipid bi-layer. I would suggest cryo-EM of non-stained samples. I do see a couple problems in your neg-staining setup: you will not see the part, incorporated into the lipid because water solution will not penetrate into lipids, so you may see only extra-membrane portion of the protein. I afraid, plastic film, you are using is too thick for such job, you better use thin carbon film. Neg staining in general deformed the liposome (they becomes flat), so there is no way to do morphometry and so on such samples. Neg-staining is OK just to check liposomes quality (size, homogenesity, concentration etc). Sergey
At 12:13 PM 5/5/2003, 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
-----Original Message----- } From: Monson, Frederick C. Sent: Tuesday, May 06, 2003 8:15 AM To: 'Wil Bigelow'
Listers:
I just recei ved an inquiry from a safety engineer asking about the nature of the oil that was used in the high voltage tank of the RCA EML and EMU-3 TEMs. It seem that their university has one of these instruments that has been standing unused for a long time, and which now is to be disposed of, and which he has the task of doing the disposing. His problem is that he doesn't know what the character of the several gallons of oil in the high voltage tank might be, and what the proper disposal procedures are for it.
As I recall the high voltage tanks on some TEMs of that vintage were filled with polychlorinated biphenyl fluids. Then, after these fluids fell into disfavor highly refined hydrocarbon oils were used. However, I don't know what RCA was using at the time (circa 1955).
Any reliable info anyone can offer will be much appreciated. Incidentalyy, if anyone would like to have the instrument for a museum piece (it is now nearly 50 years old and therefore officially an antique) let me know before the disposal process goes too far.
Best regards, Wil Bigelow -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Lista de E-MAILS DE COLOMBIA 255.000 Direcciones Empresariales
Hemos terminado la actualización de direcciones E-mail empresariales de Colombia. 255.000 direcciones Verificadas a Mayo 1 del 2003. Son direciones Empresariales de mas de 35.000 sitios de empresas Colombianas o directamente relacionadas a Colombia Garantizados Se entrega en Archivo con formato Excel® o archivo plano listo para importar a su software de envio de correo. Para compra en Pesos en Colombia. envíe sus datos a: colpe3-at-netscape.net Para compra en US$ con tarjeta de credito, envie sus datos a: coldol3-at-netscape.net Precio Económico. Por menos de lo que usted invierte en un aviso de prensa, obtenga el medio y uselo convenientemente. Disponibles tambien listas de correo E-Mail de Mexico, Chile, Costarica, Guatemala. Para mas información envie sus datos a: otrospais-at-netscape.net Opcional : Software para envío masivo de E-Mail
Rick: To add to Sergey's comments, projections of vitrified liposomes will give you nice bilayer definition but you still may want to consider a gold-conjugated Ab to an exposed part of your protein to confirm its presence. Don
Donald Gantz Dept. Physiology & Biophysics Center Advanced Biomedical Research Boston Univ. School of Medicine Boston, MA 02118 Email: gantz-at-biophysics.bumc.bu.edu Phone: 617-638-4017 (voice mail)
I goofed this morning while updating the Junk Mail filters trying to get rid of the Spammer from Colombia. Sorry gang, but nearly all mail today was rejected.
If you were rejected, please try reposting your message. I hopefully got things fixed and things are back to some semblance of normal operation.
:-(
Nestor Your Friendly Neighborhood (and Ocassionaly Human) SysOp
Disclaimer to all: if this subject has been breached and burned out before - I apologize.
We are currently looking to purchase a system(s) that will meet our sample requirements for both metal deposition carbon evaporation. differences with regards to cost.
What I could use is feedback from users of various systems. I am interested in the reliability, repeatability and deposition consistency. The equipment will be used to } support EMPA and SEM analysis.
Thank you, Evelyn
Scripps Institute of Oceanography University of California, San Diego
I suppose you need to establish, if you haven't already, how the vibration is getting to the TEM. If the vibration is being transmitted through the water line (rather than being transmitted from the chiller through the floor), then isolating the chiller body won't help - you need to dampen the pulses in the water resulting from the rotor vanes in the pump.
Since water doesn't really compress (or expand) under the kind of pressures we're dealing with I'm not sure that putting restrictors on the line would help much, although it will reduce the flow too, which may be bad. Gary Brown suggests any extra length of flexible tubing - another quite elegant way of achieving the same thing is by mounting an ordinary filter housing upside down, minus filter, in line with the output of your chiller. This looks slightly odd but it works well, as the few litres of compressible air trapped in the top of the housing adsorbs much of the pressure fluctuation. A JEOL engineer suggested this inexpensive remedy during the installation of a FE-SEM here recently, seems to work well.
Regards,
Richard
} } On Friday, May 2, 2003, at 04:24 AM, L. D. Marks wrote: } } } We want to reduce/eliminate vibration associated with some } } water cooling lines on our TEM. One suggestion has been to } } mount the water coolers (Haskris) on a platform supported } } by truck tire inner tubes (to prevent ground transmission) } } and put some constrictions in the water line to reduce } } vibration transmission. The first one sounds good; I suspect } } that some sort of in-line filter would be better to remove } } vibrations in the water. } } } } I would be interested in how others have approached this; } } I think it is a fairly general issue. =
Richard Easingwood Otago Centre for Electron Microscopy Department of Anatomy and Structural Biology School of Medical Sciences, University of Otago PO Box 913, Dunedin NEW ZEALAND Telephone: office: 0064 3 479 7301 Facsimile: 0064 3 479 7254 GSM: 0064 21 222 4759 mailto:richard.easingwood-at-stonebow.otago.ac.nz Web site: http://www.otago.ac.nz/anatomy/emunit/ or http://anatomy.otago.ac.nz/ocem/contact.html
If anyone has an old Reichert Ultracut suitable for spares that they would be willing to part with (preferably in the UK), could they kindly contact me off list.
Many thanks.
Bob Phillips
MicroServiS 11 Grafton Close, St. Ives, Cambridgeshire, PE27 3DL UK bob.phillips-at-microservis.co.uk www.microservis.co.uk
Laurence, My experience has been that the majority of the vibration comes from the return line. When you adjust the feed pressure, the system bypasses the excess flow directly to the return (assuming you have a positive displacement pump instead of a centrifugal pump).
A quick check, and sometimes permanent solution, is to remove your return line from the chiller and cap the chiller side. Then provide a direct return to the holding tank, eliminating back-pressure. The mixing will not be as good as the manufacturer's set-up, but there should be a large reduction in the pulses. The by-pass from the pressure regulator will still use their return system.
If you can't find the diaphragm units mentioned earlier, you can build a copper tube system that consists of a tee with horizontal connections to your water lines and a vertical capped section full of air. The tubing should be considerably larger than the inlet and outlet fittings. The main drawback is that you will have to recharge the air from time to time due to it dissolving in the water. The diaphragm eliminates this problem.
Ken Converse owner Quality Images third party SEM service Delta, PA
L. D. Marks wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would be grateful if you would post this position advertisement and bring it to the attention of those who might be interested.
Thanks, Susan Belfry, UNB, Fredericton, *********************************** Employment Opportunity
Electron Microscopist (Materials Science)
Microscopy and Microanalysis Facility University of New Brunswick, Fredericton, New Brunswick, Canada
The University of New Brunswick is seeking an Electron Microscopist - Materials Science Specialist to join the Microscopy and Microanalysis Facility. This is a regional facility providing research services both to the University community and to a wide range of external clients from universities, government and industry in the Maritime provinces. The Microscopy and Microanalysis Facility is currently undergoing a major upgrade to its facilities. The expanded range of instrumentation will include both transmission and scanning electron microscopes, an electron microprobe and a confocal laser scanning microscope. Analytical equipment will include EDS, WDS, and EELS. The successful applicant will join an existing team of three support personnel operating the facility.
Responsibilities: The incumbent will be responsible primarily for the application of imaging and microanalysis to the investigation of materials and duties will include the operation and maintenance of a new 200KV analytical STEM equipped with EDS, EELS, and cryo capabilities.
Requirements: Applicants should have the minimum of a university undergraduate degree in physical science or engineering, plus several years experience working with electron beam instruments, microanalytical equipment, or related instrumentation, preferably with a materials focus. Applicants should also have a sound knowledge of electrical, electronic and vacuum systems, together with experience in troubleshooting and repairing these systems. Specific experience with EFTEM/EELS applications, scientific computer systems and the nature and composition of materials would be a considerable asset. Applicants must also possess good organizational and interpersonal skills and be proficient in verbal and written communications.
Salary: The salary range for this position will be either $31980 -$39974 or $35457 - $46086 depending on the formal qualifications and experience of the successful candidate.
Appointment Type: Full-Time (36.25 HPW) until March 31, 2007; External Funding (continuation beyond March 31, 2007 dependent upon future funding). Competition #140-02.03 May 2003 (applications will be accepted until the position is filled).
Applications: To be submitted in writing on UNB application form with resume and names of at least three referees, to: UNB, Fredericton, Human Resources. Please see the UNB website for further information: www.unb.ca/postings
Since y'all are discussing disposing of old oil from a high-voltage tank, I thought someone might have an idea where to purchase such oil for refilling a high voltage tank on an SEM. Someone drained the HV tank before they stored the microscope, thinking the oil was somehow too hazardous to leave in it. I don't know how they disposed of it.
I'm told that the brand and type for this specific instrument (1984 ISI DS-130) is Shell Diala. I found it on the web, but the smallest quantity the distributor sells is a 55 gallon drum for $174 (which is not out of the question, just wasteful if the tank only takes 7-10 gallons). Any ideas on where to get 10 gallons of suitable dielectric oil?
Thanks in advance.
Regards, Andrew T. Werner Chief Metallurgist Shaped Charge Research - Metallurgy Laboratory Schlumberger Reservoir Completions Technology Center 14910 Airline Road, Rosharon, TX 77583-1590 Voice (281) 285-5272 Fax (281) 285-5273
Enclosed is the table of contents for the May issue of Microscopy Today.
I will close the magazine's subscription list on Friday, May 9th for this issue.
MT is free for any one interested in, or connected with, microscopy anywhere in Canada, Mexico, and the USA. MT is also free for Microscopy Society of America members anywhere.
The subscription rate for those not qualifying for free subscriptions has been reduced from $80 or $110 to $35US per year.
Please subscribe via our web page: http://www.microscopy-today.com
Thank you,
Ron Anderson, Editor Microscopy Today
Cryoelectron Tomography Stephen W. Carmichael, Mayo Clinic Optimizing the Sampling Design of Morphometric Experiments John M. Basgen, University of Minnesota FTIR Hyperspectral Images of Microscopic Droplets of Splattered Blood Scott W.Huffman*, Kara B. Lukasiewicz** and Chris W. Brown*** *NIH, **Mayo Clinic, ***University of Rhode Island Setting White, Black and Gamma on Continuous Tone Grayscale and Color Images using Photoshop Jerry Sedgewick, University of Minnesota How to Recognize and Avoid AFM Image Artifacts Paul West and Natalia Starostina, Pacific Nanotechnology, Inc. Quality in Electron Microscopy Tony Bruton*, Steve Chapman** and Paul Harding*** *E.M. Unit, University of Natal, **Protrain Courses, ***Integrated Systems Management, Nissan Motor Co. Use of a New Imaging Technique to Document Deformations Recorded in the Environmental Scanning Electron Microscope Edward F. ONeil,* Hamlin M. Jennings,** and Jeffrey J. Thomas** *US Army Corps of Engineers, **Northwestern University On The Limits Of Limits Tobias I. Baskin, Division of Biological Sciences, University of Missouri Examination of Hydrated Bacteria in An Environmental Scanning Electron Microscope (ESEM) Ann Fook Yang and Miloslav Kaláb, Agriculture and Agri-Food Canada Dye is Money Christian Lohr, University of Kaiserslautern, Germany Water Recirculator (Chiller) Maintenance Owen Mills, Michigan Technological Institute EDS Spectral Artifacts / Sum Peaks: A Reminder Steven S. Hurban, Endicott Interconnect Technologies, Inc.
A colleague is looking for a used (or surplus to requirement) field cancelling system for use with an SEM.
Best regards,
Mike Wombwell
**************************************************************************** ***************************** Quorum Technologies Unit 15A, Euro Business Park New Road, Newhaven East Sussex, BN9 0DQ, UK Tel: +44(0)1273 510535 (main switch board) Tel: +44(0)1273 511063 (direct line) Mobile +44(0)7989 426 431 Fax: +44(0)1273 510536 mike.wombwell-at-quorumtech.com http://www.quorumtech.com
UNIVERSITY OF CENTRAL FLORIDA TENURE-TRACK FACULTY POSITION ADVANCED MATERIALS PROCESSING AND ANALYSIS CENTER (AMPAC)
The Advanced Materials Processing and Analysis Center (AMPAC) of the University of Central Florida has an immediate opening for a tenure-track faculty position. The successful candidate would use electron microscopy as the primary support tool for their research efforts. Experience with advanced microscopy techniques such as HRTEM, HR-ADF-STEM, EELS, and Holography is advantageous. The candidate would be expected to teach courses in an appropriate academic unit, build and maintain their own research program, and play a significant role in oversight of and training in our electron microscope facility. Desire to work in a collaborative role with expanding interdisciplinary materials programs is required. The appointment will be made at a level consistent with the experience of the applicant. A Ph.D. in Materials Science and Engineering, or a closely related discipline in the physical sciences or engineering, is a minimum requirement.
The University of Central Florida is a comprehensive university located in Orlando, Florida with a current enrollment of approximately 39,000 students. Orlando and the surrounding area is quickly becoming Florida's high tech region providing opportunities to collaborate with Lockheed-Martin, Siemens-Westinghouse, Lucent Technologies, Harris Semiconductor, NASA-KSC and others. The Advanced Materials Processing and Analysis Center (AMPAC) of the University of Central Florida is a State University System recognized center for the advancement of materials science and engineering. AMPAC joins the rank of other successful centers at UCF such as Center for Research and Education in Optics and Lasers (CREOL), Institute for Simulation and Training (IST) and the Florida Solar Energy Center (FSEC).
Interested candidates should submit a letter of application, Curriculum Vitae with the names of a minimum of three references, and a detailed research plan to: Microscopy Search, AMPAC, UCF, Orlando FL 32816-2455. Fax: 407-882-1462. E-mail: ampac-at-mail.ucf.edu. The applications will be reviewed beginning June 1 and will continue to be reviewed until the position is filled.
The University of Central Florida is an Equal Opportunity/Affirmative Action Employer As an agency of the State of Florida, UCF makes all search materials, including transcripts, available for public review upon request.
Hi gang - what is your best receipe for cells in culture on a small filter for SEM? This guy is growing astrocytes on one side of filter and on the other side he is growing endothelial cells. He wants to look at blood barriers. Any ideas will be greatly appreciated. Thanks Barb
We have discontinued making single hole Be grids for a variety of reasons. We can do a single hole 3 mm disc with holes ranging from 0.5um to 2 mm in a variety of other materials such as SS, aluminum, tantalum, moly, Pt, tungsten etc. Much depends upon the aspect ratio of the hole dia to thickness. Let me know if we can assist.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "David Henriks" {henriks-at-southbaytech.com} To: "K.N. Bozhilov" {bozhilov-at-mail.ucr.edu} Sent: Thursday, May 08, 2003 12:09 PM
I would like to know supplier of high quality Carbon rod (99.9999% or better). 1/8 inch is fine. Please advise.
Thank you,
Hiromi Konishi, Ph.D Dept. of Earth and Planetary Sciences The University of New Mexico
To: Vendors of used light microscopes. See the following message for details:
} I'm only looking for the basic frame; I'll remove the } optics so they can be busted or missing or whatever. This is } something like what I'm after: } } http://www.geocities.com/nicholl.geo/stuff/d6/aoframe.jpg } } Any scope with an X-Y mechanical stage (the only critical } part) and a basic focus mechanism, then I'm set.
Please reply to Scott Childs {scott-at-rre.net} thanks for your help, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) **********************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I do believe the smallest quantity you can buy is 55 Gallons - at least in the US. For that reason, I have a significant quantity surplus to my requirements (about 30 gallons, in fact) - but it would probably cost more to ship it to you in Texas than it would for you to but your own 55 gallons! (My oil is actually Exxon Univolt 60, but I think that and Shell Diala are equivalents).
Years ago, you could get this oil in 5 gallon containers, but they leaked (as I can attest). I was told this was the reason they were discontinued.
Tony.
At 08:20 AM 5/7/2003 -0500, Andrew Werner wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** Manager, CMSE Shared Experimental Facilities ** MIT Rooms 13-1027 or 13-3090 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
A number of years ago I was able to dispose of old oil (no PCBs) and get a 5-gallon carboy of new from our local electric company. They also had facilities for disposing of PCB-contining oils. I'm fairly sure that these days I'd have to fill out EPA forms and all, but a few years ago the electric comapany was amenable because 5 gallons was peanuts to them! You might look into official ways to do the same.
Good luck!
Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kelidrwil-at-seneca24.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, May 8, 2003 at 09:07:15 ---------------------------------------------------------------------------
Email: kelidrwil-at-seneca24.net Name: R.F.Wilson
Organization: Self
Education: Graduate College
Location: Waterloo NY USA
Question: What is the best method for mounting pollen for study with the optical microscope ?
Check with a local power company, they use the oil in high voltage transformers in the powergrid. I've been able to buy small quantities when I needed it for a project.
As a sidenote, a full tankwaggon was stolen here in Umeå a couple of years ago. It was standing outside a powerstation and somebody thought it contained diesel.... I guess a number of cars broke down that winter. :-)
Regards, Göran Axelsson
Anthony J. Garratt-Reed wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I do believe the smallest quantity you can buy is 55 Gallons - at } least in the US. For that reason, I have a significant quantity } surplus to my requirements (about 30 gallons, in fact) - but it would } probably cost more to ship it to you in Texas than it would for you to } but your own 55 gallons! (My oil is actually Exxon Univolt 60, but I } think that and Shell Diala are equivalents). } } Years ago, you could get this oil in 5 gallon containers, but they } leaked (as I can attest). I was told this was the reason they were } discontinued. } } Tony. } } At 08:20 AM 5/7/2003 -0500, Andrew Werner wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Since y'all are discussing disposing of old oil from a high-voltage } } tank, I thought someone might have an idea where to purchase such oil } } for refilling a high voltage tank on an SEM. Someone drained the HV } } tank before they stored the microscope, thinking the oil was somehow } } too hazardous to leave in it. I don't know how they disposed of it. } } } } I'm told that the brand and type for this specific instrument (1984 } } ISI DS-130) is Shell Diala. I found it on the web, but the smallest } } quantity the distributor sells is a 55 gallon drum for $174 (which is } } not out of the question, just wasteful if the tank only takes 7-10 } } gallons). Any ideas on where to get 10 gallons of suitable } } dielectric oil? } } } } Thanks in advance. } } } } Regards, } } Andrew T. Werner } } Chief Metallurgist } } Shaped Charge Research - Metallurgy Laboratory } } Schlumberger Reservoir Completions Technology Center } } 14910 Airline Road, Rosharon, TX 77583-1590 } } Voice (281) 285-5272 Fax (281) 285-5273 } } } } } ** Anthony J. Garratt-Reed } ** Manager, CMSE Shared Experimental Facilities } ** MIT Rooms 13-1027 or 13-3090 } ** 77 Massachusetts Avenue } ** Cambridge, MA 02139-4307 } ** USA } ** } ** Phone: (+) 1-617-253-4622 } ** Fax: (+) 1-617-258-6478 } ** } } }
Here's an article on the subject that might be of use.
http://www.nttworldwide.com/tech2308.htm
Good luck, your problem does not have a simple solution.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: Andrew Werner [mailto:werner-at-rosharon.oilfield.slb.com] Sent: Wednesday, May 07, 2003 9:21 AM To: Microscopy Listserver
Since y'all are discussing disposing of old oil from a high-voltage tank, I thought someone might have an idea where to purchase such oil for refilling a high voltage tank on an SEM. Someone drained the HV tank before they stored the microscope, thinking the oil was somehow too hazardous to leave in it. I don't know how they disposed of it.
I'm told that the brand and type for this specific instrument (1984 ISI DS-130) is Shell Diala. I found it on the web, but the smallest quantity the distributor sells is a 55 gallon drum for $174 (which is not out of the question, just wasteful if the tank only takes 7-10 gallons). Any ideas on where to get 10 gallons of suitable dielectric oil?
Thanks in advance.
Regards, Andrew T. Werner Chief Metallurgist Shaped Charge Research - Metallurgy Laboratory Schlumberger Reservoir Completions Technology Center 14910 Airline Road, Rosharon, TX 77583-1590 Voice (281) 285-5272 Fax (281) 285-5273
Jim Darley, at Proscitech (www.proscitech.com.au) has both graphite and carbon 1/8" rods, and ships to anywhere at good prices
I have no connection apart from being a regular satisfied customer.
cheers
rtch
To: Microscopy-at-sparc5.microscopy.com
Howdy Fred,
Thank you (and several others! - I need to be better at responding) for your help and suggestions.
David Sundin at DS Fluids will sell me 5 gallon pails but, since the remaining portion of the drum will require re-processing, cost for 10 gallons will be the same as 55 gallons. It is a viable option though - does not waste the rest.
Others have suggested the local electric utility - a possibility but finding the right person to ask would be a challenge - or donating the excess from a 55 gal drum to the Physics Dept. of the University.
One person referred me to an outfit in Houston that sells him Exxon Univolt, and I have a call in to the sales dept. If this pans out it will be the exact answer to my problem.
Thanks again to all who offered advice. If it is not wasting your time, I will post about how this turns out.
Regards, Andrew
At 03:08 PM 5/9/2003 -0400, Monson, Frederick C. wrote:
} Afternoon Andrew, } It appears that you are within hailing distance of an expert in the } area of dielectric oil replacements, etc. } } These folks certainly appear to concentrate on your problem. Perhaps they } would provide a 25Gal lot for a fellow Texan. } } http://www.dsifluids.com/dielectric_fluids_home.htm } } http://www.dsifluids.com/beta.html } } Here's an article on the subject that might be of use. } } http://www.nttworldwide.com/tech2308.htm } } Good luck, your problem does not have a simple solution. } } Cheers, } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging } Mail Drop: Geology } West Chester University } West Chester, PA, 19383 } http://darwin.wcupa.edu/casi/ } Phone/FAX: 610-738-0437
Email: daniel.smith-at-liquidminerals.com Name: Daniel Smith
Organization: Liquid Minerals Group, Inc.
Education: Graduate College
Location: Houston, TX USA
Question: I am interested in measuring particle size of metal oxide particles suspended in a process oil. The particles are on the order of 1 - 300 nanometers. I have used indirect methods (DLS, etc.), but I would really like to take photos of the particles and determine particle size, crystal structure, agglomeration and other properties.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Hiromi Konishi wrote: ================================================================== I would like to know supplier of high quality Carbon rod (99.9999% or better ). 1/8 inch is fine. Please advise. ================================================================== The purity of carbon and/or graphite rods is normally discussed in terms of so many ppm ash left over after ashing in a high tempeature muffle furnace. I understand that there is some amount of "controvery" with regard to the **exact** meaning of these terms, but generally speaking, if the carbon (or graphite) material exhibits less than 10 ppm ash, it is described as being "spectrographic" purity, and if less than 6 ppm ash, then "spectroscopic" purity.
As you might imagine, there is a difference in the cost to reduce the ash content from 10 down to 6 ppm. SPI Supplies offers both spectroscopically pure carbon and also graphite rods in "spectroscopic" purity as standard products, in 1/8" and other diameters. Several of our major competitors offer similar purity, at least in terms of graphite rods, so our product is not unique in that respect.
However, it is not clear if you really wanted (literally) **carbon** or if you wanted graphite rods (but like many of us tend to call them "carbon"). More information about our carbon and graphite rod products and their purity can be found on URL www.2spi.com\catalog\spec_prep\carbon-rods.shtml
For vacuum evaporation work, most workers seem to prefer graphite to carbon rods.
Disclaimer: SPI Supplies offers both carbon as well as grahite rods in high purity (less than 6 ppm ash).
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 ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id TAA00806 for dist-Microscopy; Sun, 11 May 2003 19:01:16 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id TAA00801 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Sun, 11 May 2003 19:00:45 -0500 (CDT) Received: from mailhost2.auckland.ac.nz (mailhost2.auckland.ac.nz [130.216.191.4]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id TAA00789 for {Microscopy-at-sparc5.microscopy.com} ; Sun, 11 May 2003 19:00:32 -0500 (CDT) Received: from glgnov2.auckland.ac.nz (glgnov2.auckland.ac.nz [130.216.59.90]) by mailhost2.auckland.ac.nz (8.12.9/8.12.9/8.12.9-ua) with ESMTP id h4BNracX021457; Mon, 12 May 2003 11:53:36 +1200 (NZST) Received: from GLGNOV2/SpoolDir by glgnov2.auckland.ac.nz (Mercury 1.48); 12 May 03 11:54:51 +1200 Received: from SpoolDir by GLGNOV2 (Mercury 1.48); 12 May 03 11:54:50 +1200 MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
This may well be so, but my old Edwards 306 and I prefer 'carbon', as the graphite ones need appreciably higher temperature to evaporate it, and the rod holders get realy hot.
Give me 'carbon' any day
cheers
rtch
To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Hiromi Konishi wrote: ================================================================== I would like to know supplier of high quality Carbon rod (99.9999% or better ). 1/8 inch is fine. Please advise. ================================================================== The purity of carbon and/or graphite rods is normally discussed in terms of so many ppm ash left over after ashing in a high tempeature muffle furnace. I understand that there is some amount of "controvery" with regard to the **exact** meaning of these terms, but generally speaking, if the carbon (or graphite) material exhibits less than 10 ppm ash, it is described as being "spectrographic" purity, and if less than 6 ppm ash, then "spectroscopic" purity.
As you might imagine, there is a difference in the cost to reduce the ash content from 10 down to 6 ppm. SPI Supplies offers both spectroscopically pure carbon and also graphite rods in "spectroscopic" purity as standard products, in 1/8" and other diameters. Several of our major competitors offer similar purity, at least in terms of graphite rods, so our product is not unique in that respect.
However, it is not clear if you really wanted (literally) **carbon** or if you wanted graphite rods (but like many of us tend to call them "carbon"). More information about our carbon and graphite rod products and their purity can be found on URL www.2spi.com\catalog\spec_prep\carbon-rods.shtml
For vacuum evaporation work, most workers seem to prefer graphite to carbon rods.
Disclaimer: SPI Supplies offers both carbon as well as grahite rods in high purity (less than 6 ppm ash).
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 ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id KAA03944 for dist-Microscopy; Mon, 12 May 2003 10:11:36 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA03939 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Mon, 12 May 2003 10:11:05 -0500 (CDT) Received: from tumor.soft-imaging.com ([67.104.115.34]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id KAA03931 for {Microscopy-at-sparc5.Microscopy.Com} ; Mon, 12 May 2003 10:10:53 -0500 (CDT) Received: from lakewood.soft-imaging.com (lakewood.soft-imaging.com [192.168.5.225]) by tumor.soft-imaging.com (8.10.2/8.10.2/SuSE Linux 8.10.0-0.3) with ESMTP id h4CF4lc19564 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 12 May 2003 09:04:47 -0600 Received: by hq-dc2.soft-imaging.com with Internet Mail Service (5.5.2656.59) id {J6F4H6QZ} ; Mon, 12 May 2003 09:04:48 -0600 Message-ID: {6127CE87B9BDD511B59D0001028A497D478685-at-hq-dc2.soft-imaging.com}
Daniel,
I think you need to be more specific. Are you looking for help regarding SEM or TEM, sample preparation, observation methods, image acquisition, further image processing, etc?
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: daniel.smith-at-liquidminerals.com [mailto:daniel.smith-at-liquidminerals.com] Sent: Friday, May 09, 2003 5:28 PM To: MicroscopyListserver
Email: daniel.smith-at-liquidminerals.com Name: Daniel Smith
Organization: Liquid Minerals Group, Inc.
Education: Graduate College
Location: Houston, TX USA
Question: I am interested in measuring particle size of metal oxide particles suspended in a process oil. The particles are on the order of 1 - 300 nanometers. I have used indirect methods (DLS, etc.), but I would really like to take photos of the particles and determine particle size, crystal structure, agglomeration and other properties.
A colleague would like to purchase a camera for use in phase contrast and fluorescence microscopy of bacteria. For some of his specimens, he expects the fluorescence may be weak.
He would like a camera with a non-proprietary (firewire) interface, and any included software should be available for the Macintosh platform.
Among the cameras he is interested in are the Qimaging Micropublisher and Qimaging Micropublisher 5.0, and the Optronics Magnafire and Microfire. He would like to have feedback from anyone who has experience with one or more of these cameras, or anyone who has evaluated one of them but has chosen another camera.
Thank you,
Ed King --
Edward J. King Department of Biology University of Utah 257 South 1400 East Salt Lake City, UT 84112
Hi, we have sprung a water leak from our 3 year old TEM. We are using mains 'tap water' to cool as this was deemed (at the time) better than installing a chiller system. Has anyone any experience/advice they can give me? Is this a common occurrence?
Gillian Brown Histology Section, Asthma Biology RI CEDD, GlaxoSmithKline UK gillian.2.brown-at-gsk.com
Do you mean carbon thread? I'm trying to figure out what is better for our samples, graphite rods or carbon thread? Any information will be appreciated. Thank You, Evelyn
At 11:54 AM 5/12/2003 +1200, Ritchie Sims wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A colleague of mine needs to cut frozen sections of adult Drosophila heads for a combination immunohistochemistry/in situ hybridization study. Any suggestions?
Thank you very much,
Joel
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 6646 http://astro.temple.edu/~jbs
The College of Physical & Mathematical Sciences at Brigham Young University (BYU) invites applications for a technical administrative staff position in support of the transmission electron microscopy laboratory. The position will be administered in the Department of Physics & Astronomy.
Facility:
The laboratory is located in a new state-of-the-art facility that will house both a 200KV ultra-high-resolution (S)TEM and a 300KV TEMs, with multiple specimen preparation support labs. Analytical capabilities include EDS and EELS.
Requirements:
An M.S. degree or comparable experience, (Ph.D. degree preferred) in physics, materials science, or a related field is required, plus at least 2 years experience in operating and maintaining transmission electron microscopes for materials applications. The successful candidate must have the ability to train users in both sample preparation and TEM use, to interact effectively and collaborate with users from advanced undergraduates through faculty from a variety of disciplines, and to maintain the instruments and interact with manufacturersservice engineers, as appropriate. BYU is sponsored by The Church of Jesus Christ of Latter-day Saints (LDS, Mormon) and is an equal employment opportunity/affirmative action employer. Strong preference is given to LDS applicants. Employees are required to abide by standards consistent with LDS values.
Application:
Contact E. Daniel Johnson, N-181 ESC, BYU, Provo, UT 84602. Applications will be accepted until July 1, 2003 or until a successful candidate is identified, whichever is later. This is a permanent, full-time, budgeted position.
On Friday, May 9, 2003, at 04:28 PM, by way of MicroscopyListServer wrote:
} Question: I am interested in measuring particle size of metal oxide } particles suspended in a process oil. The particles are on the order } of 1 - 300 nanometers. I have used indirect methods (DLS, etc.), but } I would really like to take photos of the particles and determine } particle size, crystal structure, agglomeration and other properties. } } Any help would be appreciated. } Dear Daniel, You can use TEM to image the particles, but you'll have to get rid of the oil. If it is a volatile oil, things may be as simple as putting a microliter or so on a formvar-carbon grid, evaporating the oil, and inserting the grid into the scope; however, if this can't be done, you may lose some of the particles in the separation process--e.g., the smaller ones could be trapped on a filter membrane. If the oil is soluble in something like acetone or chloroform, you might be able to add about 1ml of solvent, centrifuge the particles, resuspend them, and place an aliquot on a formvar-carbon grid. (Here, carbon is essential, since the formvar will dissolve in the solvent.) Particle size and structure should be measurable, with the caveat that, if you lose particles, the size distribution will likely be altered. Agglomeration could easily be lost, since, even if you have a volatile oil, the particles can aggregate as the oil evaporates. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Senior EM Technician position available in the Department of Pathology at George Washington University Medical Center.
Must have experience in all phases of TEM: fixation, embedding, thick & thin sectioning, scoping and scope maintenance(filament changing and alignment, mostly).
Experience with clinical samples a plus, but not neccessary.
This is a service lab mostly and handles primarily clinical samples though we do get some research samples.
Interested?
Contact by e-mail at patpxs-at-gwumc.edu
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Charles Interestingly and without any "scientific" explanation, I do find that "carbon (not graphite) rods" works better to me. Somehow the properties of the evaporated film is different. To me, films from "carbon" (not graphite) is "better"... Rational (not necessary scientific) explanation may be that we have deal with "sublimation" of carbon/graphite. During this process we generate a cloud of particles (clusters), which being condensed on the surface formed the film. We have to understand that thermal evaporation of carbon creates actually particles from a few to tens atoms in size, it's not mono-atomic cloud. Therefore, the film quality will dependent from the composition of cloud. Personally, I prefer "Electron Gun" carbon evaporation: the films are much more stable and uniform. Have a good day. Sergey
At 11:17 PM 5/10/2003, 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'm shopping around for a replacement for the inkjet printer I've been using to output my SEM images. I have been using an HP 970 very happily, but it is getting worn and cranky about feeding paper consistently. I have tried cleaning the feed rollers (have lots of green ink on them by this time), but with limited success.
My reason for writing is that the SEM data system (Hitachi S3000N) uses Windows 95 (haven't gotten funding for the overhaul to allow running Windows 2000) and it does not have USB ports available. In principle USB might work under late Windows 95, but I haven't gotten that going. So I am limited to parallel port or Ethernet connections to the computer. I also hope to get a system with more than a small amount of memory built in to help speed up the return of control to the microscope while the images print out.
Any recommendations for a good printer to use based on these limitations?
I have some choices, but none seem 100% satisfactory. I'd be very happy with an HP 990, but those are off the market and the HP 995 only does USB and wireless. A number of possibilities are Windows 98 and up only. Earlier I tried an HP CP1160, but I wasn't happy with the image tones despite aggressive adjustments to the printing and it also seemed to require frequent ink changes (small cartridges only available). Image quality was ok although it was slower than the 970. So I'm wondering about other choices (Epson, Canon) before buying a unit blind and being stuck with it like I am with the 1160.
Richard Shalvoy Arch Chemicals, Inc. 350 Knotter Drive Cheshire, CT 06410 (203) 271-4394 rbshalvoy-at-archchemicals.com
Sorry for any double posting. I'm just learning how to use this listserv.
A colleague of mine needs to cut frozen sections of adult Drosophila heads for a combination immunohistochemistry/in situ hybridization study. Any suggestions?
Thank you very much,
Joel
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 6646 http://astro.temple.edu/~jbs
We are looking for an experienced EM tech. for a busy Clinical service lab. Details are online at:http://employ.uchc.edu/ It is listed under Anatomic Pathology Search code 2003-161.
Where in the cooling system did the leak occur? I assume it wasn't a coupling or someplace obvious. If it wasn't, I would look into two possible causes: 1. You have electrolysis from improper grounding between the cooling system and/or your TEM. 2. Turbulence from air in the water lines are eroding the copper cooling lines.
We have had the latter happen to us this past year. Nothing spoils a morning more than walking into your lab and finding water all over the floor. Especially if the pumps are running and there is water between you and them, along with a few power supplies in the high kV range.
Good luck, I hope this helps.
Bill
Bill Carmichael EM Faculty Madison Area Technical College (MATC) 3550 Anderson St. Madison, Wisconsin 53562 http://www.matcmadison.edu/electronmicros wcarmichael-at-matcmadison.edu
Ladd Research produces hundreds of carbon substrates daily and this is what we've found concerning carbon and graphite.
Carbon 1. Requires less current 2. burns cooler 3. doesn't chunk as much
Graphite 1. Sparks more, especially with a poor vacuum. 2. Messier to handle 3. Easier to cut and point
Since we cut and point a great deal of rods our personnel prefer to use graphite. The resulting substrates are excellent with either product. Since we also manufacture vacuum deposition systems and high volume rod sharpeners we can handle graphite easier. Bottom line is that we feel it is customer's preference, based on the available equipment and volume.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, May 12, 2003 2:58 PM
To whom it may concern:
Does anyone know of an independent service contractor for JEOL TEM's that take care of the Los Angeles area. Our current contractor will soon be retiring.
Thank You,
Michael Pidgeon University of Southern California Dept. of Cell & Neurobiology
We have had a rash of cooling leaks in our facility in copper lines running to and from our refrigerated water recirculators and in the brass plumbing connectors. The pipes and fittings are being eaten out from the inside, resulting in pinhole leaks at unpredictable times. Fortunately, the leaks usually start small and so far we've caught them before the deluge causes major damage.
There have been two causes for these leaks. One is that, until recently, I was unaware of the presence of a shut-off valve inside our water chillers, which turns the cooling water to the chiller on and off as the compressor turns on and off. The water cooling the coils should only be running when the compressor is delivering a heat load to dissipate. (The water from the tank to the scope runs constantly, of course.) These shut-off valves wear out routinely every couple of years and should be checked once a year, at least, I now know. If they wear out, the water runs constantly and can cause major internal "etching" of the metal cooling lines. Also, one of our chillers had the flow rate to the coils set WAY too high, with the same effect. Moral: have a refrigeration person check your shut-off valves and flow rates.
The other reason is that some water supplies are simply more corrosive than others (our refrigeration guy used the term "hungry" water) and will eat your copper lines and metal fittings from the inside out over varying periods of time, regardless of flow rates. Have your water checked for hardness and other water voodoo by some qualified person with a water voodoo test kit. Regular copper lines can be replaced by hardened copper (I think it's called K-copper) or stainless steel lines, but there is an expense factor involved.
So far, our main leak detectors have been our squeaky shoes on wet floors, rather than soaked and electrocuted microscopes. We now watch our lines carefully. One consistent leak trait is that they occur over weekends, so we suspect the Leak God does its thing at 5:15 p.m. on a Friday afternoon, just as I'm ordering a Katy Trail Pale Ale down at the old Flatbranch Brewpub and Grill (no financial interest, just a satisfied customer).
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Bill Carmichael [mailto:wcarmichael-at-charter.net] Sent: Tuesday, May 13, 2003 9:54 AM To: Microscopy-at-sparc5.microscopy.com
Gillian,
Where in the cooling system did the leak occur? I assume it wasn't a coupling or someplace obvious. If it wasn't, I would look into two possible causes: 1. You have electrolysis from improper grounding between the cooling system and/or your TEM. 2. Turbulence from air in the water lines are eroding the copper cooling lines.
We have had the latter happen to us this past year. Nothing spoils a morning more than walking into your lab and finding water all over the floor. Especially if the pumps are running and there is water between you and them, along with a few power supplies in the high kV range.
Good luck, I hope this helps.
Bill
Bill Carmichael EM Faculty Madison Area Technical College (MATC) 3550 Anderson St. Madison, Wisconsin 53562 http://www.matcmadison.edu/electronmicros wcarmichael-at-matcmadison.edu
Does anyone know of an independent service contractor for JEOL TEM's that take care of the Los Angeles area. Our current contractor will be retiring very soon.
Thank You,
Michael Pidgeon University of Southern California Dept. of Cell & Neurobiology
We too have had these internal leaks in copper cooling lines. Thousands of dollars of damage has been done and many of my hours spent making repairs.
We tracked the source to impingement corrosion, particulate laden water moved through small diameter tubing at high speed. We have (no, had) street water cooled condensers on our refrigeration chillers. As Randy said, without careful monitoring and adjustment, the valves controlling the required amount of street water necessary to remove the heat load stay open all the time allowing tremendous amounts of water to flow through that circuit. At high pressure (our street water is 85 psi) and with some micron size particles in it and it will eat copper tubing at any bends in the tubing. Oh, and yes, always on weekends.
We've also seen the problem on street water cooled diffusion pumps, where the coiled copper tubing soldered to the side wall of the diff stack will perforate. Yes, this tubing can be replaced in the field in a few hours with common plumbing tools.
So the result is we no longer have refrigerant cooled chillers. With Haskris' help, I converted all to water-water cooled systems with the new "braze-pak" style heat exchangers that are resistant to this corrosion. No problems since.
RE: Water. I pulled this bit of info from this list recently posted by Andrei Shchukarev who wrote that Neslab recommends water for long term usage in the 1 to 3 megohn-cm (compensated to 25oC) range.
Speaking of Neslab, they have a nice detail on their big chillers, a level switch that shuts the system down if the tank level drops below the switch setting. With that at least you won't pump all of reservoir onto floor. Easily added to any chiller.
I have no financial interest in either company mentioned in this email.
Owen Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
At 03:55 PM 5/13/2003 -0500, Tindall, Randy D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Richard, I have been very happy with a couple of Epson inkjet printers I have. They seem to have a good life and never give me any trouble. They are expensive in ink cartridges and the right paper, but cheap to buy initially. I don't think you can put much memory in them. They just are slow. Better to buy several to speed things up. The last time I bought one (one year ago) they could use parallel or USB connection. You can also buy USB cards for your WIN95 computer that should have software. I have also used a Datalink parallel to Ethernet connector to put my laser printer on the network. Using the network, I have five printers in my lab I can print to. I use the 1200 dpi laser printer for quick SEM images and the inkjets for x-ray maps and glossy SEM photos. Good luck. I have a S3000N, too.. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Shalvoy, Richard B **CHES" {RBShalvoy-at-archchemicals.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, May 12, 2003 1:04 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (s54andrews-at-stanford.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 13, 2003 at 19:01:58 ---------------------------------------------------------------------------
Email: s54andrews-at-stanford.edu Name: Scott Andrews
Organization: Stanford University
Education: Graduate College
Location: Stanford, CA
Question: Hello,
I am trying to find out the proper configuration for aligning a condenser in a microscope. Unlike all information I can find, it is a reflective setup, i.e. the light source passes through the objective on its way to the sample. To make things more complex, in the middle of the aperture kit, there is a diffusing glass plate. This can be removed, but it also removes all apertures (aperture stop and field stop) and a intermediate lens. I am unable to tell what the intermediate lens does. It lies between the aperture stop and field stop. With the diffusive glass plate in place, I cannot image the arc poles on the lamp. The light source is a mercury arc lamp with vertical and horizontal bulb alignment knobs. There is a curved, reflective mirror behind the lamp with tilt and vertical position knobs. There is a final knob that moves the mirror along the optical axis. Can you tell me how to align these five knobs so that I achieve Koehler illumination?
Hi My colleague is looking for a universal stage for optical microscope. Petorologists used it to measure optical properties of minerals probably ~30 years ago. You can rotate thin section three dimensionally using the Universal Stage. Nowadays Leica, Zeiss, Nikon, and Olympus do not produce such stage, so I am looking for a used one. You can see the stage: http://www.gemmary.com/insts/forum/messages/8797.html
If you are selling a universal stage or you know a vender for selling such stage, please advise.
hkonishi-at-unm.edu
Hiromi Konishi, Ph.D. Dept. of earth and Planetary Sciences, The University of New Mexico
Hiromi Konishi, Ph.D Dept. of Earth and Planetary Sciences The University of New Mexico
Charles Supper Company 15 Tech Circle - Natick, MA 01760 Tel: (508) 655-4610 Fax: (508) 655-3913 Toll Free: (800) 323-9645 http://www.charles-supper.com/
Thank you, Bob Voigt (bobvoigt-at-restechimage.com) Resolution Technology, Inc. (www.restechimage.com) Phone (614)921-0045 Fax (614)921-0046
-----Original Message----- } From: Hiromi Konishi [mailto:hkonishi-at-unm.edu] Sent: Wednesday, May 14, 2003 3:07 PM To: Microscopy-at-sparc5.microscopy.com
Hi My colleague is looking for a universal stage for optical microscope. Petorologists used it to measure optical properties of minerals probably ~30 years ago. You can rotate thin section three dimensionally using the Universal Stage. Nowadays Leica, Zeiss, Nikon, and Olympus do not produce such stage, so I am looking for a used one. You can see the stage: http://www.gemmary.com/insts/forum/messages/8797.html
If you are selling a universal stage or you know a vender for selling such stage, please advise.
hkonishi-at-unm.edu
Hiromi Konishi, Ph.D. Dept. of earth and Planetary Sciences, The University of New Mexico
Hiromi Konishi, Ph.D Dept. of Earth and Planetary Sciences The University of New Mexico
The following was sent to the MSA Business Office. We are forwarding it to the microscopy listserver in the hope that the originator can find some help.
Please, DO NOT reply to me. Send your responses to the requestor directly, axelschlitt-at-gmx.net, as he apparently does not subscribe to the microscopy listserver. COPY the listserver if you want.
----- Original Message ----- } From: "Axel Schlitt" {axelschlitt-at-gmx.net} To: {BusinessOffice-at-sparc5.microscopy.com} Sent: Tuesday, May 13, 2003 3:16 PM
Hello Becky,
Have you tried Duke Scientific? Their product line may not include exactly what you need (2mm spheres are glass), but they may have something I missed or know where to send you.
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Becky Holdford [mailto:r-holdford-at-ti.com] Sent: Thursday, May 15, 2003 3:09 PM To: Microscopy ListServer
All: one of my engineers needs a source for polystyrene beads approximately 2-3 millimeters in diameter. Any suggestions?
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
We use the Leica Ultrastainers in our EM Facility. Recently we've experienced problems with a nasty precipitate on the sections.
We contacted our Leica Representative and their response was as follows:
" We have a hold on all stain. there is an issue with exporting and new EU country laws.Plus leaking bags. We have an interim solution. We can supply a glass bottle and special bracket for you to mount to the stainer. You can then make up your own stain. We hope top have the issues resolved by the middle to end of the summer."
The lot #s in particular are: 16 Oct 2004 and 20 Juni 2004. We checked some older lots from 2003, and they do not have this problem.
By sending this alert, we are hoping to save you the headache we've just gone through trying to figure out what went wrong.
It is unfortunate that Leica didn't issue a recall on these lots. It would have saved us a great deal of trouble.
I have been working on Ovary samples of fishes,but never been sucessful. The infiltration always seem to be incomplete resulting in the specimen becoming powdery on sectioning. I have also faced the same problem while doing Histology of the same and so also is a friend of mine working on amphibian ovaries.
I would be grateful if any of you can give a solution to this problem of mine.
Joston Paul (Jos)Sophisticated Analytical Instrumetation Facality Shillong, Meghalaya.India
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I am looking for a method for fixing and embedding red blood cells for TEM so that the normal shape and architecture of the cell is preserved. Using my normal TEM fix and embedding, the cells end up all different shapes. Also, does any one have a protocol for freeze substitution embedding of RBC's? I am interested in looking at membranes and cell cytoskeleton.
Thank you David ____________________
David Elliott
Yale University School of Medicine 810 LCI 333 Cedar Street New Haven, CT 06520-8022
This in reference to my previous problem on the TEM sample preparation of Fish Ovary The step-by-step procedure followed are: Primary fixation in cacodylate-buffered 2.5%Glutaraldehyde;washed in 1M Cac buffer; post-fixed in 1M OsO4 and washed in 1M cac-buffer; dehydration was carried out in grades of acetone; cleared in Propylene Oxide; infiltrated in mixtures of Propylene oxide and Spurr embedding medium;Embedding was done in the pure embedding medium and then left for polymerisation. Apart from the above steps I have tried to manipulate the timings, such as fixation, infiltration, etc but to no avail. I would always get powdery sections of the ova. SEE IF YOU CAN HELP ME OUT.
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Does anybody have a Tektronix Phaser IISDX color printer that is still in use ? I need to have some information about the availability of its parts and service.
Thank you!
Pat Sadhukhan Bridgestone/Firestone Research, LLC 330-379-7518
I seem recall from way back in my biological EM days that we fixed RBC's in glutaraldehyde prior to spinning down into a pellet. The alternative to centrifugation may be to allow the fixed RBC's to slowly settle out of the plasma and form a layer that one may be able to dehydrate and embed. I just don't remember if I tried the settling approach or not. Good luck
Cheers,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
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I am looking for a method for fixing and embedding red blood cells for TEM so that the normal shape and architecture of the cell is preserved. Using my normal TEM fix and embedding, the cells end up all different shapes. Also, does any one have a protocol for freeze substitution embedding of RBC's? I am interested in looking at membranes and cell cytoskeleton.
Thank you David ____________________
David Elliott
Yale University School of Medicine 810 LCI 333 Cedar Street New Haven, CT 06520-8022
David- WE fix the cells first in glutaraldehyde and then pellet it down and treat pellet as a bulk and proceed for further processing. Cells do appear in diffeent shapes depending on their orientation in cutting plane. The membranes are well fixed and can be further improved by using tannic acid which imprves visualzation of both membrane and cytoskeleton. Good luck. Shashi Singh CCMB Hyderabad INDIA
I am looking for a method for fixing and embedding red blood cells for TEM so that the normal shape and architecture of the cell is preserved. Using my normal TEM fix and embedding, the cells end up all different shapes. Also, does any one have a protocol for freeze substitution embedding of RBC's? I am interested in looking at membranes and cell cytoskeleton.
Thank you David ____________________
David Elliott
Yale University School of Medicine 810 LCI 333 Cedar Street New Haven, CT 06520-8022
Tel: (203) 785-7573 Fax: (203) 785-3864
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Dear Josten, There is problem with ovaries because of yolk, Did you try methacrylates or spurr resin. Give longer infilteration times with diluted grades. Like for spurr start with 1:6(Resin:alcohal/acetone)followed by 1:2, 1:1 and back to 6:1 before going to pure resin. Shashi SIngh CCMB Hyderabad INDIA
I have been working on Ovary samples of fishes,but never been sucessful. The infiltration always seem to be incomplete resulting in the specimen becoming powdery on sectioning. I have also faced the same problem while doing Histology of the same and so also is a friend of mine working on amphibian ovaries.
I would be grateful if any of you can give a solution to this problem of mine.
Joston Paul (Jos)Sophisticated Analytical Instrumetation Facality Shillong, Meghalaya.India
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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PRINCIPLES OF FLUORESCENCE TECHNIQUES COURSE LAMBS, department of Physics, University of Geno, June 11-13, 2003 The use and application of fluorescence techniques is increasing daily both in the academia and in industry. The Principles of Fluorescence Techniques course will outline the basic concepts of fluorescence techniques and the successful utilization of the currently available commercial instrumentation. The course is designed for students who utilize fluorescence instrumentation and techniques, as well as for researchers and industrial scientists who intend to deepen their knowledge of fluorescence techniques. The theoretical lectures delivered by key scientists in the field are complemented by the direct utilization of steady state and lifetime fluorescence instrumentation provided by leading companies. It is recommended that participants have at least a bachelor's degree in the life science, physical sciences or engineering before attending this course. Topics addressed in this course include:
Basic Definitions and Principles of Fluorescence Fluorescence Polarization Time-resolved Fluorescence Instrumentation Data Manipulation and Data Analysis Fiber Optic Sensors Confocal Fluorescence Microscopy Lifetime Imaging Fluorescence Correlation Spectroscopy
The Principles of Fluorescence Techniques course will be held at: Laboratory for Advanced Microscopy, Bioimaging and Spectroscopy Department of Physics, University of Genoa via Dodecaneso, 33 16146 Genova, Italy Details: www.lambs.it; diaspro-at-fisica.unige.it The number of participants to the Principles of Fluorescence Techniques Course is limited to 40 people. PLEASE RESERVE YOUR PARTICIPATION AS SOON AS POSSIBLE SENDING AN E-MAIL TO: coordinator-at-fluorescence-foundation.org. Details : www.fluorescence-foundation.org
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (michael_burnham-at-ceo.cudenver.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 20, 2003 at 09:14:34 ---------------------------------------------------------------------------
Email: michael_burnham-at-ceo.cudenver.edu Name: Michael Burnham
Organization: Kent Denver School
Education: 9-12th Grade High School
Location: Denver, Colorado
Question: My questions about an older Zeiss EM-9 TEM that I've been working on for use in our High School science program. It's now fully functional and working well. However, my most important concern is to make sure that no X-rays are being emitted from around the gun. I had assumed that a Geiger tube would detect the presence of X-rays although I'm not sure that this is true. I've tried this method with negative results, yet I do want to make 100% certain before I have students learning to use this EM.
Can you please advise??
Thank you greatly,
Michael Burnham Kent Denver School Chair Science Dept. 4000 E. Quincy Ave Englewood, CO 80110
I am planning experiments to use a polyclonal antibody against hemagglutinin (Santa Cruz Biotechnology) to localize the HA epitope at the EM level, using a post-embedding protocol on transgenic nematodes expressing an HA-tagged protein. I am uncertain how strongly I can fix the HA epitope and still get specific binding on thin sections.
Does anyone have experience with aldehyde fixations for HA-tagged proteins, either for immunofluorescence or for immunoEM? Experience with any species might be helpful in guessing how the epitope will behave. Will the HA tag survive a fixation with buffered 2.5% glutaraldehyde?
I'm planning a microwave fixation in aldehydes, dehydration through methanol, and embedding into LR Gold.
Thanks in advance for any advice you may have.
Best regards,
Dave -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
David, Gluataraldehyde is not at all good for immunostudies. Fix your specimens briefly in paraformaldehyde 4% and follow it up for dehydration and LR gold embedding as usual. Set the conditions (fixative, blocking etc)for LM before and then proceed for Immunogold. Shashi
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Dear Colleagues, we are searching for an antibody recognizing the active form of Caspase 3 (mouse) to identify apoptotic cells in cryo sections which were formaldehyde-prefixed. Any experiences, tricks, tips and recommendations are welcome.
Thanks in advance
Daniel
-- (-)-(-) --------------------------- \"/ --- Daniel Eberhard =V= Developmental Biology & Molecular Pathology University of Bielefeld D 33501 Bielefeld/Germany
I am Dr Luisa Pimentel Estrada, the wife of Joseph Ejercito Estrada,former president of Philippines. I have children with my husband Jose, Jacqueline and Jude.Two sons and a daugther.This mail may be coming to you as a surprise or an article but it is very real.I gave the mail to my daugther Jude to send the mail to any contacts she sees and may be a God fearing person will listen to our plight.I will want you as the receiver to read through it and think very well if you can help or render us any assistance. My husband Joseph Ejercito estrada was elected as the 13th President of the Philippines in May 1998 by the people of Philippines due to his popularity in the film industry made him to win the largest popularity in the history of election in Philippines.He has attain the position of Senate in 1987,then vice-president in May 1992 and later become the president 1998.My husband became mayor of his hometown, San Juan in 1969 but it was 1972 that he had a string of public successes. My husband was named one of the ten Outstanding Young Men in Public Administration. He was also named Most Outstanding Mayor and Foremost Nationalist and Most Outstanding Metro Manila Mayor. My husband is recently accused of illegal acquire some four Billion Peso ($80M) during his 31 months in office as President backed up by an uprising of mass Demonstrators and Senate Traitors. They also said that he has skimmed off tobacco excise Taxes benefitting from government business deals.Most of them benefitted from my husband's generousity when he was in office. But they just turned around to be the ones to impeach him. I have tried every possible means to get him out of Detention without success. The Despotic forces in power appear bent in deriding him, rubbishing his achievements while freezing all his known Bank accounts.He has been accused of economic plunder carrying the maximum penalty of death. To the worst of it all,all other wives of my husband especailly Guia Gomez and some of his children born outside wedlock are testifying against us.In conjunction with the PCGG set up by the recent President Arroyo Macapagal Gloria. These are some of the allegations file before my husband in the impeachment trial; 1. Gov. Luis Singson, a longtime friend of my husband, said he provided the my husband with more than $8 million in payoffs from illegal gambling and $2.7 million from tobacco taxes. 2. Witnesses testify one of an account in the Philippines third largest bank held millions of dollars in bribes collected by my husband. Equitable PCI Bank President George Go resigns. The banks senior vice president, Clarissa Ocampo, said she saw my husband sign a false name to documents withdrawing $10 million from a secret personal account. 3. On Dec 31 Five synchronized bomb attacks kill 22 people and injure more than 120 in Manila, days before the trial is to return from holiday recess. Police accuse Muslim rebels but many fear the bombs may be linked to the trial. 4. That my husband received about $8.5 million in pay-offs from illegal gambling operators. 5. That my husband participated in a real estate business controlled by me and my son Jose despite a prohibition on outside business interests while in office. In fact, My husband is suffering from bronchitis and emphysema right now and he detained at the Veterans Memorial Medical Center in Quezon City in hospital prison outside Manila where the life of my husband is in danger.I will let you know that it is political motivated by Gloria Arroyo. Meanwhile,the government has said that it may drop rebellion charges against my husband allies Senator Juan Ponce Enrile and the former ambassador to the United State Ernesto Maceda because they were in the side of my husband,both men were later jailed by the government of Gloria Arroyo that they instigated a march on the presidential palace by 50,000 supporters of my husband.These are all political.All these are done to have my husband death. Presently life has not been easy for my children and I,my travelling documents have been seized by the government and they restricted me and my children to have access to my husband. Haven noticed the way things are going with us,my husband decided to let me know that he deposited some money with some Banks .These funds are presently deposited in his private Bank accounts, Three in whole, one in Europe,one in Central America and one in the Bahamas and all deposited in our name. The Fund in question is put all together exceed One hundred Million. 45 Million USD deposited with a Bank in CommonWealth of Dominica,79. Million USD with a Bank in Spain - and the Bahamas- 35. Million USD. As a prominent person here in Philippines I have no intention of getting involved in any criminality, and as such, no matter the gain I may stand to make I will not attempt to secure the Transfer of any of these sums if the process is not legal or is avoidable due to contravention of International Banking Statutes. I have in this regard studied the present status of the said funds against the order of the Government of Philippines as supported by the government of America, The Americas and Europe to freeze and terminate all Money Transfers to or From an account in our name. And I find it impossible to ensure the safe transfer of the Money in Bahamas, hence I will not attempt to.I have also found a way to tackle the Bank in Spain by using the service of a credit Commission who assist us in the lifting of the said fund from our bank PNB to the bank in Spain.Lastly,And I have reached an understanding with a Director of the bank in Commonwealth of Dominica and we have both devised a plan ( In which he is to play no active role). The above plans I mentioned are both legal, and feascible in so far as well as we can get the participation of a Foreign entity whether individual or corporate who would participate in the execution of our intention which I can safely declare to be risk free in all absolute. Your participation in this regards will entail that you enter into a trust agreement with me, wherein, in my capacity as Administrator of My Estate I will appoint you as Trustee and I, Trustor over the said sum, deposited at the bank in Dominica and the Bank in Spain. This agreement will form a legal basis for the Transfer of the amount to you, where as you will open an account with the bank of the same branch, because this is the only way we can execute a Transfer from the fund in the account without provoking an enquiry. You will noticed that when Monies are transfered from a Bank to another bank exceeding Hundred thousand American Dollars, the banks must report such transaction to the Apex Bank and if any of the parties whose itinerary is stated in the Bill of Transfer does not have clearance from the Government or is having such status as would ensure the money is siezed and the transfer will be aborted, Terminated and the fund confisticated, but if such transfer is done within the same bank ,the bank MAY NOT report such to the government Bank. Thus, if you were to execute an agreement of Trusteeship with me and open an account with the banks.I will have the banks wire the fund in our account into yours within the same bank and you will inturn wire the money to your oversea account in your country and hold the money in Trust under the terms of the agreement we will enter for a period that would not exceed four months. And then you will transfer the fund to a Bank in Malaysia where we are making arrangements to open a special account.
Now I want to start a new life with the money.I want to invest the money outside Philippines. I will want you to appreciate the fact that we are yet unaquainted, thus in your response a brief profile of yourself will certainly promote Trust on my side. I will stop here hoping to recieve an earnest positive response from you, where as I will implore you to handle this matter with utmost confidentiality and sincerity, so that we can have many other profitable interaction in the near future. I await your response. I would want us to be in partnership in any good business you may suggest in your country. Please handle this transaction with maturity and sincerity. Best Regards, Dr Mrs Luisa Pimental Estrada __________________________________________________________ Get your Private, Free Email from HTTP://www.DmailMan.Com
I'm looking for tips on sectioning 5 to 50 micron thick polymer films. Typically samples would be polyolefins, but sometimes (including my current set) they are nylons. I have a room temp microtome available, and have had *limited* success sectioning some nylons to around 2 microns, but sometimes need to go thinner.
I would appreciate any tips?embedding medium, sectioning conditions, etc.
Commercial responses are welcome off-list, both from vendors and service labs.
thanks,
Jim Passmore Sr. Analytical Chemist Cryovac Division Sealed Air Corporation james.passmore-at-sealedair.com
HI Dave, I've done immunfluor. of HA fixing with 2% pfa in the presence of 0.075% saponin. Never tried glut. 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
On Tuesday, May 20, 2003, at 09:51 AM, by way of Ask-A-Microscopist wrote:
} Question: My questions about an older Zeiss EM-9 TEM that I've been } working on for use in our High School science program. It's now fully } functional and working well. However, my most important concern is to } make sure that no X-rays are being emitted from around the gun. I had } assumed that a Geiger tube would detect the presence of X-rays } although I'm not sure that this is true. I've tried this method with } negative results, yet I do want to make 100% certain before I have } students learning to use this EM. } } Can you please advise?? } Dear Michael, A Geiger counter--the simple, hand-held type--should be sufficient. You could also use a hand-held ionization-chamber detector (one brand is called QT-pi), or you could put a film badge on the gun as a continuous monitor. Be sure to check all around to eliminate the possibility of directional x-ray leaks; e.g., between pieces of lead shielding. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
To keep the resin from splitting away from the film, you will need to section a small piece of the film that is surrounded by the resin. To do this, cut your polymer film into small triangles and then glue a thin wire with super glue to back end of the triangle. Put the pointed end into a beam capsule and wrap the end of the wire around the little piece of plastic that holds the cap on and try to wrap it tight enough to hold securely. You may have to bend the wire so that the pointed end is positioned properly for sectioning. Add some medium grade LRWhite and polymerize. We have cut both thick and thin sections from these blocks. Use formvar coated grids for thin sections.
Phoebe J. Doss Manager/Adjunct Instructor Electron Microscope Lab Department of Physiological Sciences 264 McElroy Hall Oklahoma State University Stillwater, OK 74078 Phone: 405-744-6765 Fax: 405-744-8263 http://www.cvm.okstate.edu/research/Facilities/EMLAB
In the footsteps of the ImageJ-mailinglist, I started a new mailinglist today for all Zeiss KS300/400 users among us. So if you use this program, and you would like to exchange your ideas/knowledge about the program and macro-editing in it, or you might have questions about the program or writing the macro's for image-analysis, I hereby invite you to join this mailinglist.
To subscribe, send a message to: KS300-subscribe-at-topica.com or click: http://www.topica.com/lists/KS300/subscribe/?location=listinfo
To send a message: KS300-at-topica.com
If you don't use the program, but you know some people who do, please forward this email to them. Just as the ImageJ-mailinglist, I think it might be a very helpful tool in improving the image analysis and knowledge and to get to know other persons who use these programs or to announce courses, workshops etc.
Also Zeiss LSM-macro-editors, or Axioplan-users are welcome to join the list.
I'm looking forward to your response. Best regards,
Sven Terclavers
LM/CLSM Microscopist Research Assistent Center for Transgene Technology and Gene Therapy Campus Gasthuisberg O&N Herestraat 49 3000 Leuven Belgium Tel: +32 (0)16 34 61 73 Fax: +32 (0)16 34 59 90 Email: Sven.Terclavers-at-med.kuleuven.ac.be Web: www.kuleuven.ac.be/mcm
The Lehigh Microscopy School. The Lehigh Short Courses on Electron Microscopy will be held this year (as they have for over 30 years) in June.
There are still vacancies in all of the courses. Please contact : Sharon Coe (610) 758 5133 Fax 610 758 4244 sharon.coe-at-lehigh.edu Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195
The Courses:
June 9 to 13 Scanning Electron Microscopy and X-ray Microanalysis A basic SEM course (With a one-day elementary introduction on June 8) June 16 to 20 A choice of 6 advanced courses on: Advanced SEM (Imaging) Advanced SEM (Analysis) Analytical TEM Characterization of Nanostructures Characterization of Particles AFM and other probe microscopies
Further details are at: http://www.lehigh.edu/~inmatsci/Microscourses.html or http://www.lehigh.edu/~inmatsci/shortcourses/Microscourses.html -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I was hoping someone might be able to help me, as we have just lost our only spare domed glass bell jar (305mm x 356mm) for our Edwards coating unit, and we are being quoted $2747 to get a replacement. Does anyone have one they no longer need that we can buy from the at a reasonable cost.
Thanks
Steve Parry -- Steve Parry Centre for Microscopy & Microanalysis, (M010) The University of Western Australia, 35 Sterling Highway, Crawley, WA 6009, Australia.
MY DISCLAIMER!} If what follows does not stimulate a storm of protest from the physicists on the list, then it likely is almost accurate. I am a biologist, so there is always a danger that I have missed something that is important - especially when I make a suggestion for a 'cheap' survey as below.
In Pennsylvania, and in Colorado, I bet, one can get free copies of the state regulations, usually from the state "EPA", for radiation testing and responsibility. These regs will contain sections on radioactive materials as well as three types of radiation emitting equipment: medical X-Ray equipment, analytical X-ray equipment (such as x-ray diffractometers that use "high energy" x-ray generation equipment), and electron microscopes. Before you can determine your responsibility and that of your school, you have to get those relevant state regulations - even if they are difficult to understand.
In Colorado there is common sense in place. All you have to do is ascertain from the school district what IT will consider a proper "survey". However, it is clear that if the scope fails a formal (usually by a state-registered radiation physicist) survey that you will be in for some down time and likely a continuing survey program. On the other hand, you could preempt the problems by performing your own test first (as below) to determine that the scope is without a problem and THEN go thru the formal process to provide protection for all and limit cost to a one-time affair.
Seems to me you could do a satisfactory survey with Kodak 4489 film or strips of 35mm B&W film (that you had placed in a light-tight home made cardboard envelope with a piece of Al foil wrapped around half of the film. Place one or more of these light-tight packs over each of the column joints, at the top of the column where the cable joins, one over each of the viewing chamber windows and at the 'weak' points' of the camera system [which is situated in the most dangerous place for you [unless you are as old as I], AND most importantly around the high voltage tank on each face and at weak points on and around the top. I would run the scope for 24 hrs at the highest kV with the filament on. Oh yes. If there is a filament current monitor on the 9, you can expect more x-ray emissions the higher the gun dark current, especially if it exceeds 1uA (microampere). Also, if there had been or is a serious oil dispersion thru the system, that could contribute to higher emission currents and x-rays.
Most surveys that I do here are performed at 6" from the instrument. If you are clean up close, you should be clean elsewhere. Finally, the Al foil can overcome the problem of penetrance of high energy x-rays that may not register on the film by attenuating the power, or generating lower energy Al x-ray energies sufficient to permit formation of a latent image. In the absence of these 'harder' types of x-rays the Al protected film will serve, for each sheet/strip, as a negative control.
I have a small device named the "Radiation Alert INSPECTOR", manufactured by S.E. International, INC., PO box 39, 436 Farm Rd., Summertown, TN, 38483-0039. URL: http://www.seintl.com. This meter is marketed for the home owner who wants to monitor his/her basement for Radon, thus it is perfect for our purposes as well. [NO relationship beyond customer!]
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: michael_burnham-at-ceo.cudenver.edu [mailto:michael_burnham-at-ceo.cudenver.edu] Sent: Tuesday, May 20, 2003 12:52 PM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (michael_burnham-at-ceo.cudenver.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 20, 2003 at 09:14:34 ---------------------------------------------------------------------------
Email: michael_burnham-at-ceo.cudenver.edu Name: Michael Burnham
Organization: Kent Denver School
Education: 9-12th Grade High School
Location: Denver, Colorado
Question: My questions about an older Zeiss EM-9 TEM that I've been working on for use in our High School science program. It's now fully functional and working well. However, my most important concern is to make sure that no X-rays are being emitted from around the gun. I had assumed that a Geiger tube would detect the presence of X-rays although I'm not sure that this is true. I've tried this method with negative results, yet I do want to make 100% certain before I have students learning to use this EM.
Can you please advise??
Thank you greatly,
Michael Burnham Kent Denver School Chair Science Dept. 4000 E. Quincy Ave Englewood, CO 80110
Hello Else, As you have probably noticed the inside walls of the ion mill get coated by a film deposit over periods of use. These deposits can flake off and find there way in the whisper lock mechanism. The piston rod or more likely the seals in the whisper lock mechanism might be dirty. We periodically dismantled the whisper lock mechanism, cleaned it and replaced the seals. This kept the rotation smooth and corrected vacuum leaks that would develop. You might try to see if that would fix the problem. You could also inspect the motor/gears for possible damage while disassembled. Tyrone Daulton
"Else Breval (by way of MicroscopyListserver)" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } } I am a beginner, I recently subscribed. } } I have a problem, but I do not know whom to ask. } } I am ion millimg on a GATAN Dual Ion Mill, Model 600A. } The machine is very old, but good, when it works. } } I have the problem that the stage, which usually rotates as soon as it } is brought in the lower position, does not rotate as it should. } } Can anyone give me a tip why? and what can I do about it? } } Else Breval } Penn State University } Materials Research Institute
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Facility Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Ambient temperature microtomy of polyolefins is a difficult task and one that gives generally poor results at best. Although apparently acceptable sections may be obtained for optical microscopy, one should still be very conservative in the interpretation of morphology.
I recommend that you spring for a cryogenic ultramicrotome if you plan to continue such work. This is the only way to get good quality sections from polyolefins. When sectioning polymers, the temperatures of the sample and knife need to be below the glass transition temperature of all components in the film to avoid deformation during cutting. Polypropylene should be cut below -10C; polyethylenes of any type below -120C.
Feel free to contact me off-line to discuss this further.
Cheers,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"James.Passmore-at-sea ledair.com" To: microscopy-at-sparc5.microscopy.com cc: Subject: polymer microtomy 05/21/03 08:58 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello everyone.
I'm looking for tips on sectioning 5 to 50 micron thick polymer films. Typically samples would be polyolefins, but sometimes (including my current set) they are nylons. I have a room temp microtome available, and have had *limited* success sectioning some nylons to around 2 microns, but sometimes need to go thinner.
I would appreciate any tips?embedding medium, sectioning conditions, etc.
Commercial responses are welcome off-list, both from vendors and service labs.
thanks,
Jim Passmore Sr. Analytical Chemist Cryovac Division Sealed Air Corporation james.passmore-at-sealedair.com
Our Zeiss 10C TEM has a problem to take pictures. After I started to take picture, even the screen was down in a normal position no beam light was seen for a while (say 3-7 seconds). The "exposured film" was nothing but some shadows with different darkness after development. The shadows look like you were trying to figure out the exposure time when you are printing a micrographs in the dark room. At first we thought it might be shuttle jammed, then we took it out of column, it worked fine outside of the column. However, the same problem occurred, namely no beam light was seen for a while. The shuttle problem might be excluded. We have no idea what problem could be. Any suggestions??
I appreciate your help in advance.
Haixin Xu Department of Biological Sciences University of Maryland Baltimore County Maryland USA
I've had good success using a room temperature microtome, by freezing the specimen before cutting sections. The results are not as consistent as one would expect with a cryomicrotome, but might provide suitable results, especially if a cryomicrotome is not available. I freeze the sample while it is mounted in the chuck by holding a cotton swab saturated with liquid nitrogen close to, but not touching, its surface.
James Martin Orion Analytical, LLC A Materials Analysis and Consulting Firm www.orionanalytical.com 413-458-0233
----- Original Message ----- } From: {"James.Passmore-at-sealedair.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, May 21, 2003 9:58 AM
} Pacific Northwest Microscopy Society announces agenda for its upcoming annual meeting at PNNL in Richland, WA on May 29 and 30. } } Thursday May 29, 2003 } (Registration and badging starting at 11 am in EMSL lobby) } 1:00 pm Welcome note - Alice Dohnalkova, Jim Young } 1. Focused Ion Beam (EMSL Auditorium) } 1:05 FIB - Specimen preparation for Everyting - Lucille Giannuzzi, University of Central Florida } and FEI company - MSA invited speaker } 1:45 In situ sample preparation and HR SEM-STEM analysis - Matthew Weschler, FEI Company } 2. Advances in Analytical Techniques in Material Science (EMSL Auditorium) } 2:10 Backscatter Electron Imaging of Microstructural Constituants in Steels - } James H. Steele } 2:30 Nanometer Resolution Analyses of Crack Tips in a field-emission-Gun TEM } - Larry Thomas, PNNL } 2:50 The right tool for the right job - the latest generation of x-ray detectors for } EDS - Del Redfern, EDAX } -------- Break -------- } 3:20 Use of Electron Energy-Loss Near Edge Fine Structure in the Study of Phases } from the Sr/TRU Precipitation Process - Edgar Buck, PNNL } 3:40 Projection X-ray microscopy in the SEM - Les A. Brownlow, XRT Limited } 4:00 Helical Nanowires and Nanosprings - Hai-Feng Zhang, WSU } 3. Flow Cytometry - (Conference room 1075) } (simultaneous with Material science platform) } 2:10 Basic Flow Cytometry as a Comparative Method for Imaging and } Fluorescence - Randall W. Smith, Portland State University } 2:40 Construction of the first human nonimmune scFv library for yeast display } and the utility of flow cytometric-based selections - Robert Siegel, Molecular } Biosciences, PNNL } 3:00 - 4:00 pm Demo on a practical flow cytometry application - Jeff McLean, } Environmental Microbiology, PNNL } Participants will gather in laboratory 115 in building 331; anticipated return to } EMSL at 4pm. } Poster Session (Conference room 1077) } 4:30-5:30 pm Fifteen posters on topics Advances in microscopy technologies, Fluorescence and Laser probe microscopy, Analytical applications in material science, Environmental applications, and Microscopy education will be presented. Judges will select three student posters to be awarded cash prizes. } Expo (Conference room 1077) } The exhibition of literature and selected instruments presented by companies manufacturing or marketing microscopy-related products will be held during the meeting in room 1077. Complimentary refreshment will be served. } Evening Social } 6 pm - ? } PNMS members and registered meeting participants are welcome to attend an evening reception at Gordon Brother} '} s winery in Pasco and enjoy catered dinner, local wine tasting and a talk on Geological history of the Columbia river given by Steve Reidel. The winners of the poster session will be announced and awards given to the three best presentations. Busses are leaving EMSL at 6pm and returning back to EMSL after the social. Additional tickets can be purchased at registration. } } Friday May 30, 2003 } 4. Laser Microscopy (EMSL Auditorium) } 9:00 Getting as much information as possible using nonlinear optical microscopy - } Gary Holtom, PNNL } 9:35 Imaging signal transduction in live cells of the neuroendocrine system - Anda } Cornea, OHSU } 9:55 Three Dimensional Finite Element Method Simulations of the Aperture-less, } AFM-Tip Enhanced Near-Field SERS Microscopy - M. Micic, PNNL } ------ Break ------ } 5. 3-dimensional Imaging and Reconstruction } 10:20 3-d Imaging and Cell Reconstruction by EM Tomography: a primer - Alice } Dohnalkova, PNNL } 10:40 Image Processing, Segmentation, and Feature Extraction Algorithms} } Applied to Electron Tomography Data - Harold Trease, PNNL } 11:00 High speed 3-D Visualization of Signal Transduction - Steve Wiley, PNNL } ------ Break ------ } 6. Environmental Applications (EMSL Auditorium) } 1:00 Cathodoluminescence Imaging and Spectroscopy in the SEM - Applications } from Mineralogy, Semiconductors and Biomedical Samples - Paul Mainwaring, } Gatan Inc. } 1:20 Examination of Gas Hydrates in Sediments with Environmental Scanning } Electron Microscopy - Jim Young, PNNL } 1:40 Identification of Indoor Particles by Analytical Light Microscopy - E. Russ } Crutcher, Microlab Northwest } 2:00 Probing Heterogeneous Chemistry of Individual Atmospheric Particles Using } Electron Microscopy Techniques - Alexander Laskin, PNNL } } }
Some laboratory glass blowers can make new bell jars to your specs. This should save you some money. Alternatively, check out the used equipment suppliers.
Good luck.
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Steve Parry {sparry-at-cmm.uwa.ed To: Microscopy-at-sparc5.microscopy.com u.au} (by way of cc: MicroscopyListserv Subject: Coating Unit Bell Jar er)
05/22/03 07:02 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can you help??
I was hoping someone might be able to help me, as we have just lost our only spare domed glass bell jar (305mm x 356mm) for our Edwards coating unit, and we are being quoted $2747 to get a replacement. Does anyone have one they no longer need that we can buy from the at a reasonable cost.
Thanks
Steve Parry -- Steve Parry Centre for Microscopy & Microanalysis, (M010) The University of Western Australia, 35 Sterling Highway, Crawley, WA 6009, Australia.
The schedule of abstracts for The Microscopical Society of Canada meeting in Vancouver is now on the website: http://www.emlab.ubc.ca click on MSC logo. Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility First Vice President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
Try Dunniway Stockroom, www.duniway.com for "surplus" vacuum equipment of all kinds. Phone 1-650-969-8811, email info-at-dunniway.com in Mountain View California. I have purchased at very reasonable cost both used and new vacuum equipment from them and I am a satisfied customer. Usually they have some bell jars available.
Ron Vane XEI Scientific
Disclaimer: No financial interest, just a customer.
----- Original Message ----- } From: "Steve Parry (by way of MicroscopyListserver)" {sparry-at-cmm.uwa.edu.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, May 22, 2003 5:02 AM
How did you get on with the mouldy old bread over a wet Bank Holiday? "Mouldy Old Dough" was a hit the early seventies for Lieutenant Pigeon, I believe.
Dave
On Mon, 5 May 2003 14:26:30 +0100 (BST) "Robert H. Olley" {r.h.olley-at-reading.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } This is to thank everybody who replied with a wealth of ideas concerning } Roots and SEM. If haven't had any feedback yet from our soil science } department, so I can't as yet report any progress on that front. } } However, in regard to moulds, I inadvertently (honest!) let some bread go } mouldy recently. So I've looked through all the suggestions. For fixing, } one can't slowly go up through the progressions of increasingly } concentrated ethanol, since the bread will go intractably soggy (does } anyone remember the song "Mouldy Old Dough"?) Neither do I want to hit } the mould with neat alcohol. So what I'm doing, as a first stage at } least, is to put thin mouldy crusts of the bread over methanol in a vapour } tank, and letting it stand over the long weekend (we British are having } our May Day Holiday on Monday 5th!). If anything useful results, I'll } keep you posted. } } Bye for now, } } +-----------------------------------------+ } Robert H.Olley } J.J.Thomson Physical Laboratory } University of Reading } Whiteknights } Reading RG6 6AF } England } +----------------------------------------+ } Phone:+44 (0) 118 9318572 } Fax: +44 (0) 118 9750203 } University internal extension 7867 } Email: R.H.Olley-at-reading.ac.uk } URL: http://www.reading.ac.uk/~spsolley } +----------------------------------------+ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
A position for an electron microscopy (EM) research assistant in the Shaham lab at Rockefeller University is available immediately. The successful applicant will actively participate in ongoing and new projects to understand the cellular basis of apoptosis and glia-neuron communication in C. elegans. Previous EM experience, including serial reconstructions and 3-D computer imaging is a plus, however, candidates with demonstrated interest in EM will be considered as well.
We offer an excellent benefits package, competitive salary, & tuition reimbursement. For immediate consideration, email or fax resume to The Rockefeller University at FAX (212) 327-7079 or EMAIL recruit-at-rockefeller.edu.
An Equal Opportunity/Affirmative Action Employer. The Rockefeller University appreciates all responses and will contact candidates selected for further consideration.
We have had several requests concerning the use of Mercox in plants. As far as we know our customers presently use Mercox on animals. If there is anyone out there with experience in corrosion casting with plants we would appreciate your input.
Thank you,
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
I have seen two types of Edwards 306A units with different bell jars. Do you mean you have a 306 type? Type 1 has a bell jar that is a cylinder with a gasket at both ends. At the top is a 'flip top' metal lid that opens into the interior of the glass cylinder bell jar. Type 2 is a regular dome shaped bell jar and could be removed manually. Type 3 had an implosion shield and hoist to lift up the dome shaped glass bell jar. Edwards made other type of evaporators.
Ladd or Lesker may be able to order you one. Try Lesker.com for about $800 for a 12 inch by 18 inch glass domed bell jar. You can always have a glass blower cut off a few inches to make 14 inches and then grind a sealing surface for you. http://www.lesker.com
I tried many L gaskets a few years back and Ladd had the nicest one I could find without cracks or defects in the sealing surface. They sell various Pyrex® glass bell jars too. http://www.laddresearch.com/Key_Products/VacEvap/VacEvap/Bell_Jars___Support /bell_jars___support.html
Hope this helps. I don't work at these places or have any interest in them.
Paul Beauregard Senior Research Assocaite PPG Industries Monroeville Technical Center Monroeville, PA 15146 724-325-5131
} Can you help?? } } I was hoping someone might be able to help me, as we have just lost } our only spare domed glass bell jar (305mm x 356mm) for our Edwards } coating unit, and we are being quoted $2747 to get a replacement. } Does anyone have one they no longer need that we can buy from the at } a reasonable cost. } } Thanks } } Steve Parry } -- } Steve Parry } Centre for Microscopy & Microanalysis, (M010) } The University of Western Australia, } 35 Sterling Highway, Crawley, } WA 6009, Australia. } } Fax: +61-8-9380-1087 } Email: sparry-at-cmm.uwa.edu.au } Phone: +61-8-9380-8057 [24 hour voicemail attached] } } }
Dear Listers, I've been trying to search microscopy websites for those plastic screw-top vials to store unused objectives in, but I can't seem to find them anywhere. Are they just not used anymore, or am I looking in the wrong place?
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This is a TEM job at the Burnham Institute in La Jolla, CA Please reply to Dr. Dorit Hanein (e-mail at the bottom of the announcement).
Position Open: Electron Microscopist Research Assistance, to start June, 2003. (http://www.burnham-inst.org).
Requirements include working knowledge of high resolution TEM and a BA/BS or equivalent experience in biological or material science, or bioengineering. Responsibilities include negative stain sample preparation and electron microscopy, vitreous ice sample preparation and electron cryo-microscopy, some microscope maintenance and standard biochemical analyses.
Application should include a statement of career goals and addresses for three references. Salary is competitive and commensurate with experience. EOE.
Send applications to: Dr. Dorit Hanein, e-mail dorit-at-bunham.org
The Analytical Imaging Facility of the Albert Einstein College of Medicine, a core microscopy facility, is expanding its technical staff. We have two openings for experienced biological microscopy technicians.
Position 1: Digital Flourescence and Confocal Microscopy Technician
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Details for these positions can be found in the May/June issue of Microscopy Today, page 41. Please note: The email address for responding is incorrect. For consideration send your CV to ddamiani-at-aecom.yu.edu
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http://www.msa.microscopy.com/PlacementOffice/JobListings.html **************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
Does anyone have another source for the 12mm diameter carbon conductive tabs? I used to get 16084-1 from Pella but now they are standard double thickness and not useable. They are not as sticky and do not bend.
Hi Pauline, Here's what I would try to find, though I would always remember to include sufficient size that I can also include a small pack/pillow of silica gel desiccant - I use NO substitutes for this material. BTW, to dehydrate this material (silica gel!), I place it in my EM film plate degasser for a few days - thus, any vacuum device pumped to 30in of Hg will do. No need to keep the pump on or to include P2O5, just the 30in vac is sufficient for a considerable amount. For those with confidence, for some objectives I have recycled screw-cap plastic tritium shipping containers - also other decontaminated and de-labeled radioactive containers. Mine are all blue or orange and work beautifully for my old oculars and objectives - when I, like you, can't find a REAL objective or ocular container.
Try: http://www.tri-esssciences.com/plasticjars.htm [NO knowledge of this company or relationship to it OR manufacturer, etc.]
I do not ever enclose objectives without some silica gel as a mild desiccant.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: pauline yu [mailto:pcy-at-usc.edu] Sent: Friday, May 23, 2003 5:59 PM To: Microscopy
Dear Listers, I've been trying to search microscopy websites for those plastic screw-top vials to store unused objectives in, but I can't seem to find them anywhere. Are they just not used anymore, or am I looking in the wrong place?
Please let us know what size 'objectives' you are trying to store and we'll see if we can assist.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "pauline yu" {pcy-at-usc.edu} To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 23, 2003 5:59 PM
Steve,
As Paul mentioned we have a 12" x 18" domed bell jar. It would be quite expensive to special order a 12" x 14" . If you do cut it down the L-gaskets would probably help maintain the vacuum.
John
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Beauregard" {beaurega-at-westol.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 23, 2003 5:50 PM
Hi Steve,
I have seen two types of Edwards 306A units with different bell jars. Do you mean you have a 306 type? Type 1 has a bell jar that is a cylinder with a gasket at both ends. At the top is a 'flip top' metal lid that opens into the interior of the glass cylinder bell jar. Type 2 is a regular dome shaped bell jar and could be removed manually. Type 3 had an implosion shield and hoist to lift up the dome shaped glass bell jar. Edwards made other type of evaporators.
Ladd or Lesker may be able to order you one. Try Lesker.com for about $800 for a 12 inch by 18 inch glass domed bell jar. You can always have a glass blower cut off a few inches to make 14 inches and then grind a sealing surface for you. http://www.lesker.com
I tried many L gaskets a few years back and Ladd had the nicest one I could find without cracks or defects in the sealing surface. They sell various Pyrex® glass bell jars too. http://www.laddresearch.com/Key_Products/VacEvap/VacEvap/Bell_Jars___Support /bell_jars___support.html
Hope this helps. I don't work at these places or have any interest in them.
Paul Beauregard Senior Research Assocaite PPG Industries Monroeville Technical Center Monroeville, PA 15146 724-325-5131
} Can you help?? } } I was hoping someone might be able to help me, as we have just lost } our only spare domed glass bell jar (305mm x 356mm) for our Edwards } coating unit, and we are being quoted $2747 to get a replacement. } Does anyone have one they no longer need that we can buy from the at } a reasonable cost. } } Thanks } } Steve Parry } -- } Steve Parry } Centre for Microscopy & Microanalysis, (M010) } The University of Western Australia, } 35 Sterling Highway, Crawley, } WA 6009, Australia. } } Fax: +61-8-9380-1087 } Email: sparry-at-cmm.uwa.edu.au } Phone: +61-8-9380-8057 [24 hour voicemail attached] } } }
If you want to image the particles directly you need to do transmission electron microscopy. A FEG-SEM might work but a TEM can also do some electron diffraction structural determinations on the ultimate or primary particles. If you have a mixture of phases in the nanoparticles, you should try X-Ray Diffraction on the powder to show the different phases, if present.
If you are dealing with agglomerated or aggregated finer primary particles, this will be very obvious in a TEM.
Direct observations on a TEM grid with oil present can be done. Also one can extract the oil after the dispersion is on the grid. One skilled in the art of TEM and particle dispersions will be able to extract the oil from the grid dispersion without producing flocculation effects.
Don't ignore doing nitrogen BET determinations on the powders. To extract the particles from the oil for BET, heat it up gradually to about 525°C in nitrogen and then switch to air at 525°C. A TGA savvy person can help here and can actually isolate enough material for a TEM dispersion but BET will require about 50-300 mgs net. For the BET required amounts, use a heated tube furnace and combustion boat. I am assuming the nanoparticles were made by plasma and have no molecular roughness or microporosity in them that will raise the BET surface area value. Anyway, 50 nm smooth ultimate particles with a density of, say, 3.65 will give a BET of about 33 m²/gram. 11 nms ultimates will yield a BET of about 150 m²/g. 300 nm ultimates (your top range) will give a BET of about 6 m²/g. I can email you the exact assumptions and a simplified formula separately, if you are interested.
You can determine the primary or ultimate particle size distribution of small aggregates by interactively drawing diameters across the ultimate particles on the dispersed and finest aggregates you see. Use an on-line TEM and an image analysis program for this mouse intensive work.
The presence of oil means using microtomy is probably eliminated for evaluating aggregate sizes. It's possible but doubtful. One microtome pitfall would be that you can cut the real aggregate structures in half with the knife edge. One law of microtoming says something like this. As the thickness of the thin section approaches the ultimate particle size diameter, aggregates and ultimate particles that are agglomerated tend to artificially look more dispersed and smaller.
Hope this helps you out and gives you a few more options.
Paul Beauregard Senior Research Associate Greensburg, PA USA
} } Email: daniel.smith-at-liquidminerals.com } Name: Daniel Smith } } Organization: Liquid Minerals Group, Inc. } } Education: Graduate College } } Location: Houston, TX USA } } Question: I am interested in measuring particle size of metal oxide } particles suspended in a process oil. The particles are on the order } of 1 - 300 nanometers. I have used indirect methods (DLS, etc.), but } I would really like to take photos of the particles and determine } particle size, crystal structure, agglomeration and other properties. } } Any help would be appreciated. } } --------------------------------------------------------------------------- } } }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rene.oberli-at-strasbourg.gm.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 27, 2003 at 09:15:51 ---------------------------------------------------------------------------
Organization: process enginer at general motors strasbourg france
Education: 9-12th Grade High School
Location: strasbourg France
Question: I would like to make digital pictures with Canon G2 photo camera thru a WILD M3 (6 to 40x power, no zoom 1X front lens)with a LEiCA trinocular head up to now I was not able to get a full frame picture I also tried with special lens ( 0,6x )from LEICA or NIKON between the output tube and the G2 without success; the sharp image is circular and not full frame.This assembly works fine with a NIKON coolpix 995.Why this dont work with CANON G2 with or without zooming the G2? Canon-Europe is no help from a contact with them they dont know or dont want to help!
I am trying to figure a way to determine whether there are any birefringent particles ranging in size from 100nm to 3 micrometers in a whole blood sample, maybe 1mL volume. I don't have to preserve the blood components, maintain the structural integrety of the RBCs, WBCs, platelets, etc. I can not use any solvents, only aqueous solutions and I can't have the pH go below 3. What makes this more difficult is that the particles may be few in number, very few!!!
Would there be some way to break up the cells, digest the membranes, and wash away the blood material so that I could filter out the particles? If I did, I would have to separate the buffy coat and the plasma first, then go after the RBCs. I could check the plasma for particles separately and I know that the macrophages etc would have already taken up some of the particulate but that doesn't matter. Would SEM be the best approach? I would think so to see the very small particles. The particles are also reportedly fluourescent but excited by UV (which I don't have on my confocal). Would using a gradient with centrifugation be of use to separate out the particles? But then the particles may stick to the cells. Do you know of anyone who has done this type of thing and point me to any publications. I will be doing a literature search starting tomorrow as this just came up today.
Thanks for any ideas you might be able to pass along!!!
The ones I got from SPI works fine. Here is their contact details. Sadly I do not have personal interest in this company and will continue being overworked and underpaid.
Structure Probe, Inc. / SPI Supplies ·Street address for UPS, FedEx, DHL, other couriers, trucks 569 East Gay Street West Chester, PA 19380 ·Mailing address P.O. Box 656 West Chester, PA 19381-0656 USA
Phone: 1-(800)-2424-SPI Toll-free from USA/Canada 1-(610)-436-5400 FAX: 1-(610)-436-5755 E-mail: spi3spi-at-2spi.com
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, May 27, 2003 6:22 PM To: MSA listserver
Does anyone have another source for the 12mm diameter carbon conductive tabs? I used to get 16084-1 from Pella but now they are standard double thickness and not useable. They are not as sticky and do not bend.
This is a good question and hopefully someone can answer it. We buy our tabs from ProSciTech, trade name "Leit-tabs" I believe. About 12 months ago the tabs changed, they were thicker, stiffer and less sticky, as Gary notes, and in addition nowhere near as conductive. When you rub off the "sticky" there is a stiff polymer film in the middle. Alternative supplies of sensibly conducting tabs would be welcome.
} Does anyone have another source for the } 12mm diameter carbon conductive tabs? } I used to get 16084-1 from Pella but } now they are standard double thickness } and not useable. They are not as sticky } and do not bend.
You could ask Taab or Agar Scientific Ltd. (both UK) about their current stock. I prefer the thin bendy tabs but my most recent purchase contained the thick ones. I read on this list that some thick ones contain cardboard!
Dave
On Tue, 27 May 2003 09:22:26 -0700 Gary Gaugler {gary-at-gaugler.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have another source for the } 12mm diameter carbon conductive tabs? } I used to get 16084-1 from Pella but } now they are standard double thickness } and not useable. They are not as sticky } and do not bend. } } Ideas?? } } gary g. } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
The tabs from Electron Microscopy Sciences (www.emsdiasum.com) are double thickness also so they do not bend but are still very sticky. (Cat# 77825-12) You can also try SPI supplies (www.2spi.com) Good luck! -Amanda
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, May 27, 2003 12:22 PM To: MSA listserver
Does anyone have another source for the 12mm diameter carbon conductive tabs? I used to get 16084-1 from Pella but now they are standard double thickness and not useable. They are not as sticky and do not bend.
Ya, my last batch from 2 different suppliers is as you describe below. Unfortunately I purchased a whole years supply. Incidentally, has anyone had problems with the surface cracking during/after coating. This is a recent development, and I pretty much didn't care, but a user tacked down a delicate sample and it cracked with the tab. I don't have this problem with the dwindling supply of old tabs or with the tape roll.
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Bryony James {b.james-at-auckland.ac.nz} 05/28/03 02:36AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is a good question and hopefully someone can answer it. We buy our tabs from ProSciTech, trade name "Leit-tabs" I believe. About 12 months ago the tabs changed, they were thicker, stiffer and less sticky, as Gary notes, and in addition nowhere near as conductive. When you rub off the "sticky" there is a stiff polymer film in the middle. Alternative supplies of sensibly conducting tabs would be welcome.
} Does anyone have another source for the } 12mm diameter carbon conductive tabs? } I used to get 16084-1 from Pella but } now they are standard double thickness } and not useable. They are not as sticky } and do not bend.
Hmmm. A hypotonic solution (hypotonic as compared to plasms, water or very dilute buffer) will cause the red cells (and the WBCs too I suppose) to swell and burst. Knowing the composition of the 'particles' you are looking for might help.
Geoff
Damian Neuberger wrote:
} Hi all, } } I am trying to figure a way to determine whether there are any birefringent } particles ranging in size from 100nm to 3 micrometers in a whole blood } sample, maybe 1mL volume. I don't have to preserve the blood components, } maintain the structural integrety of the RBCs, WBCs, platelets, etc. I can } not use any solvents, only aqueous solutions and I can't have the pH go } below 3. What makes this more difficult is that the particles may be few in } number, very few!!! } } Would there be some way to break up the cells, digest the membranes, and } wash away the blood material so that I could filter out the particles? If I } did, I would have to separate the buffy coat and the plasma first, then go } after the RBCs. I could check the plasma for particles separately and I } know that the macrophages etc would have already taken up some of the } particulate but that doesn't matter. Would SEM be the best approach? I } would think so to see the very small particles. The particles are also } reportedly fluourescent but excited by UV (which I don't have on my } confocal). Would using a gradient with centrifugation be of use to separate } out the particles? But then the particles may stick to the cells. Do you } know of anyone who has done this type of thing and point me to any } publications. I will be doing a literature search starting tomorrow as this } just came up today. } } Thanks for any ideas you might be able to pass along!!! } } Damian Neuberger } } } } } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I agree that these double thick tabs that have quietly replaced the thin and flexible tabs we all came to love are the pits. Glad to hear there are others that don't like them either. As an example of why they won;t work for me, I cut C tabs into thin strips and use as "ground straps" on SEM samples. Double thick are inflexible and won't work for that. I'd really love to hear from a vendor as to why we need the new thick style!
So, here's the good news. E. Fullam sells 12mm carbon tabs that are thin, flexible and reasonably sticky. The part no is 14833, I called this morning and they have 27 in stock.
SPI's #5077 is a close second, but still too thick for me.
Pella's carbon tabs are like cardboard. Unusable.
Note, this is all personal preference, I ran no tests. I am a customer of the companies I mentioned.
Owen Mills Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
At 09:22 AM 5/27/2003 -0700, Gary Gaugler wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
At ~2000g, all the cells will sediment. What about the particles? If the particles are less dense than hemoglobin, then layer a slightly less dense than particles but more dense than hemoglobin, hemolyze blood with distilled water, and spin. Your particles should 'fall' into 'dense' layer at bottom. On the other hand, your particles might be more buoyant than the cells, in which case you win that way. In the absence of answers to my questions, you are left with a manageable experiment anyway, especially if the particles start out in a jar on the shelf. If they are manufactured in the body, i.e., crystals, then you must do it the hard way. An Airfuge (Beckman) will sediment hemoglobin, but not certain mycoplasma-like organisms, and it will work with really small volumes. Even if the particles sediment faster than the hemoglobin, the hemoglobin isn't birefringent. With 1ml to start, you have a lot of flexibility.
Once you can use sedimentation, you can concentrate the stuff. In the end, by my estimate, the best bulk dehydrant of stuff in a dialysis bag is PEG (polyethylene glycol).
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: Damian Neuberger [mailto:neuberger1234-at-attbi.com] Sent: Tuesday, May 27, 2003 10:40 PM To: Microscopy-at-sparc5.microscopy.com
Hi all,
I am trying to figure a way to determine whether there are any birefringent particles ranging in size from 100nm to 3 micrometers in a whole blood sample, maybe 1mL volume. I don't have to preserve the blood components, maintain the structural integrety of the RBCs, WBCs, platelets, etc. I can not use any solvents, only aqueous solutions and I can't have the pH go below 3. What makes this more difficult is that the particles may be few in number, very few!!!
Would there be some way to break up the cells, digest the membranes, and wash away the blood material so that I could filter out the particles? If I did, I would have to separate the buffy coat and the plasma first, then go after the RBCs. I could check the plasma for particles separately and I know that the macrophages etc would have already taken up some of the particulate but that doesn't matter. Would SEM be the best approach? I would think so to see the very small particles. The particles are also reportedly fluourescent but excited by UV (which I don't have on my confocal). Would using a gradient with centrifugation be of use to separate out the particles? But then the particles may stick to the cells. Do you know of anyone who has done this type of thing and point me to any publications. I will be doing a literature search starting tomorrow as this just came up today.
Thanks for any ideas you might be able to pass along!!!
If you are examining liver for alpha-1-trypsin, then I would recommend the following text. Although it is a bit dated it has beautiful full page micrographs.
Phillips, MJ, S. Poucell, J. Patterson, P. Valencia.1987. The Liver: an ultrastructural atlas and text of liver diseases. Raven Press, NY, NY, 585 p.
} } } Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} 05/21 11:49 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have any references which show micrographs of alpha-1-trypsin as seen in the electron microscope?
Of late there has been discussion on the carbon tabs and the change in the consistency and thickness of the tabs. This is true that approximately 6-8 months ago the raw material that had been used over many years to make these tabs become obsolete. The manufacturers sourced these materials from the same manufacturers of type writer ribbon however when we all stopped using ribbon we lost the source for the carbon raw. The new material is a bit thicker than and not as sticky as the original but for most it is fine and works adequately. Alternatively we here at Electron Microscopy Sciences has found a new source from the Far East that has raw material very similar to the original which produces very thin tabs with the same sticky power as the old original. The problem is the new raw material is double the cost of what was. We do have ample stock of it for those of you whom are looking for the thin material. The good news is doing a search out there on all of the thin material our price of $60 per 200 tabs seems to be very low. Please do not hesitate to contact me if you have any questions I look forward to hearing from you. Sincerely,
Stacie Kirsch Electron Microscopy Sciences 215-646-1566
I am looking for some Mylar (polyethylene I think) sheets, clear not 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any ideas who/where sells this stuff?
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
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Hello.
I am doing some primary research of AFMs. I can cacross two scanning modes which I am unfamilaiar with, and which I cannot find a good source of information on: curren timaging and phase imaging. I have a good idea of what they do, but i wonderign if you could inform me of good descriptiuons of eahc mode or coudl tell me where I could find such information. Thank you very much for your time,
Joseph D. Matchett
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I would greatly appreciate your help in dealing with the following problem. We have a set of samples from an experiment on rats performed yesterday (Tuesday, May 27). The lung and aorta samples were divided into two parts each: one, for embedding in EMbed 12, was brought to the step of washing after osmium tetroxide staining and the other, for cryosectioning, was brought to the step of infiltration with PVP and sucrose. The first set of samples is currently in 0.1 M sodium cacodylate buffer, pH 7.4, and the other in the PVP with sucrose in phosphate buffer pH 7.0. However, there is a delay in delivery of the embedding kit and aluminum specimen pins. Most likely, both will arrive here on Monday, June 2nd. Is it safe to keep the samples at this stage of preparation until Monday? If yes, at what temperature?
I don't see big problem. Keep it refrigerated +4oC. Extended wash in cacodylate may wash out some components of the cell. You may expect some artefacts from that. If your samples for cryo-EM were fixed with formaldehyde, you know: formaldehyde fixation is reversible. So, extended wash after formaldehyde may cause "less" fixation. Sergey
At 06:13 PM 5/28/2003, 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
FWIW, Mylar and polyethylene are quite different. Anyway, If you can use it, I have some I believe to be mylar, but it is only 1/2 mil thick (0.0005").
Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Wednesday, May 28, 2003 12:58 PM To: Microscopy
Dear List:
I am looking for some Mylar (polyethylene I think) sheets, clear not 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any ideas who/where sells this stuff?
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 **********************************************
How about a couple of part numbers for the old and new tabs: Thick 12 mm: 77825-12 ? (Our order on this number was filled with thicker tabs.) Thin 12 mm: ?????
Thick 25 mm: 77825-25 ? $$ Thin 25 mm: ????? $$
Thanks,
Paul
-----Original Message----- } From: Stacie Kirsch [mailto:Stacie-at-ems-secure.com] Sent: Wednesday, May 28, 2003 11:31 AM To: Microscopy-at-sparc5.microscopy.com Cc: spb-at-mwrn.com
Dear Microscopists,
Of late there has been discussion on the carbon tabs and the change in the consistency and thickness of the tabs. This is true that approximately 6-8 months ago the raw material that had been used over many years to make these tabs become obsolete. The manufacturers sourced these materials from the same manufacturers of type writer ribbon however when we all stopped using ribbon we lost the source for the carbon raw. The new material is a bit thicker than and not as sticky as the original but for most it is fine and works adequately. Alternatively we here at Electron Microscopy Sciences has found a new source from the Far East that has raw material very similar to the original which produces very thin tabs with the same sticky power as the old original. The problem is the new raw material is double the cost of what was. We do have ample stock of it for those of you whom are looking for the thin material. The good news is doing a search out there on all of the thin material our price of $60 per 200 tabs seems to be very low. Please do not hesitate to contact me if you have any questions I look forward to hearing from you. Sincerely,
Stacie Kirsch Electron Microscopy Sciences 215-646-1566
TECHNICAL DIRECTOR FOR CRYOELECTRON MICROSCOPY NEW YORK STRUCTURAL BIOLOGY CENTER
The New York Structural Biology Center (http://www.nysbc.org) seeks a Technical Director for a high-end, state of the art facility in Cryoelectron Microscopy. The facility will include three cryoelectron microscopes at 200, and 300 kV as well as ancillary equipment required for the three main cryoEM applications: tomography, single-particles, and crystallography. The facility will be used by in-house research groups and by investigators from a consortium of ten New York academic research institutions on a broad range of biological targets.
The appointee will initially be responsible for developing specifications, testing and installing the three instruments, and later for implementing specialized technologies such as automated data collection, for supporting users and for supervising a technical staff. A strong technical background in electron microscopy is essential and familiarity with biological samples or image analysis would be a bonus. Send a biographical sketch, a statement of research experience, together with names and addresses of three references to CryoEM Search Committee, at nysbc-at-nysbc.org.
-- ------------------------------ David L. Stokes Skirball Institute NYU Medical Center 540 First Ave New York, NY 10016 tel and FAX: 212-263-1580 http://saturn.med.nyu.edu/~stokes ------------------------------
At one time I needed large Mylar sheets for HV insulation around an X-ray tube. I found "drafting fim" (mylar) in an office supply store. In "the old days" draftsmen would use their Leroy and Rapidograph pens to make their large drawings on clear film for better reproduction (originally acetate, then Mylar). In these days of AutoCAD, the film might be a bit more difficult to find.
By the way, "Mylar" is a tradename for a polyester film not a polyethylene film. Kodak film (SO-163 and 4489) uses a Mylar base.
From a DuPont web page... Mylar® is an extraordinarily strong polyester film that grew out of the development of Dacron® in the early 1950s. During the 1960s cellophane gave way steadily to Mylar® with its superior strength, heat resistance, and excellent insulating properties. The unique qualities of Mylar® made new consumer markets in magnetic audio and video tape, capacitor dielectrics, packaging and batteries possible. By the 1970s, Mylar® had become DuPonts best-selling film, despite mounting competition. Mylar® is now a product of a joint venture, DuPont Teijin Films.
Cheers, Henk Colijn
At 12:57 PM 5/28/2003 -0400, Geoff McAuliffe wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
Does anyone have a technique to visualize double stranded RNA molecules by TEM? Which way is one the best (negative staining, shadoving or combination...)?
Valentin Starovoytov The State University of NJ Division of Life Sciences Electron Imaging Facility
At 12:57 PM 5/28/2003 -0400, Geoff McAuliffe wrote: } } I am looking for some Mylar (polyethylene I think) sheets, clear not } 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any } ideas who/where sells this stuff?
Try Goodfellow Materials (www.goodfellow.com online catalogue) - they have mylar sheet in thicknesses 1,2,5 & 10 mm, 150, 300 and 600mm square.
-- Dr. Norman Charnley Department of Earth Sciences University of Oxford Oxford OX1 3PR, UK.
I asked on of my associates that deals with plastics and films. Here is his reply. Hope this helps.
..There are so many different grades of Mylar, for detailed product information and specifications for Mylar Polyester Film, try the DuPont-Teijin Film website at http://www.dupontteijinfilms.com/cntProductsMylar.htm
Also, you may call Shawn Ford, sales rep. of Plastic Films Co. at 1-800-445-0719 (I'm not sure if he is still there).
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Wednesday, May 28, 2003 12:58 PM To: Microscopy
Dear List:
I am looking for some Mylar (polyethylene I think) sheets, clear not 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any ideas who/where sells this stuff?
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 **********************************************
Just regular, old fashioned polarized light would seem to be the most direct solution. Even if the particles are small, they should respond as bright against the dark background of crossed polars. Based on the size, I would suggest higher mags (60x or even 100x). I would also make sure that you do not use plan apos but use objectives especially suited for polarized light analysis. To test if the system itself might cause a problem, put a sample in place, establish Koehler illumination, then cross the polars to get the blackest background possible. Remove the sample and see if the background stays velvety black. If you do not get velvety black at all steps, the optics are exhibiting strain and will interfer with your imaging. You may also want to inquire about the lab-level microscopes that are built specifically for polarized light analysis. Leica's have been well known for decades and Nikon came out with a new series about 3 years ago.
Re: the size of your particles The good thing about "bright against dark background" types of imaging is that it is not resolution limited: you will be able to detect the particles, even at 100nm. You may not be able to size them at that level, or to get good edge or shape information, but you should be able to detect that they are. Of course, the higher the NA on the objective (and condenser), the better the imaging in general. Oil immersion may not be a bad idea. I had my hands on a 40x/1.3NA two weeks ago which was incredible!
Re: sample prep. You may want to dilute the blood with an isotonic solution, just to avoid overcrowding from red blood cells, but unless the red blood cells are just too thick, I don't see any need to sonicate or do anything else as drastic as you have described.
Let me know how this works!
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration/ Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- At 09:40 PM 5/27/03 -0500, Damian Neuberger wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I want to model a surface one-dimensionally with respect to surface roughness. I would like to know whether I can take a segmented line where the nodes are displaced vertically from a horizontal line to approximate the RMS surface roughness and then simply take the standard deviation to give the RMS value. In effect, the segmented line would be the same as a line profile across the surface.
Secondly, what I would like to do is have a particular RMS value and randomly distribute the nodes such that the line profile would give me that RMS value.
If there is anyone that can comment on the validity of this approach or if there is already a method or model available, I would greatly appreciate it.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Does anyone have a procedure for preparing a sample to detect carbon using an electron microprobe? The particular samples are 50-100 um catalyst particles. Thanks for any help.
Greetings, I 'inherited' some old (well 1950's to 1970's) Leitz microscopes and accessories. The most interesting is an Ortholux 1 (?) with a complete fluorescence attachment that predates epi. That is, the light path is transmitted. It has an arc lamp of some kind (not carbon!) and sliding filters. There is also a box with all sorts of 'accessories', and another Lietz stand. This would be a great source of parts for collectors and afficionados. I don't want to store it any more. If anyone is interested in acquiring some or all of it, please contact me.
You mention a procedure to detect carbon, that's not so hard with a STE crystal. You should be able to make relative comparisons between your specimens. However, quantifying is another matter. I assume you be mounting and polishing the particles?
Re:carbon baseline in my probe. Due to carbin in pumping system (rp's and diff pump) I measure carbon in all samples if I leave beam on one spot long enough. To get around this I understand some have successfully used Bastin & Heijligers work. Check out their paper, "Quantitative Probe Microanalysis of Ultralight Elements (Boron-Oxygen), Scanning, Vol 12, 225-236 (1990). The authors introduce the idea of area peak factors APF. I've managed to avoid doing carbon work but I'll use APF's when it happens.
Owen
At 01:42 PM 5/29/2003 -0400, Katie.Gibbs-at-grace.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Mylar® is an oriented polyester (PET) film. The DuPont website below has info and a contact phone number in case you can't find a supplier. http://www.dupontteijinfilms.com/datasheets/mylar/overview/h67160.pdf You might also want to note that there are a variety of Mylar® options depending on your end use: http://www.dupontteijinfilms.com/FilmEnterprise/ProductLocatorResults.asp?ProductID=5&Version=US
As for who sells it, a quick "Google" search says everyone from graphics supply stores: http://www.grafixplastics.com/mylar.html to sailboat supply stores: http://store.catsailor.com/tek9.asp?pg=products&specific=jrrmdocrq (20"x27"x3mils) The catch is matching the exact characteristics and quantities you need with a possible source. Good luck with your search!
- Louise
Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-337-9529 phone 508-339-4996 fax Louise_Harner-at-albint.com
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Wednesday, May 28, 2003 12:58 PM To: Microscopy
Dear List:
I am looking for some Mylar (polyethylene I think) sheets, clear not 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any ideas who/where sells this stuff?
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
The Canon G2 has a much larger lens which contributes to the vignetting problem they are having. The Nikon 995 has a very small lens, in fact, only a 28mm threaded opening. The G2 has an external lens assembly that extends out of the front of the camera upon powering up. A lens "tube" is needed to compensate for this. Therefore, the lens of the camera and the upper element of the microscope are kept farther apart. The 995 has an internal lens assembly, which requires only a step ring and allows the camera lens to be much closer. The 995 is a much better choice for imaging through a microscope or a telescope.
A MaxView Plus adapter from klughammer gmbh system will "help" reduce the vignetting in the G2, but it will not eliminate it all together. Hope this helps!
For more information please contact me at schmaus-at-klughammer.de.
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bwoAAM} Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rene.oberli-at-strasbourg.gm.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May bwoAAM} 27, 2003 at 09:15:51 bwoAAM} ---------------------------------------------------------------------------
bwoAAM} Organization: process enginer at general motors strasbourg france
bwoAAM} Education: 9-12th Grade High School
bwoAAM} Location: strasbourg France
bwoAAM} Question: I would like to make digital pictures with Canon G2 photo camera thru a WILD M3 (6 to 40x power, no zoom 1X front lens)with a LEiCA trinocular head up to now I was not able to get a bwoAAM} full frame picture I also tried with special lens ( 0,6x )from LEICA or NIKON between the output tube and the G2 without success; the sharp image is circular and not full frame.This assembly bwoAAM} works fine with a NIKON coolpix 995.Why this dont work with CANON G2 with or without zooming the G2? Canon-Europe is no help from a contact with them they dont know or dont want to help!
At 08:02 AM 5/29/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
One of the scientists here has been asked by the local zoo for a method to preserve bacterial/fungal cultures for demonstration purposes. They are referring to culture plates, not microscope slides. The plates will probably be handled by many people so they can't be made of glass. Will some epoxy embedding chemical work or other such material?
Thanks for your assistance.
Bruce F. Ingber, Biologist Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
These are supposedly the thin ones. If they work out, they are cheaper than Fullam's units. If not right, then Fullam's tabs at $90/200 is the next US-sourced option I suppose.
I should have these thin ones next week. will advise how they work.
gary g.
At 05:30 AM 5/29/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am presently working on reconfiguring an ISI DS 130C SEM to perform electron beam lithography. This involves using a high-definition 3 channel arbitrary waveform generator to power the X and Y scan coils and also the blanking plates to deflect the beam on and off the sample at high frequencies. The hardware setup required is in place now but we have encountered an issue.
The problem I am having presently is that our DS 130C SEM has been unused for several months and was moved to two different rooms and therefore (inevitably) the schematics have gone missing. External inputs to the scanning coils are normally given through a 14 pin terminal on the back labeled 'PGT' but the pin configuration of this input terminal is unknown. Finding literature that talks about this unit has also been a problem due to age issues.
I was hoping that someone could help me out on this by providing a schematic or pinout for that terminal. The connector at this terminal is a miniature ribbon style 14 pin metal shell bail-latching female connector.
The service contract for this machine is operational but our service guys don't do too many DS 130s and they keep all schematics at the customer sites, so our copy was one of the few they have access to.
Thanks a lot for any assistance possible. I can be reached at: chety-at-asu.edu
Chetan Prasad ------------------------------------ Research Associate Nanostructures Research Group Department of Electrical Engineering Arizona State University Tempe, AZ 85287-5706 Phn: 480-965-4097 Fax: 480-965-8058 ------------------------------------
Thanks to all who have sent me ideas to try with this project. They are all good and will probably end up trying them all in one fashion or another or in combination. I must say that this listserver is a tremendous resource especially when time constraints are very tight.
Thanks again and still looking for any other ideas that come across.
DEAR FRIEND, I AM OBLIGED TO CONTACT YOU FOLLOWING INFORMATION I GOT ABOUT YOUR COUNTRY. I WISH TO GO INTO BUSINESS RELATIONSHIP WITH YOU. I AM THE ONLY SON OF LATE MR. AND MRS.AKONAM MY MOTHER DIED IN 1986 WHEN I WAS FOUR YEARS OLD. THEN, MY LATE FATHER TOOK ME SO DEAR BECAUSE I WAS THE ONLY CHILD. MY LATE FATHER WAS A POLITICIAN, AN ACCOMPLISHED, WIDELY KNOWN AND RECOGNISED MILLIONAIRE FARMER IN THIS PART OF THE REGION. HE WAS ONE OF THE GREATEST SUPPLIERS OF CATTLE, BEEF AND OTHER DIARY PRODUCTS IN THIS PART OF THE COAST. UNFORTUNATELY, HE WAS MURDERED LAST YEAR BY HIS BUSINESS ASSOCIATES WHILE ATTENDING BUSINESS SEMINAR IN BOUAKE. BEFORE HE FINALLY DIED IN THE HOSPITAL, HE CALLED ME AND INFORMED ME ABOUT SOME WEALTH HE LEFT IN THE PRIVATE VAULT OF A BANK HERE IN COTE DIVOIRE IN FORM OF A BARE BOND AND THAT HE USED MY NAME AS THE ONLY CHILD FOR THE NEXT OF KIN TO DEPOSIT THE MONEY. MY LATE FATHER EXPLAINED TO ME THAT HE TOLD THE BANK THAT HIS FOREIGN BUSINESS ASSOCIATE OWNED THE MONEY. HE ALSO TOLD ME THAT THIS MONEY WAS THE CAUSE OF HIS DEATH. HE ADVISED ME IF HE DIES THAT I SHOULD MAKE URGENT ARRANGEMENT TO TRANSFER THE MONEY TO A COUNTRY WHERE I LIKE AND USE THE MONEY FOR INVESTMENT. HE ALSO ADVISED ME TO LEAVE COTE DIVOIRE AND RUN FOR MY LIFE TO PREVENT HIS ENEMIES COMING FOR ME. PLEASE IT IS ON THIS PREMISE I WRITE YOU. I AM CONTACTING YOU BECAUSE I WANT YOU TO CONFIRM TO ME ASAP YOUR POSSIBILITY OF PRESENTING YOURSELF TO THE BANK AS MY FATHERS BUSINESS ASSOCIATE AND CLAIM THE MONEY ON MY BEHALF SINCE HE HAD DECLARED THAT THE MONEY BELONGED TO HIS FOREIGN BUSINESS PARTNER AND AS A BARE BOND, THE MONEY HAS NO SPECIFIED BENEFICIARY, HENCE ANY FOREIGNER CAN PRESENT HIM/HERSELF AS THE BENEFICIARY PROVIDED YOU PRESENT ALL THE DOCUMENTS ISSUED AS AT THE TIME OF DEPOSIT, WHICH I WILL FORWARD TO YOU. YOU SHALL ALSO BE REQUIRED TO ASSIST ME IN INVESTMENTS IN YOUR COUNTRY. I HAVE PLANS TO INVEST THIS MONEY ON REAL ESTATE AND INDUSTRIAL PRODUCTION. I WILL ALSO WANT YOU TO PROVIDE A BANK ACCOUNT WHERE THE MONEY WILL BE TRANSFERED TO BESIDES, YOU WILL BE MY GUARDIAN, TRUSTEE AND THE CUSTODIAN OF THIS MONEY. I HAVE NO KNOWLEDGE ABOUT INVESTMENT. I AM A MAN OF 20 YEARS. YOU WILL AGAIN HELP ME COME OVER TO YOUR COUNTRY WHERE I PLAN TO RESIDE AND FINISH MY EDUCATION ALSO. I TRUST YOU AS A GOD FEARING PERSON WHO WILL NOT SIT ON THIS MONEY WHEN YOU CLAIM IT, RATHER ASSIST ME PROPERLY. I PROMISE TO GIVE YOU 20% OF THE TOTAL MONEY FOR HELPING ME. THIS WE WILL WRITE AN AGREEMENT. I SPOKE TO THE BANK MANAGER YERSTERDAY AND HE TOLD ME THAT THE BANK WILL CONCLUDE THE TRANSFER OF THE MONEY TO YOUR ACCOUNT WITHIN 9 BANK WORKING DAYS AS SOON AS YOU CONTACT THEM AS THE OWNER OF THE MONEY. I WILL LET YOU KNOW THE NAME OF THE BANK AND SEND YOU THE DOCUMENTS FOR THE DEPOSIT AS SOON AS I HEAR FROM YOU PLEASE INDICATE YOUR INTEREST TO ASSIST ME BY SENDING YOUR PHONE AND FAX NUMBERS FOR IMMEDIATE CONTACT. THANKS AND GOD BLESS YOU. NKEM AKONAM.
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Dear Halina, Absolutely no problem, You can store tissue fixed for resin embedding at 4 deg and for cryosectioning at RT.
Shashi
I would greatly appreciate your help in dealing with the following problem. We have a set of samples from an experiment on rats performed yesterday (Tuesday, May 27). The lung and aorta samples were divided into two parts each: one, for embedding in EMbed 12, was brought to the step of washing after osmium tetroxide staining and the other, for cryosectioning, was brought to the step of infiltration with PVP and sucrose. The first set of samples is currently in 0.1 M sodium cacodylate buffer, pH 7.4, and the other in the PVP with sucrose in phosphate buffer pH 7.0. However, there is a delay in delivery of the embedding kit and aluminum specimen pins. Most likely, both will arrive here on Monday, June 2nd. Is it safe to keep the samples at this stage of preparation until Monday? If yes, at what temperature?
Thank you, very much.
Sincerely,
Halina Witkiewicz, PhD
Senior Research Scientist
Division of Vascular Biology and Angiogenesis
SIDNEY KIMMEL CANCER CENTER
11835 Altman Row
San Diego, CA 92121
(858) 450 5990 x 326
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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I have been asked to process adult mouse bone, a new tissue for me. I would appreciate any protocols or suggestions for processing this tissue from fixation through TEM plastic embedding, including decalcification steps. Also, should I be concerned about damage to my diamond knife during thin sectioning? Thanks in advance. Marilyn Levy
You may use both methods for visualization of dsRNA. Negative staining is a little tricky. Shadowing works better. You can use Kleinschmidt's method of spreading dsRNA with cytochrome c on buffer (water) surface or direct mounting method using spermidin buffer and addsorbtion on to glow charged carbon supports.
If you have more questions, please, fill free to email me.
Good luck! Udachi!
Alexander Makhov
Lineberger Comprehensive Cancer Center University of North Carolina at Chapel Hill Chapel Hill, NC 27599
----- Original Message ----- } From: "Starovoytov" {Starovoytov-at-nel-exchange.rutgers.edu}
} I was hoping that someone could help me out on this by providing a schematic } or pinout for that terminal. The connector at this terminal is a miniature } ribbon style 14 pin metal shell bail-latching female connector. } } The service contract for this machine is operational but our service guys } don't do too many DS 130s and they keep all schematics at the customer sites, } so our copy was one of the few they have access to. }
ISI DS130, J44 connector aka. "PGT" or "EXT" . Centronics Style (Amphenol 57-30140). External Scan Control Connector.
Pin 1 (Input) External Scan Control ( 0v internal, +5 to +12v external). Pin 2 (Input) External Brightness Control (+5 internal, 0v external, TTL). Pin 3 (Input) External Horiz Ramp ( +5 to -5 volts). Pin 4 (Input) External Vert Ramp ( +4 to -4 volts). Pin 5 not used. Pin 6 (Input) External Brightness (+14v (black) to -14v (white). Pin 7 (Input) External Blanking (+5 blanked, 0v unblanked, TTL). Pin 8-14 (Gnd) 0v.
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
What level of carbon? If it is greater than 1%, than embedded polished particles will do fine. You will not want to use sticky tape or carbon paint to mount your particles. For low carbon content you should use liquid nitrogen trap, and the trap is essential for quantification.
If your particles are not conductive, you can coat them with thin layer of C or Au, but if you coat with C than the reference point ('zero') will be intensity from substrate.
Good luck,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} } Does anyone have a procedure for preparing a sample to detect carbon } using an electron microprobe? The particular samples are 50-100 um } catalyst particles. Thanks for any help. } } } Katie Gibbs } } e-mail: katie.gibbs-at-grace.com } } }
If you're forced to store half-processed tissue for TEM, it's better to keep it at the buffer stage after glutaraldehyde and before OsO4. Since it's too late for this, you might find it a good idea to repeat the osmium step when your materials arrive and you can continue. Meanwhile, keep it at +4oC, as Sergey says.
Lesley Weston.
on 29/05/2003 12:58 AM, Sergey Ryazantsev at sryazant-at-ucla.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I don't see big problem. Keep it refrigerated +4oC. Extended wash in } cacodylate may wash out some components of the cell. You may expect some } artefacts from that. If your samples for cryo-EM were fixed with } formaldehyde, you know: formaldehyde fixation is reversible. So, extended } wash after formaldehyde may cause "less" fixation. Sergey } } At 06:13 PM 5/28/2003, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I would greatly appreciate your help in dealing with the following } } problem. We have a set of samples from an experiment on rats performed } } yesterday (Tuesday, May 27). The lung and aorta samples were divided into } } two parts each: one, for embedding in EMbed 12, was brought to the step of } } washing after osmium tetroxide staining and the other, for cryosectioning, } } was brought to the step of infiltration with PVP and sucrose. The first } } set of samples is currently in 0.1 M sodium cacodylate buffer, pH 7.4, and } } the other in the PVP with sucrose in phosphate buffer pH 7.0. However, } } there is a delay in delivery of the embedding kit and aluminum specimen } } pins. Most likely, both will arrive here on Monday, June 2nd. Is it safe } } to keep the samples at this stage of preparation until Monday? If yes, at } } what temperature? } } } } } } } } Thank you, very much. } } } } } } } } Sincerely, } } } } } } } } Halina Witkiewicz, PhD } } } } Senior Research Scientist } } } } Division of Vascular Biology and Angiogenesis } } } } SIDNEY KIMMEL CANCER CENTER } } } } 11835 Altman Row } } } } San Diego, CA 92121 } } } } (858) 450 5990 x 326 } } } } HWitkiewicz-at-SKCC.org } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } }
I'd suggest you contact one of the interferometry companies to validate this method. My customer support contact at Zygo is John Roth: 860-347-8506. My old contacts at Wyko (now Veeco, in Tucson) are gone, but I would also suggest that you give them a call.
Caveat: MME has no financial interest in this issue.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration/ Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
At 01:54 PM 5/29/03 -0400, Walck, Scott D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have adapted a Nikon Coolpix 5000 to our microscope with trinocular head. The adaptation is not specific to the microscope, and we are having a problem with focus. That is, we can elevate the stage for vision's focus, or we can elevate the stage for the camera's focus ... but neither are coincident ... and we imagine this is because the adaptation is not specific to the microscope (the adapter being quite expensive for this particular brand & model). Judging from playing with the adapter, its tube-length is slightly too long.
I suppose we were hoping the camera could be manually focussed, so that both could be made coincident, but for the Coolpix we cannot find a way of manually focussing. Or, it may be impossible to focus because the adapter's tube-length is too long rather than too short. Is this the correct assumption? What have others done?
In a message dated 5/30/2003 11:14:55 AM Eastern Standard Time, michael-at-shaffer.net writes:
} We have adapted a Nikon Coolpix 5000 to our microscope with trinocular } head. The adaptation is not specific to the microscope, } and we are having a } problem with focus.
You don't mention how you adapted the camera to the scope and whether there are optical elements in your adapter, but it sounds like you just have a straight tube with no optics in it.
For the Nikon cameras (and all other consumer/prosumer cameras with non-removable lenses), you need intermediate optics to form an image plane that can be projected through the camera's lens and onto the CCD.
The easiest way to do this is to use an eyepiece of some kind. You can verify this by holding the camera in front of one of the scope's viewing eyepieces, and you'll see that it makes a nice image through the camera.
If your adapter is homemade, you may have to modify it so that it will hold either an eyepiece or a photoeyepiece in the trinocular port. You'll also need to get the front of the camera's lens as close as possible to the eyepiece. Set the camera at "infinity" and don't use the autofocus, and you should be able to get fairly good images.
Note also, if you see a lot of vignetting (clipping of the image at the corners) you may have to use a little of the camera's digital zoom to completely fill the field of view.
Biophotonics International magazine is planning to coordinate an article that is a practical guide for research-grade light microscopy users (brightfield, epifluorescence, confocal and multiphoton). We would like your input on what is important to include in this article. This could be information that you teach in a microscopy course, common problems faced in your lab or a core facility, or perhaps issues that constantly frustrate you as a user.
If you would like to participate in this exchange please e-mail your questions directly to biophotonics-at-laurin.com with "Microscopy Guide" in the subject line. If you have suggestions on the best person to contact (company or researcher) for more information on the subjects you think are important, please include that in the e-mail as well. We may also contact you for more information on the subject.
If you are not familiar with the magazine, we cover the latest research related to biological applications of light-based techniques including microscopy. We will not print anything directly from the e-mail without first requesting your permission. For more information on the magazine or to sign up for a free subscription, please visit www.photonics.com/bio.
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When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu grids, I always find Cu peaks present no matter where the beam shoots. Sometimes they can even be more intense than the peaks from my specimen when I am working on PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu signals? If the EDS spectrum shows signals from Cu grids which is more than a micron away from the beam, how can one be sure about the probe size or spatial resolution then?
Thanks in advance for any comments.
Ling
------------------- Ling Pan, Ph.D. 248 MRL Building Materials Research Institute The Pennsylvania State University University Park, PA 16802 Tel: (814)865-6115 Fax: (814)865-2326
I agree with Lesley: ideally, it's better to break procedure at the after GA fixation and store in the buffer refrigerated. Sergey
At 07:23 AM 5/30/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
We have been discussing the cryogens normally used for plunge freezing - things like propane and ethane and (formerly) Freons of various stripe. All have some rather significant downside, like potential environmental damage with Freons and the potentially explosive condensation of liquid oxygen in the hydrocarbons. This makes a rather routine and quite useful protocol fraught with drama, especially when it must be done in underground labs.
Do any of you materials folks out there know of a compound or class of compounds that have a rather large temperature spread between the freezing and boiling points and which remain liquid at or near liquid nitrogen temperatures (-196C) and which are also essentially inert with regard to oxidation potential and which (as long as I'm dreaming) have no real effect on human health? We were thinking that probably a whole bunch of stuff has been synthesized in the last 20 or so years that fit the bill (that is, since the pioneers of cryofixation did their initial search for likely fluids), but that the people who know about these compounds don't know why we would want to use them, and those of us who need new, safe cryogens don't really know about the possibility of these new substances.
Any leads or hints (or outright answers) are welcome. Thanks in advance for advice.
William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
Ling, The Cu peaks may not be generated from the sample grid, but from the TEM itself. Backscattered electrons can cause x-rays from within the column. Try varying parameters such as accelerating voltage, detector position, spot size, etc. If that doesn't help, maybe contact your EDS and/or TEM manufacturer. There may be some things they can do to help to shield the detector from the Cu x-rays. Bill
Bill Carmichael Electron Microscopy Instructor Madison Area Technical College 3550 Anderson St. Madison, WI 53704
-----Original Message----- } From: LING PAN [mailto:lup2-at-psu.edu] Sent: Friday, May 30, 2003 3:09 PM To: Microscopy-at-sparc5.microscopy.com
When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu grids, I always find Cu peaks present no matter where the beam shoots. Sometimes they can even be more intense than the peaks from my specimen when I am working on PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu signals? If the EDS spectrum shows signals from Cu grids which is more than a micron away from the beam, how can one be sure about the probe size or spatial resolution then?
Thanks in advance for any comments.
Ling
------------------- Ling Pan, Ph.D. 248 MRL Building Materials Research Institute The Pennsylvania State University University Park, PA 16802 Tel: (814)865-6115 Fax: (814)865-2326
The camera can be set to manually focus, but that will not help your problem. You need to adjust the adapter to be in focus when the microscope binoculars are, although this is just for ease of use. You should focus the image using the camera before taking pictures and a separate larger monitor will help this. In general the little monitor on the camera is too small and cumbersome to use to focus with. Here is a link that may be helpful. A search on the web will turn up several other useful sites with good information...
http://www.microdapt.com/
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 30, 2003 9:26 AM
You will always see the Cu peaks with Cu grids in a TEM. This problem is well documented and discussed in several reference texts. I suggest that you find the Williams and Carter book on TEM for Materials Science. Nestor Zaluzec also wrote an excellent review article for MSA Bulletin a number of years ago and he has a presentation based on that for XEDS analysis in the AEM. I know this because he once directed me to his presentation and I have it somewhere. Unfortunately, I couldn't find it in the archives. I suggest that you contact him directly to see what he has.
The origin of the peaks are due to hard X-rays from the apertures coming down the column and striking your sample and from X-rays generated within your sample fluorescing the Cu grids. If you are not using an analytical sample holder, you will also get X-rays from that. You should also be using top hat apertures that are essentially thicker and help to reduce the hard x-rays. You will have to take what is known as a hole count and subtract that background signal from your data. This is done by putting the probe in a open area and collecting a spectrum. I also suggest getting rid of the Cu grids and using Be grids. You will not see a Be signal.
There is also a NiO test sample that can help you see how your system is working. Here is a web site that discusses it and talks a little about the spurious X-rays that you are seeing in your studies. http://www.tedpella.com/calibrat_html/TEM7.htm
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
-----Original Message----- } From: LING PAN [mailto:lup2-at-psu.edu] Sent: Friday, May 30, 2003 5:09 PM To: Microscopy-at-sparc5.microscopy.com
When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu grids, I always find Cu peaks present no matter where the beam shoots. Sometimes they can even be more intense than the peaks from my specimen when I am working on PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu signals? If the EDS spectrum shows signals from Cu grids which is more than a micron away from the beam, how can one be sure about the probe size or spatial resolution then?
Thanks in advance for any comments.
Ling
------------------- Ling Pan, Ph.D. 248 MRL Building Materials Research Institute The Pennsylvania State University University Park, PA 16802 Tel: (814)865-6115 Fax: (814)865-2326
The 4th ASEAN Electron Microscopy Conference Organizing Committee
SPECIAL ANNOUNCEMENT Hanoi, 29th May 2003 Dear Sir or Madam, As you may aware, the 4th ASEAN Electron Microscopy Conference (ASEANMC4) will be held on 9-10 Oct 2003 in Hanoi. We have received some scientific reports from participants worldwide. Thank you so much. Currently, because of the SARS outbreak in the world, especially in Vietnam neighbored countries is still effected and after having the approval of the members of ASEAN Electron Microscopy Society, we decide to temporarily delay the Conference until a convenient time. Thank you for your consideration. Regards,
Nguyen Van Man M.D., D.M.Sc Chairman of the Organizing Committee
In the previous answers regarding the CU peaks in STEM/EDS, the effects of the Cu grid and that of the hard X-ray coming out from the column have been explained. However not that of the backscattered electrons arising from the sample itself, which will hit the grid but more surely some parts of the specimen holder which frequently contains copper (just check that). The "heavier" the thin foil, the most pronounced the spurious Cu peak. Pt-Ru (heavy atomic species) are good candidates to enhance this artefact. It is very spectacular to 'sign' a 'heavy' precipitate within a 'light' matrix with the Cu peak with EDS maps. This effect cannot be explained by fluorescence arising from hard X-ray generated within the column of direct 'column photons' analyzed... Our experience comes from a JEOL 2010F. On the double-tilt specimen holder copper comes from the small 'cap' that is screwed on top of the sample. We did machine such a cap in titanium (which spares quite a lot of money compared with the solution to buy a Be-analytical holder...), and no more Cu peak (of course, a Ti one - but smaller - instead...)
Regards
Thierry EPICIER DR CNRS GEMPPM, umr CNRS 5510 Bât. B. Pascal INSA de Lyon F-69621 Villeurbanne Cedex Tel: +33 (0)4 72 43 84 94 Fax: +33 (0)4 72 43 88 30
First, has anyone noticed that the new tabs don't seem to be very conductive? I'm using Agar G3347 12mm conductive carbon tabs.
I've taken a multimeter to a tab on a stub and the resistance is off the scale. If I place a tab between two stubs and press them together as hard as possible, I still have a resistance of several megohms between the two Al stubs. Unfortunately I don't have any of the old, thinner ones to compare.
Second, has anybody else seen cases where the adhesive seems to have penetrated or collapsed a porous structure? We look at polymer films about 10-30 microns thick with holes ~100nm in diameter. On the thinner films especially, the bits of the film firmly in contact with the tabs are always less porous or non-porous than the bits that are suspended above. Could it be that the adhesive is that thick and that it's penetrating our films?
I am greatful to all of those who responded to my Query related to the problem I faced. I have alredy tried out one of the suggestions, and was able to get the desired blocks. I am planning on trying out the other suggestions too and work out on the one that gives me the best result.
Once again, Thank you for you timely assistance.
Joston P. Nongkynrih (Jos) Sophisticated Analytical Instrumentation Facility North-Eastern Hills University Shillong, Meghalaya India
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I know that there is concern in the community about whether what is being published as FRET is really FRET or not. I think we are seeing FRET but want to know where I could be misleading myself. We have expressed the same actin binding domain in cells from two different plasmids. One is a fusion with GFP and one with mRFP-1. The population is not uniform in expression, so that there are cells expressing green only, cells expressing red only and cells expressing both. Examining the cell population with a Leica SP2, we tuned the two channels to acquire green and red signals. The green channel is tuned to see green cells only when the 488 laser is on and under these conditions there is no signal in the red channel for those green only cells. With only the 547 laser on, we see the red only cells and nothing in the green channel. When both lasers are on simultaneously, we see green, red and yellow cells. Now when we image with the 488 laser only and look in the red channel, the cells that were expressing both show a signal that is similar to but not identical to either red or green alone, and the single expressing cells show nothing. As far as I understand things, this must be FRET. Am I missing any controls to verify this? Thanks-Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
Dear Tamara- I am presuming the samples are fixed and sort of infilterated with sucrose-PVP,they are better left at RT. This is before they are cryofixed for sectioning. Shashi
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Ling Pan wrote: ============================================================================ = When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu grids , I always find Cu peaks present no matter where the beam shoots. Sometimes they can even be more intense than the peaks from my specimen when I am working on PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu signals? If the EDS spectrum shows signals from Cu grids which is more than a micron away from the beam, how can one be sure about the probe size or spatial resolution then? ============================================================================ = There are several "solutions" to this problem:
1] Use Be grids, see URL www.2spi.com/catalog/grids/beryl.shtml
and eliminate the copper lines. If you are working in an institution where Be is not permitted, we hope to have our diamond (made from diamond) mesh grids available again later this year, after having been temporarily taken out of production.
2] Silicon nitride membrane window TEM grids http://www.2spi.com/catalog/instruments/silicon-nitride.shtml
These grids, especially at the 30 nm thickness level, should be contributing very little to any extraneous secondary fluorescence effects. I won't say zero but the problem will be far less bothersome than with a copper grid, especially if you are doing your work near the center of the window.
Disclaimer: SPI Supplies has been a long time global supplier of both beryllium TEM grids and also, silicon nitride membrane window TEM grids.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Chris Blanford wrote: ============================================================================ ====== First, has anyone noticed that the new tabs don't seem to be very conductive? I'm using Agar G3347 12mm conductive carbon tabs.
I've taken a multimeter to a tab on a stub and the resistance is off the scale. If I place a tab between two stubs and press them together as hard as possible, I still have a resistance of several megohms between the two Al stubs. Unfortunately I don't have any of the old, thinner ones to compare.
Second, has anybody else seen cases where the adhesive seems to have penetrated or collapsed a porous structure? We look at polymer films about 10-30 microns thick with holes ~100nm in diameter. On the thinner films especially, the bits of the film firmly in contact with the tabs are always less porous or non-porous than the bits that are suspended above. Could it be that the adhesive is that thick and that it's penetrating our films? ============================================================================ ======= At SPI Supplies, over the years, we have been contacted by customers who have had one problem or another with the use of not only their double sided conductive carbon tabs (we call them discs), but also essentially the same product in the form of either "carbon tape" or "carbon sheets". As a side comment, it has always been somewhat of an amazement to me that so many users of these products seem to forget where they actually did make their purchase, and once we press for further information, it turns out that the product of their frustration did not even necessarily have an SPI Supplies progeny.
But one aspect of these kinds of products (all of them, irrespective of the form, or vendor) is that they do have a finite shelf-life, meaning specifically, that with the passing of time, there is a loss of properties, the most noticeable one being loss of tac. This is also accompanied with a loss of electrical properties (the material becomes more resistive). So if you are interested in extending the shelf life of your stock of such products, they should be stored refrigerated since this will slow down significantly the changes (storing at 10 C deg. lower will slow down the ageing rate by roughly a factor of ten times). A carbon disc that is past its prime, because of the higher resistivity, can even seem to be "bubbling" or experiencing other strange behavior. But this is not a matter of thin vs. thick, but one of age.
As an aside, for some, especially those using these products in UHV systems, the off-gassing characteristics are as important if not more so than the resistivity characteristics. There are indeed differences in the vacuum compatibility between these types of products from different sources. I will avoid making comment about where the SPI Supplies origin products stand in this respect, but only to make the point that we have arrived at a system that is optimized for a number of properties other than just electrical conductivity.
Unfortunately, for the typical user, unless they are making their purchase from the "ultimate source", and they have some reason to believe they are indeed receiving "fresh" material, the important variable of remaining shelf -life is introduced. We have seen instances over the years where a customer purchases material from a stocking distributor but unbeknown to them, the material had sat in that inventory for some number of years. So while it might be a "fresh" purchase so far as the customer is concerned, the material they are actually receiving is far from being fresh. Should the problems someone might be having be with a product of SPI Supplies origin, send us the lot number on the maroon SPI Supplies label and at least as a starting point, we can establish how old or fresh is that particular material.
We have ourselves directly received reports supposedly comparing the so- called thinner material vs. the so-called thicker material. We have not ourselves actually run controlled experiments side by side to compare the thin vs. thick material but only because we have not had any reason to do so . The SPI offered material has always been what one would call the "thick" material, and we are not aware of anyone who has really had problems with the SPI product. Naturally if one was having some kind of problem, we would like to know about it so that we could better understand the cause of such a problem.
But when the material eventually gets into the hands of the final end user, in addition to the potential variation in remaining shelf life, other differences can be introduced such as for the die cut discs (when the die cutting involved too much pressure), some of the silicone release liner can be transferred to the disc, at times so much so that the disc becomes non- conductive (from an SEM perspective) and it could also result in "bubbling". One way to test out if this is indeed a problem you are experiencing, compare the result you get with the disc vs. the result you get with carbon tape (making sure it is of the same material). If the carbon tape "works" but the disc does not, all other factors such as age being the same, then it would be our own prediction that the problem is the result of release liner transferral to the disc during the die cutting.
So my point is, it is clear that this could be much more than just "where it came from" or whether it is "thin" vs. "thick" material. If you have had favorable experience with the carbon tabs from a particular vendor, then you are getting fresh and not stale dated material. If you want to extend the useful lifetime of your carbon adhesive products, store them refrigerated. If you have had a less-than-satisfactory experience with your double sided conductive carbon products, and are looking to make a change, consider the range of products on URL http://www.2spi.com/catalog/spec_prep/cond_adhes.html
If you have a UHV system, consider the 6 mm wide tape product, make expressly to minimize the amount of organics put into the vacuum system.
Disclaimer: SPI Supplies was one of the first in the world to offer double sided conductive carbon products for SEM applications so we would naturally have a vested interest in calling our range of products to everyone's attention.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
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Dear Sir/Madam,
NB: PLEASE REPLY ONLY TO ( mtakar-at-k.ro )
I am Engr Mamud Takar, Chairman of the Tender Committee of the Nigerian National Petroleum Corporation (NNPC). I know you may have come across such letter, but I plead that you pay attention to this very proposal as I am willing to satisfy any of your doubt.
My Committee is principally concerned with payment of all contract awarded from 2000 to date, in order of priority as regard capital projects of the NNPC.
We are positive and convince that you would provide us with solution to a money transfer deal valued at US$31 Million (Thirty One Million United States Dollars) and subsequently a joint business venture.
In the course of our duties as values, and project inspectors for the on-going liquefied Natural Gas (LNG) project, we have over-invoiced the value of some jobs done by foreign contractors for the NNPC to the tune of US$31M.
As follows: -
(1) Computer optimization and Installation 16,000.000.00.
(2) Installation of 250,000.00 Monax Turbine$10,000.000.00
(3) Turn Around Maintenance $5,000,000.00
This money is not related to drug or other violent crime. Our aim of over-invoicing this payment is to divert the excess amount to a discrete account abroad. This fund is now floating in suspense account at the Central Bank of Nigeria (CBN). This is the fund my colleagues and I have decided to transfer into your account since we, as civil servants are not allowed to operate or own foreign account.
The money will be shared as follows after transfer, 30% for you (Account Owner) 60% for me and my colleagues 10% to off-set both local and international expenses that would be incurred in the course of this transaction.
To be able to claim the funds, we will be purporting your company to be the original contractor / beneficiary of the funds so all procedures for international transfer shall be strictly followed, as we have worked out all modalities for a swift and risk free transfer.
If this proposal satisfies you, please call me immediately on +234-803-3433 283.
This transaction will last for 14 working days from the time we submit the required information, as all modalities concerning this transaction have been worked out and it is completely 100% risk free. Please be informed that this subject is classified sensitive. Therefore treat the transaction with utmost confidentiality and urgency.
NB: PLEASE REPLY ONLY TO ( mtakar-at-k.ro )
Yours Faithfully,
Engr Mamud Takar. Tel: +234-803-3433 283
----------
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.. And don't forget to focus the camera using the lowest power mag on your nosepiece. Then it will automatically be in focus for all other objectives.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration. Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
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Abidjan cote d'Ivoire. West Africa.
Dear Sir, I am miss Susan Buma, from Sierra Leone a country in west africa,I am 2oyrs old and a high school student.Please I would like you toassist me urgently as you can before they kill me please.
My father is a wealthy cocoa merchant.When Trouble began early last year my father's business associates start suspecting that my father is not giving proper accounts of all the tons of cocoa being cultivated by local farmers in some villages. As a result of this discovery and the enviness of the money ,he was poisioned by his business associates.At his hospital bed ,he revealed to me the reasons of his present predicament.He directed me where to get the documents of the said deposit, that he made me the next of kin and advice me to move immediately to Abidjan West Africa,where he deposited the money in a local bank.
Being the only child.He advise me that if he did not survival this illness,that I should not stay in the country because his business associates will equally distroy my life hence my mother died five years ago of breast cancer.
He directed me that I should look for a foreign partner who will help me to transfer the money into the person's or companies account and the person will help me to invest the money in their country. After all these,he finally died on 5th of November, my his genle soul rest in perfect peace Amen. And since then I was droped from school (University) and went into hiding for my life in Abidjan, because of my father's business associates. All the relevant documents of the $10.5 (ten million five hundred thousand dollars) that was deposited in the bank are with me now .
I am ready to give you 14% of this fund ,if you can help me transfer this fund into your account overseas for onward investment in your country.
Please contact me immediately with the above E-mail address and include your telephone and Fax number to enable me to forward it to the banker for the immediate transfer of the fund into your normited account, then you make an arrangement to remove me out from this country before they kill me.
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Abidjan cote d'Ivoire. West Africa.
Dear Sir, I am miss Susan Buma, from Sierra Leone a country in west africa,I am 2oyrs old and a high school student.Please I would like you toassist me urgently as you can before they kill me please.
My father is a wealthy cocoa merchant.When Trouble began early last year my father's business associates start suspecting that my father is not giving proper accounts of all the tons of cocoa being cultivated by local farmers in some villages. As a result of this discovery and the enviness of the money ,he was poisioned by his business associates.At his hospital bed ,he revealed to me the reasons of his present predicament.He directed me where to get the documents of the said deposit, that he made me the next of kin and advice me to move immediately to Abidjan West Africa,where he deposited the money in a local bank.
Being the only child.He advise me that if he did not survival this illness,that I should not stay in the country because his business associates will equally distroy my life hence my mother died five years ago of breast cancer.
He directed me that I should look for a foreign partner who will help me to transfer the money into the person's or companies account and the person will help me to invest the money in their country. After all these,he finally died on 5th of November, my his genle soul rest in perfect peace Amen. And since then I was droped from school (University) and went into hiding for my life in Abidjan, because of my father's business associates. All the relevant documents of the $10.5 (ten million five hundred thousand dollars) that was deposited in the bank are with me now .
I am ready to give you 14% of this fund ,if you can help me transfer this fund into your account overseas for onward investment in your country.
Please contact me immediately with the above E-mail address and include your telephone and Fax number to enable me to forward it to the banker for the immediate transfer of the fund into your normited account, then you make an arrangement to remove me out from this country before they kill me.
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Abidjan cote d'Ivoire. West Africa.
Dear Sir, I am miss Susan Buma, from Sierra Leone a country in west africa,I am 2oyrs old and a high school student.Please I would like you toassist me urgently as you can before they kill me please.
My father is a wealthy cocoa merchant.When Trouble began early last year my father's business associates start suspecting that my father is not giving proper accounts of all the tons of cocoa being cultivated by local farmers in some villages. As a result of this discovery and the enviness of the money ,he was poisioned by his business associates.At his hospital bed ,he revealed to me the reasons of his present predicament.He directed me where to get the documents of the said deposit, that he made me the next of kin and advice me to move immediately to Abidjan West Africa,where he deposited the money in a local bank.
Being the only child.He advise me that if he did not survival this illness,that I should not stay in the country because his business associates will equally distroy my life hence my mother died five years ago of breast cancer.
He directed me that I should look for a foreign partner who will help me to transfer the money into the person's or companies account and the person will help me to invest the money in their country. After all these,he finally died on 5th of November, my his genle soul rest in perfect peace Amen. And since then I was droped from school (University) and went into hiding for my life in Abidjan, because of my father's business associates. All the relevant documents of the $10.5 (ten million five hundred thousand dollars) that was deposited in the bank are with me now .
I am ready to give you 14% of this fund ,if you can help me transfer this fund into your account overseas for onward investment in your country.
Please contact me immediately with the above E-mail address and include your telephone and Fax number to enable me to forward it to the banker for the immediate transfer of the fund into your normited account, then you make an arrangement to remove me out from this country before they kill me.
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id IAA05881 for dist-Microscopy; Mon, 2 Jun 2003 08:34:21 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA05869 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Mon, 2 Jun 2003 08:33:50 -0500 (CDT) Received: from inbox.net (aaron.visual.com [209.123.16.34]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA05862 for {Microscopy-at-Sparc5.Microscopy.Com} ; Mon, 2 Jun 2003 08:33:38 -0500 (CDT) Received: (qmail 19934 invoked by uid 99); 2 Jun 2003 12:26:01 -0000
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Abidjan cote d'Ivoire. West Africa.
Dear Sir, I am miss Susan Buma, from Sierra Leone a country in west africa,I am 2oyrs old and a high school student.Please I would like you toassist me urgently as you can before they kill me please.
My father is a wealthy cocoa merchant.When Trouble began early last year my father's business associates start suspecting that my father is not giving proper accounts of all the tons of cocoa being cultivated by local farmers in some villages. As a result of this discovery and the enviness of the money ,he was poisioned by his business associates.At his hospital bed ,he revealed to me the reasons of his present predicament.He directed me where to get the documents of the said deposit, that he made me the next of kin and advice me to move immediately to Abidjan West Africa,where he deposited the money in a local bank.
Being the only child.He advise me that if he did not survival this illness,that I should not stay in the country because his business associates will equally distroy my life hence my mother died five years ago of breast cancer.
He directed me that I should look for a foreign partner who will help me to transfer the money into the person's or companies account and the person will help me to invest the money in their country. After all these,he finally died on 5th of November, my his genle soul rest in perfect peace Amen. And since then I was droped from school (University) and went into hiding for my life in Abidjan, because of my father's business associates. All the relevant documents of the $10.5 (ten million five hundred thousand dollars) that was deposited in the bank are with me now .
I am ready to give you 14% of this fund ,if you can help me transfer this fund into your account overseas for onward investment in your country.
Please contact me immediately with the above E-mail address and include your telephone and Fax number to enable me to forward it to the banker for the immediate transfer of the fund into your normited account, then you make an arrangement to remove me out from this country before they kill me.
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id IAA05960 for dist-Microscopy; Mon, 2 Jun 2003 08:36:57 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA05918 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Mon, 2 Jun 2003 08:36:26 -0500 (CDT) Received: from inbox.net (aaron.visual.com [209.123.16.34]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA05911 for {Microscopy-at-sparc5.Microscopy.Com} ; Mon, 2 Jun 2003 08:36:13 -0500 (CDT) Received: (qmail 20425 invoked by uid 99); 2 Jun 2003 12:28:37 -0000
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Abidjan cote d'Ivoire. West Africa.
Dear Sir, I am miss Susan Buma, from Sierra Leone a country in west africa,I am 2oyrs old and a high school student.Please I would like you toassist me urgently as you can before they kill me please.
My father is a wealthy cocoa merchant.When Trouble began early last year my father's business associates start suspecting that my father is not giving proper accounts of all the tons of cocoa being cultivated by local farmers in some villages. As a result of this discovery and the enviness of the money ,he was poisioned by his business associates.At his hospital bed ,he revealed to me the reasons of his present predicament.He directed me where to get the documents of the said deposit, that he made me the next of kin and advice me to move immediately to Abidjan West Africa,where he deposited the money in a local bank.
Being the only child.He advise me that if he did not survival this illness,that I should not stay in the country because his business associates will equally distroy my life hence my mother died five years ago of breast cancer.
He directed me that I should look for a foreign partner who will help me to transfer the money into the person's or companies account and the person will help me to invest the money in their country. After all these,he finally died on 5th of November, my his genle soul rest in perfect peace Amen. And since then I was droped from school (University) and went into hiding for my life in Abidjan, because of my father's business associates. All the relevant documents of the $10.5 (ten million five hundred thousand dollars) that was deposited in the bank are with me now .
I am ready to give you 14% of this fund ,if you can help me transfer this fund into your account overseas for onward investment in your country.
Please contact me immediately with the above E-mail address and include your telephone and Fax number to enable me to forward it to the banker for the immediate transfer of the fund into your normited account, then you make an arrangement to remove me out from this country before they kill me.
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Colleagues
Yes I see the spam attack. I'm trying to mitigate it and find the "hole" in the filters.
Sure, now conductivity of the tabs could be very poor. On the tabs of one of the suppliers I even see charging artifacts (rectangle scanning area is visible after lowering of the magnification). And in some of the tabs I have found a lot of sulfur.
Vladimir Dusevich
} First, has anyone noticed that the new tabs don't seem to be very } conductive? I'm using Agar G3347 12mm conductive carbon tabs. } } I've taken a multimeter to a tab on a stub and the resistance is off } the scale. If I place a tab between two stubs and press them together } as hard as possible, I still have a resistance of several megohms } between the two Al stubs. Unfortunately I don't have any of the old, } thinner ones to compare. } } Second, has anybody else seen cases where the adhesive seems to have } penetrated or collapsed a porous structure? We look at polymer films } about 10-30 microns thick with holes ~100nm in diameter. On } the thinner } films especially, the bits of the film firmly in contact with } the tabs } are always less porous or non-porous than the bits that are suspended } above. Could it be that the adhesive is that thick and that it's } penetrating our films? } } Looking forward to hearing any insights. } } Regards, } } Chris Blanford } } }
I am Mr. Raphael Maloney and my sister is Miss Sofie Maloney, we are the children of late Chief John Maloney from Sierra Leone. I am writing you in absolute confidence primarily to seek your assistance to transfer our cash of eleven Million Dollars ($11,000.000.00) now in the custody of a private Security trust firm in Europe the money is in trunk box deposited and declared as family valuables by my late father as a matter of fact the company does not know the content as money, although my father made them to under stand that the box belongs to his foreign partner.
Source of the money:
My late father Chief John Maloney , a native of Makeni District in the Northerh province of Sierra Leone, was the General Manager of Sierra Leone Mining co-operation (S.L.M.C.) Freetown . According to my father, this money was the income accrued from Mining Co-operation's over draft and minor sales.
Before the peak of the civil war between the rebels forces of Major Paul Koroma and the combined forces of ECOMOG peace keeping operation that almost
destroyed my country, following the forceful removal from power of the civilian elected President Ahmed Tejan Kabbah by the rebels. My father had already made arrangement for his family thats talking about my mother, my little sister and myself to be evacuated to Cotonou Republic of Benin with the CERTIFICATE OFDEPOSIT he made with a security firm in Europe through the aid
of U.N evacuation team.
During the war in my country, and following the indiscriminate looting of Public and Government properties by the rebel forces, the Sierra Leone mining coop. Was one of the target looted and it was destroyed. My father including
other top Government functionaries were attaked and killed by the rebels in November 2000 because of his relationship with the civilian Government of Ahmed Tejan Kabbah.
As a result of my father's death , and with the news of my uncle's involvement in the air crash in January it dashed our hope of survival. The untimely deaths caused my mother's heart failure and other related complications of which she later died in the hospital after we must have spent a lot of money on her early this year . Now my 18 years old sister and myself are alone in this strange country suffering without any care or help. Without any relation, we are nowlike refugees and orphans.
Our only hope now is in you and the box deposited in the Security Firm To this effect, I humbly solicit your assistance in the followings ways.
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For your assistance, I have agreed with my younger sister that 15% of the total amount will be for your effort and another 5 % to cover all the expenses that may incur during the business transaction, Last, I urge you to
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Please as you show your willingness, Forward to us your full name, address and Tel/ Fax numbers, to me.
earnestly awaiting your response.
TO: raphmalon-at-slucia.com (MAY REPLY HERE)
Thanks.
May God bless you as you assist us.
Mr. RAPHAEL MALONEY.
Welcome to Sudan pioneer ISP http://www.sudanet.net
It is well known that copper grids will give a background of Cu during EDX measurements.
As Walck said, you can use Be grids (best but $$) but I use Ti grids to eliminate this Cu interference peak. I run Cu grids and if my customer or I suspect we have copper in the sample, then I also make up a Ti grid. It helps to keep some Ni grid handy too. Use whatever grid does not show a Ti or Ni peak with the Cu grid spectra. Ti has very few interferences. Be grids will show nothing by EDX.
You should always use a Be specimen holder to do EDX. I have a Philips CM 12 and the regular holder has a higher ridge on it than the Be analytical holder right at the grid edge. I use top hat apertures too, FYI. If I use the regular holder, then I get a Cu peak with a slight Zn peak. This is coming from the holder because it is close to the beam position. This has nothing to do with your beam spot size within the grid opening unless you are right near the grid bar. More on that later.
To address your concerns about higher Cu peaks than the sample, this does happen. I do a lot of thin sections of polymer coatings and different thickness' produce higher Cu peaks. Now I do not see the Cu peak when I use my Ti or Ni grids. Otherwise if I did, everything I run would have 'scope Copper' in it. If the Cu is coming from the scope, then Be grids would also show Cu. Where's the iron peak from the scope's pole pieces? I do not think the Cu is coming from the scope for these reasons. The Cu peaks can be higher for another reason. If the amount of carbon or polymer is high in the sample and somewhat lower in other elements, a higher Cu peak is observed. On the other hand, some ink coatings and ink jet pigment coatings I run that have a high pigment loadings, have higher 'metals' peaks than the copper peak.
In my opinion, the height of the Cu peak can be used to tell you that you should SUSPECT the material you are examining is highly loaded with organic. If I run a sample with a high Cu peak and nothing else shows up, then I conclude I have high organics, Be, Li, B or the geometry in the TEM is a problem. Philips has a lower EDX detector take off angle and geometry can be a problem.
So how is this Cu peak generated. From MY experience I have observed that it is caused by electrons from the sample being scattered around the holder. Take a pure formvar-carbon coated grid and run an EDX on it. The Cu peak should show up and it should be noisy. Then run a pure organic thin section and notice how the Cu peak signal to noise changes. You can also put more carbon on the formvar grid with an high vacuum old style evaporator to make a thicker carbon film. Use the formula in Electron Microscopy by John Bozzola on page 125 to determine the thickness you have added of rod. Instead of his density of materials (cm2*atomic thickness), use the regular density in a CRC handbook for bulk density. Correct his formula for this bulk density by multiplying John's formula by 10^(-8). I would use about 8 mgs minimum of carbon rod at a 10 CM height.
Carbon Mass = (4*Pi*distance2*thickness*rho)*(10^-8)/(sin beta) The units are grams, cm, A, gm/cm3, degrees; respectively. This can be derived by the integration of the surface area of revolution of a thin film of an atoms at distance, d, from a point source. For overhead evaporations, sin beta is 1.
You will see that the signal is proportional to the thickness of the carbon. Now this suggests to ME that the electrons impinge on your sample and are sprayed all around the areas of the holder. The thicker the thin film, the more electrons are scattered due to the increased path the electrons must travel. Some electron pass right through and hit your viewing screen. Right? Back scattered ones bounce right back up from the sample to your BS detector. Now there must be situations where the intermediate scattering exists and electrons do not miss the atoms or hit them dead center. These are the electrons that are scattered off to the sides of the sample and hit the holder and the grid material. They generate Cu, Ti, Ni or Be grid characteristic X-rays. These are the electron scatterings and interactions that are causing the metals to be seen in your spectra except Be will not show up.
Don't forget to remove your objective aperture during spectral collection. If you don't, the transmitted electrons will 'back scatter' and Pt will show up from the aperture material.
I hope my humble 'observations of the phenomena' help you understand what is going on here. You probably understand a lot of this but I wanted to make sure I covered most of what I have seen, just in case.
Paul Beauregard Senior Research Associate PPG Industries Monroeville Technical Center 440 College Park Drive Monroeville, PA 15146 800-446-3382 ext 5131
-----Original Message----- } From: LING PAN [mailto:lup2-at-psu.edu] Sent: Friday, May 30, 2003 5:09 PM To: Microscopy-at-sparc5.microscopy.com
When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu grids, I always find Cu peaks present no matter where the beam shoots. Sometimes they can even be more intense than the peaks from my specimen when I am working on PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu signals? If the EDS spectrum shows signals from Cu grids which is more than a micron away from the beam, how can one be sure about the probe size or spatial resolution then?
Thanks in advance for any comments.
Ling
------------------- Ling Pan, Ph.D. 248 MRL Building Materials Research Institute The Pennsylvania State University University Park, PA 16802 Tel: (814)865-6115 Fax: (814)865-2326
i'm having a bit of a problem with your question. it sounds to me more like you are discussing co-localization than FRET. while my knowledge of FRET is based on the molecular side of my life, i have discussed this with our technician who runs the confocal lab and one of the post-docs in his lab group. they concur.
the key is that you are co-expressing chimeric proteins, GFP and mRFP-1, which should fluoresce naturally when they are excited with the proper wavelength. as soon as they are excited with the proper wavelengths they will glow. they are not quenched. the first problem here is based upon the fact that they will fluoresce regardless of proximity to each other.
the second problem is that no matter how tightly you try to cut your channels, the signal given off at each point in the spectrum represents standard curves. they look like modified bell curves. as a result, there is, and always will be a small area of overlap between two different channels. if you excite for one and amplify the signal a lot you will see signal in this area in the other channel. in short, when you excite for green and look at red, you will get something in red that does not really look like green or red. if this is what is happening, you do not have FRET,
unless things have been defined differently in the con-focal area, and we are not aware of such a difference, the major point about FRET is Resonant Energy Transfer. you have something that does not fluoresce when you excite it. the label on the molecule being used to signal what you are looking for is marked with an inactive moiety. no matter how you look at it you will see nothing. your second marker molecule contains an activator. when the activator binds in close enough proximity you will see a transfer of resonant energy. the demonstration of this will be fluorescence. if there is no binding, or if they are too far apart, you will not see fluorescence. in molecular detection, binding must be within1-5 nucleotides, distance depending on with whom you speak.
your controls here are problematic. the negative control is simple. you put in your first and second detectors separately and see if either fluoresces. if either or both do, something is wrong, it is not fluorescent resonant energy transfer. the positive control requires your attachment of the detector in close enough proximity. how you design that in an experiment where you are looking at the expression of fluorescent marker chimeric proteins eludes me totally.
I'm happy to announce that the Project MICRO (Microscopy In Curriculum - Research Outreach) web page has been thoroughly revised; the URL is below. The text has been updated, and the reviewed bibliography of books, videos, CD-ROMs, and websites has been converted to a searchable database (THANK YOU, Nestor!). There's been a major increase in the number of CDs listed. If you know about missing items (nothing that's "out of print", please) contact me.
Are you making plans to attend M&M in San Antonio? Please note that there will be a MICRO workshop on Sunday afternoon. It's a unique opportunity; all 10 of the "Microscopic Explorations" learning stations will be set up, and the leader will be a Texas "GEMS Associate", an experienced teacher-trainer. Although the MICRO workshop is grouped with several others that are intended for Spanish-speaking attendees, it will be presented in English and everyone will be welcome. If you're interested, please help us plan by signing up for SC-11 (no charge) on your meeting reg form. Questions? Contact me directly. -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
On Friday, May 30, 2003, at 02:08 PM, LING PAN wrote:
} When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu } grids, I } always find Cu peaks present no matter where the beam shoots. } Sometimes they } can even be more intense than the peaks from my specimen when I am } working on } PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of } the Cu } signals? If the EDS spectrum shows signals from Cu grids which is more } than a } micron away from the beam, how can one be sure about the probe size or } spatial } resolution then? } } Thanks in advance for any comments. } Dear Ling, There are several possible sources for these peaks. 1) You could have either electrons or x-rays coming down the column outside the beam, arising from scattering from apertures, etc. 2) Electrons could be back-scattered from the lower objective pole piece. 3) Electrons scattered from your specimen could hit the grid bars. If the peaks are constant regardless of how far you are from a grid bar, you are likely dealing with either of the first two sources, and if the peaks are related to the position on the grid, the third source is significant. If you put a bare Cu grid in the scope, focus the beam on the middle of a grid square, and take a spectrum, do you get any peaks--especially a Cu peak? If so, you need to improve the shielding in the microscope. You can take a spectrum from one of your particles, then take a series of spectra from points displaced from the particle by various amounts. This will give you a background that you can subtract for quantitation, but that is less satisfactory than eliminating the spurious peaks. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
On Friday, May 30, 2003, at 05:17 PM, William P. Sharp wrote:
} We have been discussing the cryogens normally used for plunge freezing } - things like propane and ethane and (formerly) Freons of various } stripe. All have some rather significant downside, like potential } environmental damage with Freons and the potentially explosive } condensation of liquid oxygen in the hydrocarbons. This makes a rather } routine and quite useful protocol fraught with drama, especially when } it must be done in underground labs. } } Do any of you materials folks out there know of a compound or class of } compounds that have a rather large temperature spread between the } freezing and boiling points and which remain liquid at or near liquid } nitrogen temperatures (-196C) and which are also essentially inert } with regard to oxidation potential and which (as long as I'm dreaming) } have no real effect on human health? We were thinking that probably a } whole bunch of stuff has been synthesized in the last 20 or so years } that fit the bill (that is, since the pioneers of cryofixation did } their initial search for likely fluids), but that the people who know } about these compounds don't know why we would want to use them, and } those of us who need new, safe cryogens don't really know about the } possibility of these new substances. } Dear William, I can set your mind at ease on at least one thing. Since the freezing point of ethane is given as -183 C in the Handbook of Chemistry and Physics, and the boiling point of oxygen is given as -182.9 C in the same source, it is not a real concern that oxygen will condense in liquid ethane, especially since one always uses warm ethane gas to melt solid ethane during freezing. There may be some real danger of ethane exploding--our safety office was very concerned--but I have never heard of an actual occurrence of this. As far as other substances with low freezing points (and, of course, good thermal conductivity), in general, the larger the molecule, the higher the freezing point, so suitable cryogens may be restricted to small molecules, which can be rather easily enumerated. This last comment is strictly a guess on my part, but I think it unlikely that any suitable substance has been synthesized only in the last few years. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Can anyone help me? I need to purchase a filter for viewing fluorescence but I don't know which is appropriate. I have a filter that transmits light to the eyepiece of our Wild-Heerbrugg M20 through which I am able to view both green and red fluorescence but I want to take digital images via the phototube so I need a filter for that too. Unfortunately I have no idea what type of filter it is. If possible, I would like to use two fluorophores simultaneously: FITC (ex495/em520 and FM 4-64 (ex560/em730). What should I buy?
Many thanks,
Patricia ---------------------- Patricia de Winter cdewi02-at-students.bbk.ac.uk
On Friday, May 30, 2003, at 05:17 PM, William P. Sharp wrote:
} We have been discussing the cryogens normally used for plunge } freezing - things like propane and ethane and (formerly) Freons of } various stripe. All have some rather significant downside, like } potential environmental damage with Freons and the potentially } explosive condensation of liquid oxygen in the hydrocarbons. This } makes a rather routine and quite useful protocol fraught with drama, } especially when it must be done in underground labs. } Do any of you materials folks out there know of a compound or class of compounds that have a rather large temperature spread between the freezing and boiling points and which remain liquid at or near liquid nitrogen temperatures (-196C) and which are also essentially inert with regard to oxidation potential and which (as long as I'm dreaming) have no real effect on human health? We were thinking that probably a whole bunch of stuff has been synthesized in the last 20 or so years that fit the bill (that is, since the pioneers of cryofixation did their initial search for likely fluids), but that the people who know about these compounds don't know why we would want to use them, and those of us who need new, safe cryogens don't really know about the possibility of these new substances.
We have long used a slurry of isopentane as a quenchant to produce amorphous samples of readily crystallizable polymers. The Tm is 113 K, so oxygen condensation should not be a problem and we found it gave a " better" quench than the refrigerant UCON-12 (Tm = 118) for material like linear polyethylene. We originally replaced the UCON with a slurry of nitrogen, but had to use liquid He or pumping to partially freeze it and the isopentane was simpler since it can be partially frozen in liquid N2. Refs. include Polymer, 20, 903 (1979), J. Macromol. Sci., Phys. B20, 37 (1981) and ibid, B21, 617 (1982) --
Phillip H. Geil; Ph. 217-333-0149 Fax 217-333-2736 Department of Materials Science and Engineering University of Illinois 1304 W. Green St. Urbana, IL 61801
I also use explosive liquid cryogens (usually propane) for freezing and would be interested in whether anyone else knows of cases of explosions occuring during use in this way. The two (anecdotal) cases of explosions I have heard were
1) an explosion when someone broke a light in the hood while using propane or ethane and
2) an explosion in a condensing coil being used to condense gaseous propane. The explanation of the latter was that oxygen condensed in the cold coil after the propane flow was turned off. I now condense the propane directly into a cold vessel set down in a styrofoam container of liquid nitrogen, which should keep the environment around the liquid propane purged of air. I thaw the propane when it freezes either with liquid propane or with a warm metal rod.
Any ideas about this? I am wondering whether there are other precautions.
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
I think static electricity may be a real problem for liquid propane/ethane. Therefore, it's safer to use metal container and needles instead glass or plastic. Container itself should not be a big problem because of water condensation but plastic tubings. If dry propane/ethane travel through plastic tubing it may be enough to generate static electricity and therefore spark. Sergey
At 12:42 PM 6/2/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I'm looking for a couple of 10 litre dewars for liquid nitrogen and wondered if anyone could recommend a supplier? Alternatively if anyone in New Zealand has any second hand ones for sale I'd be interested.
Electron Microscope Facility School of Science and Technology The University of Waikato Ruakura Satellite Campus (RS5) Ruakura Road Hamilton New Zealand
To whom it may concern. If somebody has got a diagram for the Spectrum Monitor and related diagrams, could you please fax it to me. Thank you very much. Its urgent. Erik Sorbroden, Norway Center for materials Science fax +47 22 84 06 51
I would assume that you are correct, the issue is the length of your trinocular adapter.
Take a look at our webpage which lists solutions for adapting to the cameras to all the major microscope manufacturers as well as the 'off-brands' as well via a couple of mounting options: http://www.mvia.com/Coolpix/clpxadpt.htm
Our solutions are probably less expensive than other options you have looked at. Please feel free to contact me with your microscope model if you would like to go over the various options.
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
michael shaffer wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We have adapted a Nikon Coolpix 5000 to our microscope with trinocular } head. The adaptation is not specific to the microscope, and we are having a } problem with focus. That is, we can elevate the stage for vision's focus, } or we can elevate the stage for the camera's focus ... but neither are } coincident ... and we imagine this is because the adaptation is not specific } to the microscope (the adapter being quite expensive for this particular } brand & model). Judging from playing with the adapter, its tube-length is } slightly too long. } } I suppose we were hoping the camera could be manually focussed, so that } both could be made coincident, but for the Coolpix we cannot find a way of } manually focussing. Or, it may be impossible to focus because the adapter's } tube-length is too long rather than too short. Is this the correct } assumption? What have others done? } } cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } www.micro-investigations.com
Thanks a lot for your information. I did collect "in-hole" spectra together with spectra on specimen. Actually I collected spectra both in complete hole area and on amorphous C film. The spectrum in complete hole area showed no peak at all, and the one collected on C film showed Cu peaks besides C peak. So the Cu peaks must be coming from the Cu grid instead of the column. (The double-tilt holder that I used was Be one.)
I did subtract the "in-hole" spectra from all other spectra. The Cu peaks were just a little weaker. I think mostly the heavy metals in my specimen accounted for that.
What I am thinking is, if the electrons scattering from the specimen could hit the Cu grid or whatever other grid, then they could hit other areas of the specimen too. Then how can one be sure the spectrum (except the Cu peaks) was really just the information of the beam-shooting area?
Sincerely,
Ling
On Tue, 03 Jun 2003 14:41:02, "CorneliuSarbu" wrote:
} } Dear colleague, } } yesterday I sent you a message about the "in-hole" X-EDS spectrum } acquisition. The bibliographical citation was not complete. Here is } the complete one: } Brent Fultz and James Howe, "Transmission Electron Microscopy } and Diffractometry of Materials", Springer-Verlag, Berlin and Heidelberg, } 2001. See page 209. } } Corneliu Sarbu, PhD } } } -----Original Message----- } From: LING PAN {lup2-at-psu.edu} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Saturday, May 31, 2003 1:09 AM } Subject: Cu peaks in STEM/EDS } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu } grids, I } } always find Cu peaks present no matter where the beam shoots. Sometimes } they } } can even be more intense than the peaks from my specimen when I am working } on } } PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu } } signals? If the EDS spectrum shows signals from Cu grids which is more than } a } } micron away from the beam, how can one be sure about the probe size or } spatial } } resolution then? } } } } Thanks in advance for any comments. } } } } Ling } } } } ------------------- } } Ling Pan, Ph.D. } } 248 MRL Building } } Materials Research Institute } } The Pennsylvania State University } } University Park, PA 16802 } } Tel: (814)865-6115 } } Fax: (814)865-2326 } } } } }
One of the listers wrote me off line, asking why the lowest mag rather than highest mag objective. I'm sure that many of you have the same question.
Here's the story: This is one of those weird idiosyncracies of microscopy imaging.
There are two planes of focus which are of interest: a. The depth in the specimen (properly called "depth of field") and b. The depth in the image - at the camera or detector (properly called "depth of focus")
The depth of field is directly related to the numerical aperture of the system. However, the depth of focus is directly related to the magnification(squared). Sooo... For a low power (ex: 5x scanner lens): 5(squared) = an image which is 25 "units" deep at the camera plane For a higher magnification (ex: 100x): 100(squared) = an image which is 10,000 "units" deep at the camera plane. Since "25 unit" slice is more shallow, it defines the depth of focus at the camera more precisely. That plane will always fall within the "10,000 unit" slice, so once the camera is "parfocal" (in focus for a plane matching the plane for the eyepieces) for your lowest magnification objective, it will parfocal for all other objectives.
Hope this is helpful.
Best regards, Barbara
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. And don't forget to focus the camera using the lowest power mag on your nosepiece. Then it will automatically be in focus for all other objectives.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration. Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
At 08:32 PM 5/30/03 -0700, David Burton wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Helen M Turner wrote: ======================================================= I'm looking for a couple of 10 litre dewars for liquid nitrogen and wondered if anyone could recommend a supplier? Alternatively if anyone in New Zealand has any second hand ones for sale I'd be interested. ======================================================= An excellent 10 liter dewar is shown on URL http://www.2spi.com/catalog/instruments/liquid-nitrogen-dewars.shtml of the SPI Supplies website. It can be purchased directly from SPI Supplies in the USA or via our distributor in New Zealand:
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
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Michael,
I would assume that you are correct, the issue is the length of your trinocular adapter.
Take a look at our webpage which lists solutions for adapting to the cameras to all the major microscope manufacturers as well as the 'off-brands' as well via a couple of mounting options: http://www.mvia.com/Coolpix/clpxadpt.htm
Our solutions are probably less expensive than other options you have looked at. Please feel free to contact me with your microscope model if you would like to go over the various options.
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
michael shaffer wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We have adapted a Nikon Coolpix 5000 to our microscope with trinocular } head. The adaptation is not specific to the microscope, and we are having a } problem with focus. That is, we can elevate the stage for vision's focus, } or we can elevate the stage for the camera's focus ... but neither are } coincident ... and we imagine this is because the adaptation is not specific } to the microscope (the adapter being quite expensive for this particular } brand & model). Judging from playing with the adapter, its tube-length is } slightly too long. } } I suppose we were hoping the camera could be manually focussed, so that } both could be made coincident, but for the Coolpix we cannot find a way of } manually focussing. Or, it may be impossible to focus because the adapter's } tube-length is too long rather than too short. Is this the correct } assumption? What have others done? } } cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } www.micro-investigations.com
I need to do some experiments using a FEG SEM. I am using infected cells that I am planing to immunolabel with antibodies coupled to colloidal gold.
My questions are : is immunolabelling carried out in the same way as for labelling on grids ? critical point drying or not ? metallisation or carbon coating ? All answers will be helpfull, thanks in advance danile
Here's an update on the Kodak MDS 100 digital camera ...
Works pretty well on the Olympus SZH stereomicroscope. I had no focusing problems 'cuz I also got the correct Olympus C-mount camera adapter, the TVZ-M. It's got corrections for image distance (0.55 to 1.1) and fine focus. The fine focus is a boon 'cuz the SZH only has rack-and-pinion focus.
Color I dealt with by adding a heat-absorbing glass in front of the fiber-optic stalk(s) and with an IR cutoff filter in front of the CCD element. Both from Edmund Industrial Optics. Gets the red mostly out.
Resolution isn't quite what it seems to be - the camera will record images at 1280X960, but in the back of the instruction book the image sensor is only 640X480 pixels. And one can see an odd waffle pattern within uniformly colored areas sometimes. Looks like the image is being projected onto the back side of the sheet of Masonite hardboard, which is a product made on metal wire screens. These effects disappear when the image is presented at a reasonable size - say, 3 by 4 inches.
One aspect of the less-than-optimum resolution is that the effective magnification is boosted approximately 2.5 times by the small size of the image sensor compared to the coverage of the eyepiece lens. That stretches the resolving power of the stereomicroscope's objective lens.
Color corrects OK, once I read the instructions. Still, upon going to maximum magnification with the microscope, the image becomes very blue, probably because there's a color shift with light level, which of course goes down at high magnifications.
Installation is a breeze, as the camera gets its power and input/ouput from the USB port of the PC.
The software works well; and the images can be nicely edited with a full range of gamma, brightness, contrast, tint, RGB, etc. One can also annotate the image in the margins with the Kodak software. That annotation vanishes if one later edits with other software, of course. One "trick" to enhance performance is to let the camera work by itself without competition from other programs like MSWord, Netscape, and the like, which starve it of memory. However, I can leave a large number of images open during any one session without crashing the program. Haven't lost one yet.
For a total expenditure of ~$500 it's a good value and runs circles around my previous method of hand-holding a Sony 1.3 megapixel camera over the eyepiece of the stereomicroscope. Seventy-five images in a two-hour session, for example. I can still take Polaroids if all else fails, but that's really tedious now that I've been to the Big City.
Best regards, George Langford, Sc.D. Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
Daniele Immunolabelling is the same as for grids. Gold sizes from ~5nm are suitable. Double labelling works fine, but don't overdo the size ratios. Mass increases with cube of diameter, so 25nm gold is 125 as massive as 5nm, and the supernova-like glare of the glare particles can obscure the smaller ones. Adjacent pairs in the table below should give sufficient size differential. Critical point drying is required. Coating is required either with carbon or chromium - you will need to experiment with coating thicknesses to suit your specimens and microscope parameters, but in general you will find optimised Cr coatings are thinner than C, give richer ultrastructural detail and excellent BSE signal. Noble metal coatings (Au, Au/Pd, Pt) are not suitable. Hope this helps Chris
C.E. Jeffree, H.W. McL. Rixon, G. Brown, J. Aitken, and R.J. Sugrue Distribution of the attachment (G) glycoprotein and GM1 within the envelope of mature respiratory syncytial virus filaments revealed using field emission scanning electron microscopy VIROLOGY 306 (2): 254-267 FEB 15 2003
I need to do some experiments using a FEG SEM. I am using infected cells that I am planing to immunolabel with antibodies coupled to colloidal gold.
My questions are : is immunolabelling carried out in the same way as for labelling on grids ? critical point drying or not ? metallisation or carbon coating ? All answers will be helpfull, thanks in advance danile
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Danile Spehner wrote: ============================================================================ = I need to do some experiments using a FEG SEM. I am using infected cells that I am planing to immunolabel with antibodies coupled to colloidal gold.
My questions are : is immunolabelling carried out in the same way as for labelling on grids ? critical point drying or not ? metallisation or carbon coating ? All answers will be helpfull, thanks in advance danile ============================================================================ == So far as I have always heard, it is done pretty much the way you would for labelling on grids. The sample is critical point dried.
When people talk about "metallization", they generally are talking about gold but you really can't coat with gold and expect to see with BSE the image of the gold distribution. But when one carbon coats, which has been the other alternative, you lose the sense of depth in the sample.
But now it has been shown that a thin layer of plasma deposited (from the vapor) osmium metal, in an extremely thin layer, such as can be done in the osmium plasma coaters gives you the best of all worlds: You have the needed conductivity, you get the sense of depth, while at the same time, you are not diminishing the signal from the surface tagged gold particles. This kind of deposition equipment is described on URL http://www.2spi.com/catalog/osmi-coat.html and I would call particular attention to the page URL http://www.2spi.com/catalog/osmium-plasma-coater-demonstration.html "Human astrocytoma cells, gold tagged, BSE imaging". If a picture is worth 10,000 words, I should be able to stop talking and let the micrographs speak for themselves.
Chuck
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Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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Following my recent cooling water leak on a TEM we have come off 'mains' water and have a loan chiller. However this is incredibly noisy and of course throws out heat to the room. We need get our own system installed asap and engineering wise something to go in the same spot would be good. Does anyone know a system either chiller or heat exchanger which is particularly quiet?
Gillian Brown Histology Section, Asthma Biology RI CEDD, GlaxoSmithKline
We have just noticed that there was a typo in my previous posting for the TEM job in La Jolla (San Diego), CA
So here is the posting with the correct e-mail address.
Position Open: Electron Microscopist Research Assistance, to start June, 2003. (http://www.burnham-inst.org).
Requirements include working knowledge of high resolution TEM and a BA/BS or equivalent experience in biological or material science, or bioengineering. Responsibilities include negative stain sample preparation and electron microscopy, vitreous ice sample preparation and electron cryo-microscopy, some microscope maintenance and standard biochemical analyses.
Application should include a statement of career goals and addresses for three references. Salary is competitive and commensurate with experience. EOE.
Send applications to: Dr. Dorit Hanein, e-mail dorit-at-burnham.org
} I need to do some experiments using a FEG SEM. I am using infected } cells that I am planing to immunolabel with antibodies coupled to } colloidal gold. } } My questions are : } is immunolabelling carried out in the same way as for labelling on grids ?
Probably. The cells could also be labeled in their incubation medium in the wells or wherever they're grown, including in suspension. Post-label, fix with 1 or 1.25% glutaraldehyde + 1% tannic acid (helps preserve the cell membrane), in whatever buffer is appropriate for your cells.
} critical point drying or not ?
Yes, definitely.
} metallisation or carbon coating ?
High-resolution platinum coating, 2 to 4 nm thick. We use an ion-beam coater. Thinner, IF your cells *and* the mounting method allow it. Image at 5kV. Gold lights up nicely in the BSE at this accelerating voltage. (Your BSE detector will likely control what kV you have to use, though, but on a FEG SEM, you should have a high-resolution BSE detector anyway.)
Phil
} All answers will be helpfull, } thanks in advance } danile } -------------------------------------------- } Danile Spehner, INSERM EPI 99-08 - EFS-Alsace } 10 rue Spielmann - 67065 STRASBOURG } tel : 03 88 21 25 25 - fax : 03 88 21 25 44 } e-mail : daniele.spehner-at-efs-alsace.fr } ------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
We have an Aluminum sheet composite material that consists of Aluminum sheet/ polymer layer/ paper backing. The Aluminum sheet corroded through the paper and polymer layers in some areas. The Al corroded to the point that it contains large holes. We would like to cross section these samples for SEM keeping the paper layer intact.
We are looking for ideas on how to cross section the samples for SEM so all three layers can be imaged.
Kerry N. Siebein University of Florida Particle Engineering Research Center P. O. Box 116135 Gainesville, FL 32611-6135
We're looking for a 100x objective for our Zeiss Axiovert that will also be compatible with an Olympus scope (not that that should be an issue as our present objectives are interchangeable, but just in case). This will initially be used for H&E stain imaging but with later applications to follow.
E-mails are already into some reps, but does anybody have an "objective" view of quality, price, flexibility issues with different lens. Likes, dislikes...etc.
Thank you.
Gary
Gary S. Laevsky, Ph.D. Research Associate The Scripps Research Institute 10550 N. Torrey Pines Road/IMM-24 La Jolla, CA 92037 (858) 784-9372
I am not sure that you are going to get wafers that thin from them, but I have bought sapphire disks from Insaco in Philadelphia in the past. They also have the capability of machining ceramics. Their web site is http://www.insaco.com/index.html
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Robb Westby [mailto:robb-at-ascendinstruments.com] Sent: Tuesday, June 03, 2003 5:50 PM To: Microscopy-at-sparc5.microscopy.com
Hello, I am trying to find a vendor for a piece of sapphire with the below criteria. Can anyone help in directing my search?
In our lab we routinely and effectively cryo-slice or cryo-cut papers/films. Freeze your material, and tweezers using LiqN2 in a mini dewar. To slice we use a variety of tools depending on what works best with the material in question. The tools can be micro scissors or straight blades. Be careful when using the blades, though! Be sure to freeze whatever slicing tool you choose.
EMS makes x-sect mounts for the SEM known as thin sample holders.
Good luck! -------------------------------- Jane Dowell Surface Science Labs WR Grace and Co. (T) 410-531-4115 (F) 410-531-4652 jane.e.dowell-at-grace.com
-----Original Message----- } From: ksiebein-at-erc.ufl.edu Sent: Wednesday, June 04, 2003 10:32 AM To: Microscopy-at-sparc5.microscopy.com
Check the paper below, in which SEM and immunogold is used to localize myosin on sperm cell surface.
Z Zhang, HQ Tian and SD Russell (1999) Localization of myosin on sperm-cell-associated membranes of tobacco. Protoplasma 208:123-128.
I'll be happy to send you a copy if you need.
Zhaojie Zhang Microscopy Core Facility University of Wyoming
-----Original Message----- } From: Spehner Daniele [mailto:daniele.spehner-at-efs-alsace.fr] Sent: Tuesday, June 03, 2003 5:14 PM To: Microscopy-at-sparc5.microscopy.com
Hi Listers,
I need to do some experiments using a FEG SEM. I am using infected cells that I am planing to immunolabel with antibodies coupled to colloidal gold.
My questions are : is immunolabelling carried out in the same way as for labelling on grids ? critical point drying or not ? metallisation or carbon coating ? All answers will be helpfull, thanks in advance danile
Thanks to all who replied, particularly Norm - your suggestion worked. I located the beamsplitter, unscrewed the head and dropped the filter in before the beamsplitter and hey presto - the fluorescence image was visible through both the eyepiece and the phototube. I've tried the filter with rhodamine and I can see red fluoresecence so thanks again - I don't need to buy a new filter!
Kind regards,
Patricia
---------------------- Patricia de Winter cdewi02-at-students.bbk.ac.uk
Dear listers, I received a call from an investigator at Univ. of Cinn who needs help with cryoultramicrotomy and TEM of nanotubes. He is willing to drive anywhere within a 4 or 5 hour drive from Cinn. If any of you would be able to help him please contact me. Thanks so much, Mary Gail Engle
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
Digital Image Capture and Management in Microscopy
October 16, 2003
A course on Digital imaging in light microscopy which will cover the following topics:
Optical Limitations in Light Microscopy...Photographic Imaging Strategies... Digital Imaging Strategies...Selection of Digital Capture (Camera vs. Scanner)...
Image Processing of Captured Images...Image File Formats...Printing Images... Color Management Systems...Database Management Software...Presentation Software for Oral Reports...Website Performance...Integration of Image Data with Sample Information, Calibration, Other Data & Reports...Acrobat and html Software for Written Reports and Archives...Examples of Efficient, Low Cost Image Handling Systems...
Examples of Electronic Microscopy Reports and Databases
The course instructors are Mary and John McCann of McCann Imaging.
WHEN: Thursday, October 16, 2003, from 9 A.M. to 5 P.M.
Never having done this, let me take a stab at it. This is free, so that it doesn't cost you anything.
Why don't you try to impregnate (vacuum impregnate?) the paper with epoxy and then try to polish it or cut it?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Siebein, Kerry [mailto:ksiebein-at-erc.ufl.edu] Sent: Wednesday, June 04, 2003 10:32 AM To: 'Microscopy-at-MSA.Microscopy.Com'
We have an Aluminum sheet composite material that consists of Aluminum sheet/ polymer layer/ paper backing. The Aluminum sheet corroded through the paper and polymer layers in some areas. The Al corroded to the point that it contains large holes. We would like to cross section these samples for SEM keeping the paper layer intact.
We are looking for ideas on how to cross section the samples for SEM so all three layers can be imaged.
Kerry N. Siebein University of Florida Particle Engineering Research Center P. O. Box 116135 Gainesville, FL 32611-6135
First, I must solicit your strictest confidence in this transaction, this is by virtue of it's nature as being utterly confidential and top secret .We are top officials from the Federal Ministry of Works and Housing,(FMWH),Federal Ministry of Finance and the Presidency, making up the Contract Review Panel (CRP) set up by the Federal Government of Nigeria to review contracts awarded by the past military administration. In the course of our work on the CRP, we discovered this fund which resulted from grossly over-invoiced contracts which were executed for the FMW&H during the last administration. The companies that executed the contracts have been duly paid and the contracts commissioned leaving the sum of US$35.4 Million floating in the escrow account of the Central Bank of Nigeria ready for payment. I have therefore been mandated as a matter of trust by my colleagues in the panel to look for an overseas partner to whom we could transfer the sum of US$35.4M legally subcontracting the entitlement to your company. This is bearing in mind that our civil service code of conduct forbids us from owning foreign company or running foreign account while in government service hence the need for an overseas partner. We have agreed that the funds will be shared thus after it has been paid into your account: (1) 30% of the money will go to you for acting as the beneficiary of the fund. (2) 10% has been set aside as an abstract projection for reimbursements to both parties for incidental expenses that may be incurred in the course of the transaction. (3) 60% to us the government officials (with which we wish to commence an importation business in conjunction with you). All logistics are in place and all modalities worked out for the smooth conclusion of the transaction within ten to fourteen days of commencement after receipt of the following information: Your company name, address, company's details, telephone & fax numbers. These information will enable us make the applications and lodge claims to the concerned ministries & agencies in favour of your company and it is pertinent to state here that this transaction is entirely based on trust as the solar bank draft or certified cheque draw able in any of the Central Bank of Nigeria correspondent bankers around the world is going to be made in your name.Please acknowledge the receipt of this letter using my e-mail address or the alternative email;tphector-at-ny.com, your interest and tel/fax number for further details. Yours faithfully, Mr. Hector Tunde Fax=234-9-2722017 Email:tphector-at-ny.com _________________________________________________ Votre mail gratuit ?ie avec http://lexpress.net
What are the thickness measurements of the constituent materials?
At first blush, a traditional Buehler polishing cross section should do the job. These are embedded in rigid epoxy. Very routine for microcircuit work. It may be applicable to your situation.
gary g.
At 07:31 AM 6/4/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Maybe. Have not tried it. I would worry about the impact of this on ultrastructure. Can you tell me how/why HMDS works? Does the specimen get coated in the stuff, and if so is the residue of the material visible in a FEGSEM??
I have also wondered about osmium impregnation e.g. by OTO as an alternative to metal or carbon coating for immunogold labelled specimens. However, so far our experiences with Osmium /Tannic Acid /Osmium in another context have resulted in highly intrusive decoration of the specimen, so that all parts have the same, clearly artifactual ultrastructure. This is so fine that it would probably not be resolved by conventional SEM, but FEGSEM is less forgiving.
Chris
Dr. Chris Jeffree
----- Original Message ----- } From: "Dusevich, Vladimir" {dusevichv-at-umkc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 04, 2003 6:33 PM
I am out of office until 16-06-2003. For urgent matters please contact micro-at-omnilabo.be or call 03 870 58 03
Hi, I was wondering which cover slips are in use for high resolution microscopy or has anyone reviewed cover slips that are commercially available? As it is possible for a user to inadvertently introduce error into a well-corrected system just by selecting the wrong cover slip, I have been instructing people who use the microscopes to utilise the correct thickness of the cover glass (0.17 millimeters) and try and move away from plastics.
Unfortunately with high quality cover glasses having a tolerance of +/- 10 micrometers, the FWHM is detrimentally altered by more than a factor of two if a cover slip is on the extreme of this tolerance.
Are there coverslips available with a finer degree of tolerance than the standard +/- 10 ? thanks
Steve
Steve Bagley Applied Imaging Facility Paterson Institute For Cancer Research Cancer Research UK Christie Hospital, Wilmslow Rd, Manchester. M20 9BX, UK
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Never had any trouble with HMDS affecting ultrastructure for SEM (even FEGSEM). As far as the OTOTO method goes, there was a paper about 20 years ago that discussed the need to find a consistent source for the TCH--they are not all created equal. Second, the authors recommended washing small amounts of the TCH crystals (can't remember the details) several times to remove impurities; this leaves TCH crystals that are white/semi-opaque, rather than the grayish color as received. Third, rigorous, thorough washing between steps is critical. I use 5x in dH2O between each and every step. This gives excellent impregnation and conductivity (plus secondary electron signal) as well as fine surface structure preservation in the FEGSEM. I have to look up the details for washing the TCH--I'll get back to you on that (it's obviously been awhile since I have done this).
Roger Moretz, Ph.d. Dept of Toxicology BI Pharmaceuticals, Inc. Ridgefield, CT -- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Maybe. Have not tried it. I would worry about the impact of this on } ultrastructure. } Can you tell me how/why HMDS works? Does the specimen get coated in } the stuff, } and if so is the residue of the material visible in a FEGSEM?? } } I have also wondered about osmium impregnation } e.g. by OTO as an alternative to metal or carbon coating for } immunogold labelled specimens. However, so far our experiences with } Osmium /Tannic Acid /Osmium in another context have resulted in highly } intrusive decoration of the specimen, so that all parts have the same, } clearly artifactual ultrastructure. This is so fine that it would
} probably not be resolved by conventional SEM, but FEGSEM is less } forgiving. } } Chris } } Dr. Chris Jeffree } } ----- Original Message ----- } } From: "Dusevich, Vladimir" {dusevichv-at-umkc.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, June 04, 2003 6:33 PM } Subject: RE: SEM and Immunolabelling } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ---. } } } } } } } } } } critical point drying or not ? } } } } } } Yes, definitely. } } } } What about HMDS? } } } } Vladimir } } } }
Kerry, I hope the University has a decent metallographic lab. There you will find the resources you need. I've often prepared similar samples for study.
If the sample is large, I would start by buttering the porous area with a prepared mixture of two-part epoxy. This epoxy is available from any metallographic supply house (Struers, Buehler, Leco, Lapmaster, MetLab, Mager), has excellent wetting properties, low viscosity for sample impregnation, and should be allowed to cure overnight. This epoxy will "fix" the porous and loose material in place for subsequent sectioning.
After the buttered area is hard, section slightly to the side of the area of interest. Rough cut the rest of the sample to fit in a metallographic mount. Mount the area - again using epoxy. After the mount has cured, rough polish the area of interest to remove cutting damage.
At this point, the sample should be "frozen" in place with a bit of porosity at the cut surface. Again, put some epoxy on the cut and rough polished surface. Use vacuum techniques to draw air out of the porosity and drive epoxy into the sample. After curing, prepare the sample metallographically for an optically flat surface. After sputter coating to make the sample conductive, it is ready for viewing in the SEM.
It takes a couple of days to prepare the sample, but this is the routine I use for samples of this type. If imaging is all you need and don't need EDS information, you may forego sputter coating and SEM examination and simply use a metallograph for your study.
Stu Smalinskas Metallurgist SKF USA Plymouth, Michigan (734) 414-6862 stu.smalinskas-at-skf.com
(I have no financial interest in the metallographic supply houses. I'm just a satisfied customer.)
We have an Aluminum sheet composite material that consists of Aluminum sheet/ polymer layer/ paper backing. The Aluminum sheet corroded through the paper and polymer layers in some areas. The Al corroded to the point that it contains large holes. We would like to cross section these samples for SEM keeping the paper layer intact.
We are looking for ideas on how to cross section the samples for SEM so all three layers can be imaged.
Kerry N. Siebein University of Florida Particle Engineering Research Center P. O. Box 116135 Gainesville, FL 32611-6135
You do not say how thick the material is but I have had success with a "cryo" method on similar materials.
The SEM is a clever tool, cut a material and it sees the cut, not the material interfaces! The only alternatives are embedding + mechanical sectioning + polishing or "cryo".
For cryo - cut a piece of the material into an egg timer shape being 1/2inch wide at top and bottom but only 1/4 inch wide in the neck. Lower this into liquid nitrogen and wait for the bubbling to subside. Remove with light weight pliers and, holding each end with the pliers, CRACK the material by bending it slightly. If the material does not crack do not repeat the bending but try again with another piece of material but make the neck 1/8 inch wide.
Hope this helps?
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "Siebein, Kerry" {ksiebein-at-erc.ufl.edu} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 04, 2003 3:31 PM
Hi
I am on the point of buying a digital camera for microscope work.
I have seen a Japanese-made adapter for the Olympus Camedia C-3040 camera in which the end away from the camera was stepped in three different diameters, to suit a trinocular port plus the two common eyepiece tube diameters. The adapter had an internal lens, and was apparently made by a small non-English- speaking Japanese company without electronic communications, and was expensive. It would be useful for my purposes as the same adapter/camera can be easily and quickly put onto any one of a number of microscopes.
Adapter manufacturers' websites that I have seen seem to describe only adapters that suit only one microscope mounting point ie either C-mount OR 23mm eyepiece tube OR 30mm eyepiece tube.
Can someone either point me in the direction of someone who makes a similarly universal adapter or explain to me why such a device doesn't work as well as a less-flexible system does?
Also, does anyone have good grounds for a preference, for this kind of work, for the Coolpix 4500 over the Olympus 3040 or vice- versa? They seem pretty similar to me on specifications.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
I sent out an email for 'help' regarding our aging 20 year-old Hitachi H-600 TEM on the 9th of April this year. We thought our HT cable had died and really could not justify spending in excess of $30,000 on a new gun/cable assembly as we were applying through a grant for a new TEM. I thought it may be beneficial to summarise the situation for Listserver members (see below). I would also like to extend a very BIG thank-you to all those who responded especially Joel McClintock, Steve Chapman,William Mushock, Terry Fitton, John Brealey, Sally Stowe, Noni Hudson and George Theodossiou. I really felt like I had a team helping me through this. Can you all now pray that my grant application is successful and we can replace the old girl sooner rather than later?
The symptoms were;
1. Unstable voltage readouts. When we set the voltage to, say, 50 Kv, it did not read 50 Kv but read 75Kv. 2. There was a 'hissing' sound when we tried to change from 75Kv to 100Kv. The hissing appeared to be coming from behind the gun. 3. Our filaments, when they broke, were showing signs of over-heating as evidenced by a round blob of tungsten on each of the broken ends. They were not being over-saturated. 4. When we opened the gun, it smelt. 5. When we disconnected the cable from the HT tank, the HT tank read-outs were OK.
Some of the people who responded to my cry for help suggested that the symptoms may indeed by caused by a dirty gun and/or poor vacuum.
What did we do? We cleaned the gun thoroughly. We did not dismantle the gun from the microscope but cleaned it in situ. We used diamond paste (wenol) sparingly on lint-free cloths to scrub off the pale toffee colored discoloration of the porcelain until it was white and shiny. We wiped any paste residue off with 10% quadralene (ammonia based) followed by water then ethanol (very dry) then acetone (very dry) . We then allowed it to pump down over the weekend. It was not a difficult procedure and one that, had I known how straight forward it would be, should have been done a lot earlier.
On Monday, we had trouble stabilising the HV at 100KV but after 3 goes, reached and held 100KV. I have taken a while to present this summary to the Listserver members as I wanted to make sure the dirty gun was the culprit. Now, 2 months down the track we remain clear of all the original symptoms. Of course there are other small problems that have sprung up associated with using a TEM that is 20 years old but they are all 'fixable'.
Again, thank-you list-server members
cheers
Sarah Ellis
Manager, Microscopy Imaging and Research Core Facility Peter MacCallum Cancer Centre Locked Bag #1 A'Beckett Street East Melbourne 8006
-----Message d'origine----- De : Dusevich, Vladimir [mailto:dusevichv-at-umkc.edu] Envoyé : mercredi 4 juin 2003 19:33 À : Microscopy-at-sparc5.microscopy.com Objet : RE: SEM and Immunolabelling
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} } critical point drying or not ? } } Yes, definitely.
Unless there is some way to adjust for tube length by a lens in the adapter, I would think you would have trouble establishing parfocality by using an adapter with a stepped outside to the tube. There have been another thread or two talking about adapters that allow for that. I know we don't have one of them.
I have posted some pictures on our ftp server to show what we had to go through with our Olympus Stereoscope. I modified the phototube so that I could use the same relay lens that worked just fine on our other two Olympus scopes (reflected and transmitted. I had to remove an adapter on the top of our tube that was used as the mechanical support for the Olympus camera. It prevented the relay lens from sliding in as far as it needed to. I then had to make a tube spacer to set the relay lens back off to the right length to obtain parfocality with the eyepieces.
I would suppose that you might be better off having your shop make some bushings for your larger phototubes to accomodate an adapter for the smaller tube. It might take a little doing to make sure you maintain the proper path length, but any machinist worth their pay should be able to rig up something. (Or maybe Edmund already stocks such bushings.) I just hope you don't run into any vignetting issues.
ftp://www.marl.iastate.edu/LabPix/Stereo_adapter/
Warren
At 02:15 PM 6/6/2003 +1200, you wrote:
} Hi } } I am on the point of buying a digital camera for microscope work. } } I have seen a Japanese-made adapter for the Olympus Camedia } C-3040 camera in which the end away from the camera was } stepped in three different diameters, to suit a trinocular port plus } the two common eyepiece tube diameters. The adapter had an } internal lens, and was apparently made by a small non-English- } speaking Japanese company without electronic communications, } and was expensive. } It would be useful for my purposes as the same adapter/camera } can be easily and quickly put onto any one of a number of } microscopes. } } Adapter manufacturers' websites that I have seen seem to } describe only adapters that suit only one microscope mounting } point ie either C-mount OR 23mm eyepiece tube OR 30mm } eyepiece tube. } } Can someone either point me in the direction of someone who } makes a similarly universal adapter or explain to me why such a } device doesn't work as well as a less-flexible system does? } } Also, does anyone have good grounds for a preference, for this } kind of work, for the Coolpix 4500 over the Olympus 3040 or vice- } versa? They seem pretty similar to me on specifications. } } cheers } } rtch } } -- } Ritchie Sims Ph D Phone : 64 9 } 3737599 ext 87713 } Microanalyst Fax : 64 9 } 3737435 } Department of Geology email : } r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand
I have acquired a surplus turbo pumped JEOL 845 SEM, but the documentation I have received doesn't seem to cover the turbo modification. Would anyone out there operating a turbo pumped JEOL 840 or 845 like to share their cold start instructions and operating instructions with me?
Sincerely,
Scott
F. Scott Miller, Ph.D. Advanced Materials Characterization Lab University of Missouri-Rolla 223 McNutt Hall Rolla, MO 65409 USA fax: 573 341 6934 voice: 573 341 4727
As Philip suggested a 2 - 4nm coating of platinum will allow you to examine the sample at 5Kv using the backscatter detector. In addition, it has been my experience that under those conditions the lower Se- detector provides some atomic number imaging, at least that's the case in my instrument. While I have not tried this technique on gold labeled cells I have had success looking for metal contaminants in organic crystal samples. The advantages offered by using the Se- detector certainly makes it worth a try with your equipment/samples.
John A. Robson Boehringer Ingelheim Pharmaceuticals, Inc. PO Box 368 900 Ridgebury Rd Ridgefield, CT 06877
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Wednesday, June 04, 2003 9:46 AM To: Microscopy-at-sparc5.microscopy.com
Daniele,
see below
} I need to do some experiments using a FEG SEM. I am using infected } cells that I am planing to immunolabel with antibodies coupled to } colloidal gold. } } My questions are : } is immunolabelling carried out in the same way as for labelling on grids ?
Probably. The cells could also be labeled in their incubation medium in the wells or wherever they're grown, including in suspension. Post-label, fix with 1 or 1.25% glutaraldehyde + 1% tannic acid (helps preserve the cell membrane), in whatever buffer is appropriate for your cells.
} critical point drying or not ?
Yes, definitely.
} metallisation or carbon coating ?
High-resolution platinum coating, 2 to 4 nm thick. We use an ion-beam coater. Thinner, IF your cells *and* the mounting method allow it. Image at 5kV. Gold lights up nicely in the BSE at this accelerating voltage. (Your BSE detector will likely control what kV you have to use, though, but on a FEG SEM, you should have a high-resolution BSE detector anyway.)
Phil
} All answers will be helpfull, } thanks in advance } danile } -------------------------------------------- } Danile Spehner, INSERM EPI 99-08 - EFS-Alsace } 10 rue Spielmann - 67065 STRASBOURG } tel : 03 88 21 25 25 - fax : 03 88 21 25 44 } e-mail : daniele.spehner-at-efs-alsace.fr } ------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
I suggest you take a look at our webpage for adapting the Coolpix series cameras to a microscope: http://www.mvia.com/Coolpix/clpxadpt.htm
1.) If you want to go through 23mm eyepieces, our optics adapter is a straight drop in.
2.) If you want to go through 30mm eyepieces, you use the exact same optics, and we have a sleeve to increase the diameter of the adapter to 30mm.
3.) If you want to go through a trinocular port, you use the exact same optics, and we have several clamps to fit to various microscopes.
4.) If you want to go through an existing 1X C-mount on your microscope, you use the exact same optics, and we have a clamp and thread adapter to convert our optics to a C-mount fitting.
In all of these cases, we are using the exact same optics system, the only thing we change is the clamp, sleeve, or thread fitting that will connect it to your microscope. All parts are interchangeable and part of our universal system.
We have an optics system specific to the Coolpix series cameras (4500 included) that will definitely work with the Coolpix 4500.
On the other hand, we also have a universal optics adapter for fitting to digital cameras, but the Olympus 3040 is an untested model for us and I am not sure how well it will work.
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Ritchie Sims wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi } } I am on the point of buying a digital camera for microscope work. } } I have seen a Japanese-made adapter for the Olympus Camedia } C-3040 camera in which the end away from the camera was } stepped in three different diameters, to suit a trinocular port plus } the two common eyepiece tube diameters. The adapter had an } internal lens, and was apparently made by a small non-English- } speaking Japanese company without electronic communications, } and was expensive. } It would be useful for my purposes as the same adapter/camera } can be easily and quickly put onto any one of a number of } microscopes. } } Adapter manufacturers' websites that I have seen seem to } describe only adapters that suit only one microscope mounting } point ie either C-mount OR 23mm eyepiece tube OR 30mm } eyepiece tube. } } Can someone either point me in the direction of someone who } makes a similarly universal adapter or explain to me why such a } device doesn't work as well as a less-flexible system does? } } Also, does anyone have good grounds for a preference, for this } kind of work, for the Coolpix 4500 over the Olympus 3040 or vice- } versa? They seem pretty similar to me on specifications. } } cheers } } rtch } } -- } Ritchie Sims Ph D Phone : 64 9 } 3737599 ext 87713 } Microanalyst Fax : 64 9 } 3737435 } Department of Geology email : } r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand
Unless specimen is mounted in direct contact with the underside of the cover glass, use No. 1 cover glasses.
Gary Gill
-----Original Message----- } From: Steve Bagley [mailto:SBagley-at-picr.man.ac.uk] Sent: Thursday, June 05, 2003 3:42 AM To: microscopy-at-sparc5.microscopy.com
Hi, I was wondering which cover slips are in use for high resolution microscopy or has anyone reviewed cover slips that are commercially available? As it is possible for a user to inadvertently introduce error into a well-corrected system just by selecting the wrong cover slip, I have been instructing people who use the microscopes to utilise the correct thickness of the cover glass (0.17 millimeters) and try and move away from plastics.
Unfortunately with high quality cover glasses having a tolerance of +/- 10 micrometers, the FWHM is detrimentally altered by more than a factor of two if a cover slip is on the extreme of this tolerance.
Are there coverslips available with a finer degree of tolerance than the standard +/- 10 ? thanks
Steve
Steve Bagley Applied Imaging Facility Paterson Institute For Cancer Research Cancer Research UK Christie Hospital, Wilmslow Rd, Manchester. M20 9BX, UK
This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.
Applied Precision produces measurement and analysis hardware and software for the semiconductor and biotechnology industries.
We are currently seeking a success-oriented Applications Scientist to support Applied Precision's Life Science Sales Team. This is a technical support position both pre- and post- sale.
This person will be responsible for providing scientific and technical support during product demonstrations, tradeshow events, and workshops. Training users on hardware and software included in bio-imaging products is also a major constituent of the position.
The successful candidate will have excellent communication, organizational, and project management skills and be able to travel extensively. Must have a B.S. or higher in Cell or Molecular Biology or related field and must have expertise in several of the following areas: Widefield/Deconvolution or Confocal Microscopy, Fluorescence Imaging, Microarray Technology, Digital Image Analysis, UNIX based operating systems.
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Kerry N. Siebein wrote: ======================================================================== We have an Aluminum sheet composite material that consists of Aluminum sheet/ polymer layer/ paper backing. The Aluminum sheet corroded through the paper and polymer layers in some areas. The Al corroded to the point that it contains large holes. We would like to cross section these samples for SEM keeping the paper layer intact.
We are looking for ideas on how to cross section the samples for SEM so all three layers can be imaged. ======================================================================== Over the years we have had systems that sound similar, where the aluminum layer could have been as thin as aluminum foil and as thick as an old style aluminum lithographic printing plate. We have found that the following will work:
1] You need to have some kind of embedding to keep the paper intact but you won't want the paper to be swollen with the embedding resin to the extent that it loses all of it original dimensions. We like to apply a passivation layer of sputtered gold (if we did this now, we would use osmium metal in the osmium coater since you can get a much thinner effective barrier layer); the purpose of the passivation layer is to limit the amount of embedding resin infiltration and also to act as a decoration layer to more easily define the paper/embedding resin interface.
2] We have always found that our own (or that offered by at least some of our competitors) version of Epon 812 (ours is called SPI-Pon 812) works better than any of the other alternatives. It also gives reasonably good adhesion with the opposite aluminum layer.
3] The next step is to diamond knife thin section the block as if you were going to be doing TEM as well. We have found that when SEM views are really what are needed, we know that the best SEM "faced-off-piece" will be obtained when the best possible TEM sections are being taken. So even if TEM is not the main interest, we still evaluate the sections coming off to make sure they are of the quality needed to result in the best possible "faced-off-piece" for SEM.
4] At this point, and in order to give the faced-off-piece a bit more contrast, we subject it to a very slight plasma etching treatment in our SPI Plasma Prep II plasma etcher. Typically 10-20 seconds might be all that is needed but that is enough to etch down so that the aluminum layer "stands up" like a little micro wall. The etched structure of the polymer vs. paper is quite distinctively different. Inorganic additives are also prominently seen, often times like little "mesas" in Arizona.
5] The diamond knife ultramicrotomy is done using an SPI Supplies Materials Science Diamond Knife. To use anything "better" (e.g. a life science knife) is a waste of money since the inorganics typically found in the paper and polymer layers will impart striations to the knife edge with the first pass of the knife over the sample. We find that knifes with 45° angle are optimum and that if 55° angle knives are used, there are significant compression effects in the sections and the face-off-piece quality suffers as well.
Using this approach, one can indeed get a good view of the aluminum foil layer but it is not as good of a view if you are looking for incipient signs of aluminum corrosion (for that, the TEM is much better). The SEM view is good in terms of looking at the degree of penetration of the polymer into the paper structure or just looking at the uniformity of layer thickness of all three of the layers (because typically by SEM one is looking at a longer length of cross-section.
Disclaimer: SPI Supplies offers most of the products mentioned in this posting, as part of our SPI Supplies product line, including the embedding resin, plasma etcher, and diamond knives. We would have a vested interest in seeing more people viewing these kinds of samples this way since it uses up more of our products.......
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
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Listers,
} From time to time we get a request here at Microscopy Today for an article that someone wants to read so as to learn about some aspect of microscopy.
Here's one: ====== Dear MT Editor,
I've been exchanging emails with a friend, and we've both decided we're mystified by coating. The physics of thin coats, how they actually form, what their real structures are, why are carbon coats conductive, those things, at the physical level. We both know how to use sputter coaters, vacuum evaporators and the like, and basically what they're doing, how they work, but ... what's *really* happening and why? One or more articles for MT? I don't know who to suggest, but with your connections, there has to be someone you know who could write them. And aren't evaporated C coats mostly bucky balls and nanotubes? ========
Is there anyone out there willing to write such an article? About 1,500 words plus pictures written in a tutorial style for a general, non-specialist, microscopy audience.
Please e-mail me off the listserver
Ron Anderson, Editor Microscopy Today microtoday-at-attglobal.net
The secondary electron detector does show the gold nicely, also. The gold particles don't light up with the SE like they do with the BSE, but they do stand out from the background of biological samples. I don't think this is atomic number imaging, though, as much as it is just that the gold particles are very good sources of secondary electrons, and much denser than the rest of the sample. If the instrument allows it, the best way to go is simultaneous acquisition of SE and BSE images.
Phil
} As Philip suggested a 2 - 4nm coating of platinum will allow you to examine } the sample at 5Kv using the backscatter detector. In addition, it has been } my experience that under those conditions the lower Se- detector provides } some atomic number imaging, at least that's the case in my instrument. } While I have not tried this technique on gold labeled cells I have had } success looking for metal contaminants in organic crystal samples. The } advantages offered by using the Se- detector certainly makes it worth a try } with your equipment/samples. } } John A. Robson } Boehringer Ingelheim Pharmaceuticals, Inc. } PO Box 368 } 900 Ridgebury Rd } Ridgefield, CT 06877 } } Phone (203)798-5640 } Fax (203)798-5698 } e-mail jrobson-at-RDG.boehringer-ingelheim.com } } } -----Original Message----- } } From: Philip Oshel [mailto:peoshel-at-wisc.edu] } Sent: Wednesday, June 04, 2003 9:46 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: SEM and Immunolabelling } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bappooh-at-myexcel.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, June 7, 2003 at 21:15:39 ---------------------------------------------------------------------------
Email: bappooh-at-myexcel.com Name: Tammy
Organization: Northside Christian School
Education: K-8 Grade Grammar School
Location: City, State, Country Louisville,Ky.u.s.a.
Question: Can you use microscopy looking at human blood cells and determine any nutritional defiencies ?
Dear Sir As an exporter in China, We Ali trade Inc. can supply 12,000 kinds of products. Currently we are selling: pro audio connectors rca connectors audio equipments and mic leads Audio plugs&jacks wire cable electronic plug-in units fuse computer microphone multimedia sound sound/megaphone/switch indoor antenna piezo tweeter and horn available package demonstration manual reset circuit breakers fused power distribution blocks power distribution block braided screen power cables amplifier installation wiring kit series rac interconnect series optical fiber cables rca plugs/jacks cable drums for stage performance speakers&speaker boxes siren driver horn microphone headphone&mic multimedia speaker system woofer car audio amplifier car speaker component speaker package dome tweeter hi fi system boom box and etc. Fax:0086-574-87317472 Mr Qiu
Dear Sir As an exporter in China, We Ali trade Inc. can supply 12,000 kinds of products. Currently we are selling: pro audio connectors rca connectors audio equipments and mic leads Audio plugs&jacks wire cable electronic plug-in units fuse computer microphone multimedia sound sound/megaphone/switch indoor antenna piezo tweeter and horn available package demonstration manual reset circuit breakers fused power distribution blocks power distribution block braided screen power cables amplifier installation wiring kit series rac interconnect series optical fiber cables rca plugs/jacks cable drums for stage performance speakers&speaker boxes siren driver horn microphone headphone&mic multimedia speaker system woofer car audio amplifier car speaker component speaker package dome tweeter hi fi system boom box and etc. Fax:0086-574-87317472 Mr Qiu
Dear Sir As an exporter in China, We Ali trade Inc. can supply 12,000 kinds of products. Currently we are selling: pro audio connectors rca connectors audio equipments and mic leads Audio plugs&jacks wire cable electronic plug-in units fuse computer microphone multimedia sound sound/megaphone/switch indoor antenna piezo tweeter and horn available package demonstration manual reset circuit breakers fused power distribution blocks power distribution block braided screen power cables amplifier installation wiring kit series rac interconnect series optical fiber cables rca plugs/jacks cable drums for stage performance speakers&speaker boxes siren driver horn microphone headphone&mic multimedia speaker system woofer car audio amplifier car speaker component speaker package dome tweeter hi fi system boom box and etc. Fax:0086-574-87317472 Mr Qiu
Dear Sir As an exporter in China, We Ali trade Inc. can supply 12,000 kinds of products. Currently we are selling: pro audio connectors rca connectors audio equipments and mic leads Audio plugs&jacks wire cable electronic plug-in units fuse computer microphone multimedia sound sound/megaphone/switch indoor antenna piezo tweeter and horn available package demonstration manual reset circuit breakers fused power distribution blocks power distribution block braided screen power cables amplifier installation wiring kit series rac interconnect series optical fiber cables rca plugs/jacks cable drums for stage performance speakers&speaker boxes siren driver horn microphone headphone&mic multimedia speaker system woofer car audio amplifier car speaker component speaker package dome tweeter hi fi system boom box and etc. Fax:0086-574-87317472 Mr Qiu
} } } } Subject: re-freezing tissue in OCT for cryo-sectioning } } } } From: Angela welford {awelford-at-salud.unm.edu} } } } } To: microscopy-at-sparc5.microscopy.com } } } } Content-Transfer-Encoding: 7bit } } } } Message-Id: {121E4BB6-96D7-11D7-81B6-000A959AEFF4-at-salud.unm.edu} } } } } X-Mailer: Apple Mail (2.552) } } } } Content-Length: 660 } } } } } } } } } } } I have some unfixed kidney tissue that is currently in OCT at -70C. } } } } The pieces are quite large, as prepared for frozen sections for } } } } histology. My need is to cryo-section a portion of the tissue for } } } } immunolabelling for EM. Is there a recommended method to handle the } } } } tissue to minimize any damage from thawing and re-freezing, or do I } } } } need to be concerned about that since it is in OCT? My plan was to } } } } cut } } } } off a piece, let it thaw and cut it down for fixation in PF, then } } } } infiltrate with sucrose before freezing (plunge in LN2). If anyone } } } } has } } } } a protocol they can recommend, I would appreciate it! } } } } Angela Welford } } } } UNM, Albuquerque, NM } } } } (505)272-1445 } } } } } } } }
Edinburgh City Council are proposing to install a tramline network and I understand that this could have a severe impact upon the operation of our FEG SEM. The route for this network is still out to consultation but I'd be grateful of any other users' experience regarding proximity, magnetic fields, vibration etc so that I'm fully informed should it become an issue.
Thanks,
Frieda Christie, Electron Microscopist, Scientific & Technical Services, Royal Botanic Garden, 20A Inverleith Row, Edinburgh, EH3 5LR
I would suggest you touch base with Dougal Mcculloch {dougal.mcculloch-at-rmit.edu.au} at RMIT in Melbourne.
He had exactly this problem and as I recall he installed field canceling coils in one FEGTEM room which was closest to the TRAM line. As I vaguely remember in a second room the coils were not needed as the field was sufficiently low as not to affect the instrument.
Nestor Your Friendly Neighborhood SysOp
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The box said ... "This program requires Win 2K/XP or better..." So I bought a G3 Mac !
Someone has asked me to explain the way a sputtered coating of metal bonds to a surface.
I am a biologist, so the details are a little sketchy for me. In general terms, what is the mechanism for the bonding, and could it ever be considered 'covalent' bonding?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
At the request of one of the list members, herewith is a summary via cut and paste of all the replies I got about this subject.
try ultra darkfield first - I think Olympus makes an ultra darkfield condenser . Do you expect there to be some slight density differences? Maybe can you centrifuge them out in a sucrose gradient?
At ~2000g, all the cells will sediment. What about the particles? If the particles are less dense than hemoglobin, then layer a slightly less dense than particles but more dense than hemoglobin, hemolyze blood with distilled water, and spin. Your particles should 'fall' into 'dense' layer at bottom. On the other hand, your particles might be more buoyant than the cells, in which case you win that way. In the absence of answers to my questions, you are left with a manageable experiment anyway, especially if the particles start out in a jar on the shelf. If they are manufactured in the body, i.e., crystals, then you must do it the hard way. An Airfuge (Beckman) will sediment hemoglobin, but not certain mycoplasma-like organisms, and it will work with really small volumes. Even if the particles sediment faster than the hemoglobin, the hemoglobin isn't birefringent. With 1ml to start, you have a lot of flexibility.
Once you can use sedimentation, you can concentrate the stuff. In the end, by my estimate, the best bulk dehydrant of stuff in a dialysis bag is PEG (polyethylene glycol).
A hypotonic solution (hypotonic as compared to plasms, water or very dilute buffer) will cause the red cells (and the WBCs too I suppose) to swell and burst. Knowing the composition of the 'particles' you are looking for might help.
Interesting problem. SEM would indeed show your particles, but there are lots of things in the given size range, and it won't tell you which ones are birefringent (or even necessarily which ones are yours). Polarized light microscopy will, so that would be a good way to go. An old geological light microscope will do this. There might even be one of these around with a black light, so you could check for fluorescence. If your budget allows, McCrone Research in Chicago can do this -- they're big on polarized light microscopy. Then again ... thinking ... get a couple of 50 - 55 mm diameter polarization filters for a 35mm camera. Linear filters, not circular. Put one above the slide with your sample and the other below, and cross them. This would show birefringence. (Polarized sunglasses lenses would do this, too.) But I wonder ... what else in blood is birefringent? I think you're right about centrifugation, but why not filter first and eliminate everything larger than 3 microns, instead of lysing cells and all that. Then get tricky. What is the composition of the particles? Could they stand up to bleach, NaOH, enzyme detergents? If yes, then after filtering why not put the filtrate into something like one of these to digest the remaining cells and gunk, then spin down and wash the particles. Mind, if these are fluorescent, then I'd just throw the filtrate under a fluorescent microscope. Mind, I don't know if there is much in blood that is autofluorescent. Baxter doesn't have an ordinary fluorescent 'scope? ... our confocal uses a Nipkow disc and Hg lamps, so we can do UV easily. And we have DIC on it (meaning we have the polarizers, the Wollensten filters just need to be taken out of the light path). 8-)
This may be simplistic, but could you just dilute the blood with a very hypotonic solution - dilute saline for example - so the blood cells burst, and presumably things like macrophages would burst too. Then you could spin down the particles, I guess 100 nm particles would need a fairly high g spin, to concentrate them a bit. On a microscope with strain-free objectives, etc for polarised light you will be able to detect birefringent particles of this size and up. cheers,
Rupture the blood cells with osmotic pressure by adding distilled water and then centrifuging or filtering the results to get the partials. I doubt your can see and reliably be able to say what it is less than 500 nm with a light microscope.
I am not sure what sub wavelength birefringent partials do in polarized light but I wouldn't count on them acting like partials } 2 wavelengths in size. Unless you have some pretty good prior research you need see what happens with know bifringent specimens over these size rangers. Preferable the same compounds as you find in blood.
I would talk to your local hospital laboratory's hematology department. Thy will have an instrument for doing automated blood counts. This instrument requires a solution that lyses the red cells. They would very likely be willing to sell or maybe even give you a small amount of it, or you can get a hold of Beckman Coulter, their product is called Coulter Lyse. They also have a whole blood lysing kit in their flow cytometry e-catalog. The URL is http://www.beckman.com/products/instrument/flowcytometry/ecatalog/ProductDet ail.asp?partnumber=6602764. After you lyse the cells you could try filtering the blood through a 5um filter and dialyze.
Just regular, old fashioned polarized light would seem to be the most direct solution. Even if the particles are small, they should respond as bright against the dark background of crossed polars. Based on the size, I would suggest higher mags (60x or even 100x). I would also make sure that you do not use plan apos but use objectives especially suited for polarized light analysis. To test if the system itself might cause a problem, put a sample in place, establish Koehler illumination, then cross the polars to get the blackest background possible. Remove the sample and see if the background stays velvety black. If you do not get velvety black at all steps, the optics are exhibiting strain and will interfer with your imaging. You may also want to inquire about the lab-level microscopes that are built specifically for polarized light analysis. Leica's have been well known for decades and Nikon came out with a new series about 3 years ago.
Re: the size of your particles The good thing about "bright against dark background" types of imaging is that it is not resolution limited: you will be able to detect the particles, even at 100nm. You may not be able to size them at that level, or to get good edge or shape information, but you should be able to detect that they are. Of course, the higher the NA on the objective (and condenser), the better the imaging in general. Oil immersion may not be a bad idea. I had my hands on a 40x/1.3NA two weeks ago which was incredible!
Re: sample prep. You may want to dilute the blood with an isotonic solution, just to avoid overcrowding from red blood cells, but unless the red blood cells are just too thick, I don't see any need to sonicate or do anything else as drastic as you have described.
There you have it. Thanks again for all the suggestions.
Covalent bonding refers to actual chemical bonding or sharing of electrons between the substrate and the coating (e.g. epoxy). A coating of metal on a non-metal surface would stick because of adhesive forces. The strength of this bonding by adhesive forces is depending on surface roughness, percent of the surface in direct contact with the coating, and the relative surface energy of the surface substrate & coating (see contact angle measurements).
The physicists in our group can explain the details of adhesive forces in terms of quantum mechanics.
J. Roy Nelson Material Testing Lab. (609) 730-0575
Jon Krupp wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi: } } Someone has asked me to explain the way a sputtered coating of metal bonds } to a surface. } } I am a biologist, so the details are a little sketchy for me. In general } terms, what is the mechanism for the bonding, and could it ever be } considered 'covalent' bonding? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
} ... } } Adapter manufacturers' websites that I have seen seem to } describe only adapters that suit only one microscope mounting } point ie either C-mount OR 23mm eyepiece tube OR 30mm } eyepiece tube. } } Can someone either point me in the direction of someone who } makes a similarly universal adapter or explain to me why such a } device doesn't work as well as a less-flexible system does? } } ...
Along these same lines, and with regard to my wanting to purchase something similar, how does a fixed adapter accommodate the the difference in stage 'Z' position, while one person will focus on the sample (specific for an interocular binoc setting, e.g., 62mm) and for another person's optical focus (e.g., interocular binoc setting, e.g., 70mm). It would seem, given trinocular head, for making camera focus coincident with optical focus, each user would first, personalize the the binocular, ... and second, fine tune the adapter with camera focus at infinity(???)
It seems to be a truism that subsidized academic SEM labs and microscopy core facilities flourish while unsubsidized labs generally languish and/or die.
Can those of you happily ensconsed in the subsidized facilities please let me know where your funding for staff salary and service contracts comes from? For my lab, at the moment, it's all got to come out of the user fees, helped somewhat by a small institutional supplement per billing hour.
Many thanks, Dee
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
Johns Hopkins University in Baltimore Maryland has an opening for an Electron Microscopy Technician. If you are interested please contact me utilizing the indicated information.
Lois Anderson Johns Hopkins University Dept. of Pathology Laboratory Manager Electron Microscopy/Immunofluorescence 600 N. Wolfe Street/Pathology 709 A Baltimore, MD 21287 (410) 955-2861/fax (410) 614-7110 landers-at-jhmi.edu
I am looking for an independent service contractor for microtomes. I have Leica UCT (with cryo), Ultracut E, LKB III, Sorvall MT-2, and microtomes for paraffin. It will be nice if the contractor can also take care of knife maker and cryostat. Our current service contracts are nearing the end.
Thanks,
Gang (Greg) Ning Director, Electron Microscopy Facility Huck Institute for Life Sciences The Pennsylvania State University 1 South Frear Lab University Park, PA 16802 Phone: 814-863-0994 Fax: 814-863-1357 Email: gxn7-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
A=sheet thickness B=thickness of sheet + pull tab + sticky tab C=Teflon pull tab thickness D=sticky tab thickness (calculated) E=resistance of tab from top to bottom sides F=resistance of tab from left side to right side on top (resistance readings taken under compression)
New Pella tab is stiff as a board. EMS is very similar to old film style Pella tab. Curiously, none of the new tabs are very conductive. Any reading lower than Infinity must be done under compression.
It may be worth comparing the Fullam thin film tab. Although, they are about $30 more per 200 than the EMS tabs.
I am running fluorescence time lapse investigations using a Zeiss Axiovert200M, with an oil immersion x63 lens (with PIFOC) and full environmental chamber. The time frame for collection of planes of focus are every three minutes for a period of twelve to sixteen hours.
My problem is that over this time frame there is a reduction of oil on the objective due to evaporation, hence the images deteriorate over time. To resolve the lens is lowered, addition of more oil, and then reposition the lens hoping that you have not knocked the slide in the process.
Is there an objective heater or similar device available, where oil can be put onto the objective with a minimum of disruption via a pump?
Many thanks,
Steve
Steve Bagley Associate Scientist Applied Imaging Facility Paterson Institute For Cancer Research Cancer Research UK Christie Hospital, Wilmslow Rd, Manchester. M20 9BX, UK
This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.
Hi Dee, At the present time, my salary (split 50-50 between the EM and optical microscopy core facilities) and that of a half-time technician are paid by the medical college, from its research environment funding. I must generate enough income in the facility to cover costs of supplies, service contracts and other equipment maintenance costs, and other misc. expenses (phone, computer networking, etc). I do not know how long this relative Shangri La will last, but I will enjoy it while it does. 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
Can anyone tell me how to measure SEM/STEM beam diameter ("spot size") in the SEM?
I did this twenty years ago and still have the photo records but not the method! The method involved photographing latex spheres at e.g. x30,000.
I recall measuring edge fringes? But, the little grey cells don't want to make the connection anymore! Would it be simply the width of any blurred edge or fringes?
We want to get a "handle" on this in connection with x-ray microanalysis of compartments in frozen unicells in a cryoSEM. Then I want to try some Monte Carlo models (if applicable to biological specimens re. matrix etc) to get some idea of the volume from which the x-rays are derived. We are getting similar results for adjacent areas on area scans (to reduce damage? Ha!). I'm hoping to calculate and to reduce the excited volume - looking for strontium in pulse chase feeding experiments.
Thanks in advance for anything you can offer
Keith Ryan Marine Biological Association Plymouth, UK
I measured the beam diameter on my Dual-Beam FIB using a TEM type sample and a modified SEM mount. The sample is essentially a "low-background" version of the usual SEM resolution standard.
Take a C-coated formvar TEM grid and sputter a few seconds worth of gold on it so that you have gold islands.
SEM mount - Take a ~1/4 inch carbon rod (~3/4 inch long) and drill out the center. Use a 1/8 inch end mill to make a little recess in the end of the rod for the TEM grid. Attach the other end of the C-rod to an SEM stub using silver paint. You now have a vertical C-rod with a recess for the TEM grid. Drilling out the center of the rod makes an effective "Faraday cup" so that you don't have bulk scattering affecting your measurements.
I imaged the sample used the SE detector at up to 800kX (in-lens mode) and was able to look at the intensity profile across the edge of the Au particle. The profile will give you your beam diameter. It was pretty easy to show 1.5nm beam diameter (20% - 80% points on the profile). If I remember correctly, I used Dave Bright's "Lispix" software to extract the profile from the saved images.
cheers, Henk
At 04:54 PM 6/11/2003 +0100, K.P.Ryan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
We have a JEOL 100S TEM scope (1974) that is in working condition and free to any taker. The scope has been under service contract since the beginning and is currently operating in our lab. Taker must pay any and all relocation fees and assume responsibility for the units upkeep. Please contact me at the e-mail address below if you are interested. Contact Terry at JEOL directly at 847-823-0306 if you want to know about costs of moving and assembly of the scope. Thanks!
Craig M. Klotz B.S., CT (ASCP) Neuromuscular Pathology cklotz-at-mcw.edu EM/Lab Tech. "Chance favors the prepared mind" - L. Pasteur
I think you will find that the spot size is not the critical dimension if you are doing x-ray analysis on thick samples. I would concentrate on a Monte Carlo simulation of the excitation volume given your matrix and beam conditions.
I routinely tell people that the volume is on the order of a micron at 20kV. Of course it is larger for low-Z materials like organics, but at least they get the idea that the volume is _much_ larger than the incident spot size.
Warren
At 04:54 PM 6/11/2003 +0100, you wrote:
} Hello Listers } } Can anyone tell me how to measure SEM/STEM beam diameter ("spot size") in } the SEM? } } I did this twenty years ago and still have the photo records but not the } method! The method involved photographing latex spheres at e.g. x30,000. } } I recall measuring edge fringes? But, the little grey cells don't want to } make the connection anymore! Would it be simply the width of any blurred } edge or fringes? } } We want to get a "handle" on this in connection with x-ray microanalysis of } compartments in frozen unicells in a cryoSEM. Then I want to try some Monte } Carlo models (if applicable to biological specimens re. matrix etc) to } get some idea of the volume from which the x-rays are derived. We are } getting similar results for adjacent areas on area scans (to reduce damage? } Ha!). I'm hoping to calculate and to reduce the excited volume - looking for } strontium in pulse chase feeding experiments. } } Thanks in advance for anything you can offer } } Keith Ryan } Marine Biological Association } Plymouth, UK
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Does anyone have experiences using Evans Blue as a counterstain for Red Blood Cell in a cell suspension for confocal microscopy?
I am trying to locate bacterial cell in the infected red blood cells. Bacteria are stained with Alexa-488 similar to FITC. I tried to counterstain RBC with Evans Blue but I can not see them. Is this because the blood has been washed after Evans Blue staining?
I would particularly like to know the diluent, working concentration, length of staining. In addition, is it necessary to wash the RBC after Evans blue staining?
Are there any alternatives recommended for use as a counterstain for RBC?
I have an Anatech Hummer VII sputter coater that is being replaced. This unit is not working (no HV). Pump is OK and control electronics are OK. I suspect that one of the drive oscillator transistors are bad. The main feed voltage circuitry is OK.
I'm replacing this unit with a Denton Desk II and would like to find someone who can fix and use the Hummer. I have the Operation Manual and full schematics. This version is the better one with a single HV transformer rather than two. The HV driver transistors are TIP19 (TO-220). The pain is getting the main PC board out to access the transistors.
The unit comes with Au/Pd, Pt and Al targets (for plating and etching). I also can include a spare Edwards E2M1 pump. Seals are new, glass is perfect. This was recently acquired as a core exchange for an earlier unit that failed.
I can tell you how to "try" to measure it. Warren and others have done pretty much the same.
the method involves doing a line scan across a perfectly sharp edge into a bottomless pit (Faraday cup). I used a Pt aperture but found that it is not really all that sharp. I wound up using a #1 cover slip that was coated with about 200A of Au/Pd. The thickness is not all that important. The goal is to get a line that has good SE at the coating and nothing at the pit. Position the edge at the center of the screen and focus on it. Then up the mag to 200K or so. Then do a line scan and capture. You will get an output like:
----- \ \ \ \ \ \ \ ------- |-----| 20/80%
Do a measurement of 20% down from the top and 20% up from the bottom. Based on the mag, this 20/80% slope will represent the probe passing over the edge...or thereabouts.
Based on the SEMs I have tested, at 10A to 50A rez, I think I have to take the maker's word for this. Really tough to measure!!
gary g.
At 08:54 AM 6/11/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone know a source (vendor, private person, researcher) for a Nikon 4.0 phase objective: Nikon Plan 4 DL 0.1 160/- PhL
Thank you very much! Michal
Dr. Michal Opas Department of Laboratory Medicine and Pathobiology University of Toronto 1 King's College Circle Medical Sciences Building, room 6326 Toronto, Ontario, M5S 1A8 Canada -------------- phone: (416) 978-8947 (laboratory) (416) 971-2140 (office) fax: (416) 978-5959 email: m.opas-at-utoronto.ca
I'd like to thank all those who passed on their thoughts regarding the possible effects on FEG SEM due to the proximity of tram lines. There certainly appear to be issues that may need consideration if our City Councillors go ahead with their plans.
Frieda Christie, Electron Microscopist, Scientific & Technical Services, Royal Botanic Garden, 20A Inverleith Row, Edinburgh, EH3 5LR
Our facility purchased a basic model XL30-TMP ESEM (tungsten filament) over two years ago. The Peltier cryo attachment was added last year and it appears we may (?) be adding an EDX system later this year.
Please send me your experiences with the different models of energy-dispersive X-ray systems integrated with ESEM microscopes offline. Thank you in advance!
Bruce F. Ingber, Biologist Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
This is a question about doing surface analysis on a large telescope mirror.
The coating on the mirror is deteriorating after some 10 years in place. It will have to be recoated. Before recoating, we would like to look at the old coating to see if there are any clues as to why it is getting flakey. The mirror is too big to just slip into the SEM.
We can strip the coating off with 'Scotch' tape, but that leaves it 'upside down' for putting into an instrument like an SEM to see the 'top' of the coating. We suspect that there may be some elements, eg sulfur or others that should not be there, but we don't know how to check it out.
I thought of stripping the film with acetate replica tape and dissolving the tape to mount the film right side up and then doing some EDS in our SEM. Think it will work?
Alternatively, does any one have some ideas about some kind of portable device we could take to the mirror and get an idea of the elements present. I thought about a handheld XRF unit, but have no experience with such things.
Thanks, now put your thinking caps on.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Hi Liststers, I have a SEM project to do that involves making replicas of teeth. The replicas I have made were made with acetate replicating tape. The guy who gave me the teeth sez it is a kind of powder that one makes into a mixture that one coats the teeth and hardens much like a latex peel. Any advice as to how to make these and which would be better for SEM?
Barbara Plowman Univ. of the Pacific School of Dentistry ph: 415-929-6692 email: Bplowman-at-sf.uop.edu
Jonathan Krupp wrote: ====================================================== Dear Brainiacs:
This is a question about doing surface analysis on a large telescope mirror.
The coating on the mirror is deteriorating after some 10 years in place. It will have to be recoated. Before recoating, we would like to look at the old coating to see if there are any clues as to why it is getting flakey. The mirror is too big to just slip into the SEM.
We can strip the coating off with 'Scotch' tape, but that leaves it 'upside down' for putting into an instrument like an SEM to see the 'top' of the coating. We suspect that there may be some elements, eg sulfur or others that should not be there, but we don't know how to check it out.
I thought of stripping the film with acetate replica tape and dissolving the tape to mount the film right side up and then doing some EDS in our SEM. Think it will work?
Alternatively, does any one have some ideas about some kind of portable device we could take to the mirror and get an idea of the elements present. I thought about a handheld XRF unit, but have no experience with such things
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bop00rav-at-sheffield.ac.uk) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, June 12, 2003 at 15:30:13 ---------------------------------------------------------------------------
Email: bop00rav-at-sheffield.ac.uk Name: BOB VASEY
Organization: university of sheffield
Education: Graduate College
Location: sheffield, uk
Question: I am using LR White for light microscopy of plant roots. I want to stain with safranin then counterstain with either astra blue or fast green. I am interested in looking at cell walls/lignin etc.
I can stain with safranin ok, but trying to counterstain with Astra Blue or Fast Green does not work. Can anyone tell me why the counterstaining is not working in LR White - it is normally fine in parafin wax.
Does anyone have a protocol for staining with safranin then counterstaining with Astra Blue/Fast Green. Are there any other stains I could counterstain with.
I would be grateful for any suggestions anyone might have, as there is nobody in my lab who can help me.
Although I'm definitely no brainiac {grin} , I've worked with many telescope mirrors in the past as an amateur telescope maker (emphasis on "amateur", by the way). However, I've never encountered one that had the coating "get flakey". Generally what happens is that the coating loses some of its reflectivity (appears dull) due to microscopic pitting which results from long term exposure to air pollution. For Newtonian, Dobsonian, or Cassegrainian telescopes located in or near large urban areas it's not at all uncommon for the primary mirror to need to be recoated after 10 years. A Schmidt-Cassegrain will usually fare better due its design.
EDXS analysis of the coating will most likely yield a wide variety of elements, so it might be difficult to identify the actual culprit (AES or XPS techniques might be more useful). Today's aluminized mirrors typically have multilayer coatings consisting of titanium dioxide, silicon dioxide (or silicon monoxide), followed by magnesium fluoride as the outermost layer. For EDXS work, you might try to remove some of the flakes by using the adhesive side of a 3M "Sticky Note". For general EDXS work these are not considered to leave any significant residue. If you can remove some of the flakes this way then it should be easy to transfer them "right side up" to a piece of carbon tape, etc. It would also be a good idea to examine the underside of the flakes since this is where adhesion appears to be deteriorating, if I understand you correctly. I would not be surprised at all to detect sulfur on the flakes due to air pollution.
I'm also curious as to whether the aluminum is flaking off, too, or just the protective coatings? Either way, this is undesirable of course, but after 10 years of use in California I'm not so sure it would be considered particularly unusual (I'm assuming the telescope is located at or near the University).
I hope this helps! I'd love to hear more about the telescope and your analysis results if you'd like to write to me off-list.
Best regards,
Sue Kent Senior Staff Engineer Motorola, Inc. 21440 W. Lake Cook Road Deer Park, IL 60010 847-862-0216
Jonathan Plastic stripping could work. The traditional method of Juniper and Bradley (1958) Journal of Ultrastructure Research 2, 16-27 for stripping replicas from leaves (it works for other surfaces too) could be modified to suit your requirements. In this method, an acetone-soluble resin (Bedacryl - is it still available?) was employed as a separation layer between acetone-insoluble formvar-supported replica and the adhesive tape. The sequence was replica - formvar - bedacryl - tape - strip - dissolve bedacryl in acetone - dissolve formvar in chloroform.
To reverse the replica onto a stub you could do: bedacryl - tape - strip - formvar support the back of the stripped replica - dissolve bedacryl in acetone - attach formvar supported replica to stub.
However, a) the offending contaminant may be soluble b) the stripping method will only succeed in removing material that has poor adhesion to the glass. Your problem may be a tightly adherent contaminant film on the surface of the glass to which the coating doesn't stick. Stripping will leave it behind. On the other hand, if there is a contaminant layer that sticks to the coating but not too well to the glass your Scotch tape method puts it exactly where you want it.
Have you considered using Raman spectroscopy?
Chris
} } Dear Brainiacs: } } This is a question about doing surface analysis on a large telescope } mirror. } } The coating on the mirror is deteriorating after some 10 years in place. It } will have to be recoated. Before recoating, we would like to look at } the } old coating to see if there are any clues as to why it is getting } flakey. } The mirror is too big to just slip into the SEM. } } We can strip the coating off with 'Scotch' tape, but that leaves it } 'upside } down' for putting into an instrument like an SEM to see the 'top' of } the } coating. We suspect that there may be some elements, eg sulfur or } others } that should not be there, but we don't know how to check it out. } } I thought of stripping the film with acetate replica tape and dissolving } the tape to mount the film right side up and then doing some EDS in our } } SEM. Think it will work? } } Alternatively, does any one have some ideas about some kind of portable } device we could take to the mirror and get an idea of the elements present. } I thought about a handheld XRF unit, but have no experience with such } things. } } Thanks, now put your thinking caps on. } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
I would think the interface of interest with a front surface coated mirror would be the one between the glass and the coating, particularly since it is flaking. Therefore, if you remove it using tape, that surface will be right- side up, as would be preferable. Since you mentioned that the surface, which I assume is aluminum, is flaking, can it simply be picked off with a pair of tweezers and both surfaces analyzed?
Regards,
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Thursday, June 12, 2003 2:57 PM To: Microscopy-at-sparc5.microscopy.com
Dear Brainiacs:
This is a question about doing surface analysis on a large telescope mirror.
The coating on the mirror is deteriorating after some 10 years in place. It will have to be recoated. Before recoating, we would like to look at the old coating to see if there are any clues as to why it is getting flakey. The mirror is too big to just slip into the SEM.
We can strip the coating off with 'Scotch' tape, but that leaves it 'upside down' for putting into an instrument like an SEM to see the 'top' of the coating. We suspect that there may be some elements, eg sulfur or others that should not be there, but we don't know how to check it out.
I thought of stripping the film with acetate replica tape and dissolving the tape to mount the film right side up and then doing some EDS in our SEM. Think it will work?
Alternatively, does any one have some ideas about some kind of portable device we could take to the mirror and get an idea of the elements present. I thought about a handheld XRF unit, but have no experience with such things.
Thanks, now put your thinking caps on.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
One has to consider both external and internal sources of potential destructive agents and you cannot exclude the glass itself. I assume this is not a hobbyist scope and that it is valuable enough to have a documented fabrication? Before proceeding too far you need to assemble the information about the materials that were used and any related experience on other scopes. You haven't indicated what the metallization is, whether is monometallic or layered, etc. The glass composition and whether other mirrors were made of the same lot, and how they have performed are all important. You should consider that the glass may be a source of either ionic contaminants or of alkali that may move the corrosion potential of the metal significantly away from the area of stability that would apply in their absence.
With the role of the glass in mind, you should consider that you might need to perform "control" analyses on non-metallized portions of the mirror. Defects related to glass breakdown may only be obvious where the metallization makes them stand out but they may be occurring everywhere.
Bear in mind that ionics often have a significant solublility in organic solvents. Therefore, attempts to replicate the defects, although a wise idea, may remove the causative agents.
Solvents themselves may also serve to introduce traces of ionics. This is particularly true of chloroform, mentioned in Chris' scheme to dissolve formvar, which can develop free chlorides if not properly stored (dry and dark). For this reason, if you expect to perform electron probe analyses on any replicas you probably should make the effort to use fresh, electronic grade solvents rather than reagent grades.
John Twilley Conservation Scientist
Jon Krupp wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Brainiacs: } } This is a question about doing surface analysis on a large telescope mirror. } } The coating on the mirror is deteriorating after some 10 years in place. It } will have to be recoated. Before recoating, we would like to look at the } old coating to see if there are any clues as to why it is getting flakey. } The mirror is too big to just slip into the SEM. } } We can strip the coating off with 'Scotch' tape, but that leaves it 'upside } down' for putting into an instrument like an SEM to see the 'top' of the } coating. We suspect that there may be some elements, eg sulfur or others } that should not be there, but we don't know how to check it out. } } I thought of stripping the film with acetate replica tape and dissolving } the tape to mount the film right side up and then doing some EDS in our } SEM. Think it will work? } } Alternatively, does any one have some ideas about some kind of portable } device we could take to the mirror and get an idea of the elements present. } I thought about a handheld XRF unit, but have no experience with such } things. } } Thanks, now put your thinking caps on. } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
Your replicating idea and then dissolving the acetate film is probably the best bet. An XRF unit could give you some information, but the X-ray have a relatively long penetration depth and so most of the signal will come from the substrate for a thin film. You would need to measure substrate/film and subtract substrate only to get what your film is and there would be problems with statistics. Portable units are EDS type and I am not sure what the range of elements that can be analyzed with them. I thought that the range is limited, but I may be wrong.
Actually, using the Scotch(R) adhesive tape might be the best approach for analyzing the failure. The failure is probably at the interface of the substrate/film or at an interface of one of the layers. By pulling the film off with the tape, you have revealed the interface that has failed. You should then apply a surface analysis technique such as X-ray photoelectron spectroscopy or Auger electron spectroscopy to look at the chemistry of the surface. If it is peeling, see if you can get a sufficiently large piece without the tape or replicating tape.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Thursday, June 12, 2003 2:57 PM To: Microscopy-at-sparc5.microscopy.com
Dear Brainiacs:
This is a question about doing surface analysis on a large telescope mirror.
The coating on the mirror is deteriorating after some 10 years in place. It will have to be recoated. Before recoating, we would like to look at the old coating to see if there are any clues as to why it is getting flakey. The mirror is too big to just slip into the SEM.
We can strip the coating off with 'Scotch' tape, but that leaves it 'upside down' for putting into an instrument like an SEM to see the 'top' of the coating. We suspect that there may be some elements, eg sulfur or others that should not be there, but we don't know how to check it out.
I thought of stripping the film with acetate replica tape and dissolving the tape to mount the film right side up and then doing some EDS in our SEM. Think it will work?
Alternatively, does any one have some ideas about some kind of portable device we could take to the mirror and get an idea of the elements present. I thought about a handheld XRF unit, but have no experience with such things.
Thanks, now put your thinking caps on.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
KeyMaster has a new handheld XRF system that goes down to Fl - see http://www.keymastertech.com
Quoting "Garber, Charles A." {cgarber-at-2spi.com} :
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jonathan Krupp wrote: } ====================================================== } Dear Brainiacs: } } This is a question about doing surface analysis on a large telescope mirror. } } The coating on the mirror is deteriorating after some 10 years in place. It } will have to be recoated. Before recoating, we would like to look at the old } coating to see if there are any clues as to why it is getting flakey. The } mirror is too big to just slip into the SEM. } } We can strip the coating off with 'Scotch' tape, but that leaves it 'upside } down' for putting into an instrument like an SEM to see the 'top' of the } coating. We suspect that there may be some elements, eg sulfur or others } that should not be there, but we don't know how to check it out. } } I thought of stripping the film with acetate replica tape and dissolving the } tape to mount the film right side up and then doing some EDS in our SEM. } Think it will work? } } Alternatively, does any one have some ideas about some kind of portable } device we could take to the mirror and get an idea of the elements present. } I thought about a handheld XRF unit, but have no experience with such things }
I have been using polystyrene beads (10-200 nm) to study uptake by cells. The product from polysciences has worked well, but these beads are almost impossible to see by TEM. Are there any suggestions as to what I can use in this size range that are electron dense? Is there a way to stain the polystyrene? I do not think gold is appropriate since it is hydrophobic and may interact in ways that I do not want.
Thanx
David ____________________
David Elliott
Yale University School of Medicine 810 LCI 333 Cedar Street New Haven, CT 06520-8022
A colleague of mine did her PhD thesis on the analysis of fossil teeth and discovered a super technique. Her name is Gina Semprebon and she is a professor at Bay Path College in Longmeadow, MA. I've left her a voice mail, asking for her email, so if you contact me off line early next week, I'll try to put the two of you together.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration. Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
At 03:10 PM 6/12/03 -0700, Barbara Plowman wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Our group is looking for a simple, conventional manual Rotation Microtome using metal blades, for paraplast and acrylat-embedded sections.
We are looking for an economic purchase, such as a second hand, demontration model, stock fade-out, etc. The microtome should be in good condition, requiring no or only minimal repairs.
If you can help us, reply to Matyas Buzgo, buzgo-at-systbot.unizh.ch Botany Department, University of Florida Gainesville 32611-8526 USA
I have a glass- nono-crystallites composite. Nanocrystallites may be ferroelectric in nature and I need to prove that nanocrystallites in the glassy phase are ferroelectric.
How can i prove using TEM that these nanocrystallites(probably single domain) are indeed ferroelectric?
Thanks Pradyumna N Gupta 5 East Packer Avenue, Whitaker Lab, MSE Lehigh University, Bethlehem, PA 18015 610 758 5590 Lab
Pradyumna N Gupta 5 East Packer Avenue, Whitaker Lab, MSE Lehigh University, Bethlehem, PA 18015 610 758 5590 Lab
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I have a glass- nono-crystallites composite. Nanocrystallites may be ferroelectric in nature and I need to prove that nanocrystallites in the glassy phase are ferroelectric.
How can i prove using TEM that these nanocrystallites(probably single domain) are indeed ferroelectric?
Pradyumna N Gupta MSE Lehigh University
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I am obtaining results on what influences people's purchasing decisions when they purchase scanning probe microscopes. If you have a moment, could you please fill out the following survey and return it to me via email? Your responses will remain anonymous. Thank you for your help.
Please, press reply, and on the following questions, please put the x near to the number, or bold or circle the number that most closely reflects your opinion when you purchase scanning probe microscopes and accessories. My e-mail address is Mickey-at-pnl.gov
How important to you is/are: Extremely Extremely Unimportant Important Price 1 2 3 4 5 Terms of delivery 1 2 3 4 5 Simple and intuitive user interface 1 2 3 4 5 Ergonomic instrument design 1 2 3 4 5 Open architecture and open software code 1 2 3 4 5 Igor Pro software Interface 1 2 3 4 5 Visual Basic interface 1 2 3 4 5 C/C++ library/interface 1 2 3 4 5 LabView interface 1 2 3 4 5 1 yr. warranty 1 2 3 4 5 3 yr. warranty 1 2 3 4 5 Annual service agreement 1 2 3 4 5 Detailed instruction manual (printed) 1 2 3 4 5 Multimedia training materials (DVD, web, video...)1 2 3 4 5 Prompt service and technical support 1 2 3 4 5 Knowledgeable customer and technical support 1 2 3 4 5 Instrument customization help and support 1 2 3 4 5 Web-based customer support 1 2 3 4 5 On-site product demonstration 1 2 3 4 5 Off-site product demonstration 1 2 3 4 5 On-site training 1 2 3 4 5 Off-site training 1 2 3 4 5 Web-based training 1 2 3 4 5 Product brochure 1 2 3 4 5 Previous publications utilizing particular product1 2 3 4 5 Instrument upgrade's capabilities 1 2 3 4 5 Reputation and previous experience 1 2 3 4 5 Resolution 1 2 3 4 5 Scan size 1 2 3 4 5 Environmental control 1 2 3 4 5 Integration with optical microscopes 1 2 3 4 5 STM 1 2 3 4 5 Contact mode AFM 1 2 3 4 5 AFM force spectroscopy 1 2 3 4 5 Tapping mode AFM or equivalent 1 2 3 4 5 Shear-Force mode 1 2 3 4 5 Pulsed-force mode or equivalent 1 2 3 4 5 Magnetic force mode 1 2 3 4 5 NSOM mode 1 2 3 4 5 E-chem SPM 1 2 3 4 5
On the following questions, please put the x near to the number, or bold or circle the number that most closely reflects your opinion.
How similar is: Extremely Extremely Similar Dissimilar
Veeco/DI to Asylum 1 2 3 4 5 Veeco/DI to Molecular Imaging 1 2 3 4 5 Veeco/DI to Omicron 1 2 3 4 5 Veeco/DI to WiTec 1 2 3 4 5 Veeco/DI to MT NDT 1 2 3 4 5 Asylum to Molecular Imaging 1 2 3 4 5 Asylum to Omicron 1 2 3 4 5 Asylum to WiTec 1 2 3 4 5 Asylum to MT NDT 1 2 3 4 5 Molecular Imaging to Omicron 1 2 3 4 5 Molecular Imaging to WiTec 1 2 3 4 5 Molecular Imaging to MT NDT 1 2 3 4 5 Omicron to WiTec 1 2 3 4 5 Omicron to MT NDT 1 2 3 4 5 Witec to MT NDT 1 2 3 4 5
Finally, please respond to the following questions:
Almost Almost Never Always
When I purchase SPM, I buy Veeco/DI 1 2 3 4 5 When I purchase SPM,, I buy Molecular Imaging 1 2 3 4 5 When I purchase SPM, I buy Asylum 1 2 3 4 5 When I purchase SPM, I buy Omicron 1 2 3 4 5 When I purchase SPM, I buy WiTec 1 2 3 4 5 When I purchase SPM, I buy MT NDT 1 2 3 4 5
Thank you for your time! The survey results will be available in three weeks !
Miodrag "Mickey" Micic mickey-at-pnl.gov ========================================================================== Miodrag Micic, Ph.D. Pacific Northwest National Lab, M/S K8-88, P.O.Box. 999 3335 Q. Avenue, Richland, WA 99352-999, USA. phone: 1-509-376-5394 fax: 1-509-376-6066 E-mail: Miodrag.Micic-at-pnl.gov
"But the fact that some geniuses were laughed at does not imply that all who are laughed at are geniuses. They laughed at Fulton, they laughed at the Wright brothers. But they also laughed at Bozo the Clown". Carl Sagan =========================================================================
We've used SEM of replicas for analysis of microwear on fossil teeth here. I was not involved in the specimen prep, but below is a reply from the researcher that was. (If you have further questions, let me know).
} Kevin, } } I don't know anything about the type of replicas described below. But for dental microwear analysis of fossil teeth, we use Colténe President Jet Plus (a polyvinylsiloxane material) to make tooth molds and then pour casts with epoxy. The epoxy is centrifuged to remove bubbles. The replicas look pretty precise in the SEM, even at thousands of times magnification.
------------------------------------------------ Kevin Frischmann Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 03:10 PM 6/12/03 -0700, Barbara Plowman wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
OK...I too have been thinking about this interesting problem.
It seems to me that giving EDS a shot is a good start. Since the issue is more of a film or surface issue, EDS may give way to WDS or SIMS. Anyway, a first method is based on EDS. The problem (among others) is to reduce the volumetric interaction of a sample. I would suggest (naively?) that you use a SEM sample stub with a sticky tab to pull off some of the crud from the glass. Not elegant but potentially useful.
You can use an Aluminum stub or a Carbon stub. If you expect to find either of these elements in the material on the glass, volumetric interaction must be reduced. This can be done by coating the stub with a material that would not be thought to exist on the glass surface. One could use Au, Au/Pd or Pt. A reasonably thick coating would stop the beam from penetrating much into the substrate. A Monte Carlo simulation can be used to get a good idea of KV and film thickness relative to penetration. It is unlikely, I would think, that Al would be found on the glass. If so, an Al stub would work fine. Coat it with, say, Pt and then put the sticky tab on top. Do an EDS baseline of the stub and tab. Then, collect material from the glass. Re-do EDS and subtract the baseline from the specimen reading. That should allow C to be in the picture.
When you do the EDS analysis, also subtract/cancel out the noble metal(s) such that the contaminants are the only ones left.
Very interesting problem. let us know how it turns out!
gary g.
} Dear Brainiacs: } } } This is a question about doing surface analysis on a large telescope mirror. } } } The coating on the mirror is deteriorating after some 10 years in place. It } will have to be recoated. Before recoating, we would like to look at the } old coating to see if there are any clues as to why it is getting flakey. } The mirror is too big to just slip into the SEM. } } } We can strip the coating off with 'Scotch' tape, but that leaves it 'upside } down' for putting into an instrument like an SEM to see the 'top' of the } coating. We suspect that there may be some elements, eg sulfur or others } that should not be there, but we don't know how to check it out. } } } I thought of stripping the film with acetate replica tape and dissolving } the tape to mount the film right side up and then doing some EDS in our } SEM. Think it will work? } } } Alternatively, does any one have some ideas about some kind of portable } device we could take to the mirror and get an idea of the elements present. } I thought about a handheld XRF unit, but have no experience with such } things. } } } Thanks, now put your thinking caps on. } } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
Here are some additonal explnataion of my survey request. It is not intented for commercial purposes, and I appologise to Dr. Zaluzec for not informing him in advance. Besides being a postdoc, I am a part-time graduate student at the Washington State University, Tri-Cities campus, in the master of technology management program, see the link http://www.tricity.wsu.edu/business/mtm.html . This survey is for final project in my marketing class, and it do not have any commercial intention. Again,I do not have any product, neither I am promoting any product . This particular survey, is designed to provide 3 answers:
1. perceptual map of the current scanning probe microscopes brands (questions like compare, for example brand A with brand B), and are neutral, i.e. it cant be used to promote or degrade any mark, as I only ask similar/dissimilarity questions, to get clustering data.
2. attributes that drive consumer decision (how important it is to you)
3. correlation matrix questions, (when I buy AFM, I buy _____), which are use to correlate with the perceptual map, from the question 1.
Furthermore, I will make survey results publiclly available in a matter which is not promoting or derogating any particular product or brand. I will be glad to disclose results to you and subsequently the the list.
I am looking forward to hearing from you, and thanking you in advance for your participation,
I would kindly ask you to complete my survey quantiative survey on AFM, which is now on-line at http://www.AdvancedSurvey.com/default.asp?SurveyID=6098 . It takes about 4 minutes to complete it, and you can see current results at the end of the survey.
I would kindly ask you to complete my survey quantiative survey on AFM, which is now on-line at http://www.AdvancedSurvey.com/default.asp?SurveyID=6098 . It takes about 4 minutes to complete it, and you can see current results at the end of the survey.
Please take note of the updated page of The International School on Electron Microscopy of Powdered Nanostructured Materials to be hold in Wroc³aw (Poland), 19-20 September 2003. You will find there the program and the registration form. For complete information, please look at: http://celtam.int.pan.wroc.pl/ISEM/ Or Email: kepinski-at-int.pan.wroc.pl
Looking forward to seeing you in Wroc³aw, Organizing Committee,
Contact: Dr. Leszek Kepinski Institute of Low Temperature and Structure Research, Polish Academy of Sciences, P.O.Box 1410, 50-950 Wroclaw, Poland http://www.int.pan.wroc.pl/
Dear List, I will be attending MSA, and I would like to see if anyone would be willing to share a room with me to cut down on expenses. Please reply off-list. TIA. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Dear Pradyumna The best way to detect and image ferroelectric inclusions in paraelectric matrix is to use a scanning probe microscopy techniques, namely piezoresponse force microscopy. This technique is specifically designed to distinguish piezoelectric and non-piezoelectric materials, and hence is probably the best choice in this case. It will be sensitive only to crystallites exposed on the surface or relatively shallow within the glass matrix (~10 nm) and will allow imaging of both in-plane and out-of-plane polarization components. You can find some examples in paper A.Y. Borisevich, S.V. Kalinin, D.A. Bonnell, and P.K. Davies, Analysis of phase distributions in the Li2O-Nb2O5-TiO2 system by piezoresponse imaging, J. Mater. Res. 16(2), 329-332 (2001). The extension of PFM, piezoresponse spectroscopy normally allows spatially resolved electromechanical hysteresis loop measurements, e.g. within single 50 nm grain, but it might not work for the crystal in the dielectric matrix. Yours Sergei
-- Sergei V. Kalinin, Oak Ridge National Laboratory, 1 Bethel Valley Rd, Bldg. 3025, MS6030, Oak Ridge, TN 37831 Phone: (865) 241-0236 FAX: (865) 574-4143 URL: sergei2.kalininweb.com
Dear Pradyumna The best way to detect and image ferroelectric inclusions in paraelectric matrix is to use a scanning probe microscopy techniques, namely piezoresponse force microscopy. This technique is specifically designed to distinguish piezoelectric and non-piezoelectric materials, and hence is probably the best choice in this case. It will be sensitive only to crystallites exposed on the surface or relatively shallow within the glass matrix (~10 nm) and will allow imaging of both in-plane and out-of-plane polarization components. You can find some examples in paper A.Y. Borisevich, S.V. Kalinin, D.A. Bonnell, and P.K. Davies, Analysis of phase distributions in the Li2O-Nb2O5-TiO2 system by piezoresponse imaging, J. Mater. Res. 16(2), 329-332 (2001). The extension of PFM, piezoresponse spectroscopy normally allows spatially resolved electromechanical hysteresis loop measurements, e.g. within single 50 nm grain, but it might not work for the crystal in the dielectric matrix. Yours Sergei
-- Sergei V. Kalinin, Oak Ridge National Laboratory, 1 Bethel Valley Rd, Bldg. 3025, MS6030, Oak Ridge, TN 37831 Phone: (865) 241-0236 FAX: (865) 574-4143 URL: sergei2.kalininweb.com
----- Original Message ----- } From: "Pradyumna N Gupta" {microprady-at-lehigh.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, June 13, 2003 4:05 PM
Hi, we have a problem with gun alignment of Cameca su-30...At such } mags 3000 or higher we can not get clear SEM images, we try to make } gun alignment in wobb mode by using the diaphragms...Does anyone know } the procedure which we have to follow(wobb, diaphragms, } astigmatism....)? Do we need to clean the apertures? But how? thanks...
Dear listers, Is there any way to quantify the lowest detection limit of SEM and compare it to ppm range?
I have a customer who would like to find out if chlorine from an ink marker diffused into the structure of 17-4 PH alloy? If it did, how deep? The customer is asking what is my detection limit for chlorine 100ppm? 200ppm?
Any advice is appreciated
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
We plan to use some internal fund to purchase a 2nd hand CM120 scope or the similar as soon as practical. Should you have one or know related info on this, please let me know. Thanks much!
**************************************** Chaoying Ni The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (venu-at-purdue.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June 16, 2003 at 17:45:28 ---------------------------------------------------------------------------
Email: venu-at-purdue.edu Name: Venu
Organization: Purdue University
Education: Graduate College
Location: City, State, Country
Question: Could you please tell me the imaging requirements (tools needed) and the proper technique to observe 30-40 nm gold particles using light microscopy?
This e-mail concerns long-wave ultraviolet stereomicroscope illumination and eye protection.
I recently inspected a long-wave UV ringlight stereomicroscope accessory (StockerYale Model 18 Superlight ringlight) at a trade show. Visual results for specimen inspection and preparation were remarkable, but the ringlight was not fitted with a barrier filter to block long-wave UV wavelengths from observation and did not appear sufficiently bright to tolerate a barrier filter. What eye safety precautions, if any, would be recommended or required to use such an illuminator?
We have being doing EM on neurons that were cultured on various coverslips, recently for a specific experiment, we have to use the poly-D-lysine coated coverslips. The problem is that it is very difficult to remove this type of coverslips as comparing to matrigel-coated ones at the end of polymerization, and we can not use Thermanox for this culture. The embedding medium we used is polybed/812 and we normally use liquid nitrogen to separate cells from the coverslips, and it works very well with matrigel-coverslips.
Any suggestions and thoughts will be highly appreciated.
Xinran
Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center at Dallas Phone: 214-648-1830 Fax: 214-648-1801 E-mail: xinran.liu-at-utsouthwestern.edu
After waiting for a long time, we must now choose in hurry a new EDS system for our TEM. Our old Kevex died ((un)fortunately ?!) and after a few cycle monney-no monney-monney-no monney etc, we are again in a "monney" phase, and we must spend it fast ! So fast that we have no time to do test.
So any advises are wanted about the Noran Six, the PGT Avalon, the Oxford INCA and the EDAX Genesis EDS systems, to fit on a 200 keV TOPCON 02B TEM, and to be used in a material research lab, on magnetic materials, catalysts, bio-materials, etc. We know a little bit the PGT and the Oxford, I use personnaly the Noran Vantage, but the Six is new (only the GUI or the electronics too ?), and we know nothing (other than the manufacturer's doc) about the EDAX Genesis.
Any advise too about reparing/upgrading our old KEVEX detector, or buying a new one (much more monney). The old one worked fine before it died, and it is bellow seald, with the length and the transfer mechanism adapted to the TEM.
Answer please on the list, direct to me or to Jacques.Werckmann-at-ipcms.u-strasbg.fr
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
I want order a LaB6 cathode from FEI Beam Tech and cannot get through to a person. Their automated phone system requires that you know who you want to talk to, no operator!! Duh!
Does anyone know a contact person there?
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Hi, everyone: An investigator here is looking at the changes in ribosomes with different chemical treatments. Can anyone recommend a fixation and/or staining procedure that will enhance the contrast of ribosomes at ultrastructural level? Thank you in advance and I look forward to hearing from you.
Hong Yi Emory School of Medicine EM Tel: (404) 727-8692 (Office), (404) 712-8491 (Lab) Fax: (404) 727-3157
It sounds like you may be already doing this but when we "pop" our cells off ECM coated glass coverslips in liquid nitrogen, we find that it is important to only polymerize for 6 hrs (if we go longer, they don't pop off well)- when we first take the coverslips out of the oven they can be slightly gooey but after they cool for 15-20 min, they are "tacky" but no longer gooey. we cross-hatch the surface with a razor blade and slowly immerse in liquid nitrogen. we have used this for matrigel but not in many years.
At 10:48 AM 6/17/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
FEI is experiencing growing pains. They are in the process of moving out from one campus into their new facility. You can reach them at this number: 503-640-7500. Since they are local to us, I called and confirmed this is the number to reach them. Hope this helps!
John Quintero MikroMasch USA http://www.spmtips.com 7086 SW Beveland St. Portland, OR 97223 john-at-mikromasch.com Office: 503-598-9828 Fax: 503-598-9721
Dear Yi Ribosome (RS) is about 28 nm in diameter and wery fagile. You may not see the structure of RS on the conventional ultrathin sections with standard processing. You also could not see fine structure on the chemically fixed ribosomes. The only one way known to me to investigate the fine structure of the free ribosomes in vitro is cryo-EM with 3D-reconstruction. Please, made search for Joachim Frank works. Ribosome is extremely sensitive and complicated "machine": you have to perform state of the art biochemistry to isolate them intact. Usually we are using RNAase-free strains or thermophilic bacteria to isolate ribosomes. As far as I know, there is just a few places in US, where people is able to isolate intact ribosomes. Of coarse, "intact" is very subjective word. You may check Harry Noller works for RS isolation. Best wishes, Sergey
At 12:07 PM 6/17/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I'd say about 0.1% ie 1,000 ppm in the absence of any complicating factors, or 5000ppm with more confidence.
cheers
rtch
} From: "AtcSEM" {atcsem-at-sbcglobal.net} To: {MIcroscopy-at-sparc5.microscopy.com}
To all TEM facilities:
I need someone to help me with plant viruses/TEM research. I do have TEM background but do not have an electron microscope(none in Alaska). I need to look at purified viruses and viruses in plant tissue. I am looking for a EM facility that either contracts research projects or allows one to use their facility for a short segment of time. My immediate need is to have grids of purified virus or leaf dips looked at for virus particles.
Thank you,
Nancy Robertson
Nancy L. Robertson Research Plant Pathologist USDA, ARS 533 E. Fireweed Ave. Palmer Alaska 99645 907-746-9465 office 907-746-2898 lab 907-746-2677 fax ffnlr-at-uaf.edu
} } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? }
I don't think you have any chance with LM, but I'll be interested to hear if there is a way.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
If you mean EDS analysis, then you can't get any lower then 2-3 %. ppm level isn't for EDS at all.
Regards,
Vladimir Igoshev, Ph.D. Toronto, Canada ----- Original Message ----- } From: "AtcSEM" {atcsem-at-sbcglobal.net} To: {MIcroscopy-at-sparc5.microscopy.com} Sent: Tuesday, June 17, 2003 8:49 AM
I have several EDAX and 2 Oxford systems, and like Oxford the better.
**************************************** Chaoying Ni, PhD The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
On Tue, 17 Jun 2003, Faerber Jacques wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi All } } After waiting for a long time, we must now choose in hurry a new EDS } system for our TEM. Our old Kevex died ((un)fortunately ?!) and after a } few cycle monney-no monney-monney-no monney etc, we are again in a } "monney" phase, and we must spend it fast ! So fast that we have no time } to do test. } } So any advises are wanted about the Noran Six, the PGT Avalon, the Oxford } INCA and the EDAX Genesis EDS systems, to fit on a 200 keV TOPCON 02B TEM, } and to be used in a material research lab, on magnetic materials, } catalysts, bio-materials, etc. We know a little bit the PGT and the } Oxford, I use personnaly the Noran Vantage, but the Six is new (only the } GUI or the electronics too ?), and we know nothing (other than the } manufacturer's doc) about the EDAX Genesis. } } Any advise too about reparing/upgrading our old KEVEX detector, or buying } a new one (much more monney). The old one worked fine before it died, and } it is bellow seald, with the length and the transfer mechanism adapted to } the TEM. } } Answer please on the list, direct to me or to } Jacques.Werckmann-at-ipcms.u-strasbg.fr } } Many thanks } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } }
I have a question regarding grids for TEM studies of nanoparticles (ferrites) with sizes from 40 - 100 nm. Can anyone help me with what kind of grids/films I can use to study my particles ? or if anyone knows if there is any good literature that I should take a look at? The particles are dispersed in ethanol and then a droplet of the ethanol with particles will be put placed on the grid/film. However, if there are other better methods, I am very interested.
Dear Faerber The important thing is that you can afford your choice and that the interfacing is being done well. What I mean is that the detector actually look at the sample. Our EDAX on the Technai 12 is a stuffed job and both EDAX and fei are keeping quiet hoping that one miracle morning the interfacing will work. Some manufacturers loos their flexibility in upgraded software versions. (Programmers write the software and are not actual operators!) Test drive the system and have a serious look at backup, downtime and poor service is expensive. The quality of after sale service differs at different parts of the world and a survey in your areas will be more helpful in that regard. Happy hunting.
-----Original Message----- } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] Sent: Tuesday, June 17, 2003 5:54 PM To: Microscopy Society of America
Hi All
After waiting for a long time, we must now choose in hurry a new EDS system for our TEM. Our old Kevex died ((un)fortunately ?!) and after a few cycle monney-no monney-monney-no monney etc, we are again in a "monney" phase, and we must spend it fast ! So fast that we have no time to do test.
So any advises are wanted about the Noran Six, the PGT Avalon, the Oxford INCA and the EDAX Genesis EDS systems, to fit on a 200 keV TOPCON 02B TEM, and to be used in a material research lab, on magnetic materials, catalysts, bio-materials, etc. We know a little bit the PGT and the Oxford, I use personnaly the Noran Vantage, but the Six is new (only the GUI or the electronics too ?), and we know nothing (other than the manufacturer's doc) about the EDAX Genesis.
Any advise too about reparing/upgrading our old KEVEX detector, or buying a new one (much more monney). The old one worked fine before it died, and it is bellow seald, with the length and the transfer mechanism adapted to the TEM.
Answer please on the list, direct to me or to Jacques.Werckmann-at-ipcms.u-strasbg.fr
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
If you say SEM do you mean imaging with SE or BSE , EDS or WDS? If it is EDS I would say 0.5 atomic percent in a metal matrix at 25kV. Since it is a light element 15 kV would be sufficient to "see" all your elements in your alloy as well as Cl. The limit would be different since your interaction volume is different therefore determining the distance your EDS can see Cl (assuming you look at a cross section of the area of interest eg a fracture or a crack. Are the Cl real or from greasy hands handling the sample even the atmosphere close to the sea? Are you using theoretical standards, pure elements or a "real" 17-4 PH alloy standard that was verified by other more accurate means eg. a probe? All these factors do influence your detection limit and accuracy. The distance detectable in a trace scan is limited by your interaction volume thus density and kV combination. Remember that Cl is often driven away by the beam and the analysis should preferably not be carried out in spot mode and on a fresh area defeating the purpose of using a line or spot trace.
As You can see, this is not a simple question with a simple answer.
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, June 18, 2003 1:47 AM To: AtcSEM; MIcroscopy-at-sparc5.microscopy.com
I'd say about 0.1% ie 1,000 ppm in the absence of any complicating factors, or 5000ppm with more confidence.
cheers
rtch
} From: "AtcSEM" {atcsem-at-sbcglobal.net} To: {MIcroscopy-at-sparc5.microscopy.com}
I appreciate all the answers I have received.
I have looked on the cross section of a sample and ran EDS and EDS line scan, but could not see anything. I did suggest the SIMS since they are looking to see how deep did the Cl diffused. Now I am trying to setup Cl standard for WDS and check the cross section, but I do not think that I would be successful.
Does any one knows if defused chlorine would cause any problems with 17-4PH, especially if the customer removes 3 mils from the surface that the parts were exposed to Cl (?) containing ink marker?
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
----- Original Message ----- } From: "Bill Miller" {microbill-at-mohawk.net} To: "AtcSEM" {atcsem-at-sbcglobal.net} Sent: Wednesday, June 18, 2003 7:23 AM
Ritchie writes ...
} I'd say about 0.1% ie 1,000 ppm in the absence of any } complicating factors, or 5000ppm with more confidence.
regarding ...
} } Is there any way to quantify the lowest detection limit of SEM and } } compare it to ppm range? } } } } I have a customer who would like to find out if chlorine from an ink } } marker diffused into the structure of 17-4 PH alloy? If it did, how } } deep? The customer is asking what is my detection limit for chlorine } } 100ppm? 200ppm?
I would think 1000ppm is a good approximation, assuming we are talking about SEM/EDX. However, I'd also comment ... a detection limit is difficult to evaluate for EDX, and indeed many EDX softwares do not even provide for it. The EDX background is relatively straight-forward with modern softwares, but the error associated with it is not ... and may also depend on parameters used for background modelling ... e.g., the absorption in the low energy end, any parameter used for the exponential high energy end, whether parameters are fixed or automatic (and an iterative function of composition), or if ROIs are used.
Not that EDX sensitivities are impossible, but rather impossible to address if a lot more is not known. It sounds like this "customer" wants not only quantitative wt%, but also wants the error quantified as well (my definition of "quantitative analysis"). This SEM operator, Mr. Pavel Lozovyy, should consult specifically with his EDX & software support.
We are only interested to see if there is Cl diffusion or not and how deep (if possible). We do not need to quantify it.
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com ----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Ritchie Sims" {r.sims-at-auckland.ac.nz} ; "AtcSEM" {atcsem-at-sbcglobal.net} ; {MIcroscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 18, 2003 8:20 AM
Pavel;
I agree with Ricth Sims about the lower limit of detection. However, there are some caveats you should be aware of such as excitation volume/interaction volume and the matrix of elements the Cl is in. The software program "Electron Flight Simulator" by Small World Systems, Inc. can give you an approximation of just how deep the interaction volume is for a given accelerating voltage. Also, you should know if any of the other elements in the matrix will absorb and of the Cl photon emissions [absorption edges].
If you send me the matrix off-line I'd be happy run the simulation for you.
If you are after a depth profile, it would be best to user AES [Auger Spectroscopy] or SIMS [Scanning Ion Mass Spectroscopy] SIMS would have the best sensitivity.
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Tuesday, June 17, 2003 8:50 AM To: MIcroscopy-at-sparc5.microscopy.com
Dear listers, Is there any way to quantify the lowest detection limit of SEM and compare it to ppm range?
I have a customer who would like to find out if chlorine from an ink marker diffused into the structure of 17-4 PH alloy? If it did, how deep? The customer is asking what is my detection limit for chlorine 100ppm? 200ppm?
Any advice is appreciated
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
You need diffraction optics of some sort. Differential Interference Contrast (aka Normarski, DIC), Asymmetric Illumination Contrast (AIC), possibly Hoffman Modulation Contrast, although I haven't tried that one. Phase Contrast *might* work, although again, I haven't tried it. The idea is that these sub-resolution particles can be detected, although they cannot be resolved (or imaged). The "inflated diffraction image" is reasonably easy to see.
Phil
} Email: venu-at-purdue.edu } Name: Venu } } Organization: Purdue University } } Education: Graduate College } } Location: City, State, Country } } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
You might also try "Lyman, Roger" {RLyman-at-FEICO.com} I haven't tried to contact anyone for some months, or anyone at all except Joe Race since the last round of buyouts (by Veeco [or whoever it was], if that's still the last round of buyouts), but I assume they're still there.
Phil
} Hi, } } I want order a LaB6 cathode from FEI Beam Tech and cannot get } through to a person. Their automated phone system requires that you } know who you want to talk to, no operator!! Duh! } } Does anyone know a contact person there? } } Owen } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
} } I'd say about 0.1% ie 1,000 ppm in the absence of any } complicating factors, or } 5000ppm with more confidence. } } cheers } } rtch
I agree with this estimate, but nevertheless it is worth a try to examine the specimens with EDS. I suspect that a customer has a pitting corrosion of a stainless steel (Cl is a usual cause of a pitting). In that case Cl in the pits could (but not must) be detectable.
Vladimir
} } } } From: "AtcSEM" {atcsem-at-sbcglobal.net} } To: {MIcroscopy-at-sparc5.microscopy.com} } Subject: Fw: SEM detection limit of Cl } Date sent: Tue, 17 Jun 2003 08:49:33 -0400 } } } } ---------------------------------------------------------------------- } } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } } -. } } } } } } Dear listers, } } Is there any way to quantify the lowest detection limit of SEM and } } compare it to ppm range? } } } } I have a customer who would like to find out if chlorine from an ink } } marker diffused into the structure of 17-4 PH alloy? If it did, how } } deep? The customer is asking what is my detection limit for chlorine } } 100ppm? 200ppm? } } } } Any advice is appreciated } } } } Pavel Lozovyy } } ATC SEM Lab } } Phone: 216-692-6637 } } E-mail: atclabs-at-sbcglobal.net } } web: www.atclabs.com } } } } } } -- } Ritchie Sims Ph D Phone : 64 9 } 3737599 ext 87713 } Microanalyst Fax : } 64 9 3737435 } Department of Geology email : } r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
Many thanks to all who responded to my query about subsidized SEM labs. As we all know, this is a chronic problem. Here's a rundown of the answers, edited for brevity and anonymity:
1. We get two full time staff positions, a faculty director and $10,000 per year from the Dean. The rest of our budget comes from user fees.
2. My salary (split 50-50 between the EM and optical microscopy core facilities) and that of a half-time technician are paid by the X college from its research environment funding. I must generate enough income in the facility to cover costs of supplies, service contracts and other equipment maintenance costs, and other misc. expenses (phone, computer networking, etc).
3. My salary is covered by the department. Our user fees pay for the service contract on our TEM and SEM. The user fee also covers incidental parts, gas, and LN2. Our greatest subsidy comes from outside users. They use our SEM more than 50% of the time.
4. In our case the director and one part time staff salary come from X college . Our computer specialist's 15% salary comes from the lab revenues . Our other part time staff's 50% salary comes from revenues and the other half comes from X college. The service contract for both SEM and TEM are fully paid by a pool of money coming from X, Y, and Z Colleges. We do not have service contracts for ancillary equipment.
5. I used to have my salary and the service contract funded by departmental contributions, in exchange for free scope time and technical assistance. There was some talk of subsidizing the facility solely through user fees, but we simply wouldn't survive that way, and many potential users couldn't afford the fees we would need to charge to keep going. For the upcoming year, I'll have to recover 1/3 of salary and contract through user fees, but I think that is do-able, and we will be able to keep user fees fairly low. The rest will come from the X Dept.
6. All the money - scope service contracts, my salary etc.- comes from (the lab's founder's) grant.
7. I would not survive here at my institution were it not for the commitment of the college. I now have a bare bones operation but I do not think I could ever recover all the costs to make these unsubsidized facilities.
8. Until this year state subsidies covered our salaries, now that this has been cut the University expects us to generate the money from recharge rates (user fees). As you know this doesn't cover the cost of running a lab. Could you please share any information you receive from 'those that are happily ensconced in subsidized facilities'! It would be wonderful to figure out how to keep our facilities alive.
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
Venu, Ritchie Any fluorescence microscope equipped with a epi-polarization cube (Chroma #33001 or a 50/50 beam splitter and two polarizers from Omega) should work quite well. Tom
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Tuesday, June 17, 2003 7:38 PM To: venu-at-purdue.edu; MicroscopyListserver
} } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? }
I don't think you have any chance with LM, but I'll be interested to hear if there is a way.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
I dont see hwo the differential interface contrast can work beyond diffraction limit ?
I will say only TEM or AFM can be used to succesfully image this small particle.
Just to detect them (like in the case of single molecular spectroscopy/imaging) a confocal imaging may work well, but you do not image them, just you see brighter spot, on the size of the half of the wavelenght.
Best regards,
Mickey
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Wednesday, June 18, 2003 6:30 AM To: Microscopy-at-sparc5.microscopy.com
Venu,
You need diffraction optics of some sort. Differential Interference Contrast (aka Normarski, DIC), Asymmetric Illumination Contrast (AIC), possibly Hoffman Modulation Contrast, although I haven't tried that one. Phase Contrast *might* work, although again, I haven't tried it. The idea is that these sub-resolution particles can be detected, although they cannot be resolved (or imaged). The "inflated diffraction image" is reasonably easy to see.
Phil
} Email: venu-at-purdue.edu } Name: Venu } } Organization: Purdue University } } Education: Graduate College } } Location: City, State, Country } } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Here is a simple method using cellulose acetate, possibly the kind of powder your "guy" had mentioned:
You add acetone (99%) to the cellulose acetate and stir quickly to make a finger nail polish thickness slurry. Then you simply paint it on the surface to be replicated and it will dry almost immediately. Then you peel it off and you have the replica - actually a "negative" of the surface.
You can then trim & mount that onto a SEM stub and sputter coat as usual.
We've done this here on plant leaf samples, for various reasons, sometimes to actually pick up fungal structures from the surface in order to look at them from underneath.
You could make a positive of the replica by coating it with a water based latex like bathroom caulking, then peel that off the replica. I've never tried it but what ever you do it must be water based material or you might dissolve or distort the acetate replica.
But most important in this case, before you start, don't forget to brush! - or not?
Good luck!
-- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Bera
} Hi Liststers, } I have a SEM project to do that involves making replicas of teeth. The } replicas I have made were made with acetate replicating tape. The guy who gave } me the teeth sez it is a kind of powder that one makes into a mixture that one } coats the teeth and hardens much like a latex peel. Any advice as to how to } make these and which would be better for SEM? } } Barbara Plowman } Univ. of the Pacific } School of Dentistry } ph: 415-929-6692 } email: Bplowman-at-sf.uop.edu
it's interesting that figures between about 0.1 to 0.5% and 2 are 3% are quoted for detectability in EDX and this appears to reinforce the fact that you can only be sure when you try it.
Just to muddy the water a bit further I would be concerned if the chloride layer was much less that 1um thick because it could then potentially represent a smaller part of the interaction volume. I also understand that chlorine compounds can be a bit volatile in the beam and so some loss may occur.
But detectable levels can of course also be affected by other parameters such as the energy of the incident electrons, the amount of time of analysis, the relative composition of the matrix, whether the sample has to be coated and whether you believe the peak exists (and probably which way the wind's blowing).
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: Vlad Igoshev {vladig-at-tht.net}
Dear Pavel, Detection limit in the EDS is not a simple issue and it varies considerably from element to element. I usually tell my customers that it is 0.5 wt. %, but it is better than that for the K lines of the metals and worse for L and M lines. Any overlapping element lines will worsen the detection limit considerably, as will large ZAF correction factors. I have detected and quantified 0.2 wt.% Mn in an aluminum alloy. For Cl it should be similar, but it is difficult to say for sure without testing it on a standard of known chloride content. 0.2 wt % is 2000ppm. Depth of penetration is another matter and is best answered by using one of the freeware Monte Carlo simulation programs available. Try the MSA Web site. To see the depth of the diffusion of the chloride, you will probably have to cross-section the metal and look for Cl from the surface down into the bulk, bearing in mind the size of the x-ray volume from the Monte Carlo. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "AtcSEM" {atcsem-at-sbcglobal.net} To: {MIcroscopy-at-sparc5.microscopy.com} Sent: Tuesday, June 17, 2003 5:49 AM
Dear Colleagues Does anybody know what electrolyte to use for making TEM foils of TiNiPt alloys using jet polishing? TIA Anita
in addition to the suggestions, 'dark field' reflection modes may do the trick. Reflection Contrast Microscopy and Epipolarisation Microscopy, the latter using polarized light in epi-mode, used to detect lowl levels of silver in autoradiography and later on for detecting silver enhanced gold particles (from subnanometer to 10nm size as a particle to start of with before the enhancement). It is a very simple technique to use. I don't know exactly about the lower size limit for the particles, but we can figure that out fast.
Hope this helps, I would be interested to know what works best for you. Cheers.
================================ Jan Leunissen Aurion - http://www.aurion.nl present address: EM-Unit Dept. Anatomy and Struct Biology University of Otago PO Box 913 Dundedin, New Zealand
-- Jan Leunissen PhD Aurion Managing Director/Director R&D current address: EM-Unit Otago University Dunedin, New Zealand --
List, Any fluorescence microscope equipped with a epi-polarization cube (Chroma #33001 or a 50/50 beam splitter and two polarizers from Omega) should work quite well. Tom
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Wednesday, June 18, 2003 8:30 AM To: Microscopy-at-sparc5.microscopy.com
Venu,
You need diffraction optics of some sort. Differential Interference Contrast (aka Normarski, DIC), Asymmetric Illumination Contrast (AIC), possibly Hoffman Modulation Contrast, although I haven't tried that one. Phase Contrast *might* work, although again, I haven't tried it. The idea is that these sub-resolution particles can be detected, although they cannot be resolved (or imaged). The "inflated diffraction image" is reasonably easy to see.
Phil
} Email: venu-at-purdue.edu } Name: Venu } } Organization: Purdue University } } Education: Graduate College } } Location: City, State, Country } } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Pavel, Too often customers do not know what really do they need. Is this project a part of failure analysis? Than profiling with SIMS will be an overkill and a little bit too expensive.
} I appreciate all the answers I have received. } } I have looked on the cross section of a sample and ran EDS } and EDS line } scan, but could not see anything.
You do not need to do a line scan if spectra acquisition did not show a Cl peak. To get the best sensitivity possible with EDS you have to use the right geometry (WD and tilt angle) and highest possible current, so that dead time is about 30-50% with acquisition time 10 min (or more).
} I did suggest the SIMS since they are looking to see how deep } did the Cl } diffused. } } Now I am trying to setup Cl standard for WDS and check the } cross section, } but I do not think that I would be successful.
For the first step you do not need any standards. Just (again) use right geometry and high current and time for spectra acquisition. You will see whether you have a Cl peak.
} Does any one knows if defused chlorine would cause any } problems with 17-4PH, } especially if the customer removes 3 mils from the surface } that the parts } were exposed to Cl (?) containing ink marker?
I have not worked with exactly this steel, but I can assume that it could have the usual problems for this class of materials in chlorine environment - pitting and stress corrosion. Both these processes are surface initiated and I will not advice a customer from nonacademic organization to look for Cl diffusion profile.
Good luck,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} Pavel Lozovyy } ATC SEM Lab } Phone: 216-692-6637 } E-mail: atclabs-at-sbcglobal.net } web: www.atclabs.com } } } } ----- Original Message ----- } } From: "Bill Miller" {microbill-at-mohawk.net} } To: "AtcSEM" {atcsem-at-sbcglobal.net} } Sent: Wednesday, June 18, 2003 7:23 AM } Subject: Re: Fw: SEM detection limit of Cl } } } } sounds like a job for SIMS particularly if you want depth } profiles - low } Z } } detection in a high Z environment with EDX is tricky at best... } } } } At 08:49 AM 6/17/2003 -0400, you wrote: } } } } ------------------------------------------------------------- } ----------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } ------------------------------------------------------------- } ----------. } } } } } } } } } Dear listers, } } } Is there any way to quantify the lowest detection limit of SEM and } compare } } } it to ppm range? } } } } } } I have a customer who would like to find out if chlorine } from an ink } marker } } } diffused into the structure of 17-4 PH alloy? If it did, } how deep? The } } } customer is asking what is my detection limit for chlorine 100ppm? } 200ppm? } } } } } } Any advice is appreciated } } } } } } Pavel Lozovyy } } } ATC SEM Lab } } } Phone: 216-692-6637 } } } E-mail: atclabs-at-sbcglobal.net } } } web: www.atclabs.com } } } } } } }
Uranyl acetate at pH 2.5 or below should help, it gives less general contrast but will be more specific towards nucleic acids. 2% solution en bloc for 3 h or overnight in the fridge, after usual GA-Os fixation and before dehydration. Then on sections 5-10 minutes, *not longer*, I used the same 2%, but 5% low pH may work even better. I would try with and without lead staining, depending on the specimen. Maybe even without Os, depending again on the purpose of investigation. And, of course, this calls for very thin sections, 40 nm and under.
Hope this is your only trouble now :) Best wishes, Vlad -- ------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
Hi, everyone: An investigator here is looking at the changes in ribosomes with different chemical treatments. Can anyone recommend a fixation and/or staining procedure that will enhance the contrast of ribosomes at ultrastructural level? Thank you in advance and I look forward to hearing from you.
Hong Yi Emory School of Medicine EM Tel: (404) 727-8692 (Office), (404) 712-8491 (Lab) Fax: (404) 727-3157
Dear Pavel, Did you detect Cl in the ink? The problem with Cl in stainless steel is that it may cause corrosion. You would see this on the cross-section, where it will cause intergranular cracks. Stainless steels with Mo added are more resistant to chloride corrosion. If you don't see any sign of intergranular cracking, a small amount of diffused Cl probably won't cause problems. It is relatively easy to calculate the detection limit of an element in WDX. I use the formula: C(l) = C(St)x(3 x square root B/P-B) where C(l) is detection limit in mass%, C(St) is mass concentration of the element of interest in the standard, P is the total peak counts in the standard and B is the total background counts in the standard. I use a mounted and polished KCl or NaCl crystal as a standard. Once you run the standard you can calculate the detection limit. It is usually much lower than in EDS. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "AtcSEM" {atcsem-at-sbcglobal.net} To: "Bill Miller" {microbill-at-mohawk.net} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 18, 2003 4:51 AM
I would be very surprised if chlorine could diffuse into the stainless steel matrix. The diffusion coefficient for elements like carbon and nitrogen are extremely slow at temperatures below 1000F, and the chlorine atoms will be bigger and slower.
If the chlorine is present as a chloride ion, corrosion could occur. Then you may have pits that could penetrate into the stainless steel. Metallurgical changes due to corrosion by the chlorides should be readily visible in your cross sections.
Hope this helps.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
Hello Richard I am biological science guy, but it seems to me the grids with "holey film" covered with thin carbon film is sort of universal solution for high-resolution work. I think, major EM suppliers sells it. Personally, I am using my own home-made stuff. The biggest issue with hi-res work is stability and drift, so "holey film" should be permanently attached to the EM grid. My "holey film" is carbon "glued" to the 150 mesh copper Hex grid. The size of holes is about 1-2 mkm. The important things about "holey film" - the "bridges" between the holes should be uniform in thickness. It provides stability and eliminates the drift. Just before sample preparation I mount some carbon film over the grid. I usually use 1.2-1.8 nm thick carbon film evaporated on freshly cleaved mica by "electron gun" in the oil-free environment. Such films show extreme stability under the beam and they attached nicely to the carbon "holey film". You may not glow-discharge those films because of their thickness. From another hand, because those films produced in the very clean environment, I do find that they do work well in many applications where glow-discharge mandatory for ordinary films (DNA adsorption for instance). The advantage of thin film is that you have better noise-signal ratio and "electron gun" produced film is really stable. Have a good day, Sergey
At 08:29 PM 6/17/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
The EDS limit is around 2% and the accelerating voltage doesn't affect it. Of cause, if you are dealing with a layer, then you can play with the voltage and the tilt, but the limit will be the same. I had to deal with a similar problem but a 'better" element - Ag. Its content in the alloy is around 2 - 2.5% and the alloy contains Sn, which makes everything much worse. I knew that the EDS limit around 2-3% but asked EDAX for a "formal" backup and they came back with the same numbers.
Regards,
Vladimir Igoshev, PhD.
Toronto, Canada ----- Original Message ----- } From: "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} To: {atcsem-at-sbcglobal.net} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 18, 2003 6:01 AM
Dee,
Thanks for posting the summary of response on facility funding. I notice that no-one has explicitly mentioned depreciation. We have to bring enough from user fees to cover 1.75 full time salaries, running costs and depreciation. We don't see the depreciated funds again, they dissapear to the University Centre to be dispensed across the board. Each instrument is depreciated over ten years, so on top of fighting for funding for the purchase cost we have to find an additional 10% of the that cost for each year of the next decade. This situation makes it impossible to break even each year let alone imagine replacing some of our aging instruments. One could become rather jaded.
right the last sentence On Mercoledì, giu 18, 2003, at 15:29 Europe/Rome, Philip Oshel wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Venu, } } You need diffraction optics of some sort. Differential Interference } Contrast (aka Normarski, DIC), Asymmetric Illumination Contrast (AIC), } possibly Hoffman Modulation Contrast, although I haven't tried that } one. Phase Contrast *might* work, although again, I haven't tried it. } The idea is that these sub-resolution particles can be detected, } although they cannot be resolved (or imaged). The "inflated } diffraction image" is reasonably easy to see. } } Phil } } } Email: venu-at-purdue.edu } } Name: Venu } } } } Organization: Purdue University } } } } Education: Graduate College } } } } Location: City, State, Country } } } } Question: Could you please tell me the imaging requirements (tools } } needed) and the proper technique to observe 30-40 nm gold particles } } using light microscopy? } } } } ---------------------------------------------------------------------- } } ----- } } -- } Philip Oshel } Supervisor, BBPIC microscopy facility } Department of Animal Sciences } University of Wisconsin } 1675 Observatory Drive } Madison, WI 53706 - 1284 } voice: (608) 263-4162 } fax: (608) 262-5157 (dept. fax) } } } ....................................................................... ........................ Alberto Diaspro, Deptartment of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480/309 fax 010314218 e-mail: diaspro-at-fisica.unige.it URL: http://www.lambs.it ....................................................................... .....................
Is sulfur a bigger issue here? I'd be concerned about it more than chlorine. But that is just me. Stainless steel is differentiated by sulfur it seems to me, rather than chlorine. But of course, your SS may vary.
gary g.
} Date: Wed, 18 Jun 2003 17:25:24 -0500 } From: Larry Hanke {hanke-at-mee-inc.com} } Organization: Materials Evaluation and Engineering, Inc. } User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.0.2) } Gecko/20030208 Netscape/7.02 } X-Accept-Language: en-us, en } To: Microscopy-at-sparc5.microscopy.com } Cc: AtcSEM {atcsem-at-sbcglobal.net} } Subject: Re: Fw: SEM detection limit of Cl } X-Spam-Status: No, hits=0.7 required=5.0 } tests=NOSPAM_INC,OUTLOOK_FW_MSG,RCVD_IN_RFCI,REFERENCES, } SIGNATURE_SHORT_DENSE,SPAM_PHRASE_00_01,USER_AGENT, } USER_AGENT_MOZILLA_UA,X_ACCEPT_LANG } version=2.43 } X-Spam-Level: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I made a service referral to someone out west who is having an Amray 1200C installed and who is now asking if I have a source for the operator's manual and schematics for it. I don't currently have those in my collection (I try to keep paper and digitized copies of old SEM documents handy for those who need them).
So, I thought I'd pass the request on to the kind citizens here. Can anyone help?
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
I was not able to detect any chlorine on the cross section that I was analyzing. WDS showed higher background than the peak. Of coarse it does not mean that we do not have any chlorine diffusion. It is entirely possible that I am not looking at the right plane of polish or in slightly different location, but there is no way of knowing where the chlorine would like to diffuse. The 17-4 PH parts marked with an ink, that possibly contains chlorine (according to my customer), were heat treated and do not meet a MIL spec due to the markings. My customer is trying to make a case that the chlorine containing ink on the parts ( value ~ $30K) would not have any effect on the parts, especially that they machine couple mils off.
I think for them to have a better case, they would have to go to SIMS.
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
----- Original Message ----- } From: "Mary Mager" {mager-at-interchange.ubc.ca} To: "AtcSEM" {atcsem-at-sbcglobal.net} Cc: "Microscopy" {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 18, 2003 4:44 PM
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Hi Barbara,
Here is a simple method using cellulose acetate, possibly the kind of powder your "guy" had mentioned:
You add acetone (99%) to the cellulose acetate and stir quickly to make a finger nail polish thickness slurry. Then you simply paint it on the surface to be replicated and it will dry almost immediately. Then you peel it off and you have the replica - actually a "negative" of the surface.
You can then trim & mount that onto a SEM stub and sputter coat as usual.
We've done this here on plant leaf samples, for various reasons, sometimes to actually pick up fungal structures from the surface in order to look at them from underneath.
You could make a positive of the replica by coating it with a water based latex like bathroom caulking, then peel that off the replica. I've never tried it but what ever you do it must be water based material or you might dissolve or distort the acetate replica.
But most important in this case, before you start, don't forget to brush! - or not?
Good luck!
-- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Bera
} Hi Liststers, } I have a SEM project to do that involves making replicas of teeth. The } replicas I have made were made with acetate replicating tape. The guy who gave } me the teeth sez it is a kind of powder that one makes into a mixture that one } coats the teeth and hardens much like a latex peel. Any advice as to how to } make these and which would be better for SEM? } } Barbara Plowman } Univ. of the Pacific } School of Dentistry } ph: 415-929-6692 } email: Bplowman-at-sf.uop.edu
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Hi Pavel,
If you mean EDS analysis, then you can't get any lower then 2-3 %. ppm level isn't for EDS at all.
Regards,
Vladimir Igoshev, Ph.D. Toronto, Canada ----- Original Message ----- } From: "AtcSEM" {atcsem-at-sbcglobal.net} To: {MIcroscopy-at-sparc5.microscopy.com} Sent: Tuesday, June 17, 2003 8:49 AM
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Dear Pavel, Detection limit in the EDS is not a simple issue and it varies considerably from element to element. I usually tell my customers that it is 0.5 wt. %, but it is better than that for the K lines of the metals and worse for L and M lines. Any overlapping element lines will worsen the detection limit considerably, as will large ZAF correction factors. I have detected and quantified 0.2 wt.% Mn in an aluminum alloy. For Cl it should be similar, but it is difficult to say for sure without testing it on a standard of known chloride content. 0.2 wt % is 2000ppm. Depth of penetration is another matter and is best answered by using one of the freeware Monte Carlo simulation programs available. Try the MSA Web site. To see the depth of the diffusion of the chloride, you will probably have to cross-section the metal and look for Cl from the surface down into the bulk, bearing in mind the size of the x-ray volume from the Monte Carlo. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "AtcSEM" {atcsem-at-sbcglobal.net} To: {MIcroscopy-at-sparc5.microscopy.com} Sent: Tuesday, June 17, 2003 5:49 AM
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Thank you for the reply.
We are only interested to see if there is Cl diffusion or not and how deep (if possible). We do not need to quantify it.
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com ----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Ritchie Sims" {r.sims-at-auckland.ac.nz} ; "AtcSEM" {atcsem-at-sbcglobal.net} ; {MIcroscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 18, 2003 8:20 AM
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Ritchie writes ...
} I'd say about 0.1% ie 1,000 ppm in the absence of any } complicating factors, or 5000ppm with more confidence.
regarding ...
} } Is there any way to quantify the lowest detection limit of SEM and } } compare it to ppm range? } } } } I have a customer who would like to find out if chlorine from an ink } } marker diffused into the structure of 17-4 PH alloy? If it did, how } } deep? The customer is asking what is my detection limit for chlorine } } 100ppm? 200ppm?
I would think 1000ppm is a good approximation, assuming we are talking about SEM/EDX. However, I'd also comment ... a detection limit is difficult to evaluate for EDX, and indeed many EDX softwares do not even provide for it. The EDX background is relatively straight-forward with modern softwares, but the error associated with it is not ... and may also depend on parameters used for background modelling ... e.g., the absorption in the low energy end, any parameter used for the exponential high energy end, whether parameters are fixed or automatic (and an iterative function of composition), or if ROIs are used.
Not that EDX sensitivities are impossible, but rather impossible to address if a lot more is not known. It sounds like this "customer" wants not only quantitative wt%, but also wants the error quantified as well (my definition of "quantitative analysis"). This SEM operator, Mr. Pavel Lozovyy, should consult specifically with his EDX & software support.
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Guys,
The EDS limit is around 2% and the accelerating voltage doesn't affect it. Of cause, if you are dealing with a layer, then you can play with the voltage and the tilt, but the limit will be the same. I had to deal with a similar problem but a 'better" element - Ag. Its content in the alloy is around 2 - 2.5% and the alloy contains Sn, which makes everything much worse. I knew that the EDS limit around 2-3% but asked EDAX for a "formal" backup and they came back with the same numbers.
Regards,
Vladimir Igoshev, PhD.
Toronto, Canada ----- Original Message ----- } From: "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} To: {atcsem-at-sbcglobal.net} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 18, 2003 6:01 AM
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right the last sentence On Mercoledì, giu 18, 2003, at 15:29 Europe/Rome, Philip Oshel wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Venu, } } You need diffraction optics of some sort. Differential Interference } Contrast (aka Normarski, DIC), Asymmetric Illumination Contrast (AIC), } possibly Hoffman Modulation Contrast, although I haven't tried that } one. Phase Contrast *might* work, although again, I haven't tried it. } The idea is that these sub-resolution particles can be detected, } although they cannot be resolved (or imaged). The "inflated } diffraction image" is reasonably easy to see. } } Phil } } } Email: venu-at-purdue.edu } } Name: Venu } } } } Organization: Purdue University } } } } Education: Graduate College } } } } Location: City, State, Country } } } } Question: Could you please tell me the imaging requirements (tools } } needed) and the proper technique to observe 30-40 nm gold particles } } using light microscopy? } } } } ---------------------------------------------------------------------- } } ----- } } -- } Philip Oshel } Supervisor, BBPIC microscopy facility } Department of Animal Sciences } University of Wisconsin } 1675 Observatory Drive } Madison, WI 53706 - 1284 } voice: (608) 263-4162 } fax: (608) 262-5157 (dept. fax) } } } ..................................................................... ...................... Alberto Diaspro, Deptartment of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480/309 fax 010314218 e-mail: diaspro-at-fisica.unige.it URL: http://www.lambs.it ..................................................................... ...................
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Venu, Ritchie Any fluorescence microscope equipped with a epi-polarization cube (Chroma #33001 or a 50/50 beam splitter and two polarizers from Omega) should work quite well. Tom
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Tuesday, June 17, 2003 7:38 PM To: venu-at-purdue.edu; MicroscopyListserver
} } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? }
I don't think you have any chance with LM, but I'll be interested to hear if there is a way.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
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} } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? }
I don't think you have any chance with LM, but I'll be interested to hear if there is a way.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
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Hi,
in addition to the suggestions, 'dark field' reflection modes may do the trick. Reflection Contrast Microscopy and Epipolarisation Microscopy, the latter using polarized light in epi-mode, used to detect lowl levels of silver in autoradiography and later on for detecting silver enhanced gold particles (from subnanometer to 10nm size as a particle to start of with before the enhancement). It is a very simple technique to use. I don't know exactly about the lower size limit for the particles, but we can figure that out fast.
Hope this helps, I would be interested to know what works best for you. Cheers.
================================ Jan Leunissen Aurion - http://www.aurion.nl present address: EM-Unit Dept. Anatomy and Struct Biology University of Otago PO Box 913 Dundedin, New Zealand
-- Jan Leunissen PhD Aurion Managing Director/Director R&D current address: EM-Unit Otago University Dunedin, New Zealand --
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I appreciate all the answers I have received.
I have looked on the cross section of a sample and ran EDS and EDS line scan, but could not see anything. I did suggest the SIMS since they are looking to see how deep did the Cl diffused. Now I am trying to setup Cl standard for WDS and check the cross section, but I do not think that I would be successful.
Does any one knows if defused chlorine would cause any problems with 17-4PH, especially if the customer removes 3 mils from the surface that the parts were exposed to Cl (?) containing ink marker?
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
----- Original Message ----- } From: "Bill Miller" {microbill-at-mohawk.net} To: "AtcSEM" {atcsem-at-sbcglobal.net} Sent: Wednesday, June 18, 2003 7:23 AM
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I have several EDAX and 2 Oxford systems, and like Oxford the better.
**************************************** Chaoying Ni, PhD The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
On Tue, 17 Jun 2003, Faerber Jacques wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi All } } After waiting for a long time, we must now choose in hurry a new EDS } system for our TEM. Our old Kevex died ((un)fortunately ?!) and after a } few cycle monney-no monney-monney-no monney etc, we are again in a } "monney" phase, and we must spend it fast ! So fast that we have no time } to do test. } } So any advises are wanted about the Noran Six, the PGT Avalon, the Oxford } INCA and the EDAX Genesis EDS systems, to fit on a 200 keV TOPCON 02B TEM, } and to be used in a material research lab, on magnetic materials, } catalysts, bio-materials, etc. We know a little bit the PGT and the } Oxford, I use personnaly the Noran Vantage, but the Six is new (only the } GUI or the electronics too ?), and we know nothing (other than the } manufacturer's doc) about the EDAX Genesis. } } Any advise too about reparing/upgrading our old KEVEX detector, or buying } a new one (much more monney). The old one worked fine before it died, and } it is bellow seald, with the length and the transfer mechanism adapted to } the TEM. } } Answer please on the list, direct to me or to } Jacques.Werckmann-at-ipcms.u-strasbg.fr } } Many thanks } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } }
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I would be very surprised if chlorine could diffuse into the stainless steel matrix. The diffusion coefficient for elements like carbon and nitrogen are extremely slow at temperatures below 1000F, and the chlorine atoms will be bigger and slower.
If the chlorine is present as a chloride ion, corrosion could occur. Then you may have pits that could penetrate into the stainless steel. Metallurgical changes due to corrosion by the chlorides should be readily visible in your cross sections.
Hope this helps.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
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I'd say about 0.1% ie 1,000 ppm in the absence of any complicating factors, or 5000ppm with more confidence.
cheers
rtch
} From: "AtcSEM" {atcsem-at-sbcglobal.net} To: {MIcroscopy-at-sparc5.microscopy.com}
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Pavel;
I agree with Ricth Sims about the lower limit of detection. However, there are some caveats you should be aware of such as excitation volume/interaction volume and the matrix of elements the Cl is in. The software program "Electron Flight Simulator" by Small World Systems, Inc. can give you an approximation of just how deep the interaction volume is for a given accelerating voltage. Also, you should know if any of the other elements in the matrix will absorb and of the Cl photon emissions [absorption edges].
If you send me the matrix off-line I'd be happy run the simulation for you.
If you are after a depth profile, it would be best to user AES [Auger Spectroscopy] or SIMS [Scanning Ion Mass Spectroscopy] SIMS would have the best sensitivity.
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Tuesday, June 17, 2003 8:50 AM To: MIcroscopy-at-sparc5.microscopy.com
Dear listers, Is there any way to quantify the lowest detection limit of SEM and compare it to ppm range?
I have a customer who would like to find out if chlorine from an ink marker diffused into the structure of 17-4 PH alloy? If it did, how deep? The customer is asking what is my detection limit for chlorine 100ppm? 200ppm?
Any advice is appreciated
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
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Pavel, Too often customers do not know what really do they need. Is this project a part of failure analysis? Than profiling with SIMS will be an overkill and a little bit too expensive.
} I appreciate all the answers I have received. } } I have looked on the cross section of a sample and ran EDS } and EDS line } scan, but could not see anything.
You do not need to do a line scan if spectra acquisition did not show a Cl peak. To get the best sensitivity possible with EDS you have to use the right geometry (WD and tilt angle) and highest possible current, so that dead time is about 30-50% with acquisition time 10 min (or more).
} I did suggest the SIMS since they are looking to see how deep } did the Cl } diffused. } } Now I am trying to setup Cl standard for WDS and check the } cross section, } but I do not think that I would be successful.
For the first step you do not need any standards. Just (again) use right geometry and high current and time for spectra acquisition. You will see whether you have a Cl peak.
} Does any one knows if defused chlorine would cause any } problems with 17-4PH, } especially if the customer removes 3 mils from the surface } that the parts } were exposed to Cl (?) containing ink marker?
I have not worked with exactly this steel, but I can assume that it could have the usual problems for this class of materials in chlorine environment - pitting and stress corrosion. Both these processes are surface initiated and I will not advice a customer from nonacademic organization to look for Cl diffusion profile.
Good luck,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} Pavel Lozovyy } ATC SEM Lab } Phone: 216-692-6637 } E-mail: atclabs-at-sbcglobal.net } web: www.atclabs.com } } } } ----- Original Message ----- } } From: "Bill Miller" {microbill-at-mohawk.net} } To: "AtcSEM" {atcsem-at-sbcglobal.net} } Sent: Wednesday, June 18, 2003 7:23 AM } Subject: Re: Fw: SEM detection limit of Cl } } } } sounds like a job for SIMS particularly if you want depth } profiles - low } Z } } detection in a high Z environment with EDX is tricky at best... } } } } At 08:49 AM 6/17/2003 -0400, you wrote: } } } } ------------------------------------------------------------- } ----------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } ------------------------------------------------------------- } ----------. } } } } } } } } } Dear listers, } } } Is there any way to quantify the lowest detection limit of SEM and } compare } } } it to ppm range? } } } } } } I have a customer who would like to find out if chlorine } from an ink } marker } } } diffused into the structure of 17-4 PH alloy? If it did, } how deep? The } } } customer is asking what is my detection limit for chlorine 100ppm? } 200ppm? } } } } } } Any advice is appreciated } } } } } } Pavel Lozovyy } } } ATC SEM Lab } } } Phone: 216-692-6637 } } } E-mail: atclabs-at-sbcglobal.net } } } web: www.atclabs.com } } } } } } }
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I am a bit lost.
If you say SEM do you mean imaging with SE or BSE , EDS or WDS? If it is EDS I would say 0.5 atomic percent in a metal matrix at 25kV. Since it is a light element 15 kV would be sufficient to "see" all your elements in your alloy as well as Cl. The limit would be different since your interaction volume is different therefore determining the distance your EDS can see Cl (assuming you look at a cross section of the area of interest eg a fracture or a crack. Are the Cl real or from greasy hands handling the sample even the atmosphere close to the sea? Are you using theoretical standards, pure elements or a "real" 17-4 PH alloy standard that was verified by other more accurate means eg. a probe? All these factors do influence your detection limit and accuracy. The distance detectable in a trace scan is limited by your interaction volume thus density and kV combination. Remember that Cl is often driven away by the beam and the analysis should preferably not be carried out in spot mode and on a fresh area defeating the purpose of using a line or spot trace.
As You can see, this is not a simple question with a simple answer.
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, June 18, 2003 1:47 AM To: AtcSEM; MIcroscopy-at-sparc5.microscopy.com
I'd say about 0.1% ie 1,000 ppm in the absence of any complicating factors, or 5000ppm with more confidence.
cheers
rtch
} From: "AtcSEM" {atcsem-at-sbcglobal.net} To: {MIcroscopy-at-sparc5.microscopy.com}
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Dear Pavel, Did you detect Cl in the ink? The problem with Cl in stainless steel is that it may cause corrosion. You would see this on the cross-section, where it will cause intergranular cracks. Stainless steels with Mo added are more resistant to chloride corrosion. If you don't see any sign of intergranular cracking, a small amount of diffused Cl probably won't cause problems. It is relatively easy to calculate the detection limit of an element in WDX. I use the formula: C(l) = C(St)x(3 x square root B/P-B) where C(l) is detection limit in mass%, C(St) is mass concentration of the element of interest in the standard, P is the total peak counts in the standard and B is the total background counts in the standard. I use a mounted and polished KCl or NaCl crystal as a standard. Once you run the standard you can calculate the detection limit. It is usually much lower than in EDS. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "AtcSEM" {atcsem-at-sbcglobal.net} To: "Bill Miller" {microbill-at-mohawk.net} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 18, 2003 4:51 AM
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Dee,
Thanks for posting the summary of response on facility funding. I notice that no-one has explicitly mentioned depreciation. We have to bring enough from user fees to cover 1.75 full time salaries, running costs and depreciation. We don't see the depreciated funds again, they dissapear to the University Centre to be dispensed across the board. Each instrument is depreciated over ten years, so on top of fighting for funding for the purchase cost we have to find an additional 10% of the that cost for each year of the next decade. This situation makes it impossible to break even each year let alone imagine replacing some of our aging instruments. One could become rather jaded.
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To all TEM facilities:
I need someone to help me with plant viruses/TEM research. I do have TEM background but do not have an electron microscope(none in Alaska). I need to look at purified viruses and viruses in plant tissue. I am looking for a EM facility that either contracts research projects or allows one to use their facility for a short segment of time. My immediate need is to have grids of purified virus or leaf dips looked at for virus particles.
Thank you,
Nancy Robertson
Nancy L. Robertson Research Plant Pathologist USDA, ARS 533 E. Fireweed Ave. Palmer Alaska 99645 907-746-9465 office 907-746-2898 lab 907-746-2677 fax ffnlr-at-uaf.edu
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Hi Hong,
Uranyl acetate at pH 2.5 or below should help, it gives less general contrast but will be more specific towards nucleic acids. 2% solution en bloc for 3 h or overnight in the fridge, after usual GA-Os fixation and before dehydration. Then on sections 5-10 minutes, *not longer*, I used the same 2%, but 5% low pH may work even better. I would try with and without lead staining, depending on the specimen. Maybe even without Os, depending again on the purpose of investigation. And, of course, this calls for very thin sections, 40 nm and under.
Hope this is your only trouble now :) Best wishes, Vlad -- ------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
Hi, everyone: An investigator here is looking at the changes in ribosomes with different chemical treatments. Can anyone recommend a fixation and/or staining procedure that will enhance the contrast of ribosomes at ultrastructural level? Thank you in advance and I look forward to hearing from you.
Hong Yi Emory School of Medicine EM Tel: (404) 727-8692 (Office), (404) 712-8491 (Lab) Fax: (404) 727-3157
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Is sulfur a bigger issue here? I'd be concerned about it more than chlorine. But that is just me. Stainless steel is differentiated by sulfur it seems to me, rather than chlorine. But of course, your SS may vary.
gary g.
} Date: Wed, 18 Jun 2003 17:25:24 -0500 } From: Larry Hanke {hanke-at-mee-inc.com} } Organization: Materials Evaluation and Engineering, Inc. } User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.0.2) } Gecko/20030208 Netscape/7.02 } X-Accept-Language: en-us, en } To: Microscopy-at-sparc5.microscopy.com } Cc: AtcSEM {atcsem-at-sbcglobal.net} } Subject: Re: Fw: SEM detection limit of Cl } X-Spam-Status: No, hits=0.7 required=5.0 } tests=NOSPAM_INC,OUTLOOK_FW_MSG,RCVD_IN_RFCI,REFERENCES, } SIGNATURE_SHORT_DENSE,SPAM_PHRASE_00_01,USER_AGENT, } USER_AGENT_MOZILLA_UA,X_ACCEPT_LANG } version=2.43 } X-Spam-Level: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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} } I'd say about 0.1% ie 1,000 ppm in the absence of any } complicating factors, or } 5000ppm with more confidence. } } cheers } } rtch
I agree with this estimate, but nevertheless it is worth a try to examine the specimens with EDS. I suspect that a customer has a pitting corrosion of a stainless steel (Cl is a usual cause of a pitting). In that case Cl in the pits could (but not must) be detectable.
Vladimir
} } } } From: "AtcSEM" {atcsem-at-sbcglobal.net} } To: {MIcroscopy-at-sparc5.microscopy.com} } Subject: Fw: SEM detection limit of Cl } Date sent: Tue, 17 Jun 2003 08:49:33 -0400 } } } } ---------------------------------------------------------------------- } } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } } -. } } } } } } Dear listers, } } Is there any way to quantify the lowest detection limit of SEM and } } compare it to ppm range? } } } } I have a customer who would like to find out if chlorine from an ink } } marker diffused into the structure of 17-4 PH alloy? If it did, how } } deep? The customer is asking what is my detection limit for chlorine } } 100ppm? 200ppm? } } } } Any advice is appreciated } } } } Pavel Lozovyy } } ATC SEM Lab } } Phone: 216-692-6637 } } E-mail: atclabs-at-sbcglobal.net } } web: www.atclabs.com } } } } } } -- } Ritchie Sims Ph D Phone : 64 9 } 3737599 ext 87713 } Microanalyst Fax : } 64 9 3737435 } Department of Geology email : } r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
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Dear listeners,
I have a question regarding grids for TEM studies of nanoparticles (ferrites) with sizes from 40 - 100 nm. Can anyone help me with what kind of grids/films I can use to study my particles ? or if anyone knows if there is any good literature that I should take a look at? The particles are dispersed in ethanol and then a droplet of the ethanol with particles will be put placed on the grid/film. However, if there are other better methods, I am very interested.
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List, Any fluorescence microscope equipped with a epi-polarization cube (Chroma #33001 or a 50/50 beam splitter and two polarizers from Omega) should work quite well. Tom
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Wednesday, June 18, 2003 8:30 AM To: Microscopy-at-sparc5.microscopy.com
Venu,
You need diffraction optics of some sort. Differential Interference Contrast (aka Normarski, DIC), Asymmetric Illumination Contrast (AIC), possibly Hoffman Modulation Contrast, although I haven't tried that one. Phase Contrast *might* work, although again, I haven't tried it. The idea is that these sub-resolution particles can be detected, although they cannot be resolved (or imaged). The "inflated diffraction image" is reasonably easy to see.
Phil
} Email: venu-at-purdue.edu } Name: Venu } } Organization: Purdue University } } Education: Graduate College } } Location: City, State, Country } } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
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Hi Barbara,
Here is a simple method using cellulose acetate, possibly the kind of powder your "guy" had mentioned:
You add acetone (99%) to the cellulose acetate and stir quickly to make a finger nail polish thickness slurry. Then you simply paint it on the surface to be replicated and it will dry almost immediately. Then you peel it off and you have the replica - actually a "negative" of the surface.
You can then trim & mount that onto a SEM stub and sputter coat as usual.
We've done this here on plant leaf samples, for various reasons, sometimes to actually pick up fungal structures from the surface in order to look at them from underneath.
You could make a positive of the replica by coating it with a water based latex like bathroom caulking, then peel that off the replica. I've never tried it but what ever you do it must be water based material or you might dissolve or distort the acetate replica.
But most important in this case, before you start, don't forget to brush! - or not?
Good luck!
-- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Bera
} Hi Liststers, } I have a SEM project to do that involves making replicas of teeth. The } replicas I have made were made with acetate replicating tape. The guy who gave } me the teeth sez it is a kind of powder that one makes into a mixture that one } coats the teeth and hardens much like a latex peel. Any advice as to how to } make these and which would be better for SEM? } } Barbara Plowman } Univ. of the Pacific } School of Dentistry } ph: 415-929-6692 } email: Bplowman-at-sf.uop.edu
You might also try "Lyman, Roger" {RLyman-at-FEICO.com} I haven't tried to contact anyone for some months, or anyone at all except Joe Race since the last round of buyouts (by Veeco [or whoever it was], if that's still the last round of buyouts), but I assume they're still there.
Phil
} Hi, } } I want order a LaB6 cathode from FEI Beam Tech and cannot get } through to a person. Their automated phone system requires that you } know who you want to talk to, no operator!! Duh! } } Does anyone know a contact person there? } } Owen } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
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Hello
it's interesting that figures between about 0.1 to 0.5% and 2 are 3% are quoted for detectability in EDX and this appears to reinforce the fact that you can only be sure when you try it.
Just to muddy the water a bit further I would be concerned if the chloride layer was much less that 1um thick because it could then potentially represent a smaller part of the interaction volume. I also understand that chlorine compounds can be a bit volatile in the beam and so some loss may occur.
But detectable levels can of course also be affected by other parameters such as the energy of the incident electrons, the amount of time of analysis, the relative composition of the matrix, whether the sample has to be coated and whether you believe the peak exists (and probably which way the wind's blowing).
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: Vlad Igoshev {vladig-at-tht.net}
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Phil and Venu,
I dont see hwo the differential interface contrast can work beyond diffraction limit ?
I will say only TEM or AFM can be used to succesfully image this small particle.
Just to detect them (like in the case of single molecular spectroscopy/imaging) a confocal imaging may work well, but you do not image them, just you see brighter spot, on the size of the half of the wavelenght.
Best regards,
Mickey
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Wednesday, June 18, 2003 6:30 AM To: Microscopy-at-sparc5.microscopy.com
Venu,
You need diffraction optics of some sort. Differential Interference Contrast (aka Normarski, DIC), Asymmetric Illumination Contrast (AIC), possibly Hoffman Modulation Contrast, although I haven't tried that one. Phase Contrast *might* work, although again, I haven't tried it. The idea is that these sub-resolution particles can be detected, although they cannot be resolved (or imaged). The "inflated diffraction image" is reasonably easy to see.
Phil
} Email: venu-at-purdue.edu } Name: Venu } } Organization: Purdue University } } Education: Graduate College } } Location: City, State, Country } } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
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Dear Yi Ribosome (RS) is about 28 nm in diameter and wery fagile. You may not see the structure of RS on the conventional ultrathin sections with standard processing. You also could not see fine structure on the chemically fixed ribosomes. The only one way known to me to investigate the fine structure of the free ribosomes in vitro is cryo-EM with 3D-reconstruction. Please, made search for Joachim Frank works. Ribosome is extremely sensitive and complicated "machine": you have to perform state of the art biochemistry to isolate them intact. Usually we are using RNAase-free strains or thermophilic bacteria to isolate ribosomes. As far as I know, there is just a few places in US, where people is able to isolate intact ribosomes. Of coarse, "intact" is very subjective word. You may check Harry Noller works for RS isolation. Best wishes, Sergey
At 12:07 PM 6/17/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
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Venu,
You need diffraction optics of some sort. Differential Interference Contrast (aka Normarski, DIC), Asymmetric Illumination Contrast (AIC), possibly Hoffman Modulation Contrast, although I haven't tried that one. Phase Contrast *might* work, although again, I haven't tried it. The idea is that these sub-resolution particles can be detected, although they cannot be resolved (or imaged). The "inflated diffraction image" is reasonably easy to see.
Phil
} Email: venu-at-purdue.edu } Name: Venu } } Organization: Purdue University } } Education: Graduate College } } Location: City, State, Country } } Question: Could you please tell me the imaging requirements (tools } needed) and the proper technique to observe 30-40 nm gold particles } using light microscopy? } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
It depends on what you mean by "can work". It can't image the particles, or resolve 2 particles close together, but the way the particles diffract light does make them detectable. They appear to be larger than they really are.
Phil
} Phil and Venu, } } I dont see hwo the differential interface contrast can work beyond } diffraction limit ? } } I will say only TEM or AFM can be used to succesfully image this } small particle. } } Just to detect them (like in the case of single molecular } spectroscopy/imaging) a confocal imaging may work well, but you do } not image them, just you see brighter spot, on the size of the half } of the wavelenght. } } Best regards, } } Mickey } } -----Original Message----- } } From: Philip Oshel [mailto:peoshel-at-wisc.edu] } Sent: Wednesday, June 18, 2003 6:30 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Ask-A-Microscopist:LM 30 nm gold } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In response to the request for information about procedures for operating a turbo pump on a JEOL 845 microscope. If you can't obtain explicit instructiuons for this particular instrument, you might refer to Section 6.18 in my book, 'Vacuum Methods in Electron Microscopy' (see:http://www.2spi.com/catalog/books/book48.html/) for a description), where general operating procedures for instruments with turbo pumps are discussed. This discussion should give you the necessary information to devise the necessary procedures for yopur instrument. Even if you do obtain such instructions, Chapter 6 contains a rather detailed description of the construction and operating characteristics of turbomolecular pumps that should help you in understanding the performance of your instrument.
Good luck , WCB
-- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
That rather sounds like double-dipping. If you received an instrumentation grant from outside the university to purchase the equipment, it seems that depreciation should not be an issue. However, if you are fighting internally for part of a university-wide pot of money, then I could see how the university might insist on the depreciation. It seems that they never really gave you the money for the instrument, they are letting you "buy" it according to the depreciation schedule. That way, they can replenish the pot for the next pieces of instrumentation.
Is that a fair assessment? If not, please clarify.
Warren
At 01:27 PM 6/19/2003 +1200, you wrote:
} Dee, } } Thanks for posting the summary of response on facility funding. I } notice that no-one has explicitly mentioned depreciation. We have to } bring enough from user fees to cover 1.75 full time salaries, running } costs and depreciation. We don't see the depreciated funds again, they } dissapear to the University Centre to be dispensed across the board. } Each instrument is depreciated over ten years, so on top of fighting } for funding for the purchase cost we have to find an additional 10% of } the that cost for each year of the next decade. This situation makes it } impossible to break even each year let alone imagine replacing some of } our aging instruments. } One could become rather jaded. } } Regards } Bryony
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Scanning electron microscopy, x-ray analysis, and image analysis of materials Computer applications and networking
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That rather sounds like double-dipping. If you received an instrumentation grant from outside the university to purchase the equipment, it seems that depreciation should not be an issue. However, if you are fighting internally for part of a university-wide pot of money, then I could see how the university might insist on the depreciation. It seems that they never really gave you the money for the instrument, they are letting you "buy" it according to the depreciation schedule. That way, they can replenish the pot for the next pieces of instrumentation.
Is that a fair assessment? If not, please clarify.
Warren
At 01:27 PM 6/19/2003 +1200, you wrote:
} Dee, } } Thanks for posting the summary of response on facility funding. I } notice that no-one has explicitly mentioned depreciation. We have to } bring enough from user fees to cover 1.75 full time salaries, running } costs and depreciation. We don't see the depreciated funds again, they } dissapear to the University Centre to be dispensed across the board. } Each instrument is depreciated over ten years, so on top of fighting } for funding for the purchase cost we have to find an additional 10% of } the that cost for each year of the next decade. This situation makes it } impossible to break even each year let alone imagine replacing some of } our aging instruments. } One could become rather jaded. } } Regards } Bryony
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Scanning electron microscopy, x-ray analysis, and image analysis of materials Computer applications and networking
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Thanks for the reply,
my material is a glass containing nano-size ferroelectric crystallites 100- 200 nm. I want to make sure that these nano-crystallites are ferroelectic. I read your reply to use PFM and i have some questions after reading some papers on PFM.
QUE1- What is the minimum d33 coefficient to get a recognizable contrast because i read that the piezoresponse depends upon the d33 coefficient? the d33 coefficient of the single crystals same as nanocrystallites in my material is 1.3 pm/V, which is very small compared to BaTiO3.
Question2- In response to Dr Sergei's letter (below), He said that since crystallites are inside the dielectric medium this technique might not work. My question is-- in this technique, should upper and lower both srfaces of the crystallites should touch conductive surface?
QUE2- Is there any book describing this technique?
Thanks Pradyumna
Quoting "Sergei V. Kalinin" {sergei2-at-ornl.gov} :
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Pradyumna } The best way to detect and image ferroelectric inclusions in } paraelectric matrix is to use a scanning probe microscopy techniques, } namely piezoresponse force microscopy. This technique is specifically } designed to distinguish piezoelectric and non-piezoelectric materials, } and hence is probably the best choice in this case. It will be } sensitive only to crystallites exposed on the surface or relatively } shallow within the glass matrix (~10 nm) and will allow imaging of both } in-plane and out-of-plane polarization components. You can find some } examples in paper } A.Y. Borisevich, S.V. Kalinin, D.A. Bonnell, and P.K. Davies, } Analysis of phase distributions in the Li2O-Nb2O5-TiO2 system by } piezoresponse imaging, J. Mater. Res. 16(2), 329-332 (2001). } The extension of PFM, piezoresponse spectroscopy normally allows } spatially resolved electromechanical hysteresis loop measurements, e.g. } within single 50 nm grain, but it might not work for the crystal in the } dielectric matrix. } Yours } Sergei }
Hi, } } I have a glass- nono-crystallites composite. Nanocrystallites may be } ferroelectric in nature and I need to prove that nanocrystallites in the } glassy phase are ferroelectric. } } How can i prove using TEM that these nanocrystallites(probably single domain) } are indeed ferroelectric?
} -- } Sergei V. Kalinin, } Oak Ridge National Laboratory, } 1 Bethel Valley Rd, } Bldg. 3025, MS6030, } Oak Ridge, TN 37831 } Phone: (865) 241-0236 } FAX: (865) 574-4143 } URL: sergei2.kalininweb.com } } }
610 7585590 Lab Pradyumna N Gupta MSE Lehigh University
610 7585590 Lab Pradyumna N Gupta MSE Lehigh University
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} } Guys, } } The EDS limit is around 2% and the accelerating voltage
OK. To get real numbers from real system I just performed test on XL30 with field emission gun at 15 kV, 40-45% dead time, 10 min acquisition time. I measured Cl peak from Au/Pd coated crystals of table salt and background in Cl peak position on a stainless steel razor blade. With ZAF correction detection limit was 0.09%. If Cl could diffuse in steel it would be very convenient element for trace analysis.
} doesn't affect it.
It does affect. It changes background, peak, correction coefficients.
} Of cause, if you are dealing with a layer, then you can play with the } voltage and the tilt, but the limit will be the same. I had } to deal with a } similar problem but a 'better" element - Ag. Its content in } the alloy is } around 2 - 2.5% and the alloy contains Sn, which makes everything much } worse. I knew that the EDS limit around 2-3% but asked EDAX } for a "formal" } backup and they came back with the same numbers.
I am very surprised. But did you asked service technician or application lab?
Vladimir Dusevich
} Regards, } } Vladimir Igoshev, PhD. } } Toronto, Canada } ----- Original Message ----- } } From: "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} } To: {atcsem-at-sbcglobal.net} } Cc: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, June 18, 2003 6:01 AM } Subject: RE: SEM detection limit of Cl } } } } } -------------------------------------------------------------- } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } } } } I am a bit lost. } } } } If you say SEM do you mean imaging with SE or BSE , EDS or WDS? } } If it is EDS I would say 0.5 atomic percent in a metal } matrix at 25kV. } Since it is a light element 15 kV would be sufficient to } "see" all your } elements in your alloy as well as Cl. The limit would be } different since } your interaction volume is different therefore determining } the distance your } EDS can see Cl (assuming you look at a cross section of the } area of interest } eg a fracture or a crack. Are the Cl real or from greasy } hands handling the } sample even the atmosphere close to the sea? } } Are you using theoretical standards, pure elements or a } "real" 17-4 PH } alloy standard that was verified by other more accurate means } eg. a probe? } All these factors do influence your detection limit and accuracy. The } distance detectable in a trace scan is limited by your } interaction volume } thus density and kV combination. } } Remember that Cl is often driven away by the beam and the } analysis should } preferably not be carried out in spot mode and on a fresh } area defeating the } purpose of using a line or spot trace. } } } } As You can see, this is not a simple question with a simple answer. } } } } CHEMICAL COMPOSITION of an typical 17-4 PH alloy } } } } CARBON. 0.07% MAX. } } CHROMIUM. 15.O - 17.5%. } } MANGANESE. 1.00% } } MAX. NICKEL. 3.00- 5.00%. } } PHOSPHORUS. 0.04% MAX. } } COPPER. 3.00- 5.00%. } } SULPHUR. 0.03% MAX. } } } } -----Original Message----- } } } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] } } Sent: Wednesday, June 18, 2003 1:47 AM } } To: AtcSEM; MIcroscopy-at-sparc5.microscopy.com } } Subject: Re: Fw: SEM detection limit of Cl } } } } } } } -------------------------------------------------------------- } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } } } } } } I'd say about 0.1% ie 1,000 ppm in the absence of any complicating } factors, or } } 5000ppm with more confidence. } } } } cheers } } } } rtch } } } } } } } } } From: "AtcSEM" {atcsem-at-sbcglobal.net} } } To: {MIcroscopy-at-sparc5.microscopy.com} } } Subject: Fw: SEM detection limit of Cl } } Date sent: Tue, 17 Jun 2003 08:49:33 -0400 } } } } } } ---------------------------------------------------------------------- } } } -- The Microscopy ListServer -- Sponsor: The Microscopy } Society of } } } America To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } ---------------------------------------------------------------------- } } } -. } } } } } } } } } Dear listers, } } } Is there any way to quantify the lowest detection limit of SEM and } } } compare it to ppm range? } } } } } } I have a customer who would like to find out if chlorine } from an ink } } } marker diffused into the structure of 17-4 PH alloy? If } it did, how } } } deep? The customer is asking what is my detection limit } for chlorine } } } 100ppm? 200ppm? } } } } } } Any advice is appreciated } } } } } } Pavel Lozovyy } } } ATC SEM Lab } } } Phone: 216-692-6637 } } } E-mail: atclabs-at-sbcglobal.net } } } web: www.atclabs.com } } } } } } } } } } -- } } Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 } } Microanalyst Fax : 64 9 3737435 } } Department of Geology email : r.sims-at-auckland.ac.nz } } The University of Auckland } } Private Bag 92019 } } Auckland } } New Zealand } } } } } } }
We have a SEM Phillips XL30. All the images taken at extra high resolution (3232 x 2420 pixels) come with faint vertical bands spaced 11 pixels. The bands are about 4 pixels wide, and they are apparent by having a greater variance in gray levels (i.e., they look more grainy).
Does anyone have an idea of why this occur or how to solve it?
Martin
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
A DAKO Autostainer and a Shandon Consul Automatic Coverslipper are available for a reasonable price. These automated systems were purchased 3 years ago and have been maintained under service contract through the manufacturer. Since the equipment was primarily designed for labs routinely with large volume of histology and immunohistochemistry/cytochemistry work, they have never been heavily used in our EM research lab. In another word, they are almost new and in a very good condition. The EM Facility will provide manual and maintenance records, and training if required.
Please contact Jocelyn Torcolini in the Electron Microscopy Facility at Penn State (814-865-0212 or jmt175-at-psu.edu) for more information.
Gang (Greg) Ning Director, Electron Microscopy Facility Huck Institute for Life Sciences The Pennsylvania State University 1 South Frear Lab University Park, PA 16802 Phone: 814-863-0994 Fax: 814-863-1357 Email: gxn7-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Well, here are another numbers. I collected an EDS spectrum from 1010 steel (Jeol/Kevex/tungsten, 20 kV, 30% dead time, 2 minutes), forced the software to account for Cl and Ar peaks and ran quantification. What I've got was 0.11at%Cl and 0.26at%Ar. I like the "trick" and use it once in a while to demonstrate my customers that if you don't see a peak then the EDS isn't the right technique for the analysis and some other methods should be used (like SIMS), particularly for light elements. Of cause, if you know the element is there then it's easier to "believe" the numbers.
Regards,
Vladimir ----- Original Message ----- } From: "Dusevich, Vladimir" {dusevichv-at-umkc.edu} To: "Vlad Igoshev" {vladig-at-tht.net} ; {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, June 19, 2003 5:39 PM
Did you try your question on the XL30 list?
gary g.
At 06:19 AM 6/20/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I could use a little help with overlaying images. On our Zeiss LSM-310, we can scan a reflected laser image of a sample (computer chip, in our case) into the Green plane, and then maybe an OBIC image into the Red plane, and then select the RGB mode. This will display our OBIC image overlaid on the reflected light image, showing where the OBIC signal is coming from on the sample.
Now the problem. We have a Roper Scientific CCD mounted on the Zeiss, which we use for reflected light images, and for capturing images of the light emitted from the computer chip while it is powered up (photo emission microscopy - PEM). We are using the WinView32 software that came with the CCD, and I have tried all of the math functions, etc., that I can find in WinView32. I have been unable to do an "overlay" of two images.
Does anyone know how to do a simple overlay with WinView32, or if there is another application that will work? (Hopefully cheap or free) I don't need a lot of image processing/analysis power. Just the ability to overlay. WinView32 can do pseudo coloring of the images to emulate the Red and Green planes of the Zeiss.
Thanks for your attention, Darrell IBM Microelectronics
I don't know WinView32 at all, but Adobe Photoshop has a variety of techniques for merging and overlaying images either using Layers with effects such as muliplication, difference or exclusion, or by pasting images into separate RGB or CMYK colour channels.
Dr. Chris Jeffree Inveresk Cottage 26, Carberry Road Inveresk Musselburgh Midlothian EH21 8PR Tel: +44 131 665 6062 FAX +44 131 653 6248 Mobile 07710 585 401 ----- Original Message ----- } From: "Darrell Miles" {milesd-at-US.ibm.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, June 21, 2003 1:54 AM
You can either use a conventional photo program such as photoshop, or look into ImageJ, a pc port of NIH-Image (http://rsb.info.nih.gov/ij/). Both of these can do overlays. If you have to adjust the sizes to match, both programs do have ways of doing that, as well.
If you need specifics, feel free to contact me off list. Joel
You should be able to do it with ImagJ http://rsbweb.nih.gov/ij/ It is pubic domain so the price is right. It is written in Java and runs on virtually any computer and has support for user written plugins and a very large collection growing of them. There is an active users group on developing plugins.
I think ImageJ is a tool everyone that works with images should have in their tool box since it an open ended growing project and not tied to any platform, computer of software. You can have familiar tools on Windows, Mac, Unix and Linux.
It sounds like all you need to do is use ImageJ's combine RGB plugin on the files you have. you might have to massaged he files a little first to get them in the correct format.
If I can help let me know.
Good luck Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. ----- Original Message ----- } From: "Darrell Miles" {milesd-at-US.ibm.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, June 20, 2003 7:54 PM
On Wednesday, June 18, 2003, at 04:51 AM, AtcSEM wrote:
} I have looked on the cross section of a sample and ran EDS and EDS line } scan, but could not see anything. } Dear Pavel, Something that has not yet been mentioned is that, since Cl is volatile, working at low temperature will be an advantage. If you have cryo-SEM available to you, try it. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Paint Sho Pro from Jasc Software (www.jasc.com) has similar basic features to those in Photosphop for lower price (around $100). The layer function will alloy you to overlay two images and adjust the relative intensity of each.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
} Hi all... } } } } I could use a little help with overlaying images. On our Zeiss } } LSM-310, we } } } } can scan a reflected laser image of a sample (computer chip, in our } } case) } } } } into the Green plane, and then maybe an OBIC image into the Red } } plane, } } } } and then select the RGB mode. This will display our OBIC image } } overlaid } } } } on the reflected light image, showing where the OBIC signal is } } coming } } } } from on the sample. } } } } } } Now the problem. We have a Roper Scientific CCD mounted on the } } Zeiss, } } } } which we use for reflected light images, and for capturing images of } } the } } } } light emitted from the computer chip while it is powered up (photo } } emission } } } } microscopy - PEM). We are using the WinView32 software that came } } with } } } } the CCD, and I have tried all of the math functions, etc., that I } } can find } } } } in } } } WinView32. I have been unable to do an "overlay" of two images. } } } } } } Does anyone know how to do a simple overlay with WinView32, or if } } there } } } } is another application that will work? (Hopefully cheap or free) I } } don't } } } } need a lot of image processing/analysis power. Just the ability to } } } overlay. } } } WinView32 can do pseudo coloring of the images to emulate the Red } } and } } } } Green planes of the Zeiss. } } } } } } Thanks for your attention, } } } Darrell } } } IBM Microelectronics }
If you can't actually see the peak then you can't trust what the software says.
I bet the detection limit of 0.09% was generated by the software from consideration of counting statistics. These may well be OK for WDS, but for EDS it's not the counting stats that limit detectability, it's the overlaps and other spectral interferences.
Even if you do 10 replicate measurements and look at the stats of the actual wt % results you're not getting the full story as precision is not the same as accuracy.
However, the 2% figure is unnecessarily pessimistic and I find it difficult to believe that it originated from an EDS detector manufacturer.
If anyone wishes to email me, I will send them the spectrum of volcanic glass containing 0.36% Cl, obtained on my JEOL 840/PGT Prism/Moran Scientific system in 130 seconds.
cheers
rtch
} } Well, here are another numbers. I collected an EDS spectrum from 1010 } steel (Jeol/Kevex/tungsten, 20 kV, 30% dead time, 2 minutes), forced } the software to account for Cl and Ar peaks and ran quantification. } What I've got was 0.11at%Cl and 0.26at%Ar. I like the "trick" and use } it once in a while to demonstrate my customers that if you don't see a } peak then the EDS isn't the right technique for the analysis and some } other methods should be used (like SIMS), particularly for light } elements. Of cause, if you know the element is there then it's easier } to "believe" the numbers. }
} } } } } } The EDS limit is around 2% and the accelerating voltage } }
} } OK. To get real numbers from real system I just performed } } test on XL30 with field emission gun at 15 kV, 40-45% dead time, 10 } } min acquisition time. I measured Cl peak from Au/Pd coated crystals } } of table salt and background in Cl peak position on a stainless } } steel razor blade. With ZAF correction detection limit was 0.09%.
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
I want to embed a layer of epithelium growing on a lens explant. The whole thing is only a thin collagenous matrix with a single layer of cells on top, about 4mm in diameter. Rather fragile. Does anyone have a suggestion as to the best way to keep this flat without losing it or mashing it? It's currently sitting in fixative. The aim is to cut sections perpendicular to the plane of growth.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sfontouris-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June 23, 2003 at 08:00:10 ---------------------------------------------------------------------------
Question: I would appreciate it very much if someone explained the differences between i)phase contrast, ii)bright field, iii) Normanski and iv) Hoffmann optics. Many thanks in advance
If you are EXTREMELY careful, and check with your Life Safety office first, you can do what I do...I etch the glass away with HF. It is extremely hazardous and you must be very careful about handling it, etc., but it works beautifully. A couple of hours on a stir plate in HF and all the glass is gone. You then retrieve the blocks (with plastic forceps) and wash them like mad. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I have an old JEOL JXA-840 SEM, late 80s vintage in storage. I need to clear out all storage areas to prepare for upcoming renovations to the building. I am offering the SEM free to a good home. It was working when it was replaced with a newer, donated JSM-840A about 2 years ago. The old scope has occasionaly been used as a source of parts for troubleshooting the newer scope. At this time all of the parts have been returned, but it has not been powered up in two years. One module in the newer scope (magnification power amp) had a hard failure and was replaced with the module from the old scope. The power amp will probably require repair or replacement by JEOL. The water chiller runs too warm and needs a refrigerant recharge. There is no EDS or computer image acquisition with the scope, as the one we had was transferred to the newer scope.
Preference given to academic labs. If you want it, you need to make arrangements for shipping, or pick it up in Medford, MA. Contact me offline for further information.
Dave Wilbur
-- __________________________________ David J. Wilbur, Ph.D. Instrumentation Specialist Department of Chemistry Tufts University 62 Talbot Ave. Medford, MA 02155 voice: 617-627-2163 Fax: 617-627-3443 email: david.wilbur-at-tufts.edu __________________________________
On Sunday, June 22, 2003, at 02:11 PM, Ritchie Sims wrote:
} If you can't actually see the peak then you can't trust what the } software } says.
} I bet the detection limit of 0.09% was generated by the software from } consideration of counting statistics. These may well be OK for WDS, but } for EDS it's not the counting stats that limit detectability, it's the } overlaps } and other spectral interferences. } Dear Richie, Overlaps should not be a problem for Cl and steel. When I was looking for Cl in sediment, I saw what were very small peaks that were only slightly larger than the noise. However, in several examples there were small amounts of Ti such that the Ti k-alpha peaks were clearly present and the k-beta peaks were the same size as the Cl k-alpha. Since I knew that the Ti k-beta peaks were real, that gave me confidence that the Cl peaks were also. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
We would like to match two images, ideally pixel-to-pixel, obtained from a single sample under two different conditions. The sample may have changed a little (small tears, streaks) between the acquisition of the two images and the re-loading onto the microscope may change the orientation slightly. The extent of differences is expected to be less than 10% of the total number of pixels.
I'll be grateful if anyone could please suggest an algorithm we could use to program these tasks. Commercial software that can accomplish this would also be useful and pointers to those are welcome.
Thanks for your help, Rohit
Rohit Bhargava National Institutes of Health Building 5, B1-38W LCP/NIDDK Bethesda, MD 20892-0520
Greetings, All microscopies involve a probe, a sample, and a detector. They generate contrast by virtue of a specific interaction between the probe and the sample. In the case of light microscopies, the probe is light. The different types of microscopy you list differ in the property of the sample used to generate contrast.
1) Brightfield would be much more usefully called 'absorption contrast': Contrast is generated based on differences in absorption. That is why it is good for stained samples, which absorb specfic wavelengths strongly.
2) Phase contrast depends on the optical path through the sample (refractive index times thickness). It is tricky for generating contrast based on optical path differences and F. Zernike (spelling??) got a Nobel prize for figuring out how.
3) Nomarski (also called differential interference contrast) depends on the gradient of optical path in a sample (this was invented by George Nomarski who should have gotten a Nobel prize for this work too).
4) I am not sure about Hoffman.
You can find further details in any good book about microscopy.
Tobias Baskin
} Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (sfontouris-at-hotmail.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, } June 23, 2003 at 08:00:10 } --------------------------------------------------------------------------- } } Email: sfontouris-at-hotmail.com } Name: Yannis Sfontouris } } Organization: University of Nottingham } } Education: Graduate College } } Location: Nottingham, UK } } Question: I would appreciate it very much if someone explained the } differences between i)phase contrast, ii)bright field, iii) } Normanski and iv) Hoffmann optics. } Many thanks in advance } } Yannis } } ---------------------------------------------------------------------------
Please put the answer on the list server for those of us who have been so immersed in EM of various types. I've always wanted a better explanation than I got way back in high school. Thanks Ken Converse owner Quality Images third party SEM service Delta, PA
by way of Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pgan-at-ap.ansell.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, June 24, 2003 at 05:58:42 ---------------------------------------------------------------------------
Email: pgan-at-ap.ansell.com Name: Phay Fang Gan
Organization: Ansell Shah Alam Sdn Bhd
Education: Graduate College
Location: Shah Alam,Selangor, Malaysia
Question: We are having a unit of Hitachi S3000N SEM in our lab.
It would be nice if anyone out there who can tell me where I could get local agent to provide Preventive Maintenance on our SEM in Malaysia.
The article in Microscopy Today, May/June 2003 by O'Neil et al. appears to be exactly what you want. Title: "Use of a New Imaging Technique to Document Deformations Recorded in the ESEM."
I see that you have a subscription. The article is on page 36.
Ron Anderson, MT Editor
-----Original Message----- } From: Rohit Bhargava [mailto:rxb29-at-po.cwru.edu] Sent: Monday, June 23, 2003 5:02 PM To: Microscopy-at-sparc5.microscopy.com
Hello:
We would like to match two images, ideally pixel-to-pixel, obtained from a single sample under two different conditions. The sample may have changed a little (small tears, streaks) between the acquisition of the two images and the re-loading onto the microscope may change the orientation slightly. The extent of differences is expected to be less than 10% of the total number of pixels.
I'll be grateful if anyone could please suggest an algorithm we could use to program these tasks. Commercial software that can accomplish this would also be useful and pointers to those are welcome.
Thanks for your help, Rohit
Rohit Bhargava National Institutes of Health Building 5, B1-38W LCP/NIDDK Bethesda, MD 20892-0520
On Lunedì, giu 23, 2003, at 23:49 Europe/Rome, Tobias Baskin wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Greetings, } All microscopies involve a probe, a sample, and a detector. They } generate contrast by virtue of a specific interaction between the } probe and the sample. In the case of light microscopies, the probe is } light. The different types of microscopy you list differ in the } property of the sample used to generate contrast. } } 1) Brightfield would be much more usefully called 'absorption } contrast': Contrast is generated based on differences in absorption. } That is why it is good for stained samples, which absorb specfic } wavelengths strongly. } } 2) Phase contrast depends on the optical path through the sample } (refractive index times thickness). It is tricky for generating } contrast based on optical path differences and F. Zernike (spelling??) } got a Nobel prize for figuring out how. } } 3) Nomarski (also called differential interference contrast) depends } on the gradient of optical path in a sample (this was invented by } George Nomarski who should have gotten a Nobel prize for this work } too). } } 4) I am not sure about Hoffman. } } You can find further details in any good book about microscopy. } } Tobias Baskin } } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (sfontouris-at-hotmail.com) from } } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June } } 23, 2003 at 08:00:10 } } ---------------------------------------------------------------------- } } ----- } } } } Email: sfontouris-at-hotmail.com } } Name: Yannis Sfontouris } } } } Organization: University of Nottingham } } } } Education: Graduate College } } } } Location: Nottingham, UK } } } } Question: I would appreciate it very much if someone explained the } } differences between i)phase contrast, ii)bright field, iii) Normanski } } and iv) Hoffmann optics. } } Many thanks in advance } } } } Yannis } } } } ---------------------------------------------------------------------- } } ----- } } } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ Biology } Department } / / / / \ \ \ University of } Massachusetts } /_ / __ /__ \ \ \__ Amherst, MA, 01003 } / / / \ \ \ / / } / \ \ \ Voice: 413 - 545 - 1533 / } / ___ / \ \__/ \ ____ Fax: } 413 - } http://www.biosci.missouri.edu/Baskin/baskin_lab__home_page.htm } } } ....................................................................... ........................ Alberto Diaspro, Deptartment of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480/309 fax 010314218 e-mail: diaspro-at-fisica.unige.it URL: http://www.lambs.it ....................................................................... .....................
Dear colleagues, in our lab of Electron Microscopy, we have an "Ultramicrotomy TOP 170 Pabish". Until nobody has used it, but we want to try preparation for TEM observations with it. Unfortunately we have lost its manual, please could anybody with the same equipment help us and send us a copy of the manual. My e-mail is marilena.re-at-brindisi.enea.it Thanks in advance Marilena Re
Our address is: ENEA C.R. Brindisi - Lab. of Electron Microscopy S.S. 7 - Appia km 712,700 72100 Brindisi - Italy
I have just received the good news that I have been approved for new SEM equipment to replace our old SEM. The new SEM will go in the room where the old SEM is now.
We have had problems with stray fields in the past and would like to remedy this situation before installation of the new SEM.
The 2 solutions to deal with the stray fields problem, of which I am aware, are either a field cancellation device or room shielding.
I need to determine the cost of eliminating the stray fields from the room and submit it to Management within the next day or so.
Could any companies which deal with either field cancellation devices or room sheilding materials please contact me offline as soon as possible?
Thanks in advance for your help.
Paula.
Paula M. Allan-Wojtas Research Scientist - Food Microstructure / Chercheur scientique - microstructure des aliments Food Safety and Quality Team / Salubrité et qualité des aliments Agriculture and Agri-Food Canada /Agriculture et Agroalimentaire Canada Telephone / Téléphone: 902-679-5566 Facsimile / Télécopieur: 902-679-2311 32 Main Street / 32 rue Main Kentville, Nova Scotia / Kentville (Nouvelle-Écosse) B4N 1J5 allanwojtasp-at-agr.gc.ca
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Here's my quick and dirty attempt to work through this.
First, all three contrast systems are based on the phenomenon that light can interact with regions of different refractive index by (a) slowing down, and (b) changing its path. (Some of the people I have talked to suggest that the change in path is really a diffraction effect, rather than refraction. Maybe theyre actually the same phenom.)
The three microscope systems you mention take advantage of this in different ways. All, however, have to face the challenge of how to distinguish the altered light from the wash of unaffected light -the background.
This is accomplished by constricting the illumination. For phase contrast, a hollow cone of light is projected through the sample onto a ring in the objective lens. For Hoffman, a narrow beam is projected through the sample obliquely to a small region on the side of the objective lens. For Nomarski, the system is a bit more complex. The illuminating system is set up between crossed polarizers, so that no undeviated light should get through at the end. The light passes through the first polarizer, and then through a special prism system (Wollaston) that splits the light into two beams whose vibrations are perpendicular. This pair of beams passes through the sample.
Now, what happens in the sample? I am referring only to the light that interacts, not the background. In the phase contrast system, the light is bent out of the path of the background light, and ends up passing through a different part of the ring in the objective. For Hoffman, the light is again bent to a different part of the Hoffman plate in the objective. In Nomarski, if (and this is critical) the two beams interact differently, they end up out of phase with each other.
As we follow the light up in the microscope, we reach the back focal plane of the objective lens. This is where the Phase and Hoffman systems do their dirty work. In the phase contrast system, there is a phase plate to which the cone of light is projected. Remember that the background light is projected to the ring itself. The light from the sample will be bent away from this ring, and elsewhere on this plate. Ziernike realized that not only was the light bent, but it had been slowed down so that it was 1/4 wavelength delayed compared to the background. He reasoned that if that difference could be expanded to 1/2 a wavelength, the light from the two sources could be made to interfere destructively --i.e. cancel each other out, and one would see a dark region--this would emphaize contrast.
In the Hoffman system, the deviated light passes through a region that is different in transmission from the plate to which most of the light is projected. As a result, it appears bright, compared to the background.
For Nomarski, the two perpendicular beams passs through a second Wollaston prism, which serves to reunite them. However, if one of the two beams is shifted in phase compared to the other, the resulting beam has an altered polarization, and passes through the final polarizer, while the background does not. --or vice versa, depending on how the system is set up.
The result of all three systemss is that one obtains contrast from refractive index differences within a sample. The consequence is that stains are not necessary, and details of living cells can be visualized.
You can get much more detail, with beautiful illustrations and animations at the microscopy web site:
http://micro.magnet.fsu.edu Look for the "primer" section.
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (sfontouris-at-hotmail.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June } 23, 2003 at 08:00:10 } ---------------------------------------------------------------------- } ----- } } Email: sfontouris-at-hotmail.com } Name: Yannis Sfontouris } } Organization: University of Nottingham } } Education: Graduate College } } Location: Nottingham, UK } } Question: I would appreciate it very much if someone explained the } differences between i)phase contrast, ii)bright field, iii) Normanski } and iv) Hoffmann optics. Many thanks in advance } } Yannis } } ---------------------------------------------------------------------- } ----- }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
the easiest way to match two similar images would be to use a cross correlation between the two images. It uses Fourier algorithms to come up with a "most likely" match of the two images by allowing a shift in X and Y (typically no rotation).
In your case it might already be sufficient. In case the images need to further adjusted manually, I would suggest looking at fluorescence software. Many of the available programs allow to shift two or more images against each other manually to compensate for small shifts between acquisition of the various fluorescence channels.
If you want further information how our analySIS software does this, or if you want to send me a couple of images for testing, please contact me off-line.
Thanks.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Rohit Bhargava [mailto:rxb29-at-po.cwru.edu] Sent: Monday, June 23, 2003 3:02 PM To: Microscopy-at-sparc5.microscopy.com
Hello:
We would like to match two images, ideally pixel-to-pixel, obtained from a single sample under two different conditions. The sample may have changed a little (small tears, streaks) between the acquisition of the two images and the re-loading onto the microscope may change the orientation slightly. The extent of differences is expected to be less than 10% of the total number of pixels.
I'll be grateful if anyone could please suggest an algorithm we could use to program these tasks. Commercial software that can accomplish this would also be useful and pointers to those are welcome.
Thanks for your help, Rohit
Rohit Bhargava National Institutes of Health Building 5, B1-38W LCP/NIDDK Bethesda, MD 20892-0520
Below are some rather useful resources when trying to get some light microscopy answers. Here are some short answers from me (please understand the complexity of these systems, these are VERY abbreviated answers);
There are three basic ways that light interacts with non fluorescent samples. They are; 1) absorption of light 2) phase shift of light 3) diffraction of light
Only the first can be seen by the eye. There are different modes of microscopy that permit us to take advantage of 2 and 3.
i) When light passes through something with a different optical density, the phase of the light is changed. (More dense samples will slow the light down more). The human eye can not see these changes in phase. In a phase contrast system, these changes in phase are changed into changes in brightness (contrast), which the eye can see. The image then becomes a gray scale image where the differences in intensity are representations in the differences in the optical density of the sample. This is done with a phase ring (in the objective) and a phase annulus (in the condenser). This system is lower resolution than bright field, but permits viewing samples that absorb almost no light (such as living cells). ii) Bright field is the most common way to look at a sample slide. The information is generated by differences in the absorption of light. This is seen as color and darkness. This is normally used for samples that have great differences in the amount of light absorbed (non biological samples) or samples that have been stained. Bright field also takes advantage of diffracted light. The condenser focuses light on the sample in such a way that the light diffracted by the sample is focused by the objective. This diffracted information dramatically increases the resolution of the system. A stained cell preparation on a slide would be a standard biological sample. iii) Nomarski (or DIC for Differential Interference Contrast microscopy) is a method that used differences in optical density to create differences in brightness. Phase contrast shows the observer the absolute optical density of the sample. DIC shows the local CHANGES in optical density. This is done by an optical system that employs two closely spaced beams of light that start in phase with each other. They then pass through the sample close to each other. The beams are then recombined in such a way that any differences in the optical path is seen as a difference in intensity. This is accomplished with polarized light and a pair of prisms that split the light before the sample and recombine the light after the sample. iv) Hoffmann will give results like DIC but is cheeper to implement and can be used to image through plastic. It is lower resolution (I assume that this will be argued by the company reps. that sell the system :-) but has made great strides recently.
I recommend the book "Fundamentals of Light Microscopy and Electronic imaging" by Doug Murphy. This is a great book that starts from simple principals and goes into great depth in all of these questions and more. If running a class, this should be the text. It is available on Amazon. I have no financial interest in the book or the publisher, I just think it is the best.
On Monday, June 23, 2003, at 09:05 AM, by way of Ask-A-Microscopist wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (sfontouris-at-hotmail.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June } 23, 2003 at 08:00:10 } ----------------------------------------------------------------------- } ---- } } Email: sfontouris-at-hotmail.com } Name: Yannis Sfontouris } } Organization: University of Nottingham } } Education: Graduate College } } Location: Nottingham, UK } } Question: I would appreciate it very much if someone explained the } differences between i)phase contrast, ii)bright field, iii) Normanski } and iv) Hoffmann optics. } Many thanks in advance } } Yannis } } ----------------------------------------------------------------------- } ---- } }
____________________
David Elliott
Yale University School of Medicine 810 LCI 333 Cedar Street New Haven, CT 06520-8022
I would like to thank those of you who kindly shared the experience regarding removing coverslips off EM blocks.
Since some of you are also interested in this issue, here I post the following summary of the responses.
1. Use chemicals, such as: hydrofluoric acid or hydrogen fluoride, the potential problem is that it may also cause damage to the cell or remove osmium, and they are hazardous.
2. Play with a hot plate instead of liquid nitrogen.
3. Use a jeweler's saw to carefully remove glass coverslips.
4. This method maybe is the cleanest way to get rid of the glass off blocks. I have tried myself and it worked quite well. Polymerize the coverslips with a glass slide together and place on a hotplate or dip into liquid nitrogen. The coverslip will come off with glass slide and leave the cells on the top of the blocks.
We are doing this type of things on a routine basis, so if someone needs more information, please let me know.
Xinran
Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center at Dallas Phone: 214-648-1830 Fax: 214-648-1801 E-mail: xinran.liu-at-utsouthwestern.edu
Does anyone on the list have possession of a Cressington 208HR or Emitech K575X high resolution sputter coater for FEG-SEM with a working IRIDIUM target?
Thanks in advance for any help.
Paul Beauregard PPG Industries Senior Research Associate Monroeville Technical Center 440 College Park Drive Monroeville, PA 15146
I was referring in a general sense to detection limits and/or precision estimates that are delivered by the EDS spectrum-processing software.
My point is that while statistical considerations may well give a realistic estimate of the precision of determination of higher concentrations, they don't RELIABLY provide meaningful detection limits for EDS, in which overlaps and other spectral artefacts such as coincidence peaks and escape peaks can play a large part.
Do you really believe the quoted 'detection limit' of 0.09%?
Even without entering the discussion of exactly what is meant by 'detection limit'.
It would be interesting to hear something on this subject from the EDS system manufacturers. My personal suspicion is that a lot of 'quantitative' results, obtained from standardless EDS packages, are quoted to unjustifiable precision, even to two decimal places!
cheers
rtch
} Dear Richie, } Overlaps should not be a problem for Cl and steel. When I was } looking } for Cl in sediment, I saw what were very small peaks that were only } slightly larger than the noise. However, in several examples there } were small amounts of Ti such that the Ti k-alpha peaks were clearly } present and the k-beta peaks were the same size as the Cl k-alpha. } Since I knew that the Ti k-beta peaks were real, that gave me } confidence that the Cl peaks were also. Yours, Bill Tivol EM Scientist } and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code } 114-96 California Institute of Technology Pasadena CA 91125 (626) } 395-8833 tivol-at-caltech.edu }
} } } On Sunday, June 22, 2003, at 02:11 PM, Ritchie Sims wrote: } } } If you can't actually see the peak then you can't trust what the } } software says. } } } I bet the detection limit of 0.09% was generated by the software } } from consideration of counting statistics. These may well be OK for } } WDS, but for EDS it's not the counting stats that limit } } detectability, it's the overlaps and other spectral interferences. } }
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
Hi all, I have an investigator whose lab is in another building, making it inconvenient for them to come over to use our light microscopes routinely. They have an older Zeiss fluorescent microscope and a Coolpix 900 digital camera that they want to set up to quick-check samples. Do you know of any software that would let them preview the images on a computer screen and capture directly to the computer? Using the small camera view screen is not nearly as desirable an option.
Another, probably more expensive option, would be to purchase a new camera. Please let me know if anyone has a camera option for under $2000. Remember that this is for screening only and is not intended to replace the more sophisticated microscope-camera systems available.
Thanks in advance, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nmorehouse-at-asu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, June 24, 2003 at 14:54:17 ---------------------------------------------------------------------------
Question: Hi! I'm looking to measure 100-150 nm features (microstructures on the surface of butterfly wing scales) in a low Z number matrix using a secondary electron signal on a JEOL 840A SEM (LaB 6). The specimens are dehydrated but not fixed. Any suggestions on specimen prep and/or techniques for conducting spatial measurements?
Dear Colleagues, The Organizing Committee has the honor to announce that:
1. The 4th ASEAN Microscopy Conference and The 3rd Vietnam Conference on Electron Microscopy will be continuously held in Hanoi on 05-06 January 2004.
2. The deadline for article submission will be on 30 August 2003.
3. The submitted and approved articles will be put in the Proceedings of the Conference.
Chairman of the Organizing Committee
Nguyen Van Man M.D., D.M.Sc
If you need more informations, please contact at: Assoc. Prof Nguyen Kim Giao Electron Microscopy Unit National Institute of Hygiene and Epidemiology 1- Yersin Str - HaiBaTrung Distr - HaNoi-VietNam Tel: 84.4.9715434 Fax: 84.4.8210853 Email: emlad-at-hn.vnn.vn or emunihe-at-vol.vnn.vn
We do not have either of those instruments, but we routinely sputter iridium with our IBS/e system. Is there something we can do to help or perhaps refer you to one of our customers for some assistance?
Please let me know.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
"Beauregard, Paul A." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } } Does anyone on the list have possession of a Cressington 208HR or Emitech K575X high resolution sputter coater for FEG-SEM with a working IRIDIUM target? } } Thanks in advance for any help. } } Paul Beauregard } PPG Industries } Senior Research Associate } Monroeville Technical Center } 440 College Park Drive } Monroeville, PA 15146
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you are right. The detection limits of an EDX spectrometer depends not only from statistic. Main cause to change the detection limits to higher values are peak overlaps and spectra distortions (escape, shelf, tail, pile-up, diffraction peaks ..).
But the basics of calculation of detection limits are signal to noise (peak to background) ratios. The background with electron probe microanalysis is well defined with Bremsstrahlung (99% of all background contributions in most cases). The Bremsstrahlung and its absolute relation to the excited characteristic lines depends from physics only, mainly from primary electron energy and critical excitation energies of used line series. It's true in region of 10 { Z { 30 it can be achieved 0.1% (K-radiation) and for Z } 80 only 0.3% as detection limits (mean atomic number about 26, 60 s acquisition time, 2000 cps). The probability of Bremsstrahlung generation rises with higher mean atomic number of specimen. The detection limit in a weak matrix is much higher compared to a heavy matrix (it can be factors of 3 and more). A detection limit of 0.05% (mass units) for iron is possible in a matrix of glass or in biological specimens, but not in Pb.
You will find at the given link an image with detection limits curves, which can give you some support. These calculated curves give you an idea about 'best case' detection limits in EPMA without overlaps etc.:
http://www.microanalyst.net/index_e.phtml
.. hit [Information] and then scroll down (last image)
Frank Eggert
Ritchie Sims wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Yes. } } I was referring in a general sense to detection limits and/or precision } estimates that are delivered by the EDS spectrum-processing software. } } My point is that while statistical considerations may well give a realistic } estimate of the precision of determination of higher concentrations, they } don't RELIABLY provide meaningful detection limits for EDS, in which } overlaps and other spectral artefacts such as coincidence peaks and } escape peaks can play a large part. } } Do you really believe the quoted 'detection limit' of 0.09%? } } Even without entering the discussion of exactly what is meant by 'detection } limit'. } } It would be interesting to hear something on this subject from the EDS } system manufacturers. My personal suspicion is that a lot of 'quantitative' } results, obtained from standardless EDS packages, are quoted to } unjustifiable precision, even to two decimal places! } } cheers } } rtch } } } Dear Richie, } } Overlaps should not be a problem for Cl and steel. When I was } } looking } } for Cl in sediment, I saw what were very small peaks that were only } } slightly larger than the noise. However, in several examples there } } were small amounts of Ti such that the Ti k-alpha peaks were clearly } } present and the k-beta peaks were the same size as the Cl k-alpha. } } Since I knew that the Ti k-beta peaks were real, that gave me } } confidence that the Cl peaks were also. Yours, Bill Tivol EM Scientist } } and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code } } 114-96 California Institute of Technology Pasadena CA 91125 (626) } } 395-8833 tivol-at-caltech.edu } } } } } } } } } On Sunday, June 22, 2003, at 02:11 PM, Ritchie Sims wrote: } } } } } If you can't actually see the peak then you can't trust what the } } } software says. } } } } } I bet the detection limit of 0.09% was generated by the software } } } from consideration of counting statistics. These may well be OK for } } } WDS, but for EDS it's not the counting stats that limit } } } detectability, it's the overlaps and other spectral interferences. } } } } } -- } Ritchie Sims Ph D Phone : 64 9 3737599 } ext 87713 } Microanalyst Fax : 64 9 3737435 } Department of Geology email : } r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand
Couple of days ago we had received an error message on my INCA Wave 1394 "Error 308"; "Failed to write Pha Threshold GSR location 7B8"; "Failed to create object FwMipConnector.Comm Settings"
I have checked Fire Wire devices and they work properly. The reset button on the INCA Wave box sends signal to the WDS detector and it seems to operate properly. Is there any suggestions how to fix it? Did any body have had the same problems?
Your help is very appreciated! Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
We have had a consistent problem on our two Noran Voyager x-ray analysis systems. Often when users attempt to start an x-ray linescan or x-ray map in Voyager, the Voyager hardware begins to take control of the SEM scan, then aborts. The error message (from Voyager console) suggests that the software thinks that some other process is already operating. I have checked with the users and been through this several times myself, making sure that no other functions are operating. Someone suggested that there may be some Voyager command which can be invoked from the console to kill any other processes. Unfortunately, Noran have not been able to provide any answers to the problem, so our frustration continues. Has any Voyager user out there had any similar problems, or could offer constructive advice?
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-cam.ac.uk
} } Yes. } } I was referring in a general sense to detection limits and/or } precision } estimates that are delivered by the EDS spectrum-processing software. } } My point is that while statistical considerations may well } give a realistic } estimate of the precision of determination of higher } concentrations, they } don't RELIABLY provide meaningful detection limits for EDS, in which } overlaps and other spectral artefacts such as coincidence peaks and } escape peaks can play a large part. } } Do you really believe the quoted 'detection limit' of 0.09%?
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We have just purchased a Canon Powershot G5 that lets us preview and focus the image on the computer monitor and save directly to the computer. It is 5 Mp at a cost of ca. $750, including cables, AC adapter, etc. The G3 will do the same (4 Mp) at { $550. You still need an adapter to mount on the scope (which we are waiting for). The preview image on my 20 " monitor is ca. 3 x 4 inches and I can't (yet) find a way to enlarge until I record it. The image is quickly renewed when focussing.
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Phillip H. Geil; Ph. 217-333-0149 Fax 217-333-2736 Department of Materials Science and Engineering University of Illinois 1304 W. Green St. Urbana, IL 61801
I am thinking about buying an Epson 2200 ink jet printer for printing hardcopies of my digital microscopy images. I am aware of the great reviews this $600 printer gets for printing on high end photographic type paper. But I will also want to print images on "regular" paper, e.g., when I embed them within a grant application. Can anyone comment on this usage? Thanks, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I used to work with epithelial cells grown on collagen mats. For processing for EM, we would actually pin the sample down to a piece of dental wax. I would carry out fixation through infiltration of the specimen with drops placed onto the tissue (in a fume hood). To embed, I would cut the piece into strips and flat embed, collagen down, in order to get sections across cells & collagen.
Hope this helps? Good luck! Angela Welford awelford-at-salud.unm.edu
On Sunday, June 22, 2003, at 07:57 PM, Diana van Driel wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } I want to embed a layer of epithelium growing on a lens explant. The } whole thing is only a thin collagenous matrix with a single layer of } cells on top, about 4mm in diameter. Rather fragile. Does anyone have } a suggestion as to the best way to keep this flat without losing it or } mashing it? It's currently sitting in fixative. The aim is to cut } sections perpendicular to the plane of growth. } } Thanks, as always. }
CombineZ (freeware, see http://www.hadleyweb.pwp.blueyonder.co.uk/CombineZ/combinez.htm) works with displacements and magnification, but not with rotation.
Somebody know anything about the announcement: "CombineZ4 will shortly be available from Microscopy-UK?
Martin
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
Thanks to the many people who responded to a request for information on image matching on the list and off it as well. While I appreciate the many references to commercial software, if possible we would really like to program it into our own lab software to integrate with instrumentation built in-house.
For those who may be curious: The best solution appears to be two separate procedures. One is to use coarse resolution edges to find the correct rotation angle and then a regression to match pixel for pixel by optimizing over only the better fit pixels (rejecting 10-20%) to account for artifacts. This would also identify "hot" pixels where some change may have occured. I must stress that this procedure suits us because we know what changes/artifacts are likely to result, is somewhat limited in the range of problems that can be handled and may not be an optimal approach for all cases.
Thanks,
Rohit
Rohit Bhargava National Institutes of Health Building 5, B1-38W LCP/NIDDK Bethesda, MD 20892-0520
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We use Noran Voyager 3 and I can only say that I know the problem much too well. We commonly experience exactly the same error when we want to start a linescan or an x-ray map. I know that there are other users here in Denmark who have the same experience. Norans advice to us is to shut down the system and start up again - this works, but it is rather frustrating and time consuming in the long run.
I, too, shall be very interested if anybody have found a solution to the problem.
Yours sincerely Henning Sund Sørensen Materials and Process Consultant
Danfoss A/S Central Service - Danfoss Technology Centre Nordborgvej 81, L7-S40 6430 Nordborg Denmark
Ph. 7488 2309 Fax 7488 2670 E-mail henning.s-at-danfoss.com Internet www.teknologicenter.danfoss.dk
-----Original Message----- } From: David Vowles [mailto:djv23-at-msm.cam.ac.uk] Sent: 25. juni 2003 15:41 To: Microscopy Listserver
We have had a consistent problem on our two Noran Voyager x-ray analysis systems. Often when users attempt to start an x-ray linescan or x-ray map in Voyager, the Voyager hardware begins to take control of the SEM scan, then aborts. The error message (from Voyager console) suggests that the software thinks that some other process is already operating. I have checked with the users and been through this several times myself, making sure that no other functions are operating. Someone suggested that there may be some Voyager command which can be invoked from the console to kill any other processes. Unfortunately, Noran have not been able to provide any answers to the problem, so our frustration continues. Has any Voyager user out there had any similar problems, or could offer constructive advice?
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-cam.ac.uk
We have two Noran Voyagers here. Both of them Voyager 4 systems, running version 4.3.1 of the software on 170MHz Sparc V machines. (Well ok so it sounded impressive years ago.) We also encounter this problem occasionally. In regard to Henning's message the problem may be worse in earlier versions of the Voyager software/hardware combinations than later versions, since we definitely saw an improvement in the overall stability and usability of our systems when we upgraded ours from Voyager 3 to 4. That said, that improvement definitely didn't come without some hassles at the time though, including the problems you describe.
The immediate cause does sound like an "orphaned" process -- if that's the right terminology? The orphaned process is possibly actually running on the other CPU down inside the main box of stuff under the table and not on the main workstation. If some process still has control of the hardware down there as a result of some previous problem then attempting to start a new map or line profile etc will fail because it is already under the spell of the previous copy of the process still running (I think). If this is the case the problem won't go away if you just reboot the workstation alone, even if you do a completely cold re-boot of the workstation alone, because the offending thing is still alive in another part of the creature's body. As Noran suggests the cleanest solution is to do a proper "power off" shutdown of the workstation and then kill the power to the entire system -- unit under the table included -- wait at least 30 seconds -- then turn the power back on again to the entire system. If it is "only" a software problem then that is all it should take to make the problem go away.
The next question is that even if this fixes the problem in the short term, does it then come back with such frequency that it makes the system useless? In our experience the problem you describe could arise for example if the Image Display program crashes for some reason before an acquisition has been completed. I now recall (after sending a message to David earlier) that we used to encounter this situation if we cranked up the beam current too high when setting up do X-ray maps. When the X-ray count rates were too high it would crash the acquisition, leading to the situation that you describe. We worked around this by training our users and inserting alerttool dialog boxes to remind users not to exceed a certain count rate. (Your experiences re actual maximum count rates may differ however).
Also, if the Image Display program gets forced to quit without exiting the acquisition properly for other reasons as well, then the process(es) operating in the background in the unit under the table, that are actually controlling the SEM, won't necessarily be in a position to know when to cease execution and let go. It/they'll keep on running. If you then just re-launch new copies of the Image Display program -- even after starting a new session, or even from a cold re-boot of just the workstation alone, you still won't be able to do an acquisition. You are left with the Noran "solution" of complete shutdown and restart.
These systems behave like chickens with their heads cut off sometimes -- or maybe a better analogy is like dinosaurs with two brains. If the problem is occurring with such frequency that it is rendering the system unusable then maybe you have a hardware fault, or maybe it is just one of your users trying to screw too many counts out of it or shutting down the software/their session in an "incorrect" manner.
First thing I'd try after the restart is to see if it all works fine with no beam/no X-rays in the detector. So, with no beam, start an X-ray map acquisition then stop it via the software and see if you can start another one with no hassles. If so, launch the PHA Status program and perhaps also select the HIGH2 count rate (analog pp system) or fastest "rate file" (digital pp) under the pulse processor setup dialogue, and watch the "detects" count rate. Keep cranking up the beam current, attempting to cleanly start, stop, and start a new map or line scan at each count rate. See if you can reach a count rate where it falls over and you have to restart the entire system to get it to work again. See if not exceeding this count rate for a while then stops a recurrence -- although we found that pushing the maximum a little too close for comfort still caused a crash part way through a map so we backed it off even further. In the end you still should be able to get high enough count rates to make mapping and line scans practical if there are no actual hardware problems.
Hope this helps. We are basically still very happy with our Voyager 4 systems - now - and they work extremely well but they are idiosyncratic. We still get the sort of problem you describe 3 or 4 times a year.
Cheers, Arthur.
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I found NDS in a recipe for immunolabeling and I wonder what the function of this substance is. Is it used for pre incubation just like BSA? If so, does anyone have any guidelines for which one to choose?
Sincerely, Pernilla Nevsten
--------------------------------------------------- Pernilla Nevsten, PhD student Department of Materials Chemistry Lund University Sweden
We are using the McDowell/Trump fix at the moment for both LM/EM--and in fact for all of our current EM projects. It is a good compromise, giving very good fixation for LM and adequate to good fixation to TEM. For immersion fixation for EM, it may be one of the best. I have used similar formulations using freshly made paraformaldehyde instead of formalin with (I believe--note the faitrh thing here--I have no quantitative data) improved ultrastructural preservation as compared to the McDowell/Trump formulation. I also prefer using cacodylate buffer, since my experience has been with uranyl precipitate following use of phosphate buffers--even after extensive changes into Tris prior to en bloc staining or grid staining. I have also used PIPES buffer instead of cacodylate or phosphate with excellent results.
The McDowell/Trump fixative was designed to permit storage of tissues in the fixative over long time periods. I can't speak to the efficacy of that, since we routinely embed all of our samples.
Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals Ridgefield, CT
-- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } can anyone shed some light on concerns when going from phosphate buffer to } cacodylate buffer? } } Does anyone routinely use McDowells/Trump fix for LM/EM } any comment on this fix, is there a better dual purpose Fix? } } Gregory Argentieri } Electron Microscopy lab } gregory.argentieri-at-pharma.novartis.com } } }
Since Bremsstrahlung radiation is also called "braking radiation", is there general agreement that the braking radiation will be directly proportional to surface charge on the target sample? That is, a surface that becomes charged by the e-beam will generate more braking radiation than a surface whose charge is at zero. Also, is it fair to say that braking radiation can happen within a sold and not just during the flight of the electron down the EM column? For example, if a backscattered electron becomes slowed down by it's local environment, in a solid or any sample for that matter, it must release that energy in the form of photons i. e. Brermsstrahlung radiation, yes?
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Frank Eggert [mailto:Eggert-at-mikroanalytik.de] Sent: Wednesday, June 25, 2003 7:38 AM To: Ritchie Sims Cc: Bill Tivol; microscopy-at-sparc5.microscopy.com
Dear all,
you are right. The detection limits of an EDX spectrometer depends not only from statistic. Main cause to change the detection limits to higher values are peak overlaps and spectra distortions (escape, shelf, tail, pile-up, diffraction peaks .).
But the basics of calculation of detection limits are signal to noise (peak to background) ratios. The background with electron probe microanalysis is well defined with Bremsstrahlung (99% of all background contributions in most cases). The Bremsstrahlung and its absolute relation to the excited characteristic lines depends from physics only, mainly from primary electron energy and critical excitation energies of used line series. It's true in region of 10 { Z { 30 it can be achieved 0.1% (K-radiation) and for Z } 80 only 0.3% as detection limits (mean atomic number about 26, 60 s acquisition time, 2000 cps). The probability of Bremsstrahlung generation rises with higher mean atomic number of specimen. The detection limit in a weak matrix is much higher compared to a heavy matrix (it can be factors of 3 and more). A detection limit of 0.05% (mass units) for iron is possible in a matrix of glass or in biological specimens, but not in Pb.
You will find at the given link an image with detection limits curves, which can give you some support. These calculated curves give you an idea about 'best case' detection limits in EPMA without overlaps etc.:
http://www.microanalyst.net/index_e.phtml
. hit [Information] and then scroll down (last image)
Frank Eggert
Ritchie Sims wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Yes. } } I was referring in a general sense to detection limits and/or precision } estimates that are delivered by the EDS spectrum-processing software. } } My point is that while statistical considerations may well give a realistic } estimate of the precision of determination of higher concentrations, they } don't RELIABLY provide meaningful detection limits for EDS, in which } overlaps and other spectral artefacts such as coincidence peaks and } escape peaks can play a large part. } } Do you really believe the quoted 'detection limit' of 0.09%? } } Even without entering the discussion of exactly what is meant by 'detection } limit'. } } It would be interesting to hear something on this subject from the EDS } system manufacturers. My personal suspicion is that a lot of 'quantitative' } results, obtained from standardless EDS packages, are quoted to } unjustifiable precision, even to two decimal places! } } cheers } } rtch } } } Dear Richie, } } Overlaps should not be a problem for Cl and steel. When I was } } looking } } for Cl in sediment, I saw what were very small peaks that were only } } slightly larger than the noise. However, in several examples there } } were small amounts of Ti such that the Ti k-alpha peaks were clearly } } present and the k-beta peaks were the same size as the Cl k-alpha. } } Since I knew that the Ti k-beta peaks were real, that gave me } } confidence that the Cl peaks were also. Yours, Bill Tivol EM Scientist } } and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code } } 114-96 California Institute of Technology Pasadena CA 91125 (626) } } 395-8833 tivol-at-caltech.edu } } } } } } } } } On Sunday, June 22, 2003, at 02:11 PM, Ritchie Sims wrote: } } } } } If you can't actually see the peak then you can't trust what the } } } software says. } } } } } I bet the detection limit of 0.09% was generated by the software } } } from consideration of counting statistics. These may well be OK for } } } WDS, but for EDS it's not the counting stats that limit } } } detectability, it's the overlaps and other spectral interferences. } } } } } -- } Ritchie Sims Ph D Phone : 64 9 3737599 } ext 87713 } Microanalyst Fax : 64 9 3737435 } Department of Geology email : } r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand
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Surface charge becomes irrelevant in the creation of brehmstrahlung since the final energy of the electron winds up being zero. Whether it decelerates before the sample or within it, you will decelerate the same amount and so create the same brehmstrahlung. It is a characteristic of the acceleration of the electron. Synchrotron radiation can be considered to be a special case of brehmstrahlung where the acceleration of the electron is perpendicular to its trajectory.
That said... Surface charge WILL affect your spectrum, since the electron within the sample is at a lower energy.
Cheers, Henk Colijn
At 09:25 AM 6/26/2003 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
We have a plateout issue in one of our manufacturing facilities, on a Nickle plated tool used for making PVC cast skins (slush molding). Initially this material is not visible but after many casts, the plateout shows up on the surface of the Nickle shell tool. Not sure how thick it is. We need to ID the material. We suspect it is an organic layer coming from the PVC material, probably the internal mold release which is an organic ester. we also suspect the mold release to be a carrier for the plasticizer, also an organic ester, both of which we suspect migrating to the surface and plating out. But, we need to confirm it.
I remember years ago using a blue tape replicating technique to transfer surface features from hard substrates for TEM. I am wondering if such a method might work for FTIR/XPS, etc. The tool is sometiimes cleand with THF, Acetones, and in one plant, HCL.. It is fairly robust.
I am looking for a way to remove this plateout material without scraping the tool surface, perhaps using a tape/stick/peel method.
All suggestions appreciated.
Thanks in advance
Fred Hayes Collins & Aikman Corp IntelliMold Division Ann Arbor MI
Toxicity is the #1 concern. Otherwise, cacodylate seems to have a slight edge in that it (1) precipitates much less with UA, and (2) is more extractive, which often means a better view.
Vlad ------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
Greg wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } can anyone shed some light on concerns when going from phosphate buffer to } cacodylate buffer? } } Does anyone routinely use McDowells/Trump fix for LM/EM } any comment on this fix, is there a better dual purpose Fix? } } Gregory Argentieri } Electron Microscopy lab } gregory.argentieri-at-pharma.novartis.com } } }
When an electron is slowed on it's approach to the target, where does that energy go? If I were able to instantaneously stop an electron in flight, would it not expend it's entire incident energy as photon radiation? And would that not be "braking radiation?"
Thanks,
Peter
-----Original Message----- } From: Hendrik O. Colijn [mailto:colijn.1-at-osu.edu] Sent: Thursday, June 26, 2003 10:28 AM To: Tomic, Peter (Peter); microscopy-at-sparc5.microscopy.com
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
On Thursday, June 26, 2003, at 06:25 AM, Tomic, Peter (Peter) wrote:
} Since Bremsstrahlung radiation is also called "braking radiation", is } there } general agreement that the braking radiation will be directly } proportional } to surface charge on the target sample? That is, a surface that becomes } charged by the e-beam will generate more braking radiation than a } surface } whose charge is at zero. Also, is it fair to say that braking } radiation can } happen within a sold and not just during the flight of the electron } down the } EM column? For example, if a backscattered electron becomes slowed } down by } it's local environment, in a solid or any sample for that matter, it } must } release that energy in the form of photons i. e. Brermsstrahlung } radiation, } yes? } Dear Peter, Bremsstrahlung is not proportional to the charge--surface or bulk--on the sample. It can, be produced by neutral atoms and is proportional to a power of Z. (I don't have my reference material with me, but I think it is Z^2.) Since both energy and momentum must be conserved, bremsstrahlung must involve another particle besides the electron and photon, so it cannot be generated in a perfect vacuum; however, in addition to being generated in solids, bremsstrahlung can be generated in gasses, so, if the electron happens to pass near enough to a residual atom as it travels down the EM column, bremsstrahlung can be produced (extremely rarely). In addition to bremsstrahlung, there are several energy-loss processes that do not involve photon production, such as ionization, excitation, plasmon production, etc., so when electrons are slowed down, most of the energy released is not in the form of photons. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
We've been using Epsons for several years now. And as with ALL ink jet printers paper type/quality is absolutely key to the quality of the image. The images on "regular" paper - well diagrams, line drawings, gels, charts and grafts are fine, but microscopy images just loose too much they're dull and have poor resolution. You are much better off going to epson photo quality ink jet paper - costs a little more than "regular paper" (I think we're paying ~ $0.08 sheet) but far less than the "Photo Glossy" media.
You're going to find the same problems with any other ink jet printer as well. I suggest you go down to your local computer store or electronics store, and get them to print you out and example on "regular" paper and see what you think. (For Grey scale images in grants we use our Lexmark laser printer and are very very happy with them).
} } I am thinking about buying an Epson 2200 ink jet printer for printing } hardcopies of my digital microscopy images. I am aware of the great } reviews this $600 printer gets for printing on high end photographic type } paper. But I will also want to print images on "regular" paper, e.g., when I } embed them within a grant application. Can anyone comment on this usage? } Thanks, Tom } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
Can someone give me the recipe for osmium staining of latex 20nm-spheres for contrast enhancement in a TEM?
The microspheres that we are looking at are described as "carboxylate modified polystyrene latex". Therefore their surface should be full of carboxylate groups and be negatively charged.
I would assume that NDS stands for Normal Donkey Serum and is used as a background suppressor in the blocking and/or incubation steps when using donkey secondary immunoconjugates. Basically, you will need something like BSA or Normal Serum (same species as in the secondary immunostep) to block sticky areas in your specimen during blocking on the one hand and on the other to compete with immunoconjugates for aspecific binding during the labeling step. NDS is what you might use with Donkey immunoconjugates. Which one is better? That probably depends on the situation. I would think BSA is most widely used, which is an indication of its suitability to a certain degree. Sometimes such compounds complement each other and a combination may give the best results. Hope this helps a bit.
================================ Jan Leunissen Aurion - http://www.aurion.nl present address: EM-Unit Dept. Anatomy and Struct Biology University of Otago PO Box 913 Dundedin, New Zealand
On Thursday, June 26, 2003, at 10:00 PM, Pernilla Nevsten wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Dear Microscopy Listserver members, } I found NDS in a recipe for immunolabeling and I wonder what the } function of this substance is. } Is it used for pre incubation just like BSA? If so, does anyone have } any guidelines for which one to choose? } Sincerely, } Pernilla Nevsten } } --------------------------------------------------- } Pernilla Nevsten, PhD student } Department of Materials Chemistry } Lund University } Sweden
I think that they can simply connect their Coolpix directly to a standard video monitor (eg the AV input of a TV) and directly view, focus, etc in real time on the monitor screen.
I think that images get saved to the camera rather than direct to the computer, but that they can easily be downloaded to the computer.
Get them to check the Coolpix manual.
cheers
rtch
} } } } Hi all, } } I have an investigator whose lab is in another building, making } } it } } inconvenient for them to come over to use our light microscopes } } routinely. They have an older Zeiss fluorescent microscope and a } } Coolpix 900 digital camera that they want to set up to quick-check } } samples. Do you know of any software that would let them preview the } } images on a computer screen and capture directly to the computer? } } Using the small camera view screen is not nearly as desirable an } } option.
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
Hello Everyone The question has arisen here about drying molecular sieve. How often do you dry your molecular sieve and when you do, what temperature and for how long?
Alternatively does anyone have a better method of maintaining 100% ethanol. Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
Sorry, you're right, I put that sentence in as a hasty afterthought.
I shouldn't malign figures which support my initial guesstimate!
But I stand by my contention that EDS package software precisions calculated from counting stats are not very reliable.
And that it is wrong for standardless EDS results to be quoted to a precision of more than two significant figures.
cheers
rtch
} Your estimate was 0.1%. How far is it from 0.09%? } } Vladimir }
} } } } } Yes. } } } } I was referring in a general sense to detection limits and/or } } precision } } estimates that are delivered by the EDS spectrum-processing } } software. } } } } My point is that while statistical considerations may well } } give a realistic } } estimate of the precision of determination of higher } } concentrations, they } } don't RELIABLY provide meaningful detection limits for EDS, in which } } overlaps and other spectral artefacts such as coincidence peaks and } } escape peaks can play a large part. } } } } Do you really believe the quoted 'detection limit' of 0.09%? }
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
And while we are on the subject of drying ethanol, what is the feeling about whether this is necessary? What is the problem if your "100%" ethanol has a small (1-2%) of water in it. I never dry my ethanol. I used to only use fresh bottles but over the last couple of years have evolved into using "recently opened" (less than 2 weeks or so) bottles and see no difference. Am i losing something in morphological quality or sectioning quality that I don't notice? Tom
At 02:53 PM 6/26/2003 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Regarding the question of "why does alcohol need to be dried over sieves anyway?":
With some plastics, a trace of water (or even more than a trace) is not a problem. Some acrylics (LR White, for example) and some epoxies (Epon equivalents) can tolerate traces of water. BUT, some plastics are notoriously poor in their tolerance of even traces of water. A good example is Spurrs resin where exposure of the resin even to atmospheric humidity will result in a brittle block that is useless (been there, done that).
So, sometimes you need absolutely dry specimens while at other times this is not so critical. Like Tom Phillips we now use recently opened ethanol pints and relegate them to "95%" after they have been opened a dozen or so times (or are near the half-way point). When we use Spurrs, we always pass specimens through ethanol followed by propylene oxide rather than acetone dehydration since acetone is never as dry as ethanol. Admittedly, this is based on experience rather than hard science, but we don't tamper with success.
John ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu ##############################################################
Has any one out there found a good substitute for the mineral oil layer over your lead stock? The newer mineral oils that I have tried since my 1970 bottle of Nujol extra heavy mineral oil ran out, contaminate my lead after only one or two draws from the lead. The oil turns dark brown and clumpy. HELP! Thanks, Sharon Drew
Another thing to consider is price. Printing on cheap paper you will still use the same costly inks. Again, inkJets usually much slower than laser printers. To resolve this problem, we have central networked color laser printer (10 cents/page I believe?), which do 90% of job just perfectly and we are using inkJets for really fine photo-quality small jobs. I do agree: the media is nearly 95% of success with inkJets. Sergey At 12:33 PM 6/26/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
.. and (3) you may store it at room temperature not afraid bacterial contamination... Sergey
At 10:21 AM 6/26/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
The serum of the animal used in production of the secondary antibody (or the first in a direct labelling experiment) is usually used throughout the labelling procedure, not just during the preincubation. It helps prevent non specific binding of the antibody. The serum is present in excess, sticking to the protein binding sites which would normally attract the antibody. Because the binding is of low affinity it is wise to keep the serum present throughout. Therefore DNS would be used for a donkey secondary; goatNS for a goat secondary etc.
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Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
ImageJ http://rsb.info.nih.gov/ij/ is free and has Java interface that you might be able to incorporate into your own software.
I believe a radial or polar Fourier transform can be used to line up the axis of the two images.
ImageJ is also good for comparing images by hand and not too difficult to add your own code to.
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
} From: "Rohit Bhargava" {rxb29-at-po.cwru.edu} : : Thanks to the many people who responded to a request for information on : image matching on the list and off it as well. While I appreciate the many : references to commercial software, if possible we would really like to : program it into our own lab software to integrate with instrumentation built : in-house. : : For those who may be curious: The best solution appears to be two separate : procedures. One is to use coarse resolution edges to find the correct : rotation angle and then a regression to match pixel for pixel by optimizing : over only the better fit pixels (rejecting 10-20%) to account for artifacts. : This would also identify "hot" pixels where some change may have occured. I : must stress that this procedure suits us because we know what : changes/artifacts are likely to result, is somewhat limited in the range of : problems that can be handled and may not be an optimal approach for all : cases. : : Thanks, : : Rohit : : Rohit Bhargava : National Institutes of Health : Building 5, B1-38W : LCP/NIDDK : Bethesda, MD 20892-0520 : :
200 proof ethanol is 100% free of water. As my friend-chemist told me, for 200 proof ethanol direct synthesis from ethane utilized. There is no distillation involved. So, fresh opened bottle of such ethanol is very good source for "100% ethanol". How long already opened bottle will still be considered 100% I really don't know. It depends from so many factors including hymidity and how fast you are working. Usually I use about half of the bottle and then consider it's 95%. I stopped using molecular sieves a few years ago when completely switch to the diamond knifes. Even with every precaution, there is still chance to have particles from molecular sieve in your plastic. It may damage diamond knife. As for Elanie question, you may "activate" molecular sieve by heating 200-250oC for 18-24 hours. Use approximately 1/10-1/5 of sieve by volume. If you will not leave bottle open forever, molecular sieve will be good for 3-6 month. I did not re-use it. Sergey
At 03:25 PM 6/26/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
a surface charging influences only the primary electron energy (Eo) - including the absolute Bremsstrahlung generation - , according my knowledge and my experience. If you have a stationary charging (not changing during spectrum acquisition time), then you can determine the resulting Eo with checking the Bremstrahlung end in the spectrum. Change the Eo in your quantification procedure and you should get right results! But the charging must be stable in time. Otherwise you have a mix of different Eo and the quantitative correction program must fail!
It is a very interesting question about the energy lost during the slow-down-process caused by surface charging. I think, there is a Bremsstrahlung generation process, emitted with 100% perpendicular only . That is why you have no additional Bremsstrahlung in your spectrum and all calculation are the same like a lower Eo, taking into account a 'normal' SEM - EDX setup. But there should be effects, if you have a EDX detector with zero degree take-off angle (tilted specimen ).
Am I right? Who has some experience?
Frank
"Tomic, Peter (Peter)" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Henk; } } When an electron is slowed on it's approach to the target, where does that energy go? If I were able to instantaneously stop an electron in flight, would it not expend it's entire incident energy as photon radiation? And would that not be "braking radiation?" } } Thanks, } } Peter } } -----Original Message----- } } From: Hendrik O. Colijn [mailto:colijn.1-at-osu.edu] } Sent: Thursday, June 26, 2003 10:28 AM } To: Tomic, Peter (Peter); microscopy-at-sparc5.microscopy.com } Subject: Re: [Bremsstrahlung Radiation] } } Peter, } } Surface charge becomes irrelevant in the creation of brehmstrahlung since } the final energy of the electron winds up being zero. Whether it } decelerates before the sample or within it, you will decelerate the same } amount and so create the same brehmstrahlung. It is a characteristic of } the acceleration of the electron. Synchrotron radiation can be considered } to be a special case of brehmstrahlung where the acceleration of the } electron is perpendicular to its trajectory. } } That said... Surface charge WILL affect your spectrum, since the electron } within the sample is at a lower energy. } } Cheers, } Henk Colijn } } At 09:25 AM 6/26/2003 -0400, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Folks; } } } } Since Bremsstrahlung radiation is also called "braking radiation", is there } } general agreement that the braking radiation will be directly proportional } } to surface charge on the target sample? That is, a surface that becomes } } charged by the e-beam will generate more braking radiation than a surface } } whose charge is at zero. Also, is it fair to say that braking radiation can } } happen within a sold and not just during the flight of the electron down the } } EM column? For example, if a backscattered electron becomes slowed down by } } it's local environment, in a solid or any sample for that matter, it must } } release that energy in the form of photons i. e. Brermsstrahlung radiation, } } yes? } } } } } } Peter Tomic } } Agere Systems } } Allentown, PA } } } } -----Original Message----- } } } From: Frank Eggert [mailto:Eggert-at-mikroanalytik.de] } } Sent: Wednesday, June 25, 2003 7:38 AM } } To: Ritchie Sims } } Cc: Bill Tivol; microscopy-at-sparc5.microscopy.com } } Subject: Re: Real EDS detection limits } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear all, } } } } you are right. The detection limits of an EDX spectrometer depends not only } } from } } statistic. Main cause to change the detection limits to higher values are } } peak } } overlaps and spectra distortions (escape, shelf, tail, pile-up, diffraction } } peaks } } .). } } } } But the basics of calculation of detection limits are signal to noise (peak } } to } } background) ratios. The background with electron probe microanalysis is } } well } } defined with Bremsstrahlung (99% of all background contributions in most } } cases). } } The Bremsstrahlung and its absolute relation to the excited characteristic } } lines } } depends from physics only, mainly from primary electron energy and } } critical } } excitation energies of used line series. It's true in region of 10 { Z { 30 } } it } } can be achieved 0.1% (K-radiation) and for Z } 80 only 0.3% as detection } } limits } } (mean atomic number about 26, 60 s acquisition time, 2000 cps). The } } probability of } } Bremsstrahlung generation rises with higher mean atomic number of specimen. } } The } } detection limit in a weak matrix is much higher compared to a heavy matrix } } (it } } can be factors of 3 and more). A detection limit of 0.05% (mass units) } } for iron } } is possible in a matrix of glass or in biological specimens, but not in Pb. } } } } You will find at the given link an image with detection limits curves, which } } can } } give you some support. These calculated curves give you an idea about 'best } } case' } } detection limits in EPMA without overlaps etc.: } } } } http://www.microanalyst.net/index_e.phtml } } } } . hit [Information] and then scroll down (last image) } } } } } } Frank Eggert } } } } } } } } } } Ritchie Sims wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Yes. } } } } } } I was referring in a general sense to detection limits and/or precision } } } estimates that are delivered by the EDS spectrum-processing software. } } } } } } My point is that while statistical considerations may well give a } } realistic } } } estimate of the precision of determination of higher concentrations, they } } } don't RELIABLY provide meaningful detection limits for EDS, in which } } } overlaps and other spectral artefacts such as coincidence peaks and } } } escape peaks can play a large part. } } } } } } Do you really believe the quoted 'detection limit' of 0.09%? } } } } } } Even without entering the discussion of exactly what is meant by } } 'detection } } } limit'. } } } } } } It would be interesting to hear something on this subject from the EDS } } } system manufacturers. My personal suspicion is that a lot of } } 'quantitative' } } } results, obtained from standardless EDS packages, are quoted to } } } unjustifiable precision, even to two decimal places! } } } } } } cheers } } } } } } rtch } } } } } } } Dear Richie, } } } } Overlaps should not be a problem for Cl and steel. When I was } } } } looking } } } } for Cl in sediment, I saw what were very small peaks that were only } } } } slightly larger than the noise. However, in several examples there } } } } were small amounts of Ti such that the Ti k-alpha peaks were clearly } } } } present and the k-beta peaks were the same size as the Cl k-alpha. } } } } Since I knew that the Ti k-beta peaks were real, that gave me } } } } confidence that the Cl peaks were also. Yours, Bill Tivol EM Scientist } } } } and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code } } } } 114-96 California Institute of Technology Pasadena CA 91125 (626) } } } } 395-8833 tivol-at-caltech.edu } } } } } } } } } } } } } } } } } } } On Sunday, June 22, 2003, at 02:11 PM, Ritchie Sims wrote: } } } } } } } } } If you can't actually see the peak then you can't trust what the } } } } } software says. } } } } } } } } } I bet the detection limit of 0.09% was generated by the software } } } } } from consideration of counting statistics. These may well be OK for } } } } } WDS, but for EDS it's not the counting stats that limit } } } } } detectability, it's the overlaps and other spectral interferences. } } } } } } } } } } } -- } } } Ritchie Sims Ph D Phone : 64 9 3737599 } } } ext 87713 } } } Microanalyst Fax : 64 9 } } 3737435 } } } Department of Geology email : } } } r.sims-at-auckland.ac.nz } } } The University of Auckland } } } Private Bag 92019 } } } Auckland } } } New Zealand } } Hendrik O. Colijn colijn.1-at-osu.edu } Campus Electron Optics Facility Ohio State University } (614) 292-0674 http://www.ceof.ohio-state.edu } Time is that quality of nature which keeps events from happening all at } once. Lately it doesn't seem to be working.
Dear Nancy Did you get my e-mails? I did send it 3 times and got no reply. If you are interested in collaborative research the excess to our facility is free. That means free microscope time and no bench fees. It will stay that way till management changes it. We are equipped with a Technai 12 and a SIS camera. If you send us your samples on grids and in a box we will stain them and view them for you.
The images we will write to a CD and post it to you for interpretation and to publish the work.
To all TEM facilities:
I need someone to help me with plant viruses/TEM research. I do have TEM background but do not have an electron microscope(none in Alaska). I need to look at purified viruses and viruses in plant tissue. I am looking for a EM facility that either contracts research projects or allows one to use their facility for a short segment of time. My immediate need is to have grids of purified virus or leaf dips looked at for virus particles.
Thank you,
Nancy Robertson
Nancy L. Robertson Research Plant Pathologist USDA, ARS 533 E. Fireweed Ave. Palmer Alaska 99645 907-746-9465 office 907-746-2898 lab 907-746-2677 fax ffnlr-at-uaf.edu
Dear All I have a query. I have inherited a Meteor II board and JVC camera with a Y/C output. The board works OK as I can get a real time image in monochrome but cannot fathom out how to get it in colour. Any help appreciated. Jonathan
It just occurs to me that we may have a semantic issue here. Bremsstrahlung radiation is generated whenever a charged particle is accelerated. The continuum portion of a spectrum is also often referred to as bremsstrahlung.
You will generate bremsstrahlung radiation as the electron is being slowed down by a surface charge. This radiation will be in the forward direction and will not be picked up by the EDS detector. It will irradiate your sample producing (I suppose) a minimal amount of "x-ray fluorescence". Additional bremsstrahlung radiation will be created as the electron slows down and is scattered within the sample. This leads to the continuum background seen in the spectrum. The high energy limit of the continuum (Duane-Hunt limit) indicates the maximum energy of the electron within the sample. Surface charge will reduce the Duane-Hunt limit from the nominal beam voltage.
cheers, Henk
At 09:10 AM 6/27/2003 +0200, Frank Eggert wrote: } Henk and Peter, } } a surface charging influences only the primary electron energy (Eo) - } including the absolute Bremsstrahlung generation - , according my } knowledge and my experience. If you have a stationary charging (not } changing during spectrum acquisition time), then you can } determine the resulting Eo with checking the Bremstrahlung end in the } spectrum. Change the Eo in your quantification procedure and you should } get right results! But the charging must be stable in time. Otherwise you } have a mix of different Eo and the } quantitative correction program must fail! } } It is a very interesting question about the energy lost during the } slow-down-process caused by surface charging. I think, there is a } Bremsstrahlung generation process, emitted with 100% perpendicular only . } That is why you have no additional Bremsstrahlung in } your spectrum and all calculation are the same like a lower Eo, taking } into account a 'normal' SEM - EDX setup. But there should be effects, if } you have a EDX detector with zero degree take-off angle (tilted specimen ). } } Am I right? Who has some experience? } } Frank } } } } "Tomic, Peter (Peter)" wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Henk; } } } } When an electron is slowed on it's approach to the target, where does } that energy go? If I were able to instantaneously stop an electron in } flight, would it not expend it's entire incident energy as photon } radiation? And would that not be "braking radiation?" } } } } Thanks, } } } } Peter } } } } -----Original Message----- } } } From: Hendrik O. Colijn [mailto:colijn.1-at-osu.edu] } } Sent: Thursday, June 26, 2003 10:28 AM } } To: Tomic, Peter (Peter); microscopy-at-sparc5.microscopy.com } } Subject: Re: [Bremsstrahlung Radiation] } } } } Peter, } } } } Surface charge becomes irrelevant in the creation of brehmstrahlung since } } the final energy of the electron winds up being zero. Whether it } } decelerates before the sample or within it, you will decelerate the same } } amount and so create the same brehmstrahlung. It is a characteristic of } } the acceleration of the electron. Synchrotron radiation can be considered } } to be a special case of brehmstrahlung where the acceleration of the } } electron is perpendicular to its trajectory. } } } } That said... Surface charge WILL affect your spectrum, since the electron } } within the sample is at a lower energy. } } } } Cheers, } } Henk Colijn } } } } At 09:25 AM 6/26/2003 -0400, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Folks; } } } } } } Since Bremsstrahlung radiation is also called "braking radiation", is } there } } } general agreement that the braking radiation will be directly proportional } } } to surface charge on the target sample? That is, a surface that becomes } } } charged by the e-beam will generate more braking radiation than a surface } } } whose charge is at zero. Also, is it fair to say that braking } radiation can } } } happen within a sold and not just during the flight of the electron } down the } } } EM column? For example, if a backscattered electron becomes slowed down by } } } it's local environment, in a solid or any sample for that matter, it must } } } release that energy in the form of photons i. e. Brermsstrahlung } radiation, } } } yes? } } } } } } } } } Peter Tomic } } } Agere Systems } } } Allentown, PA } } } } } } -----Original Message----- } } } } From: Frank Eggert [mailto:Eggert-at-mikroanalytik.de] } } } Sent: Wednesday, June 25, 2003 7:38 AM } } } To: Ritchie Sims } } } Cc: Bill Tivol; microscopy-at-sparc5.microscopy.com } } } Subject: Re: Real EDS detection limits } } } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear all, } } } } } } you are right. The detection limits of an EDX spectrometer depends not } only } } } from } } } statistic. Main cause to change the detection limits to higher } values are } } } peak } } } overlaps and spectra distortions (escape, shelf, tail, pile-up, } diffraction } } } peaks } } } .). } } } } } } But the basics of calculation of detection limits are signal to noise } (peak } } } to } } } background) ratios. The background with electron probe microanalysis is } } } well } } } defined with Bremsstrahlung (99% of all background contributions in most } } } cases). } } } The Bremsstrahlung and its absolute relation to the excited } characteristic } } } lines } } } depends from physics only, mainly from primary electron energy and } } } critical } } } excitation energies of used line series. It's true in region of 10 { } Z { 30 } } } it } } } can be achieved 0.1% (K-radiation) and for Z } 80 only 0.3% as detection } } } limits } } } (mean atomic number about 26, 60 s acquisition time, 2000 cps). The } } } probability of } } } Bremsstrahlung generation rises with higher mean atomic number of } specimen. } } } The } } } detection limit in a weak matrix is much higher compared to a heavy } matrix } } } (it } } } can be factors of 3 and more). A detection limit of 0.05% (mass units) } } } for iron } } } is possible in a matrix of glass or in biological specimens, but not } in Pb. } } } } } } You will find at the given link an image with detection limits curves, } which } } } can } } } give you some support. These calculated curves give you an idea about } 'best } } } case' } } } detection limits in EPMA without overlaps etc.: } } } } } } http://www.microanalyst.net/index_e.phtml } } } } } } . hit [Information] and then scroll down (last image) } } } } } } } } } Frank Eggert } } } } } } } } } } } } } } } Ritchie Sims wrote: } } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -----------------------------------------------------------------------. } } } } } } } } Yes. } } } } } } } } I was referring in a general sense to detection limits and/or precision } } } } estimates that are delivered by the EDS spectrum-processing software. } } } } } } } } My point is that while statistical considerations may well give a } } } realistic } } } } estimate of the precision of determination of higher } concentrations, they } } } } don't RELIABLY provide meaningful detection limits for EDS, in which } } } } overlaps and other spectral artefacts such as coincidence peaks and } } } } escape peaks can play a large part. } } } } } } } } Do you really believe the quoted 'detection limit' of 0.09%? } } } } } } } } Even without entering the discussion of exactly what is meant by } } } 'detection } } } } limit'. } } } } } } } } It would be interesting to hear something on this subject from the EDS } } } } system manufacturers. My personal suspicion is that a lot of } } } 'quantitative' } } } } results, obtained from standardless EDS packages, are quoted to } } } } unjustifiable precision, even to two decimal places! } } } } } } } } cheers } } } } } } } } rtch } } } } } } } } } Dear Richie, } } } } } Overlaps should not be a problem for Cl and steel. When I was } } } } } looking } } } } } for Cl in sediment, I saw what were very small peaks that were only } } } } } slightly larger than the noise. However, in several examples there } } } } } were small amounts of Ti such that the Ti k-alpha peaks were clearly } } } } } present and the k-beta peaks were the same size as the Cl k-alpha. } } } } } Since I knew that the Ti k-beta peaks were real, that gave me } } } } } confidence that the Cl peaks were also. Yours, Bill Tivol EM } Scientist } } } } } and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code } } } } } 114-96 California Institute of Technology Pasadena CA 91125 (626) } } } } } 395-8833 tivol-at-caltech.edu } } } } } } } } } } } } } } } } } } } } } } } } On Sunday, June 22, 2003, at 02:11 PM, Ritchie Sims wrote: } } } } } } } } } } } If you can't actually see the peak then you can't trust what the } } } } } } software says. } } } } } } } } } } } I bet the detection limit of 0.09% was generated by the software } } } } } } from consideration of counting statistics. These may well be OK for } } } } } } WDS, but for EDS it's not the counting stats that limit } } } } } } detectability, it's the overlaps and other spectral interferences. } } } } } } } } } } } } } } -- } } } } Ritchie Sims Ph D Phone : 64 9 3737599 } } } } ext 87713 } } } } Microanalyst Fax : 64 9 } } } 3737435 } } } } Department of Geology email : } } } } r.sims-at-auckland.ac.nz } } } } The University of Auckland } } } } Private Bag 92019 } } } } Auckland } } } } New Zealand } } } } Hendrik O. Colijn colijn.1-at-osu.edu } } Campus Electron Optics Facility Ohio State University } } (614) 292-0674 http://www.ceof.ohio-state.edu } } Time is that quality of nature which keeps events from happening all at } } once. Lately it doesn't seem to be working.
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
Bremsstrahlung is generated in perpendicular direction (90 degree angle) to the direction of charged particles accelaration (positive and negative). Therefore the stopping of charged particles (electrons) generates Bremsstrahlung to 0 degree take off angle detector position (only in this direction, if the electrical field is homogeneous), and not in direction to specimen.
That is why you should be able to measure additional Bremsstrahlung parts in spectrum, if the detectors position is at 0 degree take off. I expect a spectrum with more Bremsstrahlung (from zero to specimen- charging in keV) than in the same specimen without charging but with a primary electron energy, which is reduced by the same energy of charging. It is an interesting question, isn't it?
The Bremsstrahlung from the specimen has an isotrope distribution because of the scattering effects (there exists all electron directions after a couple of scatter acts). But we know, Bremsstrahlung generation has a anisotrope component by bombarding a target. That part comes direct from surface (all electrons still have one direction). It is to taken into account, if you calculate the Bremsstrahlung distribution. The anisotropy is becoming more important with reducing the take off angle of the detector. You have more Bremsstrahlung with lower angles. The same processes occur with the electron beam like in direct surface of specimen. The electron beam de-accelerated in charge field of specimen (without changing of direction, if the field is homogeneous). The acceleration radiation (Bremsstrahlung) emits perpendicularly to the electron beam direction.
Are there other comments? Is there somebody, who has already measured this effect yet?
Frank
"Hendrik O. Colijn" wrote:
} Frank and Peter, } } It just occurs to me that we may have a semantic issue } here. Bremsstrahlung radiation is generated whenever a charged particle is } accelerated. The continuum portion of a spectrum is also often referred to } as bremsstrahlung. } } You will generate bremsstrahlung radiation as the electron is being slowed } down by a surface charge. This radiation will be in the forward direction } and will not be picked up by the EDS detector. It will irradiate your } sample producing (I suppose) a minimal amount of "x-ray } fluorescence". Additional bremsstrahlung radiation will be created as the } electron slows down and is scattered within the sample. This leads to the } continuum background seen in the spectrum. The high energy limit of the } continuum (Duane-Hunt limit) indicates the maximum energy of the electron } within the sample. Surface charge will reduce the Duane-Hunt limit from } the nominal beam voltage. } } cheers, } Henk } } At 09:10 AM 6/27/2003 +0200, Frank Eggert wrote: } } Henk and Peter, } } } } a surface charging influences only the primary electron energy (Eo) - } } including the absolute Bremsstrahlung generation - , according my } } knowledge and my experience. If you have a stationary charging (not } } changing during spectrum acquisition time), then you can } } determine the resulting Eo with checking the Bremstrahlung end in the } } spectrum. Change the Eo in your quantification procedure and you should } } get right results! But the charging must be stable in time. Otherwise you } } have a mix of different Eo and the } } quantitative correction program must fail! } } } } It is a very interesting question about the energy lost during the } } slow-down-process caused by surface charging. I think, there is a } } Bremsstrahlung generation process, emitted with 100% perpendicular only . } } That is why you have no additional Bremsstrahlung in } } your spectrum and all calculation are the same like a lower Eo, taking } } into account a 'normal' SEM - EDX setup. But there should be effects, if } } you have a EDX detector with zero degree take-off angle (tilted specimen ). } } } } Am I right? Who has some experience? } } } } Frank } }
Hi Sharon, Try putting your lead solution in a syringe (no needle). Put the stopper and parafilm on it. Keep in the fridge. Before using it, get rid of the first 3-4 drops. It works well. Emmanuelle
Hi again, I did not mention it in my first message but, with the syringe you do not need mineral oil or anything else. That is the advantage of that technique , no contamination by other chemicals. Emmanuelle
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, June 27, 2003 at 02:48:51 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin Andreev
Organization: 54 st"Ivan Rilski"
Education: 6-8th Grade Middle School
Location: Sofia Bulgaria
Question: Can I use liquid paraffin as immersion oil? And how can I clean up my lenses after work?
In an oven at 150 deg C or so (not really critical, just not too hot), for however long it takes for the indicator to turn blue again. Usually one to 3 hours. There is no better method to dry 100% EtOH that I know of, and to keep it dry. You've probably read the earlier posts about putting the sieve in dialysis tubing to contain the dust -- if you didn't write one of them (I forget, sorry). Phil
} Hello Everyone } The question has arisen here about drying molecular sieve. How often } do you dry your molecular sieve and when you do, what temperature } and for how long? } } Alternatively does anyone have a better method of maintaining 100% ethanol. } Elaine } } -- } Dr. Elaine Humphrey } Director, BioImaging Facility } President, Microscopy Society of Canada } University of British Columbia } 6270 University Blvd, mail-stop Botany } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-interchange.ubc.ca } website: www.emlab.ubc.ca -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
I would expect more problems with embedding resins, but ... I'm really a SEM jock, so, yes, I think -- I admit that -- that I see better drying with sieve-dried EtOH when looking at cell membranes in our high resolution FESEM. Lower magnifications, say { 10,000 X, may not show a big difference. But recall that during critical point drying, if there is free water within the specimen, then at least where the water is, you're drying from water not liquid CO2.
Phil
} And while we are on the subject of drying ethanol, what is the } feeling about whether this is necessary? What is the problem if } your "100%" ethanol has a small (1-2%) of water in it. I never dry } my ethanol. I used to only use fresh bottles but over the last } couple of years have evolved into using "recently opened" (less than } 2 weeks or so) bottles and see no difference. Am i losing something } in morphological quality or sectioning quality that I don't notice? } Tom } } } } At 02:53 PM 6/26/2003 -0700, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Would you agree with this statement and empirical observation. When I image a sample that is highly insulative, and I know the surface is partially charged toward Eo, I get a larger background noise level than when I do the same to a highly conductive specimen. Now the effective Eo is diminished by the quantity of that surface charge on the sample. This charge will slow the electron down by some amount.
Question: No matter what axis photon emission takes place and whether the EDX detector sees it, is this not braking radiation happening during the "free space" flight of the electron, prior to solid interaction? If I had the EM column loaded with detectors at every conceivable angle, wouldn't they "see" these photons?
Peter
-----Original Message----- } From: Frank Eggert [mailto:Eggert-at-mikroanalytik.de] Sent: Friday, June 27, 2003 3:11 AM To: Tomic, Peter (Peter) Cc: Hendrik O. Colijn; microscopy-at-sparc5.microscopy.com
Dear members of the list server,
Has anybody got experience with processing snail tissue for EM? I am trying to process neurons of the snail Helix pomatia for the TEM, but had only poor success so far.
- fine structural preservation is poor- the membranes are hardly visible. - there is extremely poor contrast- the tissue does not seem to take up lead citrate and uranyl acetate. - the tissue is very hard and difficult to section.
I have been using our usual method that works extremely well in insects: -Fixing in 0.1M phosphate buffer, pH 7.2 with a 2% glutaraldehyde, 0.5% paraformaldehyde-mixture -rinsing in the same buffer, -1% Osmium tetraoxide in the same buffer, -dehydration in a acetone series and embedding in TAAB epoxy resin.
Has anybody got an idea about where we have gone wrong? Is there a special osmolarity/different pH we should stick to in the snail, or should we use sodium cacodylate buffer instead of phosphate buffer? So far, I refrained from using cacodylate buffer since the tissue is being fixed in Italy and then sent to Austria by our Italian colleages, and it would be difficult to transport harmful material.
I would be grateful for help by a more experienced snail EM investigator.
Gerd Leitinger
-- Dr. Gerd Leitinger Department of Histology and Embryology Karl Franzens University of Graz Harrachgasse 21 A-8010 Graz Austria
As far as I remember, the Meteor boards came in different flavors. Perhaps you have one that can only acquire b/w?
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Biomaterials [mailto:Biomat-at-eastman.ucl.ac.uk] Sent: Friday, June 27, 2003 4:41 AM To: Microscopy-at-sparc5.microscopy.com
Dear All I have a query. I have inherited a Meteor II board and JVC camera with a Y/C output. The board works OK as I can get a real time image in monochrome but cannot fathom out how to get it in colour. Any help appreciated. Jonathan
I think, the answer is Yes and No. If you slow down or "bend" electrons in an electric field, such as in a microscope, it is an acceleration, and the electrons will emit radiation. However, the radiation is probably so low in energy that it is way beyond the detection limit of your detector. If electrons bent at macroscopic radii would emit radiation, your computer and every electric device should give off x-rays, which, to my knowledge, they don't.
On the other hand, if you put a field on your sample, you don't really slow down the electrons, you rather make them turn and slam into something else. Could be the sample holder, the stage, even the walls of the chamber. There they will still create bremsstrahlung. In other words: your signal from your sample goes down, the background radiation does not really change significantly. Result: Your S/N ratio goes up.
mike
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Sent: Friday, June 27, 2003 6:22 AM To: Mike Bode
Folks;
Since Bremsstrahlung radiation is also called "braking radiation", is there general agreement that the braking radiation will be directly proportional to surface charge on the target sample? That is, a surface that becomes charged by the e-beam will generate more braking radiation than a surface whose charge is at zero. Also, is it fair to say that braking radiation can happen within a sold and not just during the flight of the electron down the EM column? For example, if a backscattered electron becomes slowed down by it's local environment, in a solid or any sample for that matter, it must release that energy in the form of photons i. e. Brermsstrahlung radiation, yes?
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Frank Eggert [mailto:Eggert-at-mikroanalytik.de] Sent: Wednesday, June 25, 2003 7:38 AM To: Ritchie Sims Cc: Bill Tivol; microscopy-at-sparc5.microscopy.com
Dear all,
you are right. The detection limits of an EDX spectrometer depends not only from statistic. Main cause to change the detection limits to higher values are peak overlaps and spectra distortions (escape, shelf, tail, pile-up, diffraction peaks .).
But the basics of calculation of detection limits are signal to noise (peak to background) ratios. The background with electron probe microanalysis is well defined with Bremsstrahlung (99% of all background contributions in most cases). The Bremsstrahlung and its absolute relation to the excited characteristic lines depends from physics only, mainly from primary electron energy and critical excitation energies of used line series. It's true in region of 10 { Z { 30 it can be achieved 0.1% (K-radiation) and for Z } 80 only 0.3% as detection limits (mean atomic number about 26, 60 s acquisition time, 2000 cps). The probability of Bremsstrahlung generation rises with higher mean atomic number of specimen. The detection limit in a weak matrix is much higher compared to a heavy matrix (it can be factors of 3 and more). A detection limit of 0.05% (mass units) for iron is possible in a matrix of glass or in biological specimens, but not in Pb.
You will find at the given link an image with detection limits curves, which can give you some support. These calculated curves give you an idea about 'best case' detection limits in EPMA without overlaps etc.:
http://www.microanalyst.net/index_e.phtml
. hit [Information] and then scroll down (last image)
Frank Eggert
Ritchie Sims wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Yes. } } I was referring in a general sense to detection limits and/or precision } estimates that are delivered by the EDS spectrum-processing software. } } My point is that while statistical considerations may well give a realistic } estimate of the precision of determination of higher concentrations, they } don't RELIABLY provide meaningful detection limits for EDS, in which } overlaps and other spectral artefacts such as coincidence peaks and } escape peaks can play a large part. } } Do you really believe the quoted 'detection limit' of 0.09%? } } Even without entering the discussion of exactly what is meant by 'detection } limit'. } } It would be interesting to hear something on this subject from the EDS } system manufacturers. My personal suspicion is that a lot of 'quantitative' } results, obtained from standardless EDS packages, are quoted to } unjustifiable precision, even to two decimal places! } } cheers } } rtch } } } Dear Richie, } } Overlaps should not be a problem for Cl and steel. When I was } } looking } } for Cl in sediment, I saw what were very small peaks that were only } } slightly larger than the noise. However, in several examples there } } were small amounts of Ti such that the Ti k-alpha peaks were clearly } } present and the k-beta peaks were the same size as the Cl k-alpha. } } Since I knew that the Ti k-beta peaks were real, that gave me } } confidence that the Cl peaks were also. Yours, Bill Tivol EM Scientist } } and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code } } 114-96 California Institute of Technology Pasadena CA 91125 (626) } } 395-8833 tivol-at-caltech.edu } } } } } } } } } On Sunday, June 22, 2003, at 02:11 PM, Ritchie Sims wrote: } } } } } If you can't actually see the peak then you can't trust what the } } } software says. } } } } } I bet the detection limit of 0.09% was generated by the software } } } from consideration of counting statistics. These may well be OK for } } } WDS, but for EDS it's not the counting stats that limit } } } detectability, it's the overlaps and other spectral interferences. } } } } } -- } Ritchie Sims Ph D Phone : 64 9 3737599 } ext 87713 } Microanalyst Fax : 64 9 3737435 } Department of Geology email : } r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand
I think you are talking about pair production, not bremsstrahlung. If my memory serves me right, a charge can create bremsstrahlung also in vacuum. How else would a synchroton work?
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Thursday, June 26, 2003 1:30 PM To: microscopy-at-sparc5.microscopy.com
On Thursday, June 26, 2003, at 06:25 AM, Tomic, Peter (Peter) wrote:
} Since Bremsstrahlung radiation is also called "braking radiation", is } there } general agreement that the braking radiation will be directly } proportional } to surface charge on the target sample? That is, a surface that becomes } charged by the e-beam will generate more braking radiation than a } surface } whose charge is at zero. Also, is it fair to say that braking } radiation can } happen within a sold and not just during the flight of the electron } down the } EM column? For example, if a backscattered electron becomes slowed } down by } it's local environment, in a solid or any sample for that matter, it } must } release that energy in the form of photons i. e. Brermsstrahlung } radiation, } yes? } Dear Peter, Bremsstrahlung is not proportional to the charge--surface or bulk--on the sample. It can, be produced by neutral atoms and is proportional to a power of Z. (I don't have my reference material with me, but I think it is Z^2.) Since both energy and momentum must be conserved, bremsstrahlung must involve another particle besides the electron and photon, so it cannot be generated in a perfect vacuum; however, in addition to being generated in solids, bremsstrahlung can be generated in gasses, so, if the electron happens to pass near enough to a residual atom as it travels down the EM column, bremsstrahlung can be produced (extremely rarely). In addition to bremsstrahlung, there are several energy-loss processes that do not involve photon production, such as ionization, excitation, plasmon production, etc., so when electrons are slowed down, most of the energy released is not in the form of photons. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
} } Dear all, } } } } I am interested in measuring the axial and radial thermal expansion } } coefficient in fibers (diameter 7-15ìm) up to 1200 C. So I address to labs } } with heating stage equipment for SEM or TEM. To be more precise the job } } description is as follows: } } We want to measure the coefficient of thermal expansion (CTE) in three } } types of ceramic fibers (carbon, Al2O3, SiC) in both axial and radial } } direction up to 1200 C. The diameters are 7ìm for carbon, 14 ìm for Al2O3 } } and 15ìm for SiC. If anyone thinks he can undertake the job then we can } } discuss further about the cost, the time it takes and other details. We } } have attempted to do it with optical microscopy but seems impossible, } needs } } greater magnification. } } On the other hand if anyone has heating stage equipment for SEM and he } } doesn't need it for this period then we can discuss about the perspective } of } } borrowing it, or any formula that would allow us to do the job. } } In any case I would appreciate any information regarding labs having this } } kind of facilities because in the web it seems that there are not so many. } } Thank you very much for your time. } } } } } } Theodoros Loutas } } Dipl. Mechanical Engineer, Research assistant } } Applied Mechanics Lab } } Dpt. of Mechanical and Aeronautics Engineering } } University of Patras, Greece } } }
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For those of you who were interested in how to prepare replicas of teeth for analysis in the SEM, my colleague, Dr. Gina Semprebon, has just returned and would be happy to discuss this analysis with you off-line.
You can reach her at g.sempreb-at-baypath.edu
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
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All charged particles radiate Bremmstrahlung whenever they are accelerated (i.e. whenever their direction and/or velocity is changed). Digging back into my Atomic Physics (which was far more years ago than I care to reveal!), it seems to me that this is a consequence of the impossibility of conserving both energy and momentum in a general 2-body elastic interaction. The energy of the photon depends on the magnitude of the acceleration, though I can't tell you the form of the dependence. I'll guess that it is linear for illustrative purposes.
However...
The energy depends on the magnitude of the acceleration. By electron standards, the field outside a charged surface is miniscule (less than a volt per micron). Electrons approaching a charged surface will not be stopped - they will be deflected. To get an order of magnitude, we can say that the radius of curvature is of the order of the working distance, say 5 mm.
Electrons are deflected elastically by neutral atoms only if the incident electron penetrates inside the atomic electron cloud. The "size" of an atom is far less than 0.1nm - lets approximate this distance as 0.005nm. The difference between these numbers is 10^9. For a given incident electron energy, the acceleration is inversely proportional to the radius of curvature (ignoring relativistic effects). Hence, if an electron/atom interaction generates a 1KeV bremsstrahlung photon, then the electron/surface charge interaction will generate a 1 micro-eV bremsstrahlung photon, for an equal angular deviation.
Hence ...
Yes, the electron acceleration by a surface does produce bremsstrahlung, but the photon energy is so low you don't detect it (it is actually UHF RF). Since the individual photon energy is so low, the total energy loss is also negligible, so the signal is not observable even on a sensitive radio receiver.
You can think of it another way. The wavelength of the photon must be comparable to the distance over which the charge experiences an acceleration (think of the oscillating charge in a radio antenna/aeriel). The total distance over which an electron experiences the charge of the atomic nucleus is a few times the closest approach - lets say 0.02 nm or, using the units of Wavelength spectroscopy, 0.2 Angstroms. An x-ray with a wavelength of 0.2 Angstroms has an energy of roughly 60KeV - higher than typical for bremsstrahlung, but close enough to illustrate my point.
So ...
Why do you observe a degradation in the x-ray spectrum when the sample charges? Because the energy of the electrons reaching the surface is reduced. The cross-section for production of characteristic x-rays goes down faster with incident energy than the cross-section for production of bremsstrahlung (at least in typical cases in EDX analysis). So the signal-to-noise ratio of the resulting spectrum is lower than would be the case of a neutral sample surface.
Everything I have written is generalized, rough and qualitative, of course, but illustrates the point.
The angular dependence of bremsstrahlung emission is another interesting point, but I'll leave that to someone else.
Tony.
PS I have unsubscribed from the list as I am going on vacation, so I won't see anything you write in response, unless you address it to me specifically.
PPS One physicist who wrote about bremsstrahlung production (back in the '70's of the last century) was our own friendly neighborhood SysOp. There were several others, dating back to Kramers in 1923.
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
On Thursday, June 26, 2003, at 07:28 AM, Hendrik O. Colijn wrote:
} Surface charge becomes irrelevant in the creation of brehmstrahlung } since the final energy of the electron winds up being zero. Whether } it decelerates before the sample or within it, you will decelerate the } same amount and so create the same brehmstrahlung. It is a } characteristic of the acceleration of the electron. Synchrotron } radiation can be considered to be a special case of brehmstrahlung } where the acceleration of the electron is perpendicular to its } trajectory. } } That said... Surface charge WILL affect your spectrum, since the } electron within the sample is at a lower energy. } Dear Hendrik, Bremsstrahlung photons can have any energy from 0 to the energy of the incident electron (or other charged particle), E_e. The number of photons produced at a given energy E_g is proportional to E_e - E_g, so the electron only rarely winds up with 0 energy. Surface charge will have no effect on the spectrum. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
I meant DOWN. The S/N ratio goes down, not up. Sorry.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Friday, June 27, 2003 9:38 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Peter,
I think, the answer is Yes and No. If you slow down or "bend" electrons in an electric field, such as in a microscope, it is an acceleration, and the electrons will emit radiation. However, the radiation is probably so low in energy that it is way beyond the detection limit of your detector. If electrons bent at macroscopic radii would emit radiation, your computer and every electric device should give off x-rays, which, to my knowledge, they don't.
On the other hand, if you put a field on your sample, you don't really slow down the electrons, you rather make them turn and slam into something else. Could be the sample holder, the stage, even the walls of the chamber. There they will still create bremsstrahlung. In other words: your signal from your sample goes down, the background radiation does not really change significantly. Result: Your S/N ratio goes up.
mike
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Sent: Friday, June 27, 2003 6:22 AM To: Mike Bode
Folks;
Since Bremsstrahlung radiation is also called "braking radiation", is there general agreement that the braking radiation will be directly proportional to surface charge on the target sample? That is, a surface that becomes charged by the e-beam will generate more braking radiation than a surface whose charge is at zero. Also, is it fair to say that braking radiation can happen within a sold and not just during the flight of the electron down the EM column? For example, if a backscattered electron becomes slowed down by it's local environment, in a solid or any sample for that matter, it must release that energy in the form of photons i. e. Brermsstrahlung radiation, yes?
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Frank Eggert [mailto:Eggert-at-mikroanalytik.de] Sent: Wednesday, June 25, 2003 7:38 AM To: Ritchie Sims Cc: Bill Tivol; microscopy-at-sparc5.microscopy.com
Dear all,
you are right. The detection limits of an EDX spectrometer depends not only from statistic. Main cause to change the detection limits to higher values are peak overlaps and spectra distortions (escape, shelf, tail, pile-up, diffraction peaks .).
But the basics of calculation of detection limits are signal to noise (peak to background) ratios. The background with electron probe microanalysis is well defined with Bremsstrahlung (99% of all background contributions in most cases). The Bremsstrahlung and its absolute relation to the excited characteristic lines depends from physics only, mainly from primary electron energy and critical excitation energies of used line series. It's true in region of 10 { Z { 30 it can be achieved 0.1% (K-radiation) and for Z } 80 only 0.3% as detection limits (mean atomic number about 26, 60 s acquisition time, 2000 cps). The probability of Bremsstrahlung generation rises with higher mean atomic number of specimen. The detection limit in a weak matrix is much higher compared to a heavy matrix (it can be factors of 3 and more). A detection limit of 0.05% (mass units) for iron is possible in a matrix of glass or in biological specimens, but not in Pb.
You will find at the given link an image with detection limits curves, which can give you some support. These calculated curves give you an idea about 'best case' detection limits in EPMA without overlaps etc.:
http://www.microanalyst.net/index_e.phtml
. hit [Information] and then scroll down (last image)
Frank Eggert
Ritchie Sims wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Yes. } } I was referring in a general sense to detection limits and/or precision } estimates that are delivered by the EDS spectrum-processing software. } } My point is that while statistical considerations may well give a realistic } estimate of the precision of determination of higher concentrations, they } don't RELIABLY provide meaningful detection limits for EDS, in which } overlaps and other spectral artefacts such as coincidence peaks and } escape peaks can play a large part. } } Do you really believe the quoted 'detection limit' of 0.09%? } } Even without entering the discussion of exactly what is meant by 'detection } limit'. } } It would be interesting to hear something on this subject from the EDS } system manufacturers. My personal suspicion is that a lot of 'quantitative' } results, obtained from standardless EDS packages, are quoted to } unjustifiable precision, even to two decimal places! } } cheers } } rtch } } } Dear Richie, } } Overlaps should not be a problem for Cl and steel. When I was } } looking } } for Cl in sediment, I saw what were very small peaks that were only } } slightly larger than the noise. However, in several examples there } } were small amounts of Ti such that the Ti k-alpha peaks were clearly } } present and the k-beta peaks were the same size as the Cl k-alpha. } } Since I knew that the Ti k-beta peaks were real, that gave me } } confidence that the Cl peaks were also. Yours, Bill Tivol EM Scientist } } and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code } } 114-96 California Institute of Technology Pasadena CA 91125 (626) } } 395-8833 tivol-at-caltech.edu } } } } } } } } } On Sunday, June 22, 2003, at 02:11 PM, Ritchie Sims wrote: } } } } } If you can't actually see the peak then you can't trust what the } } } software says. } } } } } I bet the detection limit of 0.09% was generated by the software } } } from consideration of counting statistics. These may well be OK for } } } WDS, but for EDS it's not the counting stats that limit } } } detectability, it's the overlaps and other spectral interferences. } } } } } -- } Ritchie Sims Ph D Phone : 64 9 3737599 } ext 87713 } Microanalyst Fax : 64 9 3737435 } Department of Geology email : } r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand
} We have two elderly SEM's: a JEOL 35 and a ISI DS 130. One of the } instruments must be surplussed. Which instrument would you recomment to } keep?
In response to Pauls query, we were operating an electron-beam lithography set-up with a modified ISI DS 130 before we transitioned on to a JEOL 6000FS Nanowriter and this SEM has many useful features that come in handy. It has a very good ultra-high resolution first stage and a larger second stage for flexibility in sample sizing and types. The max mag is about 240kX in the second stage operation and I have personally seen reasonable image quality persisting upto 100 - 150kX.
Operation-wise, we haven't had too many problems requiring service over the last 5 years ... mainly just minor things such as one diffusion pump oil change and a few faulty capacitors here and there. The only issue here could be that ISI is out of business and finding parts and service might be an issue. Our present contract is through a company called Scanservice Corp somewhere in CA and they seem to be doing a pretty decent job of things.
I don't know much about the JEOL 35 but then my suggestion would be to retain the SEM that best suits your needs presently.
Chetan Prasad ------------------------------------ Graduate Research Associate Nanostructures Research Group Department of Electrical Engineering Arizona State University Tempe, AZ 85287-5706 Phone : 480-965-4097 Fax : 480-965-8058 -------------------------------------
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} As my friend-chemist told } me, for 200 proof ethanol direct synthesis from ethane utilized. } There is no distillation involved.
Is this really so in the USA?
I was under the impression that fermentation was the main source of ethanol all over the world.
In New Zealand we make methanol from natural gas, but ethanol is manufactured, by fermentation, from whey. This does, of course, involve distillation, but there's no problem industrially removing the 5% or so of water which distills over with the ethanol. It used to be removed by further distillation with benzene, but that was phased out some years ago because of the carcinogenicity of benzene. The small residue of benzene used to be a good argument against using lab alcohol in the Xmas party punch.
There's quite an industry in New Zealand making budget-priced spirits (gin, vodka, etc) from the whey alcohol.
Somehow the gin and tonic doesn't seem to slip down so well when you know it was made from milk.
BTW, 100% ethanol is not exactly 200% proof, nor is 50% ethanol exactly 100% proof. Proof spirit was originally defined as spirit with just enough ethanol in it so that standard gunpowder, when saturated with the mixture in question, would still burn. It's within a few percent of 50.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
} } Would you agree with this statement and empirical observation. When I image a sample that is highly insulative, and I know the surface is partially charged toward Eo, I get a larger background noise level than when I do the same to a highly conductive specimen. Now the effective Eo is diminished by the quantity of that surface charge on the sample. This charge will slow the electron down by some amount. }
I agree. It is the same with identical specimen and lower Eo but without charging (if specimen would be conductive).
} } Question: No matter what axis photon emission takes place and whether the EDX detector sees it, is this not braking radiation happening during the "free space" flight of the electron, prior to solid interaction?
A question of definition. It is radiation due to acceleration of charged particles. I would say 'Bremsstrahlung' too. It is Bremsstrahlung not in Coulomb field of 'single' nuclei, but in 'collective' electrical field due to specimen charging. In synchrotron the charged particle radiation due to acceleration in electrical or magnetical fields is called 'Synchrotron' radiation (different types, simple synchrotron radiation, Wigglers and so on).
} If I had the EM column loaded with detectors at every conceivable angle, wouldn't they "see" these photons?
According my opinion, the flat mounted detectors will see them (0 degree take off angle, x-ray acceptance angle perpendicular to electron beam path). But I'm not sure!
I don't know exactly, but a interesting question. As I remember, 20 keV electrons in SEM has 0.2c - relativistic too. Therefore, differences between TEM and SEM shouldn't be a question of relativistic considerations.
Electrons have very high energy and specimen are very thin in TEM, in comparison to SEM. That is why only the perpendicular acceleration due to Coulomb field of nuclei has a not zero probability. There is no chance for e.g. a full stop (not enough atoms along the electron path) and acceleration components in same direction like the electron flight path. Most electrons have no interaction in thin specimen. Electrons in SEM 'see' very thick specimens. They do interact with the specimen (100 per cent). There are high probabilities for acceleration components in flight path direction even before first interaction (Bremsstrahlung of high energy in perpendicular direction), but much more after first interaction because of all scattering processes.
May be, that is why in TEM the main components of Bremsstrahlung are into electron path direction and you are right. I'm not sure to be right in all considerations.
Cheers,
Frank http://www.microanalyst.net
"Hendrik O. Colijn" schrieb:
} Hi Frank, } } I'm a TEM guy, so I tend to think in terms of } 100kV energies. At } relativistic velocities (which is true in TEM) the radiation becomes } forward scattered in the lab frame of reference. I'm not sure how true } this is for SEM energies. } } cheers, } Henk }
I have seen postgraduate research areas where I suspect the molecular sieve is never dried. The students used LR White so it probably did not matter.
I use araldite or Spurr's resin and am a fanatic about dry ethanol. I dry the molecular sieve when the bottle is empty. I put it in a 200 degree C oven for 3hrs.
Dave
On Fri, 27 Jun 2003 08:56:17 -0500 Philip Oshel {peoshel-at-wisc.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Tom, } } I would expect more problems with embedding resins, but ... I'm } really a SEM jock, so, yes, I think -- I admit that -- that I see } better drying with sieve-dried EtOH when looking at cell membranes in } our high resolution FESEM. Lower magnifications, say { 10,000 X, may } not show a big difference. But recall that during critical point } drying, if there is free water within the specimen, then at least } where the water is, you're drying from water not liquid CO2. } } Phil } } } And while we are on the subject of drying ethanol, what is the } } feeling about whether this is necessary? What is the problem if } } your "100%" ethanol has a small (1-2%) of water in it. I never dry } } my ethanol. I used to only use fresh bottles but over the last } } couple of years have evolved into using "recently opened" (less than } } 2 weeks or so) bottles and see no difference. Am i losing something } } in morphological quality or sectioning quality that I don't notice? } } Tom } } } } } } } } At 02:53 PM 6/26/2003 -0700, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hello Everyone } } } The question has arisen here about drying molecular sieve. How } } } often do you dry your molecular sieve and when you do, what } } } temperature and for how long? } } } } } } Alternatively does anyone have a better method of maintaining 100% ethanol. } } } Elaine } } } } } } -- } } } Dr. Elaine Humphrey } } } Director, BioImaging Facility } } } President, Microscopy Society of Canada } } } University of British Columbia } } } 6270 University Blvd, mail-stop Botany } } } Vancouver, BC } } } CANADA, V6T 1Z4 } } } Phone: 604-822-3354 } } } FAX: 604-822-6089 } } } e-mail: ech-at-interchange.ubc.ca } } } website: www.emlab.ubc.ca } } } } Thomas E. Phillips, PhD } } Associate Professor of Biological Sciences } } Director, Molecular Cytology Core } } 3 Tucker Hall } } University of Missouri } } Columbia, MO 65211-7400 } } } } 573-882-4712 (office) } } 573-882-0123 (fax) } } PhillipsT-at-missouri.edu } } -- } Philip Oshel } Supervisor, BBPIC microscopy facility } Department of Animal Sciences } University of Wisconsin } 1675 Observatory Drive } Madison, WI 53706 - 1284 } voice: (608) 263-4162 } fax: (608) 262-5157 (dept. fax) }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
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All things being equal I vote for keeping the JEOL 35. I have used an ISI and a JEOL 35. My limited personal experience was more positive with the JEOL 35. It's a higher-quality instrument. Others have mentioned to me that ISI is a lower-quality instrument.
Stu Smalinskas, P.E. Metallurgist (734) 414-6862 SKF USA Plymouth, Michigan
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My late husband, Chief Vincent Kamara , a native of Mende District in the Northerh province of Sierra Leone, was the General Manager of Sierra Leone Mining Corporation (S.L.M.C.) Freetown . According to my late husband, this money was the income accrued from mining coporation's over draft and minor sales. Before the peak of the civil war between the rebels forces of Major Paul Koroma and the combined forces of ECOMOG peace keeping operation that almost destroyed my country, following the forceful removal from power of the civilian elected President Ahmed Tejan Kabbah by the rebels. My late husband had already made arrangement for us be evacuated to Abidjan, Cote d' Ivoire with the Certificate of Deposit issued to him by the security firm in Abidjan. During the war in my country, and following the indiscriminate looting of Public and Government properties by the rebel forces, the Sierra Leone Mining Corp. was one of the target looted and destroyed.
My late husband, including other top Government functionaries were attacked and killed by the rebels in November 2000 because of his relationship wi usbandŽs death dashed our hope of survival. I and my son are now alone in this strange country suffering without any care or help. Without any relation, we are now like refugees.Our only hope now is in you and the box deposited in the security firm. To this effect, I humbly solicityour assistance in the followings ways.
1.to assist me claim this box from the security firm as our beneficiary;
2.to transfer the money in your name to your country
3.to make a good arrangement for a joint business investment on our behalf in your country with you as our adviser;
4.To secure a college for my son in your country after the money has been transferred to your account in your country.
For your assistance, I have agreed with my son to give you 20% of the total amount for your efforts; 10 % to cover all the expenses that may be incurred during the business transaction. Lastly, Iurge you to keep this transaction strictly confidential as no one else knows our whereabout.
Please, as you show your willingness, forward to us your full name, address and Tel/ Fax numbers,and new email address. You can contact us with these telephone numbers for more details: 00225 07 888 660.email addersse j_kamara20032003-at-yahoo.fr
I am Mrs. JOSEPHINE KAMARA and my Son is Fred Kamara. I feel pleased contacting you personally with the hope to establish good business relationship that is strongly rooted in truth and the fear of God. I would honestly confide in you and seek your interest to transact a well conceived and nurtured business I have at hand. I am writing you in absolute confidence primarily to seek your assistance to transfer our cash of thirty Million Dollars ($30,000.000.00) now in the custody of a private Security trust firm in Abidjan, the money is in a trunk box deposited and declared as family valuables by my late husband,as a matter of fact, the company does not know the content as money, although my late husband made the management of the security company to understand that the box belongs to his foreign partner.
Source of the money:
My late husband, Chief Vincent Kamara , a native of Mende District in the Northerh province of Sierra Leone, was the General Manager of Sierra Leone Mining Corporation (S.L.M.C.) Freetown . According to my late husband, this money was the income accrued from mining coporation's over draft and minor sales. Before the peak of the civil war between the rebels forces of Major Paul Koroma and the combined forces of ECOMOG peace keeping operation that almost destroyed my country, following the forceful removal from power of the civilian elected President Ahmed Tejan Kabbah by the rebels. My late husband had already made arrangement for us be evacuated to Abidjan, Cote d' Ivoire with the Certificate of Deposit issued to him by the security firm in Abidjan. During the war in my country, and following the indiscriminate looting of Public and Government properties by the rebel forces, the Sierra Leone Mining Corp. was one of the target looted and destroyed.
My late husband, including other top Government functionaries were attacked and killed by the rebels in November 2000 because of his relationship wi usbandŽs death dashed our hope of survival. I and my son are now alone in this strange country suffering without any care or help. Without any relation, we are now like refugees.Our only hope now is in you and the box deposited in the security firm. To this effect, I humbly solicityour assistance in the followings ways.
1.to assist me claim this box from the security firm as our beneficiary;
2.to transfer the money in your name to your country
3.to make a good arrangement for a joint business investment on our behalf in your country with you as our adviser;
4.To secure a college for my son in your country after the money has been transferred to your account in your country.
For your assistance, I have agreed with my son to give you 20% of the total amount for your efforts; 10 % to cover all the expenses that may be incurred during the business transaction. Lastly, Iurge you to keep this transaction strictly confidential as no one else knows our whereabout.
Please, as you show your willingness, forward to us your full name, address and Tel/ Fax numbers,and new email address. You can contact us with these telephone numbers for more details: 00225 07 888 660.email addersse j_kamara20032003-at-yahoo.fr
Originator: "MR ANDERSON BELLO" {andersonbello-at-o2.pl} Recipients: MicroscopyFilteredEmail3-at-sparc5.microscopy.com
I am trying to look at a native silica aerogel in a Hitachi S4700 FE-SEM. I want to look at a fractured specimen in a 100 to 200 nanometer range. The problem seems to be optimal sample preparation. I can view the sample with a gold or gold/palladium sputter coat but in order to view at the magnification I'm looking for the coating is substantial enough that it fills in some of the pores or open pathways and totally coats over the primary silica particles. I have a comparison of gold sputtered onto a glass slide with a Balzer Coater, 40 ma and 1.5 inches to the top of the jar at 10, 20 and 30 seconds. The ten second sputter coating viewed by SEM at x250k and 200nm somewhat mimics photographically the channels and porosity of the native silica. At the higher sputter times of 20 and 30 seconds the pores and channels begin to close or fill in. I have also tried a carbon deposition coating. It's lighter in density not filling in pores and channels and not obliterating primary particles but doesn't seem to be conductive enough to dissipate charging in a manner that lends itself to anything but low magnification photos. I've tried mounting on carbon tape, and/or carbon paint but the isopropanol dissolves the sample. I've tried varying degrees of the carbon and gold coat. I've tried silver epoxy, I've even tried, after the coating running a thin line of the carbon paint up the side of the sample onto the area to be viewed. Of course with the native silica being so easily degraded the carbon paint was actually eating away the silica aerogel and breaking the contact of the existing coating. I've tried running a thin copper wire or copper tape from the face of the sample to the sample holder. The problem being there is no good method of adhesion to the sample. I have simply run out of ideas. Does anyone please have a soluton?
Linda
Linda S. McCorkle Ohio Aerospace Institute Materials Division, Polymers Branch NASA John H. Glenn Research Center at Lewis Field 21000 Brookpark Road Cleveland, OH 44135
Enclosed is the table of contents for the July/August issue of Microscopy Today.
I will close the magazine's subscription list on Thursday, July 3rd, for this issue.
MT is free for any one interested in, or connected with, microscopy anywhere in Canada, Mexico, and the USA. MT is also free for Microscopy Society of America members anywhere.
The subscription rate for those not qualifying for free subscriptions has been reduced from $80 or $110 to $35US per year.
Please subscribe via our web page: http://www.microscopy-today.com
Thank you,
Ron Anderson, Editor Microscopy Today
Microscopy Today July/August 2003
Taking Correlative Microscopy to a New Level Stephen W. Carmichael, Mayo Clinic Imaging of Big and Messy Biological Structures Using Electron Tomography Gina E. Sosinsky and Maryann E. Martone, University of California A New Capability for Light Microscopes: Mid Infrared Molecular Analysis Alan J. Rein, SensIR Technologies Brightfield Illumination of Large Field Sizes Theodore M. Clarke, Metallurgical Failure Analysis Consultant Special Topic: Advanced Basics of Immunostaining and Antigen Retrieval W. Gray (Jay) Jerome, Vanderbilt University How To Get Some Action (In Photoshop) Jerry Sedgewick, University of Minnesota Macro Imaging with Digital Cameras Bryan Burnett, Nyoptics, Inc./Meixa Tech and Steven Blaauw, Nyoptics,Inc. Rapid Preparation of a Polymer Fiber and a Free-Standing Polymer Film for Cross-Sectional Microtomy Andreas Taubert, University of Basel, and James H. Ferris, and Karen I. Winey University of Pennsylvania M&M 2002: Core Facility Management Session: Maintaining Major Equipment in the Core Microscopy Facility Session Chair: Debby Sherman, Purdue University Making Your Own Molds For The EM Lab Mary Mager, University of British Columbia Vascular Corrosion Casting Fred E. Hossler, East Tennessee State University Benzyldimethylamine (BDMA): Catalyst of Choice with Epoxy Embedding Media José A. Mascorro, Tulane University Health Sciences Center The Most Likely Sources of EDX Copper Peaks in Samples Run by TEM Paul Beauregard, Chemist and Electron Microscopist
I am looking for help in finding and/or designing a petrographic slide holder for our SEM. We have a Zeiss DSM960A with a 4-axis mechanical stage (100x100mm travel). We currently have a 'homemade' holder with room for one slide (26x46mm) OR one standard block (25mm diam.). I would like a top-referencing holder for use with our Oxford EDS system.
I want to come up with something similar JEOL's multi-specimen holder (model KR-6M150-58) and have considered buying this and modifying it for our machine. Our main problem in our own designs is the machining involved (we can do it in-house) and specifically, how do we top-reference a slide; springs or clamps of some sort or maybe a hinged top plate or...??
How have other users tackled this problem when a specific holder is not produced by your instrument manufacturer? I did not see any messages relating to this in the archives. Any information or advice would be tremendously appreciated.
For high magnifications I usually coat with Au/Pd at 10 ma for 20-60 sec., depending on the roughness of the specimens surface. Usually, when coating is thin ehough, I do not observe it for magnifications up to 100-150k. Sticky tape (and carbon paint) are not stable enough, so I prefer to glue specimens with Quick Gel superglue (drying it overnight).
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: Linda S. McCorkle [mailto:Linda.S.McCorkle-at-grc.nasa.gov] } Sent: Monday, June 30, 2003 9:23 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM of Native Silica Aerogel } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } I am trying to look at a native silica aerogel in a Hitachi } S4700 FE-SEM. I } want to look at a fractured specimen in a 100 to 200 } nanometer range. The } problem seems to be optimal sample preparation. } I can view the sample with a gold or gold/palladium } sputter coat but in } order to view at the magnification I'm looking for the coating is } substantial enough that it fills in some of the pores or open } pathways and } totally coats over the primary silica particles. } I have a comparison of gold sputtered onto a glass slide with } a Balzer } Coater, 40 ma and 1.5 inches to the top of the jar at 10, 20 and 30 } seconds. The ten second sputter coating viewed by SEM at } x250k and 200nm } somewhat mimics photographically the channels and porosity of } the native } silica. At the higher sputter times of 20 and 30 seconds the } pores and } channels begin to close or fill in. } I have also tried a carbon deposition coating. It's } lighter in density not } filling in pores and channels and not obliterating primary } particles but } doesn't seem to be conductive enough to dissipate charging in } a manner that } lends itself to anything but low magnification photos. I've } tried mounting } on carbon tape, and/or carbon paint but the isopropanol } dissolves the } sample. I've tried varying degrees of the carbon and gold } coat. I've tried } silver epoxy, I've even tried, after the coating running a } thin line of the } carbon paint up the side of the sample onto the area to be viewed. Of } course with the native silica being so easily degraded the } carbon paint was } actually eating away the silica aerogel and breaking the } contact of the } existing coating. I've tried running a thin copper wire or } copper tape from } the face of the sample to the sample holder. The problem } being there is no } good method of adhesion to the sample. I have simply run out } of ideas. Does } anyone please have a soluton? } } Linda } } } } Linda S. McCorkle } Ohio Aerospace Institute } Materials Division, Polymers Branch } NASA John H. Glenn Research Center at Lewis Field } 21000 Brookpark Road } Cleveland, OH 44135 } } Tel.: (216) 433-3689 } Fax: (216) 977-7132 } e-mail: Linda.S.McCorkle-at-grc.nasa.gov } } }
Before you use the expensive solutions of room shielding or field canceling, you should determine the source(s) of the field(s). In some cases, large fields can be produced by simple wiring problems that can be fixed relatively easily once they are identified. In other cases, the sources of the fields can be shielded or moved, or may even be associated with the SEM that you are replacing.
An inexpensive gauss meter can be used to locate the typical sources of magnetic fields. See the section "11. Magnetic Shielding and Gauss Meters" at "www.jcnabity.com/links.htm". (Note: I do not have any financial interest in the companies listed, but have found them to be useful.)
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Group, Is there a way to convert an RGB image to a CMMYK image without having the colors go "haywire"? I ask on behalf of a colleage who has a request to submit an image for publication in the CMMYK format, an image that he made in RGB. If you reply, please be sure to include his email (cardenas-at-bio.umass.edu) because he is not a subscriber.
What imaging conditions are you using? In my evaluations of FE-SEM's, that included the Hitachi 4700, we examined uncoated silica aerogel samples. I mounted them using carbon paint on a stub. We used a voltage of about 1 kV and a working distance of about 2 mm if I recall correctly. There were some issues with drift, but we used a continuous average mode, soaked the sample at a lower mag than what we wanted to take the micrograph for a few minutes, focused out of the area that we took the picture and brought it back in and took the picture when the image stabilized. We were able to look at all of the samples in the 50kX -150kX range on all of the systems without too much trouble.
BTW, we bought the LEO 1530.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Linda S. McCorkle [mailto:Linda.S.McCorkle-at-grc.nasa.gov] Sent: Monday, June 30, 2003 10:23 AM To: Microscopy-at-sparc5.microscopy.com
I am trying to look at a native silica aerogel in a Hitachi S4700 FE-SEM. I want to look at a fractured specimen in a 100 to 200 nanometer range. The problem seems to be optimal sample preparation. I can view the sample with a gold or gold/palladium sputter coat but in order to view at the magnification I'm looking for the coating is substantial enough that it fills in some of the pores or open pathways and totally coats over the primary silica particles. I have a comparison of gold sputtered onto a glass slide with a Balzer Coater, 40 ma and 1.5 inches to the top of the jar at 10, 20 and 30 seconds. The ten second sputter coating viewed by SEM at x250k and 200nm somewhat mimics photographically the channels and porosity of the native silica. At the higher sputter times of 20 and 30 seconds the pores and channels begin to close or fill in. I have also tried a carbon deposition coating. It's lighter in density not filling in pores and channels and not obliterating primary particles but doesn't seem to be conductive enough to dissipate charging in a manner that lends itself to anything but low magnification photos. I've tried mounting on carbon tape, and/or carbon paint but the isopropanol dissolves the sample. I've tried varying degrees of the carbon and gold coat. I've tried silver epoxy, I've even tried, after the coating running a thin line of the carbon paint up the side of the sample onto the area to be viewed. Of course with the native silica being so easily degraded the carbon paint was actually eating away the silica aerogel and breaking the contact of the existing coating. I've tried running a thin copper wire or copper tape from the face of the sample to the sample holder. The problem being there is no good method of adhesion to the sample. I have simply run out of ideas. Does anyone please have a soluton?
Linda
Linda S. McCorkle Ohio Aerospace Institute Materials Division, Polymers Branch NASA John H. Glenn Research Center at Lewis Field 21000 Brookpark Road Cleveland, OH 44135
Ritchie Yes, I do believe, the fermentation is major way to make ethanol. Contrary, 200 proof ethanol in US made using direct synthesis from ethane according my friend's information. To us it means that such alcohol is very clean from any contamination including water. In Russia we did not have "200 proof ethanol". We had "absolute ethanol", which was dehydrated with CuSO4 I believe. In the beginning, I was so picky to use "200 proof ethanol" because of possible water contamination. Nevertheless, I am using it now without any problems. I do find it very economical: I used half of the bottle as "100%" and the rest of the bottle to make 30,50,70,95% Et-OH for dehydratation steps. So, I utilized 100% of the stuff. It's a little bit risky, because you relied on unknown quality, so to be sure, you definitely need to use "molecular sieve". Sergey
At 11:11 AM 6/30/03 +1200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Linda: you didn't say what kV accelerating voltage you were using. You might try varying the accelerating voltage to see if you can find a voltage that results in a non-charging (or very minimally-charging) sample. You might try anywhere from 0.5kV to 2 kV. Another alternative is to find a colleague with a variable-pressure SEM with a secondary electron detector that functions in the VP mode (as opposed to the BSD).
"Linda S. McCorkle" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am trying to look at a native silica aerogel in a Hitachi S4700 FE-SEM. I } want to look at a fractured specimen in a 100 to 200 nanometer range. The } problem seems to be optimal sample preparation. } I can view the sample with a gold or gold/palladium sputter coat but in } order to view at the magnification I'm looking for the coating is } substantial enough that it fills in some of the pores or open pathways and } totally coats over the primary silica particles. } I have a comparison of gold sputtered onto a glass slide with a Balzer } Coater, 40 ma and 1.5 inches to the top of the jar at 10, 20 and 30 } seconds. The ten second sputter coating viewed by SEM at x250k and 200nm } somewhat mimics photographically the channels and porosity of the native } silica. At the higher sputter times of 20 and 30 seconds the pores and } channels begin to close or fill in. } I have also tried a carbon deposition coating. It's lighter in density not } filling in pores and channels and not obliterating primary particles but } doesn't seem to be conductive enough to dissipate charging in a manner that } lends itself to anything but low magnification photos. I've tried mounting } on carbon tape, and/or carbon paint but the isopropanol dissolves the } sample. I've tried varying degrees of the carbon and gold coat. I've tried } silver epoxy, I've even tried, after the coating running a thin line of the } carbon paint up the side of the sample onto the area to be viewed. Of } course with the native silica being so easily degraded the carbon paint was } actually eating away the silica aerogel and breaking the contact of the } existing coating. I've tried running a thin copper wire or copper tape from } the face of the sample to the sample holder. The problem being there is no } good method of adhesion to the sample. I have simply run out of ideas. Does } anyone please have a soluton? } } Linda } } } Linda S. McCorkle } Ohio Aerospace Institute } Materials Division, Polymers Branch } NASA John H. Glenn Research Center at Lewis Field } 21000 Brookpark Road } Cleveland, OH 44135 } } Tel.: (216) 433-3689 } Fax: (216) 977-7132 } e-mail: Linda.S.McCorkle-at-grc.nasa.gov
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
In practice, it is not possible to convert RGB into CYMK without hassle. In Photoshop, if you will go to the color palette in RGB an pick some colors close to the upper right corner of the palette, you will see "!" mark indicating that this color will not be reproduced correctly when printing. CYMK palette in general represents colors how it will be on the paper, RGB - on screen. So, if you have those colors with "!" mark in the picture - they will be translated into CYMK incorrectly. Similarly, you will see "!" in the navigator window if color does not match CYMK palette. Good luck! Sergey
At 01:59 PM 6/30/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
I was wondering wether there is a non-commercial file format for storing microscopy images with additional data (like magnification etc) attached to it in a standardized way. There is an MSA EELS format which stores EELS spectra, but I never came across an image format which has the same goal. I`ve read some comments on TIFF images on this list (which seem a good way to go to me). Is anybody working on this, or does it exist already?
Jo -- Dr. J. Verbeeck Electron Microscopy for Materials Research University of Antwerp, Belgium
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id EAA03335 for dist-Microscopy; Tue, 1 Jul 2003 04:45:37 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id EAA03330 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Tue, 1 Jul 2003 04:45:06 -0500 (CDT) Received: from unionbio-eu.com (union-biometrica-gw.cpgle.be.easynet.net [213.193.139.10] (may be forged)) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id EAA03319 for {Microscopy-at-sparc5.Microscopy.Com} ; Tue, 1 Jul 2003 04:44:53 -0500 (CDT) Received: (qmail 2936 invoked from network); 1 Jul 2003 09:38:37 -0000 Received: from linux-2.unionbio-eu.com (HELO unionbio-eu.com) (192.168.0.23) by server.unionbio-eu.com with SMTP; 1 Jul 2003 09:38:37 -0000 Sender: pvosta Message-ID: {3F01569E.AEFE9B54-at-unionbio-eu.com}
Hi,
I am looking for a 3CCD color video camera (PAL) which is capable of progressive scan and which can be used on a microscope with a C-mount adapter ?
Linda, } } Aerogel is without doubt, one of the more difficult samples to look at in } the SEM. It seems you are trying all the right things, but you did not } mention the actual microscope conditions you were using. Aerogel is very } beam sensitive and I remember some years ago having some success looking at } it at 700 volts, very low beam current. I remember also looking at as thin a } piece of it as I could and sticking it in a bed of carbon paint. } I hope this helps in some way--keep trying! } } bill riley
There are some better ways to convert to CMYK (Photoshop comes to mind), but you must remember that the CMYK color space can't reproduce many RGB colors. A pure green will never be reproduced in CMYK. This is a fact of printing on paper, rather than on a monitor. Many of the color aberrations seen when color spaces are changed are due to the monitor you are using. If it has not been specifically calibrated for print publishing, the colors are quite likely to be off. This is especially true if you are using a flat screen monitor. It may be worth your while to convert to CMYK, and then get a professional print made of your figure (Staples, or some such place) and see if you can live with the colors. David
On Monday, June 30, 2003, at 04:59 PM, Tobias Baskin wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Group, } Is there a way to convert an RGB image to a CMMYK image without } having the colors go "haywire"? I ask on behalf of a colleage who has } a request to submit an image for publication in the CMMYK format, an } image that he made in RGB. If you reply, please be sure to include his } email (cardenas-at-bio.umass.edu) because he is not a subscriber. } } Thanks, } Tobias } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ Biology } Department } / / / / \ \ \ University of } Massachusetts } /_ / __ /__ \ \ \__ Amherst, MA, 01003 } / / / \ \ \ } / / / \ \ \ Voice: 413 - } 545 - 1533 } / / ___ / \ \__/ \ ____ } Fax: 413 - } http://www.biosci.missouri.edu/Baskin/baskin_lab__home_page.htm } } ___________________
David Elliott, Ph.D. Yale University School of Medicine 810 LCI 333 Cedar St. New Haven, CT 06520
I think a Sony DXC-9100P will do the job! But the resolution is only about 768*576, is this enough? Maybe another possibility might be the DT1100 HR (1392*1040) from DuncanTech but I don't know any specifications, just the name and company... Best regards,
Sven Terclavers
-------Original Message-------
} From: pvosta-at-unionbio-eu.com
Hi,
I am looking for a 3CCD color video camera (PAL) which is capable of progressive scan and which can be used on a microscope with a C-mount adapter ?
Do you have access to an ion-beam coater? I should think there would be one in your neck of the woods -- Ohio State, or Wright Patterson AFB, if there isn't one in Cleveland. We routinely use one with a platinum target for high resolution SEM of subcellular structures, gels, etc., and I don't coating effects until 300 - 400,000X normally. Depending on thickness -- normally we use 1 - 2 nm. Phil
} I am trying to look at a native silica aerogel in a Hitachi S4700 } FE-SEM. I want to look at a fractured specimen in a 100 to 200 } nanometer range. The problem seems to be optimal sample preparation. } I can view the sample with a gold or gold/palladium sputter } coat but in order to view at the magnification I'm looking for the } coating is substantial enough that it fills in some of the pores or } open pathways and totally coats over the primary silica particles. } I have a comparison of gold sputtered onto a glass slide with a } Balzer Coater, 40 ma and 1.5 inches to the top of the jar at 10, 20 } and 30 seconds. The ten second sputter coating viewed by SEM at } x250k and 200nm somewhat mimics photographically the channels and } porosity of the native silica. At the higher sputter times of 20 and } 30 seconds the pores and channels begin to close or fill in. } I have also tried a carbon deposition coating. It's lighter } in density not filling in pores and channels and not obliterating } primary particles but doesn't seem to be conductive enough to } dissipate charging in a manner that lends itself to anything but low } magnification photos. I've tried mounting on carbon tape, and/or } carbon paint but the isopropanol dissolves the sample. I've tried } varying degrees of the carbon and gold coat. I've tried silver } epoxy, I've even tried, after the coating running a thin line of the } carbon paint up the side of the sample onto the area to be viewed. } Of course with the native silica being so easily degraded the carbon } paint was actually eating away the silica aerogel and breaking the } contact of the existing coating. I've tried running a thin copper } wire or copper tape from the face of the sample to the sample } holder. The problem being there is no good method of adhesion to the } sample. I have simply run out of ideas. Does anyone please have a } soluton? } } Linda } } } } Linda S. McCorkle } Ohio Aerospace Institute } Materials Division, Polymers Branch } NASA John H. Glenn Research Center at Lewis Field } 21000 Brookpark Road } Cleveland, OH 44135 } } Tel.: (216) 433-3689 } Fax: (216) 977-7132 } e-mail: Linda.S.McCorkle-at-grc.nasa.gov
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
} Is there a way to convert an RGB image to a CMMYK image } without having the colors go "haywire"? I ask on behalf of a colleage } who has a request to submit an image for publication in the CMMYK } format, an image that he made in RGB. If you reply, please be sure to } include his email (cardenas-at-bio.umass.edu) because he is not a } subscriber.
I am always surprised when publishers ask for CMYK. Since RGB=} CMYK conversions need CMYK (ink) definitions, how do they expect you to know what the definitions are? Or, they should at least ask you to convert using specific definitions as provided by Photoshop (e.g., Euroscale coated, U.S. sheetfed coated, ...).
As you may now know, ... since RGB=} CMYK involves CRT color definitions (or working color space definitions) going to ink+paper color definitions, there is no way to keep the colors from going "haywire" (i.e., out of gamut).
Sure, I do it all the time for Microscopy Today--thanks to Jerry Sedgewick's Photoshop book!*
Here's what to do: In RGB mode, before conversion, under view, select gamut warning (ctrl-shift-y)**. The out-of-gamut colors in the RGB image will turn gray. Open the 'image/adjust/hue and saturation' box and adjust saturation, color by color (in the drop down box) until the gray overlay disappears, bump up the lightness and then increase saturation a bit until you find the saturation and lightness compromise that doesn't effect contrast too much. Flip in and out of gamut warning to view your progress. You may have to cycle through the color drop down box more than once. Convert to CMYK when finished (Not before!). Try it with copies of images until you become skilled.
Ron
* "Quick Photoshop for Research" Kluwer Academic/Plenum ISBN: 0-306-47375-5. 2002.
** Gamut refers to the range of colors visible in any color system. There are more hues and saturations possible in RGB vs. CMYK. Colors that you see in RGB that you can't see in CMYK are "out of gamut" with respect to CMYK.
-----Original Message----- } From: Tobias Baskin [mailto:Baskin-at-bio.umass.edu] Sent: Monday, June 30, 2003 4:59 PM To: Microscopy-at-sparc5.microscopy.com Cc: cardenas-at-bio.umass.edu
Group, Is there a way to convert an RGB image to a CMMYK image without having the colors go "haywire"? I ask on behalf of a colleage who has a request to submit an image for publication in the CMMYK format, an image that he made in RGB. If you reply, please be sure to include his email (cardenas-at-bio.umass.edu) because he is not a subscriber.
there are many different formats that are being used by different programs. TIFF (or TIF, Tagged Image File Format) is one of the most flexible and seems to be a de-facto standard now. It is now in revision 6, I believe. The format allows not only for data, but also for meta-data, such as magnification, etc., to be stored. It also allows for multiple images in a file, compression, and other goodies. There is a set of "public" tags, which include magnification, and each user can define "private" tags, which are simply ignored by other programs. There are also some drawbacks. For example the tag "magnification". If you put in a large number there, some programs (Word used to do it), would interpret this as information about how big you wanted the image displayed in a document. If the tag was, let's say "1000", Word would interpret this as "the image is 1000 times as large as the original", and try to print it at the original size, resulting in an image a few pixels across. Not good. There are other image formats, some of them require royalty payments (GIF), other store only image data (BMP, JPG). As I mentioned above, TIF seems to be the de-facto standard.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: jo verbeeck [mailto:joverbee-at-ruca.ua.ac.be] Sent: Tuesday, July 01, 2003 1:16 AM To: MSA listserver
Hi,
I was wondering wether there is a non-commercial file format for storing microscopy images with additional data (like magnification etc) attached to it in a standardized way. There is an MSA EELS format which stores EELS spectra, but I never came across an image format which has the same goal. I`ve read some comments on TIFF images on this list (which seem a good way to go to me). Is anybody working on this, or does it exist already?
Jo -- Dr. J. Verbeeck Electron Microscopy for Materials Research University of Antwerp, Belgium
No relevant experience still .... Perhaps you could get access to a sputter coating unit for a metal which gives a finer coat eg chromium. Some companies have some sort of osmium "coating" device.
Dave
On Mon, 30 Jun 2003 18:24:59 -0500 Becky Holdford {r-holdford-at-ti.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Linda: you didn't say what kV accelerating voltage you were using. You might try } varying the accelerating voltage to see if you can find a voltage that results in a } non-charging (or very minimally-charging) sample. You might try anywhere from 0.5kV } to 2 kV. Another alternative is to find a colleague with a variable-pressure SEM with } a secondary electron detector that functions in the VP mode (as opposed to the BSD). } } "Linda S. McCorkle" wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } I am trying to look at a native silica aerogel in a Hitachi S4700 FE-SEM. I } } want to look at a fractured specimen in a 100 to 200 nanometer range. The } } problem seems to be optimal sample preparation. } } I can view the sample with a gold or gold/palladium sputter coat but in } } order to view at the magnification I'm looking for the coating is } } substantial enough that it fills in some of the pores or open pathways and } } totally coats over the primary silica particles. } } I have a comparison of gold sputtered onto a glass slide with a Balzer } } Coater, 40 ma and 1.5 inches to the top of the jar at 10, 20 and 30 } } seconds. The ten second sputter coating viewed by SEM at x250k and 200nm } } somewhat mimics photographically the channels and porosity of the native } } silica. At the higher sputter times of 20 and 30 seconds the pores and } } channels begin to close or fill in. } } I have also tried a carbon deposition coating. It's lighter in density not } } filling in pores and channels and not obliterating primary particles but } } doesn't seem to be conductive enough to dissipate charging in a manner that } } lends itself to anything but low magnification photos. I've tried mounting } } on carbon tape, and/or carbon paint but the isopropanol dissolves the } } sample. I've tried varying degrees of the carbon and gold coat. I've tried } } silver epoxy, I've even tried, after the coating running a thin line of the } } carbon paint up the side of the sample onto the area to be viewed. Of } } course with the native silica being so easily degraded the carbon paint was } } actually eating away the silica aerogel and breaking the contact of the } } existing coating. I've tried running a thin copper wire or copper tape from } } the face of the sample to the sample holder. The problem being there is no } } good method of adhesion to the sample. I have simply run out of ideas. Does } } anyone please have a soluton? } } } } Linda } } } } } } Linda S. McCorkle } } Ohio Aerospace Institute } } Materials Division, Polymers Branch } } NASA John H. Glenn Research Center at Lewis Field } } 21000 Brookpark Road } } Cleveland, OH 44135 } } } } Tel.: (216) 433-3689 } } Fax: (216) 977-7132 } } e-mail: Linda.S.McCorkle-at-grc.nasa.gov } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Becky Holdford (r-holdford-at-ti.com) } 972-995-2360 } 972-648-8743 (pager) } SC Packaging FA Development } Texas Instruments, Inc. } Dallas, TX } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
One of the other responders to your post mentioned the technique of moving around on the specimen to minimize localized charging on the specimen surface. This technique takes a little practice, but is the best way to manage a specimen like Silica Aerogel.
At Lawrence Berkeley Laboratory, we sometimes wrapped the specimen in aluminum foil, with a small hole approximately 2~3mm in diameter through which we would view the specimen. This worked well for Auger analysis too.
Hope this helps, Ken Gaugler
Linda S. McCorkle wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am trying to look at a native silica aerogel in a Hitachi S4700 } FE-SEM. I want to look at a fractured specimen in a 100 to 200 nanometer } range. The problem seems to be optimal sample preparation. } I can view the sample with a gold or gold/palladium sputter coat but } in order to view at the magnification I'm looking for the coating is } substantial enough that it fills in some of the pores or open pathways } and totally coats over the primary silica particles. } I have a comparison of gold sputtered onto a glass slide with a Balzer } Coater, 40 ma and 1.5 inches to the top of the jar at 10, 20 and 30 } seconds. The ten second sputter coating viewed by SEM at x250k and 200nm } somewhat mimics photographically the channels and porosity of the native } silica. At the higher sputter times of 20 and 30 seconds the pores and } channels begin to close or fill in. } I have also tried a carbon deposition coating. It's lighter in } density not filling in pores and channels and not obliterating primary } particles but doesn't seem to be conductive enough to dissipate charging } in a manner that lends itself to anything but low magnification photos. } I've tried mounting on carbon tape, and/or carbon paint but the } isopropanol dissolves the sample. I've tried varying degrees of the } carbon and gold coat. I've tried silver epoxy, I've even tried, after } the coating running a thin line of the carbon paint up the side of the } sample onto the area to be viewed. Of course with the native silica } being so easily degraded the carbon paint was actually eating away the } silica aerogel and breaking the contact of the existing coating. I've } tried running a thin copper wire or copper tape from the face of the } sample to the sample holder. The problem being there is no good method } of adhesion to the sample. I have simply run out of ideas. Does anyone } please have a soluton? } } Linda } } } } Linda S. McCorkle } Ohio Aerospace Institute } Materials Division, Polymers Branch } NASA John H. Glenn Research Center at Lewis Field } 21000 Brookpark Road } Cleveland, OH 44135 } } Tel.: (216) 433-3689 } Fax: (216) 977-7132 } e-mail: Linda.S.McCorkle-at-grc.nasa.gov } }
A while ago I posted a message on this issue (pasted below). Here's a brief of what I could get to the moment.
At least TIFF and JPG formats have more or less standardized tagged fields to store metadata (= data on the image itself). The appropriate place to store microscope and image setting data seems to be the EXIF fields. (See Digital Still Camera Image File Format Standard. Version 2.1. JEIDA-49-1998. http://www.kodak.com/global/plugins/acrobat/en/service/digCam/exifStandard.pdf. A more recent draft in www.dpnet.com.cn/download/software/exif22.pdf.)
Several programs let you view and/or access the data stored on the EXIF fields, e.g., Photostudio (freeware), IMatch (shareware, $50). Using Photostudio, I can see that our SEM FEI XL stores all the microscope settings and image data in one EXIF field (#8778).
Digital cameras use the more or less standardized EXIF fields for date, resolution, size, etc., while storing other specific data (exposure, flash, diafragm, etc.) in non-standard, proprietary fields. For accessing and managing such data, you need a filter that tells you what to look for in what field. The funny thing is that camera makers tend to keep this information secret (I presume they trade the format specifications with software makers over a commercial agreement). There are several non-official filters that will permit you to read, manage, and edit these camera-specific EXIF fields. I only know those of IMatch (http://www.photools.com/, manual in http://www.photools.net/bin/imatchdoc.zip).
Some microscope makers store all image and microscope settings in only one EXIF field, while leaving the more standardized fields empty. For example, the EXIF field 8778 in our SEM FEI XL looks like this:
Some microscopes produce one text file for every image, with all this data. For example, the Hitachi S4700 FE-SEM produces a file with the same name as the image and .txt extension, looking like this:
In either case, one could easily extract this information with a script in a text editor (i.e., building your own filter), and then load the data in your system of choice.
It seems that other microscope makers went a step beyond, and are way more cryptic. The SEM LEO 1430 VP stores in the EXIF filed 8546 something like this:
In some cases the software that controls the microscope has basic capabilities to build databases with this information. But that machine is typically heavily used by many users on a tight schedule, and it won't help if you collaborate with other researchers using different microscopes.
The microscope image managing programs have specific filters for each microscope type. For instance, the one by Soft Imaging (http://www.soft-imaging.net/) has filters for many microscope makers. If you install the freeware Soft Imaging Viewer, it seems that only the filter for the FEI is activated, the rest are blocked. You can read the metadata from each image, but not really manage it. The full version supposedly has all these filters active and permits data manipulation--but it is expensive.
Reading the EXIF fields or the text files associated with the image, copying to the clipboard, and pasting fragments in a database (or in more meaningful EXIF fields) could work for a few images, but it is not a solution for large batches and for the everyday work.
The question is how we can efficiently manage the data associated with the images, if we don't have the resources for expensive microscope image software.
First of all, knowing the specifications for metadata storage by each microscope is a great start. (For instance, do somebody know how to read the SEM LEO 1430 VP data?)
Ideally, there should be a standard format such that one knows where to look for magnification, KV, etc., in any image coming from a microscope. But if camera makers cannot converge on a standard, I doubt that microscope makers, which seem to be moving in the opposite direction, will converge ever. Perhaps the microscopy community could come to agree on a standard (I think there are such standards in the health sciences) that we all can use to properly document, transmit, and manage our data--it's a lot of work though.
Best regards,
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
} Date: Wed, 19 Feb 2003 14:17:12 -0300 } Hi all, } } I wonder if there are standard (or usual) ways for storing setting data } from electron microscopes (magnification, working distance, acceleration } V, etc.) into the image file itself, such that they can be automatically } imported to a database. Some other devices (like digital cameras) } automatically use the IPTC or EXIF fields for this. } } Any general idea about how preserve and manage these data together with } the images will be very welcome. } } Martín J. Ramírez
} Hi, } } I was wondering wether there is a non-commercial file format for storing } microscopy images with additional data (like magnification etc) attached } to it in a standardized way. } There is an MSA EELS format which stores EELS spectra, but I never came } across an image format which has the same goal. I`ve read some comments } on TIFF images on this list (which seem a good way to go to me). Is } anybody working on this, or does it exist already? } } Jo } -- } Dr. J. Verbeeck } Electron Microscopy for Materials Research } University of Antwerp, Belgium
I have a researcher who dissected out a beetle stomach, digested much of the tissue away with KOH and wants to examine the contents to identify the pollen it was feeding on. There was still a sheath of tissue around it so we ran it through alcohol, placed it and a drop of alcohol between 2 slides, and froze it in liquid nitrogen. When I snapped it apart it just fractured in cross section rather than the longitudinal plane I had hoped. I then stuck a piece of tape over and ripped it off. Basically we ended up just destroying everything, but one of the pieces looked like it might have been some kind of pollen grain at one time or other.
Has anyone done this before? The entire stomach is about 4-500um long and under the light microscope we can see several grains that look like they could be pollen. Any suggestions on how to get them out and captured for SEM examination?? I only have a couple more so experimentation is out and I am running out of time.
Thanks
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
When creating images in Photoshop in RGB space that will be converted to CMYK later, there is a Preview in CMYK mode that should be turned on. The reason is that CMYK is for four color separation ink printing and cannot reproduce many of the more saturated colors in particular, especially the greens, of RGB.
What I do is to keep the image RGB so that info isn't lost, go into the CMYK preview mode and readjust the image to something suitable but admittedly less vivid, and then convert to CMYK.
} Group, } Is there a way to convert an RGB image to a CMMYK image without } having the colors go "haywire"? I ask on behalf of a colleage who has a } request to submit an image for publication in the CMMYK format, an image } that he made in RGB. If you reply, please be sure to include his email } (cardenas-at-bio.umass.edu) because he is not a subscriber. } } Thanks, } Tobias
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
Is there anyone out there in ListserverLand that has found a way to bring TIF images into Visual Basic 6.0 and work with them? Since I'm doing this for myself, I don't want to buy a commercial package because they pretty expensive. If you have an example of programming code, it would be greatly appreciated.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
We also have an Oxford EDS, but we have a Hitachi S-2460N rather than a Zeiss.
We have made numerous holders over the years for our scope to accommodate various sample forms. It is certainly nice to have a top referencing holder. It would be one less thing to be concerned about. However, we normally cannot count on all of our samples being at the same working distance even then. We still have to do a little focusing as we move from sample to sample. But when we do the focusing, we try to do most, if not all, of it by using the Z control. We initially set the focal length of the beam at that distance best for x-ray analysis, and then bring the samples to that focal plane.
Therefore, a top referencing holder would eliminate gross adjustments in Z moving from sample to sample, but it is more of a convenience than a necessity. I would probably give up the idea of the top reference and just have the shop machine a holder for a standard and a common thickness of slide plus sample and expect to make fine adjustments in Z as I move between them.
Warren
At 10:33 AM 6/30/2003 -0500, you wrote:
} Dear Listserv members: } } I am looking for help in finding and/or designing a petrographic slide } holder for our SEM. We have a Zeiss DSM960A with a 4-axis mechanical } stage (100x100mm travel). We currently have a 'homemade' holder with room } for one slide (26x46mm) OR one standard block (25mm diam.). I would like } a top-referencing holder for use with our Oxford EDS system. } } I want to come up with something similar JEOL's multi-specimen holder } (model KR-6M150-58) and have considered buying this and modifying it for } our machine. Our main problem in our own designs is the machining } involved (we can do it in-house) and specifically, how do we top-reference } a slide; springs or clamps of some sort or maybe a hinged top plate or...?? } } How have other users tackled this problem when a specific holder is not } produced by your instrument manufacturer? I did not see any messages } relating to this in the archives. Any information or advice would be } tremendously appreciated. } } Thanks for your time. } } Jeff Thole } } Jeff Thole - Geology Laboratory Supervisor and Instructor } Geology Department, Macalester College } 1600 Grand Avenue, St. Paul, MN 55105 } (ph. 651-696-6426, fax. x6122, email thole-at-macalester.edu) } Web: http://www.macalester.edu/geology } ------------------------------------------
No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
An excellent and free library for use in Visual Basic is available through the XnView imaging package by Pierre Gougelet. Some of the features include import of ~360 graphic file formats, LUT transforms, filters and a range of tools. It is available at http://perso.wanadoo.fr/pierre.g/xnview/engfl.html
Cheers, Paul Baggethun Alcoa Technical Center
-----Original Message----- } From: Walck, Scott D. [mailto:walck-at-ppg.com] Sent: Tuesday, July 01, 2003 5:12 PM To: Microscopy (E-mail)
Is there anyone out there in ListserverLand that has found a way to bring TIF images into Visual Basic 6.0 and work with them? Since I'm doing this for myself, I don't want to buy a commercial package because they pretty expensive. If you have an example of programming code, it would be greatly appreciated.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
I believe some of the later printers try to "compensate" for the flat screen and other monitor effects. I know this is a feature of my HP7550 and it got a good review from the latest issue of popular photography magazine. Now, however, I can't say I've done it. But the same article did describe the conversion to CMYK from GB as a necessity for every photographer for the obvious reasons. Photoshop was also discussed, but it didn't allude to any real problems with the conversion. Perhaps one of the digital photography magazines can lend further insight. I just can't imagine there isn't a solution to such a widespread application, just gotta find the people who know the tricks...maybe.
Regards, Ed ----- Original Message ----- } From: "David Elliott Ph.D." {David.Elliott-at-yale.edu} To: {cardenas-at-bio.umass.edu} ; "Microscopy ListServer" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, July 01, 2003 4:26 AM
When in doubt, read the destructions! The following are extracts from Adobe Photoshop 5's online help ========================== Reproducing Colour Accurately. *see also: About calibration About ICC profiles Choosing a color management module About the RGB, Grayscale, and CMYK Setup dialog boxes Calibrating your monitor Entering RGB setup information Entering CMYK setup information Using ICC profiles to define the CMYK color space Using the Built-in option to define the CMYK color space Adjusting separation options Printing a color proof Calibrating the screen image to the proof Converting to CMYK Managing ICC profiles in files Converting the color space of open images =========================== Using ICC Profiles to define the CMYK color space.
The CMYK Setup dialog box lets you specify the CMYK color space based on the ICC profile of the printer you select. The CMM then maps the colors in the image to the profiled printer's color gamut, or range of printable colors.You can choose the method (called rendering intent) that is used to translate the colors to the printed gamut.
To use ICC printer profiles to define the CMYK color space:
1 Choose File } Color Settings } CMYK Setup. 2 For CMYK Model, select ICC. 3 Select Preview to display a preview of your changes. A flashing bar under the option indicates a preview is being created. 4 For Profile, choose the printer profile you want to use. If the printer you use is not listed in the Profile menu, contact your printer manufacturer for the appropriate printer profile or create one using third-party printer profiling software. 5 For Engine, choose the CMM you want to use. Built-in refers to Photoshop's built-in CMM.
Note: This option is not the same as choosing Built-in for CMYK Model.
6 For Intent, choose one of the following:
. Perceptual (Images) to maintain the relative color values among the original pixels as they are mapped to the printer gamut. This method preserves the relationship between colors, although the color values themselves may change. . Saturation (Graphics) to maintain the relative saturation values of the original pixels. Out-of-gamut colors are converted to colors that have the same saturation but fall just inside the gamut. . Relative Colorimetric to leave colors that fall inside the gamut unchanged. This method usually converts out-of-gamut colors to colors that have the same lightness but fall just inside the gamut.
. Absolute Colorimetric to disable white point matching when converting colors. This option is not generally recommended.
7 If desired, choose Black Point Compensation to map the darkest neutral color of the source's color space to the darkest neutral color of the destination's color space rather than to black when converting colors. 8 Click OK. ======================== About ICC profiles
One of the methods Photoshop can use to manage color is based on the use of ICC profiles. An ICC profile is a color space description. The ICC profile format was defined by the International Color Consortium (ICC) as a cross-application standard. ICC profiles help you reproduce colors accurately across different platforms, devices, and ICC-compliant applications (such as Adobe Illustrator and Adobe PageMaker®). Adobe Photoshop uses a Color Management Module (CMM) to interpret the ICC profiles that describe the RGB and CMYK color spaces you are using in your system. You can select from existing ICC profiles or create your own. These profiles can then become part of your image files. The CMM interprets the ICC profiles to automatically manage color issues among different color models as well as color issues between your monitor, other monitors, and the final print image. Although you do not have to use ICC profiles, it can greatly simplify managing color.
Important: To ensure that color management works correctly on your system, change the color management settings every time you change printing devices. ========================= hth Chris
Dr. Chris Jeffree Inveresk Cottage 26, Carberry Road Inveresk Musselburgh Midlothian EH21 8PR Tel: +44 131 665 6062 FAX +44 131 653 6248 Mobile 07710 585 401
I had just received some information on 3-CCD cameras when I saw your message (as below). DuncanTech lists some cameras at http://www.duncantech.com/area_scan_cameras.htm
Apparently DuncanTech have been acquired by Redlake ( http://www.redlake.com ) and there are 3-CCD models listed under their website. Not sure of the C-mount though.
=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= Dr Thor Bostrom Acting Director, Analytical EM Facility; and School of Physical and Chemical Sciences, Queensland University of Technology (QUT) GPO Box 2434, Brisbane, QLD 4001, Australia Ph: +61 7 3864-2351 FAX: +61 7 3864-5100 http://www.sci.qut.edu.au/aemf/ CRICOS No. 00213J =-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=
I am looking for information about which database to use to organize my pictures (from the microscope, SEM, EM, confocal, etc.). What database do you use? Why?
:-) Torsten
Torsten Fregin
Universität Hamburg - Zoologisches Institut Abt. Neurophysiologie AG Wiese - Raum 413 Martin-Luther-King-Platz 3 20146 Hamburg, Germany Telefon *49-(0)40-42838-3931 Fax *49-(0)40-42838-3937 eMail Torsten.Fregin-at-zoologie.uni-hamburg.de Torsten-at-Fregin.de
} ... TIF, Tagged Image File Format) is one of the most flexible and } seems to be a de-facto standard now. ... } There are also some drawbacks. For example the tag "magnification". } If you put in a large number there, some programs (Word used to do } it), would interpret this as information about how big you wanted } the image displayed in a document. If the tag was, let's say } "1000", Word would interpret this as "the image is 1000 times as } large as the original", and try to print it at the original size, } resulting in an image a few pixels across. Not good.
This is an interesting point. My own opinion would ask if "magnification" being anything meaningful in the first place. Of course it is in the context of the images original acquisition, but many acquistion softwares, while writing magnification to the file, pay absolutely no attention to "print size" (or print resolution). That is, "mag" and "size" should be interdependent, and there is no provision maintaining the relationship. My first experience with SEM software would write the magnification to the TIF file, but pay absolutely no attention to the size neccessarily being 4 by 5. Therefore the mag was useless, unless it was subsequently always printed correctly.
This would be an interesting plugin for Photoshop ... that is, would recognize the "mag tag", and understand (and maintain) the relationship with print size. It would also be interesting if anyone was aware of current software for maintaining this relationship ... I am not aware of any.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
} At least TIFF and JPG formats have more or less standardized } tagged fields to store metadata (= data on the image itself). } The appropriate place to store microscope and image setting } data seems to be the EXIF fields. } (See Digital Still Camera Image File Format Standard. } Version 2.1. JEIDA-49-1998. } {http://www.kodak.com/global/plugins/acrobat/en/service/digCam/exifStandard. pdf} } A more recent draft in www.dpnet.com.cn/download/software/exif22.pdf.) } } [...] } } the EXIF field 8778 in our SEM FEI XL looks like this: } } [DatabarData] } flAccV = 20000.000 } flSpot = 6.000 } [...]
} the Hitachi S4700 FE-SEM produces a file with the same } name as the image and .txt extension, looking like this: } } [SemImageFile] } InstructName=S-4700 } FileName=xxxx 371bALSleft.tif } SampleName= } DataNumber= } } [...] } } In either case, one could easily extract this information } with a script in a text editor (i.e., building your own filter), } and then load the data in your system of choice.
Thanx for this info ... I'll at least know where to explore for more info. I did notice references to "micron bar", but knowing what it meant wasn't obvious. And, I did see references to "magnification", but never did see any instance which would have implied a "print size"(?)
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
I am into my 6th box of 250 sheets of the new formulation. Up until this 6th box, everything was fine. We use nitrogen burst (2 sec every 10 sec) and Kodak HRP developer. The 6th box, however, is a nightmare. Streaking, uneveness and even darker than the other 5.
Has anybody experienced this variation among the boxes? I can't find any lot number on the boxes to see if there might be variation among the lots.
Grrrr!
Paula :-P
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
I would just like to say "Thank you" to everyone that responded to my inquiry on native silica. My biggest problem is the charging issue. When I do get a good image of it at required magnifications, there is too much drift to photograph it. I have received a lot of responses and some offers to get in touch with individuals over the phone. I'm sorting through the information and intend to try some of the suggestions and get in touch with some of the individuals. Once again Thank you Linda
Linda S. McCorkle Ohio Aerospace Institute Materials Division, Polymers Branch NASA John H. Glenn Research Center at Lewis Field 21000 Brookpark Road Cleveland, OH 44135
I am looking for input from all of you experts in quantitative SEM/EDS analyses in trying to optimize conditions for EDS analyses of soda-lime type glass. I am currently trying to estimate the compositions in high alumina (Al2O3 ~ 20 wt%) soda lime glass with B2O3 levels anywhere from 0-15wt%. I am trying to account for some recent results which have been less than satisfactory. I am open to suggestions, indications of pitfalls, and misconceptions involving reliable quantitative analyses of glass by EDS. My objective is to get an estimate of the B2O3 levels with the balance from 100% in non-normalized results. Am I expecting too much from EDS methods by trying to treat the balance of totals below 100% as an indication of levels of light elements not detectable by EDS methods? The instrumentation conditions and procedures I have followed are listed below (in no particular order).
-Utilizing EDAX DX-4 (ver 2.3) system with AmRay 1830 SEM and tungsten filament. -Samples and standards mounted in Struers epoxy/resin mount. -Samples and standards cleaned with 1-micron diamond (to remove previous carbon coat) and methanol. I stopped using acetone, as I believe it leaves a film. -Glass sample and glass standard mounts both coated with carbon at the same time to hopefully insure identical carbon coat thickness. -Beam current closely monitored. Currently 1 nanoamp at 20kv operating voltage. I know lower operating voltage is desirable, but I need to check for titanium and iron. -Unknown glass is referenced to the NBS 620 and NBS 621 glass standards as well as a high alumina, zirconia-bearing in-house standard. -Working distance is closely monitored (frequent degaussing of lenses) to insure it is the same between the sample and standards. -Use box raster scan at ~ 800x for standard and ~1500-2500x in unknown (glass pools at grain boundaries). Should these be the same? -Zero degree tilt (checked with small level) and 24 degree TO angle.
Previously my data gave totals close to 100% in non- borosilicate glass. In borosilicate glasses, the amount less than 100% was consistent with the expected B2O3 levels. Now the results are inconsistent.
Thank you in advance for your anticipated input.
Kevin Selkregg Microanalytical Vesuvius Monofrax Inc
Yes, our JEOL 5600 has a similar txt file that comes along with all the images. I wrote a little program to extract this info in batch mode for import into a spreadsheet, database, or whatever. It's posted at:
http://www.mta.ca/~jehrman/software.htm
Simple to do. I have the (Turbo Pascal, I think) source code if anybody is interested.
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Martin Ramirez writes ... } } } At least TIFF and JPG formats have more or less standardized } } tagged fields to store metadata (= data on the image itself). } } The appropriate place to store microscope and image setting } } data seems to be the EXIF fields. } } (See Digital Still Camera Image File Format Standard. } } Version 2.1. JEIDA-49-1998. } } } {http://www.kodak.com/global/plugins/acrobat/en/service/digCam/exifStandard. } pdf} } } A more recent draft in www.dpnet.com.cn/download/software/exif22.pdf.) } } } } [...] } } } } the EXIF field 8778 in our SEM FEI XL looks like this: } } } } [DatabarData] } } flAccV = 20000.000 } } flSpot = 6.000 } } [...] } } } the Hitachi S4700 FE-SEM produces a file with the same } } name as the image and .txt extension, looking like this: } } } } [SemImageFile] } } InstructName=S-4700 } } FileName=xxxx 371bALSleft.tif } } SampleName= } } DataNumber= } } } } [...] } } } } In either case, one could easily extract this information } } with a script in a text editor (i.e., building your own filter), } } and then load the data in your system of choice. } } Thanx for this info ... I'll at least know where to explore for more info. } I did notice references to "micron bar", but knowing what it meant wasn't } obvious. And, I did see references to "magnification", but never did see } any instance which would have implied a "print size"(?) } } cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } www.micro-investigations.com (in progress)
without trying to sound too commercial, but you asked specifically this question: You may want to take a look at our analySIS software.
As you mentioned (rightly so!), magnification is really a misleading quality, as it always involves the print size. Our approach is to keep track of the calibration (i.e., nm/pixel or similar). This is independent of the display or print size. Of course, if you do display the image, this can then be converted back into a magnification. For example, when you print a report in our software, you can tell it to print the magnification. This will then be the true magnification of that print.
Of course, if you put it on a copier and blow it up 2x, the magnification printed is wrong again.
A scale bar is a much better approach, which we normally use.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: michael shaffer [mailto:michael-at-shaffer.net] Sent: Wednesday, July 02, 2003 6:02 AM To: MSA listserver
Mike Bode writes ...
} ... TIF, Tagged Image File Format) is one of the most flexible and } seems to be a de-facto standard now. ... } There are also some drawbacks. For example the tag "magnification". } If you put in a large number there, some programs (Word used to do } it), would interpret this as information about how big you wanted } the image displayed in a document. If the tag was, let's say } "1000", Word would interpret this as "the image is 1000 times as } large as the original", and try to print it at the original size, } resulting in an image a few pixels across. Not good.
This is an interesting point. My own opinion would ask if "magnification" being anything meaningful in the first place. Of course it is in the context of the images original acquisition, but many acquistion softwares, while writing magnification to the file, pay absolutely no attention to "print size" (or print resolution). That is, "mag" and "size" should be interdependent, and there is no provision maintaining the relationship. My first experience with SEM software would write the magnification to the TIF file, but pay absolutely no attention to the size neccessarily being 4 by 5. Therefore the mag was useless, unless it was subsequently always printed correctly.
This would be an interesting plugin for Photoshop ... that is, would recognize the "mag tag", and understand (and maintain) the relationship with print size. It would also be interesting if anyone was aware of current software for maintaining this relationship ... I am not aware of any.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
Suggest that you contact Dr. Ken Piel in our office. He is a world class expert on Pollen. E: kenpiel-at-mme1.com P: 413-746-6931
Hope this is helpful.
Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration. Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
At 01:41 PM 7/1/03 -0400, Scott Whittaker wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Foregut, midgut, or hindgut? The exact answer will depend a little on this, but arthropod gut contents are really pretty simple. Your questions reads like the midgut was removed. Fix, dehydrate, and critical point dry the intact gut + contents. HMDS could work here, especially for pollen. Mind, I'd suggest avoiding KOH digestion anyway. Dissect and fix, or, if the critter is too small, fix and dissect. Put double sticky tape on a SEM stub (I would've said double-sticky carbon tab, but given the ones sold now aren't very sticky anymore I won't). Or else use Al foil tape -- glue the Al side down with Ag paint. Keep the paint handy. Stick the dried gut on the tape glue ("sticky tabs" or sticky "paint" aren't sticky enough). Cut (tear) a line along the gut to open it. Use a #0 insect pin for this, smaller or larger depending on the beetle. Roll the gut along the tape. The contents will be exposed as the gut is stuck to the tape. Carefully use a fine insect pin to spread the gut contents out on the tape. Not a brush or anything similar, or the sample will be lost in the brush. Take the Ag paint and using a sharpened applicator stick (or the like) paint a ring close around the gut contents, then a line from this ring to the stub. This will leave less insulting surface (tape glue) between the sample and the conductive path to ground. Gold coat. Note that the foregut is chitinous, part of the exoskeleton. So it's more robust and easier to handle than contents from the midgut or hindgut, which are often in a fine, membranous structure. I think this is what you describe, but I'm not entirely certain you mean this "bag", and not the gut itself. The method works regardless of which you have, you just have to be more careful the farther back you go in the beetle, or if you have the "bag" and not the gut itself. You might want to get a pair of flexible, "fine" point dried insect handling forceps for handling the guts. Fine Science Tools carries these, but I'm sure other companies do also (especially entomology supply houses). I used this method a fair amount to look at amphipod guts, which are smaller than the beetle stomachs you describe. Just cut back on the coffee, or have a beer for lunch, before starting.
Phil
} I have a researcher who dissected out a beetle stomach, digested much of the } tissue away with KOH and wants to examine the contents to identify the } pollen it was feeding on. There was still a sheath of tissue around it so we } ran it through alcohol, placed it and a drop of alcohol between 2 slides, } and froze it in liquid nitrogen. When I snapped it apart it just fractured } in cross section rather than the longitudinal plane I had hoped. I then } stuck a piece of tape over and ripped it off. Basically we ended up just } destroying everything, but one of the pieces looked like it might have been } some kind of pollen grain at one time or other. } } Has anyone done this before? The entire stomach is about 4-500um long and } under the light microscope we can see several grains that look like they } could be pollen. Any suggestions on how to get them out and captured for SEM } examination?? I only have a couple more so experimentation is out and I am } running out of time. } } } Thanks } } Scott Whittaker } Laboratories of Analytical Biology } Smithsonian Institution } National Museum of Natural History } PO Box 37012 MRC104 } Washington DC 20013-7012 } 202-357-1651
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
I'm still having problems with uneven negatives. It really varies a lot. There are days that the problem is not as obvious and at times really bad negatives.
I wonder if there is an alternative brand? Does Fuji or Ilford make any EM film? If so where can one purchase them?
Rajesh Patel Robert Wood Johnson Medical School Department of Pathology 675 Hoes Lane Piscataway, NJ 08854
} } Fellow microscopists. } } } } Due to the imminent departure of my colleague Corinna Wauchope for the } } Land Down Under, I am in need a a person to help me run the North } } Campus EMAL at the University of Michigan } } (http://emalwww.engin.umich.edu/emal/fset.html). QUALIFIED } } individuals } } should respond to the job posting which is at: } } http://websvcs.itd.umich.edu/jobnet/ } } job_posting.php?postingnumber=3D032406&searchwhat=3Dcurrent } } DO NOT send resumes or applications to me personally they will NOT get } } processed. Please follow the instructions on the Job Posting Web } } Site. } } } } (http://www.umich.edu/~jobs/). } } } } } } } } John Mansfield PhD MInstP } } North Campus Electron Microbeam Analysis Laboratory } } 417 SRB, University of Michigan } } 2455 Hayward, Ann Arbor MI 48109-2143 USA } } Phone: (734) 936-3352 } } FAX (734) 763-2282 } } Cell. Phone: (734) 834-3913 } } (Leaving a phone message at 936-3352 is preferable to 834-3913) } } Email: jfmjfm-at-engin.umich.edu } } URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html } } Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Just a thought - make certain that the surface is really free of 1 micron alumina. At 20kV these particles can be nearly transparent to the beam depending on the surface beneath, especially if you are working in the backscatter mode. Their presence could inflate the apparent response from aluminum, throwing off the calculation by difference.
The boron content (presumably by weight?) sounds extremely high and potentially subject to volatilization. I understand that you are rastering the beam which should reduce that problem. However, you don't indicate what the beam diameter is. You might consider spreading out the beam as an additional means of avoiding volatile losses.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am looking for input from all of you experts in quantitative SEM/EDS analyses in trying to optimize conditions for EDS analyses of soda-lime type glass. I am currently trying to estimate the compositions in high alumina (Al2O3 ~ 20 wt%) soda lime glass with B2O3 levels anywhere from 0-15wt%. I am trying to account for some recent results which have been less than satisfactory. I am open to suggestions, indications of pitfalls, and misconceptions involving reliable quantitative analyses of glass by EDS. My objective is to get an estimate of the B2O3 levels with the balance from 100% in non-normalized results. Am I expecting too much from EDS methods by trying to treat the balance of totals below 100% as an indication of levels of light elements not detectable by EDS methods? The instrumentation conditions and procedures I have followed are listed below (in no particular order). } } -Utilizing EDAX DX-4 (ver 2.3) system with AmRay 1830 SEM and tungsten filament. } -Samples and standards mounted in Struers epoxy/resin mount. } -Samples and standards cleaned with 1-micron diamond (to remove previous carbon coat) and methanol. I stopped using acetone, as I believe it leaves a film. } -Glass sample and glass standard mounts both coated with carbon at the same time to hopefully insure identical carbon coat thickness. } -Beam current closely monitored. Currently 1 nanoamp at 20kv operating voltage. I know lower operating voltage is desirable, but I need to check for titanium and iron. } -Unknown glass is referenced to the NBS 620 and NBS 621 glass standards as well as a high alumina, zirconia-bearing in-house standard. } -Working distance is closely monitored (frequent degaussing of lenses) to insure it is the same between the sample and standards. } -Use box raster scan at ~ 800x for standard and ~1500-2500x in unknown (glass pools at grain boundaries). Should these be the same? } -Zero degree tilt (checked with small level) and 24 degree TO angle. } } Previously my data gave totals close to 100% in non- borosilicate glass. In borosilicate glasses, the amount less than 100% was consistent with the expected B2O3 levels. Now the results are inconsistent. } } Thank you in advance for your anticipated input. } } Kevin Selkregg } Microanalytical } Vesuvius Monofrax Inc
Paula: We were among the first ones to have problems with the "new formulation" and as my people used to say it is a "limon" or a lemon in English. This film has never worked properly for us, even as we accommodate our processing to fit its particular needs. We go through 200 to 250 plates in a week to week and a half, so we see the differences in each box and lot. Each box of film is like a box of melted chocolates. You never know the mess you will get. We use Kodak D19 at 2:1 water/developer ratio, for developing and each time I develop, new surprises come to life. I'm trying to contain the comments that this film deserve in reality. The old formulation worked great and it was consistent from box to box. But now this film is just making life very interesting in the dark room. I'll just say, is just a matter of time and patience to develop the "new Formulation".
Omayra Velez EM Specialist Patholy Department Weill Cornell Medical College NY,NY. 10021 Phone: 212-746-6437
--- Paula Sicurello {patpxs-at-gwumc.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Listers, } } I am into my 6th box of 250 sheets of the new } formulation. Up until this 6th box, everything was } fine. We use nitrogen burst (2 sec every 10 sec) } and Kodak HRP developer. The 6th box, however, is a } nightmare. Streaking, uneveness and even darker } than the other 5. } } Has anybody experienced this variation among the } boxes? I can't find any lot number on the boxes to } see if there might be variation among the lots. } } Grrrr! } } Paula :-P } } Paula Sicurello } George Washington Univ. Medical Center } Electron Microscope Lab } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax } }
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bop00rav-at-sheffield.ac.uk) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, July 2, 2003 at 16:59:13 ---------------------------------------------------------------------------
Email: bop00rav-at-sheffield.ac.uk Name: BOB VASEY
Organization: UNIVERSITY OF SHEFFIELD
Education: Graduate College
Location: sheffield, uk
Question: Hello,
My name is Robert Vasey and I am a PhD student at the university of Sheffield, England. I am working on Arabidopsis roots. I am currently trying to make root sections to stain for lignin and cellulose. I have been using LR White resin but it does not allow my stains to penetrate (phloroglucinol for lignin). I have also tried staining with Methylene Blue followed by counterstaining with Fuchsin Red, without success.
To stain for cell wall components, specifically lignin and cellulose, can anyone advise me on:
(a) What is the appropriate resin to use (b) What stains can I use - I would be grateful for recipes and how to apply them. (c) Are there any good counterstaining techniques or metachromatic stains to stain multiple cell components (apart from Toluidine Blue).
I would be grateful for any advice as I am struggling with this.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gaston.garcia-at-123.cl) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, July 1, 2003 at 14:25:02 ---------------------------------------------------------------------------
Email: gaston.garcia-at-123.cl Name: gaston
Organization: leon prado school
Education: Graduate College
Location: santiago, chile
Question: I have a microscope with a reticle eyepiece. I want to know the scale of reticle. I see for an objetive 40X/0.65 and the scale is a 100 parts. Can you help me please?
Dear Sir or Madam: We take the liberty to write to you and will appreciate your attention on this mail. We have a foundry engaging in brake disc casting and have been in this field for years and have gained the fame for our products step by step.
With the CNC lathe from Japan, we are able to keep the run out below 0.04mm and the surface roughness to be 1.6.
Now we export about 50 containers of brake rotor and other casting parts annually to the US market, but our ability is not fully used and we are now seeking for new buyers.
Quotation by drawings or samples will be ok and we will quote in 3days after reception of your inquiry.
You can refer to our website for details: www.szautoparts.com and www.bacoengineering.com
Thanks and best regards
China machinery imp/exp co., ltd Tel: 86-512-65325936 Fax: 86-512-65321605 Korin Tian the sales manager
Dear Sir or Madam: We take the liberty to write to you and will appreciate your attention on this mail. We have a foundry engaging in brake disc casting and have been in this field for years and have gained the fame for our products step by step.
With the CNC lathe from Japan, we are able to keep the run out below 0.04mm and the surface roughness to be 1.6.
Now we export about 50 containers of brake rotor and other casting parts annually to the US market, but our ability is not fully used and we are now seeking for new buyers.
Quotation by drawings or samples will be ok and we will quote in 3days after reception of your inquiry.
You can refer to our website for details: www.szautoparts.com and www.bacoengineering.com
Thanks and best regards
China machinery imp/exp co., ltd Tel: 86-512-65325936 Fax: 86-512-65321605 Korin Tian the sales manager
Dear reader, We used for years in combination with TEM the Reflection Contrast Microscope (RCM)for a overview of the ultrathin sequentional sections (EPON,Spurr, Lowicryl or Cryo) Al you need is a fluorescence microscope with 3 extra parts: A central stop, antiflex objective and a polarization block for more infomation see the website or mail
At the NESM Woods Hole Meeting in May, the Kodak rep suggested adjusting the nitrogen burst to 2sec every 8sec (instead of 10). I have no clue if this will help.
Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals
-- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Listers, } } I am into my 6th box of 250 sheets of the new formulation. Up until this 6th } box, everything was fine. We use nitrogen burst (2 sec every 10 sec) and Kodak } HRP developer. The 6th box, however, is a nightmare. Streaking, uneveness and } even darker than the other 5. } } Has anybody experienced this variation among the boxes? I can't find any lot } number on the boxes to see if there might be variation among the lots. } } Grrrr! } } Paula :-P } } Paula Sicurello } George Washington Univ. Medical Center } Electron Microscope Lab } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax } }
Greetings, As for part a, while I have no specific experience with histo stains on LR White, I can say that using Butyl-methyl methacrylate produces blocks that section much more easily than does LR White, and certainly stain well with many things as a rule. As for part b, a typical stain for celluose is calcofluor white (sometimes called cellufluor). It is not strictly specific for cellulose as it will stain some other beta glucans, but it is easy and widely used. You can also use polarized light which avoids staining altogether and can be quantitative.
Good luck, Tobias Baskin } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (bop00rav-at-sheffield.ac.uk) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } July 2, 2003 at 16:59:13 } --------------------------------------------------------------------------- } } Email: bop00rav-at-sheffield.ac.uk } Name: BOB VASEY } } Organization: UNIVERSITY OF SHEFFIELD } } Education: Graduate College } } Location: sheffield, uk } } Question: Hello, } } My name is Robert Vasey and I am a PhD student at the university of } Sheffield, England. I am working on Arabidopsis roots. I am } currently trying to make root sections to stain } for lignin and cellulose. I have been using LR White resin but it } does not allow my stains to penetrate (phloroglucinol for lignin). I } have also tried staining with Methylene Blue followed by } counterstaining with Fuchsin Red, without success. } } To stain for cell wall components, specifically lignin and } cellulose, can anyone advise me on: } } (a) What is the appropriate resin to use } (b) What stains can I use - I would be grateful for recipes and how } to apply them. } (c) Are there any good counterstaining techniques or metachromatic } stains to stain multiple cell components (apart from Toluidine Blue). } } I would be grateful for any advice as I am struggling with this. } } Bob. } } } } ---------------------------------------------------------------------------
The reticle scale can sometimes be arbitrary. Regardless of whether it is arbitrary or not, you need to calibrate the reticle. There is an item available called a "microscope slide", which is a special glass slide that you place on the microscope stage. The slide has measureable divisions of known length appropriate for calibrating high magnifications. The calibration would need to be made for each objective lens on the microscope.
Stu Smalinskas Metallurgist SKF USA Plymouth, Michigan
-----------------------------------
Gaston wrote:
Email: gaston.garcia-at-123.cl Name: gaston
Organization: leon prado school
Education: Graduate College
Location: santiago, chile
Question: I have a microscope with a reticle eyepiece. I want to know the scale of reticle. I see for an objetive 40X/0.65 and the scale is a 100 parts. Can you help me please?
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Good day to all, I have been a member of MSA for over 5 years now - I first joined in Oklahoma. In Oklahoma, the local society was quite busy and I really enjoyed being a part of a group of people that enjoyed the same things I did! My current problem is that I live in North Carolina where the local chapter seems to have completely died - or at least no one will let me know anything about meetings etc even after repeated request! My question - is there any other state society near NC that would allow me the opportunity to join or does anyone in the national MSA society have suggestions as to how to start a local chapter or revive the one that is suppose to be in NC? Thanks a whole bunch, Connie Cummings
Connie,
The Appalacian Regional Microscopy Society (AREMS) is an active chapter of MSA and serves microscopists in VA, WV, NC, SC, TN, and GA. You can find more about AReMS on-line at www.uvawise.edu/arems/index.html or contact me if you wish to become a member. My numbers are below.
Thank you,
Rich
Richard Fiore NC State University Analytical Instrumentation Facility 2410 Campus Shore Drive 318 EGRC, Campus Box 7531 Raleigh, NC 27695 Tel: 919-515-2348 Fax: 919-515-6965 Email: rafiore-at-ncsu.edu
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I forgot to meantion that the next AReMS meeting is October 2-3, 2003 in Boone, NC at the Broyhill Inn & Conference Center.
Rich
Original Message -------- Subject: MSA help
Good day to all, I have been a member of MSA for over 5 years now - I first joined in Oklahoma. In Oklahoma, the local society was quite busy and I really enjoyed being a part of a group of people that enjoyed the same things I did! My current problem is that I live in North Carolina where the local chapter seems to have completely died - or at least no one will let me know anything about meetings etc even after repeated request! My question - is there any other state society near NC that would allow me the opportunity to join or does anyone in the national MSA society have suggestions as to how to start a local chapter or revive the one that is suppose to be in NC? Thanks a whole bunch, Connie Cummings
Connie,
The Appalacian Regional Microscopy Society (AREMS) is an active chapter of MSA and serves microscopists in VA, WV, NC, SC, TN, and GA. You can find more about AReMS on-line at www.uvawise.edu/arems/index.html or contact me if you wish to become a member. My numbers are below.
Thank you,
Rich
Richard Fiore NC State University Analytical Instrumentation Facility 2410 Campus Shore Drive 318 EGRC, Campus Box 7531 Raleigh, NC 27695 Tel: 919-515-2348 Fax: 919-515-6965 Email: rafiore-at-ncsu.edu
A position is available at Texas Instruments in Dallas TX as a Physical Failure Analyst. Details are provided below. To apply e-mail your resume to faresume-at-list.ti.com with "DMFA" in the subject line of the e-mail.
Texas Instruments Product Engineer SiTD CMOS1
You'll develop the latest technology and will have many learning opportunities in this dynamic engineering position.
As a CMOS1 Product Engineer, you'll be responsible for PFA and EFA of C021 material. Other responsibilities include: SRAM cell debug CO21 F/A of low K dielectrics Metal gate failure analysis Leading edge product failure analysis on wireless, DSP, and microprocessors Pareto creation Baseline F/A Scrap assignment
The ideal candidate will have 5-10 years of related experience. Process knowledge is a must. PFA candidates with Cu experience on 130nm or 90nm nodes are preferred. Experience with reliability fails and constructional analysis is also preferred.
Additional information follows Primary Responsibilities: PFA and EFA of C021 Products and understanding of process limitations through physical and electrical characterization Supporting & Secondary Responsibilities: Baseline pareto creation and development. Interaction with Process Integration engineers, Process Engineers, and other Product Engineers in order to ensure that the correct solutions are implemented for yield and reliability learning. Management /Organization Skills: Detail individual with a desire to derive statistically significant backup for the improvement of our baseline process, and the evaluation of new processes. Opportunities for Improvements: Electrical Failure Analysis, device physics understanding, process knowledge, micromanipulator tool knowledge, automated FIB, TEM, SEM, statistical analysis, copper interconnect, low-K dielectrics, metal gates, etc. Complex Tasks: Failure analysis of processes that have never been F/A before. Fail mechanisms that do not have any history behind them. Fail mechanisms that will push past the common failure analysis techniques. Technical Abilities: 130nm/90nm process, deprocessing techniques, electrical isolation techniques, SEM use, TEM use, FIB use Team & People Skills: Work with a team of a handful of failure analysis engineers as part of a group working on the qualification of TI's 65nm technology. Interaction with business units ranging from the wireless products to microprocessors. Projects & Deliverables: Failure pareto, constructional analysis, process characterization, design rules and yield limiters, scrap assignment, etc. Additional skills and experience (include years of required experience): 5-10 years List unique selling features of this position, team or project: Dynamic team involved on the development of TI's 65nm technology. Opportunity to gain visibility within the development team, TI's manufacturing facilities, and business units. Special Work Environment: F/A lab with chemicals, SEM, FIB, TEM, polish wheels, etc. EFA lab with all the necessary tools for electrical isolation.
E-mail your resume to faresume-at-list.ti.com with "DMFA" in the subject line of the e-mail.
Kim Christensen Texas Instruments, Inc 13560 N Central Expressway Dallas, TX 75243 Ph: 972-995-3855 E-mail: kchristen-at-ti.com
What you need to find is something you know the size of that you can look at through the microscope. You can then calibrate the grid you have for each objective size. Red blood cells are of a known size and can give you an approximation of the scale of the grid. What you really need is called a "stage micrometer". This is an accurate scale mounted on a slide which will allow you to accurately calibrate your grid. If you can, take a look at this web site for more information:
Dr. Allerton: I am the President/CEO of a company in Natick, MA that specializes in Optical coatings. We specialize in antireflection coatings of optics. I am going to have to look a little bit further in to the exact specifications of the AR coating on your microscope objectives but I believe we should be able to strip and recoat the lenses with a durable coating with specifications at least as good as the original.
Bob O'Leary - President, CEO Optical Coating Corporation 01760 6R Mercer Road Natick, MA P: 508.655.1650 F: 508.653.0729 http://www.opticalcc.com BOLeary-at-opticalcc.com
----- Original Message ----- } From: "Ron Allerton" {rsallerton-at-caribsurf.com} To: {Microscopy-at-sparc5.microscopy.com} Cc: {zaluzec-at-sparc5.microscopy.com} Sent: Tuesday, July 01, 2003 3:41 PM
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Hi listers,
Perhaps I didn`t put my question very clearly. I am aware that tiff can hold extra info like mag (and calibration and scale bar etc). But my main question is: is there a STANDARD (convention) way of naming these tags. It seems to me that a lot of tags could be defined so they are useful in TEM, SEM, STM, LM etc etc. Defining a set of rules like this could maybe stop commercial companies of inventing their own file formats (which is a bad idea for scientists in all respects except for the company monopoly)
The good thing about that format (the MSA-tiff?) is that it is still viewable with a normal image viewer but to get more info you need special software. There are good GNU imageviewers available that allow to easily include this new image format and give them the functionality (and better) of other commercial (very expensive) programs.
kind regards,
Jo
On Wed, 2003-07-02 at 19:33, James M. Ehrman wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Yes, our JEOL 5600 has a similar txt file that comes along with all } the images. I wrote a little program to extract this info in batch mode } for import into a spreadsheet, database, or whatever. It's posted at: } } http://www.mta.ca/~jehrman/software.htm } } Simple to do. I have the (Turbo Pascal, I think) source code if anybody } is interested. } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/~jehrman } } } } michael shaffer wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Martin Ramirez writes ... } } } } } At least TIFF and JPG formats have more or less standardized } } } tagged fields to store metadata (= data on the image itself). } } } The appropriate place to store microscope and image setting } } } data seems to be the EXIF fields. } } } (See Digital Still Camera Image File Format Standard. } } } Version 2.1. JEIDA-49-1998. } } } } } {http://www.kodak.com/global/plugins/acrobat/en/service/digCam/exifStandard. } } pdf} } } } A more recent draft in www.dpnet.com.cn/download/software/exif22.pdf.) } } } } } } [...] } } } } } } the EXIF field 8778 in our SEM FEI XL looks like this: } } } } } } [DatabarData] } } } flAccV = 20000.000 } } } flSpot = 6.000 } } } [...] } } } } } the Hitachi S4700 FE-SEM produces a file with the same } } } name as the image and .txt extension, looking like this: } } } } } } [SemImageFile] } } } InstructName=S-4700 } } } FileName=xxxx 371bALSleft.tif } } } SampleName= } } } DataNumber= } } } } } } [...] } } } } } } In either case, one could easily extract this information } } } with a script in a text editor (i.e., building your own filter), } } } and then load the data in your system of choice. } } } } Thanx for this info ... I'll at least know where to explore for more info. } } I did notice references to "micron bar", but knowing what it meant wasn't } } obvious. And, I did see references to "magnification", but never did see } } any instance which would have implied a "print size"(?) } } } } cheerios ... shAf :o) } } Avalon Peninsula, Newfoundland } } www.micro-investigations.com (in progress) } } } -- Dr. J. Verbeeck Electron Microscopy for Materials Research University of Antwerp, Belgium
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id VAA14149 for dist-Microscopy; Fri, 4 Jul 2003 21:23:40 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id VAA14146 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Fri, 4 Jul 2003 21:23:09 -0500 (CDT) Received: from mta7.pltn13.pbi.net (mta7.pltn13.pbi.net [64.164.98.8]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id VAA14139 for {microscopy-at-sparc5.microscopy.com} ; Fri, 4 Jul 2003 21:22:56 -0500 (CDT) Received: from nigelhomepc (adsl-64-175-40-97.dsl.pltn13.pacbell.net [64.175.40.97]) by mta7.pltn13.pbi.net (8.12.9/8.12.3) with SMTP id h652GgeS024709 for {microscopy-at-msa.microscopy.com} ; Fri, 4 Jul 2003 19:16:42 -0700 (PDT) Message-Id: {4.1.20030704190401.00ae4960-at-yellow.ucdavis.edu} X-Sender: nigel-at-yellow.ucdavis.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 4.1
POSTDOCTORAL POSITIONS IN ELECTRON MICROSCOPY
DEPARTMENT OF CHEMICAL ENGINEERING AND MATERIALS SCIENCE UNIVERSITY OF CALIFORNIA-DAVIS
Two postdoctoral positions are available in the Electron Microscopy Group at the University of California-Davis (UCD). Research in the newly formed Electron Microscopy Group focuses on the use of atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. Current research programs involve catalysts, ferroelectrics, high-Tc superconductors and optoelectronic/high-power semiconducting materials and devices. The positions that are currently available are for the study of metal-oxide interactions in heterogeneous catalysts and the vacancy mediated fatigue mechanisms in ferroelectric thin films. The experimental analyses will primarily be performed using the facilities in the National Center for Electron Microscopy at Lawrence Berkeley National Laboratory. In particular, extensive use will be made of the new 200kV monochromated FEI G2 Tecnai that has recently been installed and the Cs-corrected VG HB501 dedicated STEM that will be installed later this year. There will also be the opportunity for both postdoctoral appointees to be involved in the development of in-situ stages for the recently funded 200kV field emission TEM to be installed at UC-Davis next year. Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevant materials issues and an ambition to be part of a developing program pushing at the frontiers of interface science. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience.
Nigel D. Browning
Department of Chemical Engineering and Materials Science University of California-Davis One Shields Ave Davis, CA 95616
AND
National Center for Electron Microscopy, MS 72-150 Lawrence Berkeley National Laboratory Berkeley, CA 94720
EPSRC funded Ph.D. Research Project on the Chemistry and Biological Activity of Nanoparticles using Aberration-Corrected Scanning Transmission Electron Microscopy (SuperSTEM).
Institute for Materials Research (IMR) at the University of Leeds and Daresbury Laboratories
Supervisors: Dr Rik Brydson & Dr Andy Brown, Institute for Materials Research, University of Leeds, Leeds LS2 9JT, U.K. Please Contact mtlrmdb-at-leeds.ac.uk or a.p.brown-at-leeds.ac.uk or phone +44 (0)113 343 2348/2369 for further details.
What is SuperSTEM ? The SuperSTEM project involves the development of two aberration-corrected STEM instruments in a five-year, £4.2 million project funded as a result of the NW Science Review. The microscopes have been installed in a specially designed and purpose built facility on the Daresbury Laboratory site in Cheshire, and ultimately they will be available as a national resource. SuperSTEM provides a world-leading atomic imaging and analysis facility capable of chemical spectroscopy from single atoms and atomic columns further details may be found by visiting www.superstem.org.uk. The SuperSTEM project is led by five scientists from four partner universities [Peter Goodhew, Mick Brown, Alan Craven, Rik Brydson and Gordon Tatlock]. The full-time research team comprises a Technical Director [Dr Andrew Bleloch], two research microscopists [Dr Uwe Falke and Dr Meiken Falke], a computer specialist [Will Costello] and a technician [Peter Shiels]. This project is part of a set of six studentships recently funded by the Engineering and Physical Sciences Research Council and as such is fully funded (fees and maintenance) for both UK and EC students. The project will be based at the IMR in Leeds but will involve extensive travel to Daresbury Laboratories.
The studentship will examine a limited range of metal oxide nanoparticles of varying mean particle size and size distribution. Systems of current interest include: (a) bare, carboxylic acid stabilized and charge stabilized Fe3O4 nanoparticles produced by standard colloidal routes at Leeds. (b) g-Al2O3 and -g-Al2O3/ CeO2 catalyst supports produced at Leeds or obtained commercially.
Atomic resolution HAADF and EELS measurements using the SuperSTEM I facility will be undertaken. With an aberration corrected probe size of around 0.14 nm and below it will be possible to resolve the oxygen sub-lattice. A combination of Z contrast imaging and EELS measurements at particle surfaces (both direct and aloof beam measurements to maximize surface excitations) and within the particle core will give structural and spatially resolved chemical/ electronic information. It is intended that results will be compared with theoretical calculations, both ab-initio total energy/ electronic structure calculations as well thermodynamic predictions.
An extension of this project, pertaining directly to the life sciences, would involve an investigation of iron oxide nanoparticles in human liver biopsies in collaboration with Dr Alice Warley and Dr Jonathan Powell at King¹s College London. -- _____________________________ Dr. Rik Brydson, Leeds Electron Microscopy and Spectroscopy (LEMAS) Centre Institute for Materials Research, University of Leeds, Leeds LS2 9JT, U.K.
New MSc in Nanoscale Science and Technology Visit http://www.ee.leeds.ac.uk/nanomsc/ ______________________________
UK SuperSTEM facility - Atomic resolution Analysis http://www.superstem.org.uk
_______________________________
Bedtime reading.......... Series: Microscopy Handbooks - Electron Energy Loss Spectroscopy by Rik Brydson Published by BIOS September 2001 £29.99 ISBN: 1859961347
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===================================== EELS and X ray database : http://www.cemes.fr/~eelsdb/ =====================================
I don't think there is or will be ever a standard that can anticipate and provide tags for all possible fields. Let's face it: the imaging field is HUGE, and microscopy is a tiny part of that. You have many fields, where magnification is a meaningless quantity. For example, take all your vacation photos. You can't assign a magnification to them. Then there is the medical field. Do you want to provide a field for each disease of each bone for the X-rays? Dental images, Forensics, Space imaging, Military Imaging, etc., all with their specific requirements. TIF proides SOME standard fields, but they are metadata about the image itself (size, magnification, etc.) If you need to store more information, you need to use an archiving system (here again a pointer to our software, but other software does similar things).
If I remember corectly, the TIF tags are just numbers. Certain numbers are defined in the TIF specs and have specific meaning (for example "Number of pixels in X direction"). Other numbers are not defined and can be used by other software.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: jo verbeeck [mailto:joverbee-at-ruca.ua.ac.be] Sent: Friday, July 04, 2003 1:49 AM To: MSA listserver
My name is Bob Vasey and I am currently conducting light microscopy on Arabidopsis using LR White resin. I want to stain root sections for cell wall components, specifically lignin and cellulose. I would also like to try some counterstains to stain multiple cell wall components on a single section.
Could anyone help with the following:
(a) A protocol for Methylene Blue/Fuchsin counterstaining that works in LR White (including recipes for stains). When I try, the Fuchsin seems to wash out the Toluidine Blue. (b) Suggest any other polychromatic stains (apart from Toluidine Blue) or counterstains. (c) I have tried phloroglucinol for lignin but it does not seem to be taken up, probably because it is made up in ethanol. Are there any other stains/protocols that work for lignin in LR White. Will phloroglucinol work in epoxy resins, e.g. Spurrs or Araldite? (d) Am I using the best resin do other resins allow a greater diversity of stains to be used. Does LR White give the best quality (intensity) of stain sometimes my sections look a bit pale when Toluidine Blue is used. (e) I have heard a stain called Paragon mentioned could anyone tell me what it is and how to make and use it? (f) In the literature, some people fix in glutaraldehyde, some in formaldehyde and some in both. Could anyone tell me the difference between them and which is the best to use?
I would be grateful for any advice as I am a PhD student in a department with limited experience of conducting staining of resin sections, so I am having to work it out as I go along!
My email is { HYPERLINK "mailto:bop00rav-at-sheffield.ac.uk" }bop00rav-at-sheffield.ac.uk. The two 00 in the middle are zeros and not capital os!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bbaxter-at-westminstercollege.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, July 6, 2003 at 13:18:04 ---------------------------------------------------------------------------
Email: bbaxter-at-westminstercollege.edu Name: Bonnie K. Baxter
Organization: Westminster College
Education: Undergraduate College
Location: SLC, UT
Question: I am attempting some microscopy of microbes from Great Salt Lake. Our best scope is a Nikon inverted phase contrast. The problem I'm having is doing iol immersion with an inverted scope. ANy pointers for doing this upside down and actually keeping the bugs in focus? thanks!! Bonnie
We have a Denton Vacuum DFE-3 Freeze Etch Apparatus with DRM-1 Resistance Monitor (for precision C and C-Pt film deposition) available to anyone interested.
You must pick up or arrange for packing and shipping.
For further information contact Sue Gruber, (segruber-at-mtholyoke.edu).
Marian Rice Assoc. Dir. Labs Dept Biological Sciences Mount Holyoke College South Hadley, MA 01075
We have a Sorvall MT-2 ultramicrotome and we are missing the glass knife holder. Does somebody know of a service that provides spare parts for old microtomes? Thanks you very much. Tea
-- *************************************** Tea Meulia Research Scientist and Director Molecular and Cellular Imaging Center Ohio State University/OARDC 1680 Madison Ave. Wooster OH 44691
We need to move a JEOL 35C out of our facility. Your only cost would be shipping. It is fully functional, except it does need a turbo pump. We purchased this new in 1980 and has been under service contract with JEOL service until one year ago when we stopped using it. It was purchase loaded with about every option they had including a four crystal spectrometer. We do have five crystals for that if you are interested. Give me a call or reply to this message if you are interested.
Mark Windland Honeywell Minneapolis, Minnesota 763-954-2845 mark.j.windland-at-honeywell.com {mailto:mark.j.windland-at-honeywell.com}
We are doing immunogold labeling for electron microscopy on adult drosophila eyes. I would like to chat with someone who has done successful labeling on this tissue. Off-line replies are preferred. Thank you very much.
Hong
Hong Yi Emory EM Tel: (404) 727-8692 (Office), (404) 712-8491 (Lab) Fax: (404) 727-3157 Email: hyi-at-emory.edu
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We have a Sorvall MT-2 ultramicrotome and we are missing the glass knife holder. Does somebody know of a service that provides spare parts for old microtomes? Thanks you very much. Tea
-- *************************************** Tea Meulia Research Scientist and Director Molecular and Cellular Imaging Center Ohio State University/OARDC 1680 Madison Ave. Wooster OH 44691
Hello, I've tried a few times now to do cryo SEM after freeze fracturing and cold coating samples. Each time I end up having the fracturing removing the sample from the stage/hat/TEM grid, or ending up melting upon transfer from the freeze fracture device into the cryo SEM wand. I am working with a Hitachi S5000 SEM, a Balzers 301 Freeze fracture unit, a gatan 626 53h31 cryotransfer system, a gatan dry pumping station, a gatan 626DH cryoholder, and either clipring holders for TEM grids, or gatan specimen capsules (626.04352).
I really dislike using TEM grids with samples, fracturing and then trying to load the TEM grids into the gatan clipring holder. I want to try using either larger hats, or the gatan specimen capsules, but I'm either unsure how to mount the hats into the gatan cryoholder, or I'm unsure how to mount the gatan specimen capsules into the Balzers.
Could anyone give me some advice into doing a successful freeze fracture, transfer, and then image with the cryo sem holder? The manuals that came with the Gatan hardware are pretty vague at many many levels. Thanks for the help.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I am interested in cutting the seeds of crops such as rice, corn, cotton, and soybeans, and identifying protein, lipid, and starch components in SEM. Can you tell me what approaches and methods I could use?
Good morning We are experiencing some contamination issues in our process. These particulates are poly clothing, dust rubber compounds and other "stuff". With the exception of a few metal particles none of these items are identifiable with the SEM although it is excellent for actual physical structure identification. My question is what optical microscopy techniques do you use for identifying these particulates such as filters, polarizers etc.. I do have most filters at my disposal. I just need the right combination to get positive identification. Also the base the materials are mounted to view these particulates may be important? I am currently using a white background but this may have some effect due to the translucence of some of the materials. Is there a standard background material that you use? Thank you in advance for your input.
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com
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To all subscribers, Due to circumstances beyond my control I must downsize/close my metallurgical engineering consulting practice and move toward retirement. As a first step I am liquidating all of my metallographic laboratory equipment which includes a JEOL T-330 SEM and a full compliment of Buehler metallographic sample preparation/examination equipment. All of the items were purchased new and have been used only by myself in my practice. The T-330 SEM has always been and is currently under a manufacturers (JEOL) service agreement. For anyone interested in
a list of the items, specifications and photographs of same please contact me offline at kmuszar-at-iquest.net.
Karl E. Muszar Jr. M.E.I., Inc. P.O. Box 50366 6161 E. 75th Street Indianapolis, Indiana 46250 (O) 317-842-1906 (F) 317-595-7606
I am looking for a PC file conversion possibility of our old 1993 Voyager EDS sun system (.eds) to a ASCII or text format. Or does somebody know the file structure of data saved in .eds-format ?
Many thanks in advance,
Carsten Ronning
************************************* Dr. Carsten Ronning School of Materials Science and Engineering Georgia Institute of Technology 771 Ferst Drive Atlanta, GA 30332-0245 USA
on leave: II. Insitute of Physics University of Goettingen Bunsenstr. 7-9 37073 Goettingen, Germany ****************************************
We manufacture the JB-4, which was once a Sorvall product. Contact us and we will try to be of assistance.
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Tel: 413 786-9322 Fax: 413 789-2786 mnesta-at-ebsciences.com www.ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: tea meulia [mailto:meulia.1-at-osu.edu] Sent: Sunday, July 08, 2007 10:36 AM To: Microscopy-at-sparc5.microscopy.com
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We have a Sorvall MT-2 ultramicrotome and we are missing the glass knife holder. Does somebody know of a service that provides spare parts for old microtomes? Thanks you very much. Tea
-- *************************************** Tea Meulia Research Scientist and Director Molecular and Cellular Imaging Center Ohio State University/OARDC 1680 Madison Ave. Wooster OH 44691
Any suggestions on how to avoid getting small bubbles in mixed epoxy?
My application is in cementing ultrasonic transducers to glass. The transducers look like very thick washers, about 1-inch in diameter. I am getting a cavitation ringing in the applied voltage (10-volts, at 20-30 KHz). I suspect the bubbles are causing this, but do not know for sure.
I've tried slowly mixing the resin and catalyst beads, but the bubbles persist.
Any suggestions are welcome.
Nathan Haese Moraga, CA
__________________________________ Do you Yahoo!? SBC Yahoo! DSL - Now only $29.95 per month! http://sbc.yahoo.com
After mixing mounting medias in our lab (epoxy, OCT, ProLong, etc.), we place the container under vacuum to get the bubbles to rise out of the media. The amount of time spent under vacuum will depend on how quickly the media dries. This could possibly work for your application.
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
You could try putting the epoxy in a vacuum for a short time or curing under vacuum.
Ron L
-----Original Message----- } From: Nathan Haese [mailto:nathanhaese-at-yahoo.com] Sent: Wednesday, July 09, 2003 1:27 PM To: Microscopy-at-sparc5.microscopy.com
List members,
Any suggestions on how to avoid getting small bubbles in mixed epoxy?
My application is in cementing ultrasonic transducers to glass. The transducers look like very thick washers, about 1-inch in diameter. I am getting a cavitation ringing in the applied voltage (10-volts, at 20-30 KHz). I suspect the bubbles are causing this, but do not know for sure.
I've tried slowly mixing the resin and catalyst beads, but the bubbles persist.
Any suggestions are welcome.
Nathan Haese Moraga, CA
__________________________________ Do you Yahoo!? SBC Yahoo! DSL - Now only $29.95 per month! http://sbc.yahoo.com
Nathan: have you tried outgassing your epoxy under vacuum before you use it on the transducers? This works well for embedding and backfilling epoxies. It would depend on the cure time of your epoxy; if it cures too fast (under 15 minutes) this wouldn't be a good thing to try as the epoxy would be on its way to setting before all the bubbles were out. Let me know how it goes.
Nathan Haese wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } List members, } } Any suggestions on how to avoid getting small bubbles } in mixed epoxy? } } My application is in cementing ultrasonic transducers } to glass. The transducers look like very thick } washers, about 1-inch in diameter. I am getting a } cavitation ringing in the applied voltage (10-volts, } at 20-30 KHz). I suspect the bubbles are causing } this, but do not know for sure. } } I've tried slowly mixing the resin and catalyst beads, } but the bubbles persist. } } Any suggestions are welcome. } } Nathan Haese } Moraga, CA } } __________________________________ } Do you Yahoo!? } SBC Yahoo! DSL - Now only $29.95 per month! } http://sbc.yahoo.com
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I agree with Christopher. Be carefully for the time under vacuum vs. polymerisation time. Also be careful with the rate of pumping down. You should not have trapped gas in your samples but expanding gas can be destructive. I prefer to "see" the bubbles raising to prevent "boiling" of the resin. If this occur your whole vacuum system is contaminated. I left a sample to cure under vacuum once. Big mistake! You need it to cure in air. The reassure of the atmosphere will close the bubbles that are left over. Depending on the resin/epoxy you can heat it to 40 degrees prior to exposing it to the vacuum to decrease viscosity allowing a shorter time in the vacuum.
-----Original Message----- } From: Nathan Haese [mailto:nathanhaese-at-yahoo.com] Sent: Wednesday, July 09, 2003 7:27 PM To: Microscopy-at-sparc5.microscopy.com
List members,
Any suggestions on how to avoid getting small bubbles in mixed epoxy?
My application is in cementing ultrasonic transducers to glass. The transducers look like very thick washers, about 1-inch in diameter. I am getting a cavitation ringing in the applied voltage (10-volts, at 20-30 KHz). I suspect the bubbles are causing this, but do not know for sure.
I've tried slowly mixing the resin and catalyst beads, but the bubbles persist.
Any suggestions are welcome.
Nathan Haese Moraga, CA
__________________________________ Do you Yahoo!? SBC Yahoo! DSL - Now only $29.95 per month! http://sbc.yahoo.com
If my memory serves me correctly, there is a conversion routine in the Voyager software that converts the file to an emsa format which can then be read by programs such as Excel. I have not used the system in a long time and I can't remember exactly where the command is. The version of the software I used was 2.4.
Good luck.
Colin Veitch
Instrumentation Scientist
Late Stage Innovation Group
CSIRO Textile and Fibre Technology
PO Box 21, BELMONT, Vic. 3216. Australia.
E-mail: colin.veitch-at-csiro.au
Web: http://www.tft.csiro.au
Tel: +61 (0) 3 5246 4000
Fax: +61 (0) 3 5246 4811
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
-----Original Message----- } From: Carsten Ronning [mailto:Carsten.Ronning-at-phys.uni-goettingen.de] Sent: Thursday, 10 July 2003 2:10 AM To: Microscopy-at-sparc5.microscopy.com
Dear All,
I am looking for a PC file conversion possibility of our old 1993 Voyager EDS sun system (.eds) to a ASCII or text format. Or does somebody know the file structure of data saved in .eds-format ?
Many thanks in advance,
Carsten Ronning
************************************* Dr. Carsten Ronning School of Materials Science and Engineering Georgia Institute of Technology 771 Ferst Drive Atlanta, GA 30332-0245 USA
on leave: II. Insitute of Physics University of Goettingen Bunsenstr. 7-9 37073 Goettingen, Germany ****************************************
*From: "Christopher S. Zurenko" {czurenko-at-mail.khri.med.umich.edu} *Organization: Kresge Hearing Research Institute *To: Microscopy-at-sparc5.microscopy.com *Date sent: Wed, 09 Jul 2003 13:59:34 -0400 *Subject: Re: Epoxy: how to avoid small bubbles *Send reply to: czurenko-at-umich.edu *Priority: normal
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Well, in my practice we did once something like this and the oil in rotary pump had to been replaced. However it was big volume of epoxy so maybe in case of small one it wouldn't be so serious damage.
Best regards,
Witold Zielinski
*After mixing mounting medias in our lab (epoxy, OCT, ProLong,
*etc.), we place the container under vacuum to get the bubbles to rise *out of the media. The amount of time spent under vacuum will *depend on how quickly the media dries. This could possibly work for *your application. * *----------------------------------------------------------------------- *Christopher S. Zurenko *Research Assistant II *Kresge Hearing Research Institute, Otopathology *The University of Michigan Medical School *MSRB 3, Room 9303 *1150 W. Medical Center Dr. *Ann Arbor, MI 48109-0648 *Lab Phone: 734.763.9680 *Fax: 734.615.8111 *czurenko-at-umich.edu *http://www.khri.med.umich.edu/research/raphael_lab/index.htm * * * :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) ;)
Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
} I am looking for a PC file conversion possibility of our old 1993 } Voyager EDS sun system (.eds) to a ASCII or text format. Or does } somebody know the file structure of data saved in .eds-format ? } I have written a multi-purpose application to read all Noran Voyager (and Vantage) file formats to enable, among a lot of other features, data to be exported in ascii format. Noran do not (will not?) give any information on their file structures so this has been the result of many hours of hacking. Unfortunately, I have found that some files from other users do not read on my application. Send me some of your files (.eds, .grey, etc) and I will see if they read OK.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-cam.ac.uk
It has been my experience doing this that one needs to evacuate the container (vacuum oven, desiccator, whatever)by slowly opening the valve, allowing the bubbles rise to the top, close the valve, etc, effectively cycling the system until the valve can be completely opened to permit removal of the few remaining bubbles.
Roger Moretz, Ph.D. BI Pharmaceuticals, Inc Dept of Toxicology Ridgefield, CT
-- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } *From: "Christopher S. Zurenko" {czurenko-at-mail.khri.med.umich.edu} } *Organization: Kresge Hearing Research Institute } *To: Microscopy-at-sparc5.microscopy.com } *Date sent: Wed, 09 Jul 2003 13:59:34 -0400 } *Subject: Re: Epoxy: how to avoid small bubbles } *Send reply to: czurenko-at-umich.edu } *Priority: normal } } *------------------------------------------------------------------------ } *The Microscopy ListServer -- Sponsor: The Microscopy Society of America } *To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver
} *On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } *-----------------------------------------------------------------------. } * } Hello, } } Well, in my practice we did once something like this and the oil in } rotary pump had to been replaced. However it was big volume of epoxy } so maybe in case of small one it wouldn't be so serious damage. } } Best regards, } } Witold Zielinski } } } *After mixing mounting medias in our lab (epoxy, OCT, } ProLong, } } *etc.), we place the container under vacuum to get the bubbles to rise } *out of the media. The amount of time spent under vacuum will } *depend on how quickly the media dries. This could possibly work for } *your application. } * } *----------------------------------------------------------------------- } *Christopher S. Zurenko } *Research Assistant II } *Kresge Hearing Research Institute, Otopathology } *The University of Michigan Medical School } *MSRB 3, Room 9303 } *1150 W. Medical Center Dr. } *Ann Arbor, MI 48109-0648
If you have access to an acoustic microscope [SonooScan or Sonix] you can image through the transducer and detect voids in the epoxy non-destructively. I assume the transducer is a solid piezoelectric device.
Peter Tomic Agere Systems
-----Original Message----- } From: Nathan Haese [mailto:nathanhaese-at-yahoo.com] Sent: Wednesday, July 09, 2003 1:27 PM To: Microscopy-at-sparc5.microscopy.com
List members,
Any suggestions on how to avoid getting small bubbles in mixed epoxy?
My application is in cementing ultrasonic transducers to glass. The transducers look like very thick washers, about 1-inch in diameter. I am getting a cavitation ringing in the applied voltage (10-volts, at 20-30 KHz). I suspect the bubbles are causing this, but do not know for sure.
I've tried slowly mixing the resin and catalyst beads, but the bubbles persist.
Any suggestions are welcome.
Nathan Haese Moraga, CA
__________________________________ Do you Yahoo!? SBC Yahoo! DSL - Now only $29.95 per month! http://sbc.yahoo.com
This year there is a program for spouses and companions at M&M. (Details on page 34 of EXPO, if you have it - mine arrived today).
So far only a few people have registered for this program and we will have to cancel it unless more people sign up. If you or your companion intend to take part in this program but have not yet registered, please do so immediately.... Or else...
My apologies for the threatening and harassing tone but we have to make a decision at once.
Alwyn Eades -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
You'll need freezing methods for lipids -- cryo fixation, cryo coating and cryo SEM, or you'll have to do these by 'subtraction'. That is, do something like a chloroform extraction, and look at the result, assuming the holes are where the lipids were. Dangerous, because the holes could've been air or water, too. And you'd be assuming that the chloroform (and EtOH during dehydration) removed *all* of the lipids. Proteins can be done somewhat on morphology, but also by labeling: use gold-conjugated antibodies to the proteins of interest. Starches typically are as smooth grains, so they're pretty easy to ID by morphology. At least the ones I've looked at are. But if you need to ID "starch A" vs "starch B" ... well. There might be something in the literature, but I don't know of a method.
Phil
} I am interested in cutting the seeds of crops such as rice, corn, } cotton, and soybeans, and identifying protein, lipid, and starch } components in SEM. Can you tell me what approaches and methods I } could use? } } Thanks, } } Raja Rao
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
You might also try centrifuging the mixed epoxy, which would avoid the pump problems other listers have mentioned.
------------------------------------------------ Kevin Frischmann Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 03:17 PM 7/9/03 -0400, Ron L'Herault wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good Day. On behalf of the Local Arrangements Committee of the 2003 Microscopy and Microanalysis Meeting in San Antonio, Texas, I would like to invite you to the 2003 Golf Outing to be held Sunday, August 3 at the Pecan Valley Golf Club. Pecan Valley is a majestic golf course, located only six miles from downtown San Antonio and the Riverwalk. Pecan Valley was the site of the 50th Anniversary PGA Championship in 1968 when Julius Boros edged Arnold Palmer on the 18th hole and has also hosted three Texas Opens. It has been rated in Golf Digest¹s Top 50 Public Courses and #1 Public Golf Course in the State of Texas for 2002 (http://www.golftexas.com/pecan1.htm Rating:74.5 - Slope: 136 - Yards:7,071). The cost will be $70.00 and will include greens fees, cart, transportation to and from the Convention Center, Driving range, lunch buffet and awards banquet. We will leave the Convention Center at 6:45 - 7:00am (to beat the heat). Callaway club rental is available onsite at $45.00 per set. Another addition to this year¹s outing is the inclusion of various types of sponsorships. We are also looking for additional raffle prizes and / or giveaways for the event. Any company or individual that becomes a sponsor will be promoted in the following ways: * On the hole of their choice (for hole sponsorships) * On the lunch placemats * In each ³goodie² bag that every golfer will receive at the course
If you are interested in playing in the outing, please register on the web at http://www.microscopy.com/MSAMeetings/MMMeeting.html or contact me directly (msanders-at-cbs.umn.edu). If you have a foursome, please let me know and include all four names. If you do not have a foursome, that is fine as well we can pair you into a foursome at the course. If you are interested in becoming a sponsor, please contact me directly and I can get further details to you.
I would also like to announce that there will be 2 ³free² slots for students (preferably). These will be in the name of a former MSA member, golfer and a good friend of mine who passed away a few years ago, Joe Polak.
Contact Mark Sanders with any questions (msanders-at-cbs.umn.edu)
Thanks in advance for your consideration and participation in this matter!
At Pecan Valley you can truly ³Play where Champions Have Left Their Footprints.²
My Best Regards,
Mark Sanders MSA/MAS Golf Tournament at the Pecan Valley Golf Club
I have no suggestions from personal experience, but interestingly, shortly after reading your question, I opened up my most recent copy of Scanning, and the first paper was:
Kohler A, Høst V, Enersen G, Ofstad R: Identification of Fat, Protein Matrix, and Water/Starch on Microscopy Images of Sausages by a Principal Component Analysis-Based Segmentation Scheme Scanning 25, 3, 109-115 (2003)
It's not an SEM paper, but if you have access to a fluorescence scope, you might try it (at least for checking SEM results).
In short, samples were sectioned (10um) with a cryostat, and stained with Nile red (9-diethylamino-5H-benzophen-oxazin-5-one) dissolved in acetone (0.1 mg/ml overnight at 4 deg C). Stained sections were examined using a fluorescence microscope with a blue excitation filter (450-490nm). After staining, lipids show strong fluorescence, proteins weak fluorescence, and starch near zero fluorescence.
Granted, sausages and seeds are a bit dissimilar, but it seems like it ought to work.
------------------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 07:59 AM 7/9/03 -0500, RAO, RAJA S [AG/1000] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I don't think you can avoid getting bubbles in epoxy, but you can remove them after mixing if it's not a quick-setting epoxy. Place the mixed epoxy under a light vacuum for about five to ten minutes, or until there are no more small bubbles rising in the mix. If the vacuum is too strong, the epoxy will start to outgas - large bubbles rising from the bottom of the vessel, which interferes with the later polymerisation.
Lesley Weston.
on 09/07/2003 10:27 AM, Nathan Haese at nathanhaese-at-yahoo.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } List members, } } Any suggestions on how to avoid getting small bubbles } in mixed epoxy? } } My application is in cementing ultrasonic transducers } to glass. The transducers look like very thick } washers, about 1-inch in diameter. I am getting a } cavitation ringing in the applied voltage (10-volts, } at 20-30 KHz). I suspect the bubbles are causing } this, but do not know for sure. } } I've tried slowly mixing the resin and catalyst beads, } but the bubbles persist. } } Any suggestions are welcome. } } Nathan Haese } Moraga, CA } } } } } __________________________________ } Do you Yahoo!? } SBC Yahoo! DSL - Now only $29.95 per month! } http://sbc.yahoo.com }
My practice for years has been to mix the components less the catalyst/accelerator and place under vacuum (15-20 in Hg) overnight or for no less than 4 hr. If the room is too cold, I precondition the vacuum oven in which I have done this to about 22 degrees C (i.e., just a little warm). To prevent excessive air entrainment, I use only 2 sticks to stir the materials and I do so slowly without any attempt to 'beat' the mixture as one does with egg whites. In some cases, I have mixed by very slow rolling or tumbling for 4-6 hr. NEVER shake to mix, because that only ensures air entrainment, and remember, low temperature will exacerbate the solution of atmospheric gases. The final alternative is to place large volumes under light vacuum ( {15 in Hg) for a sufficiently long time (e.g., 24 hr) to insure that they are clear of entrained air. Then mix and expose to light vacuum again. One manufacturer suggests warming all components to 60 degrees before mixing.
These amount to an admission that I have had the same problems. The issue with your problem is that you must be particularly careful to degas the system because you appear to be using it to build a sandwich on the glass.
Finally, I believe you should use a very slow cure. Rapid curing is will known to cause bubbles. The best example is methacrylate plastics polymerized in bulk. In most cases the 'pour' is not made until the amount of monomer has been significantly reduced.
Suggestion. If you were to use a 90 min epoxy (two part available at a local hardware store) you would notice that both components exhibit a lot of viscosity. You will also note that you hardly ever get bubbles in the seal if even a little care is exercised. You might want to try a simple solution like that. If you are using EPON, I would suggest a partial cure at 45 degrees for 4-6 hr. At that temp a lot of degassing will have occurred and polymerization will be well on its way. Then transfer only what you need to your job and complete the cure at 55. In this manner, the HOT part of polymerization would likely be done during the preliminary cure and no cavities should be produced. Then, at the end, turn the oven off and let it slowly come to room temperature before you remove your devices.
Rationale/Guess? Consider the two substances, glass and transducer material, and their coefficients of expansion. Most resins remain somewhat fluid at higher temperatures, if two layers with different coefficients of expansion are connected by a flowing plastic at 60 degrees and the system is cooled rapidly, what happens when, and if, the polymer reaches its glass transition, and tries to hold two materials that are contracting at different rates? Cavities, I bet! Perhaps you want a polymer that will survive the stress at operating temperatures above its glass transition.
http://www.warmglass.com/COESummary.htm
http://www.psrc.usm.edu/macrog/tg.htm
http://www.dymax.com/pdf/SPIE_Paper_Appendix.pdf
If I have gotten it all wrong, please feel free to reply with more info about the specific epoxy you are using.
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: Nathan Haese [mailto:nathanhaese-at-yahoo.com] Sent: Wednesday, July 09, 2003 1:27 PM To: Microscopy-at-sparc5.microscopy.com
List members,
Any suggestions on how to avoid getting small bubbles in mixed epoxy?
My application is in cementing ultrasonic transducers to glass. The transducers look like very thick washers, about 1-inch in diameter. I am getting a cavitation ringing in the applied voltage (10-volts, at 20-30 KHz). I suspect the bubbles are causing this, but do not know for sure.
I've tried slowly mixing the resin and catalyst beads, but the bubbles persist.
Any suggestions are welcome.
Nathan Haese Moraga, CA
__________________________________ Do you Yahoo!? SBC Yahoo! DSL - Now only $29.95 per month! http://sbc.yahoo.com
I have a guy about to go on a cruise to collect coccolithophorids. He wants to bring them back and examine them using SEM. He asked for advice on collection and preservation techniques.
I guessed at standard SEM fixation with glutaraldehyde and keep track of the pH to prevent dissolution of the coccoliths. It is important that they be preserved intact, ie. individual cells retain all their coccoliths and not fall apart. He wants to count coccoliths/cell from different collecting stations.
Any coccolithophorid experts out there with practical experience on how to bring them back in one piece, or at least dead but still looking alive?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
The Journal of Microscopy publishes top quality review articles, original research papers, short technical notes, short communications, rapid publications and letters to the Editors, covering all aspects of microscopy and high-energy in situ beam analysis. Papers that emphasize the application of microscopical techniques or specimen preparation procedures in an investigation are also welcome.
} From August 1st, 2003 it will be possible to submit your papers online to the Journal of Microscopy, simply go to http://jmi.manuscriptcentral.com and follow the online instructions.
This system will enable us to cut down on decision and publication time, whilst ensuring the same high standards of peer review.
We look forward to receiving your contributions to this ever-growing journal.
Dr Ilaria Meliconi, Executive Editor Journal of Microscopy 37/38 St Clements, Oxford OX4 1AJ, UK Tel +44 (0)1865 248768 Fax +44 (0)1865 791237 http://www.blackwell-science.com/jmi
Note to anyone with experience in chick embryonic tissue and/or eye tissue, I recently did some TEM on chick embryonic conjunctival epithelium and mesenchyme and discovered some odd cells that need identifying.
The cells are generally in clumps, with regular edges, very very few organelles (a few mitochondria and many many free ribosomes) - no other organelles can be identified (no RER or SER or anything else). The cells within the mesenchymal tissue. We suspect the cells have recently divided and are beginning to differentiate but we are not sure into what. Anyone with insight into what these cells could be, please email me at {mailto:tfranzod-at-dal.ca} tfranzod-at-dal.ca - I can also send a few images to you if required/wanted. Dr. Tamara Franz-Odendaal, Halifax, Canada
Hello. We found a staining procedure for ruthenium hexaammine trichloride and we would like to try it. Has anybody used this chemical in their labs? I have questions regarding safety and handling of it. Is it easy to dissolve? MSDS says that it is highly reactive and air sensitive. How do you store it when a bottle is open?Label indicates that it should be stored under nitrogen, so what does this mean? How do you dispose of it? Can stock solution be made ahead of time and safely store in the fridge? Thanks Dorota
} Microscopy and Microanalysis 2003 } San Antonio } } This year there is a program for spouses and companions at M&M. } (Details on page 34 of EXPO, if you have it - mine arrived today). } Another meeting event will interest spouses who are teachers. Project MICRO will present a workshop on the use of "Microscopic Explorations" in the classroom. It's SC-11, Sunday Aug. 3,from 2-5. Unlike most Sunday courses, it's free - and it's an outstanding opportunity. Look it up! -- 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
Has anyone done EDX analysis and/or EM imaging on gold corrosion or dendritic growth in the presence of a halogen like bromine? I'd like to compare SEM's and spectra with anyone involved in this.
I have what looks like to be the complete document set (instructions, schematics, spare parts list, etc.; multiple copies of some or all) for the Philips 200 TEM. Anybody interested?
Be the first caller (well, emailer) for these fine collectibles and I'll throw in two (two!) plate magazines complete with cassettes and three (three!) receiver boxes, all from the EM 200.
But wait, there's more! At no extra charge, you also receive the vintage EM 200 "Simplified Cross-Section" poster, suitable for framing or covering up that nasty hole in your wall.
Don't wait! All of this can be yours, for a modest shipping fee (I'm guessing maybe 30 pounds worth of stuff).
Contact me off-list. If I don't have any bites in a week or so, I'm going to toss the whole lot.
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I read your email. First of all you need to state the epoxy you are using. I am sure it is not Epon. You need to test each component under vacuum to see what happens. Apply the vacuum in stages to see how the bubbles form and their sizes.
Below is a paragraph from an article I wrote for Microscopy Today about capillary forces and epon epoxy but have not sent in yet. Anyway below is the part I want you to consider. The capillary forces part would seem to be irrelevant because you are not wetting microporous membranes or nano-porous agglomerates.
Referring to the Epon Clone epoxy component DDSA for some perspective:
"Try this experiment. Pour DDSA into a long test tube and fill it about half full. Put the test tube in a beaker and pull a vacuum on both using a bell jar. You will notice the formation of small bubbles. These are entrained or dissolved air. Before they reach the surface and break, release the vacuum. Notice what happens to the air bubbles. They just shrink in place. Next immediately pull a vacuum and notice they reappear in the same place. Let them rise to the top surface, combine to form larger bubbles and then break them by applying and releasing the vacuum. Eventually only larger bubbles will form under increased rotary pump vacuum. These are not air bubbles but vaporizing DDSA or whatever other high boiler is in DDSA. After you only get the large bubbles, release the vacuum. Using a micro-spatula, stir up the DDSA and entrain some air. With the spatula, break the large bubbles that rise to the surface. Pull a vacuum on the DDSA again. Small bubbles will form again. The larger DDSA bubbles will follow these smaller bubbles. This will give you insight as to what really happens in a microporous membrane or porous agglomerated powder during vacuum impregnation." My DDSA from Pella foams under vacuum. I think all DDSA does but I haven't tried them all.
I can't know how your epoxy system will react but try this experiment. It will tell you how your epoxy components and air in your components behave under vacuum. I realize you are probably using a small amount of epoxy as a thin film. Use a larger amount of each component to determine where the bubbles are coming from in your samples.
What about the transducer surface as a source? Do you have sub-micron particles forming nucleation sites in a non-transparent epoxy? Can you try that special automotive glass adhesive used for attaching rear view mirrors? It is very thin but my not be strong enough for you. In any case, it might last long enough to show the elimination of the ringing.
I hope this gives you some help or gives you some ideas that lead you to a solution.
Sincerely,
Paul Beauregard Senior Research Associate Electron Microscopy PPG Industries
} List members, } } Any suggestions on how to avoid getting small bubbles } in mixed epoxy? } } My application is in cementing ultrasonic transducers } to glass. The transducers look like very thick } washers, about 1-inch in diameter. I am getting a } cavitation ringing in the applied voltage (10-volts, } at 20-30 KHz). I suspect the bubbles are causing } this, but do not know for sure. } } I've tried slowly mixing the resin and catalyst beads, } but the bubbles persist. } } Any suggestions are welcome. } } Nathan Haese } Moraga, CA } } } } } __________________________________ } Do you Yahoo!? } SBC Yahoo! DSL - Now only $29.95 per month! } http://sbc.yahoo.com } } }
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Dorita Wadowska wrote: ============================================================================ ==== We found a staining procedure for ruthenium hexaammine trichloride and we would like to try it. Has anybody used this chemical in their labs? I have questions regarding safety and handling of it. Is it easy to dissolve? MSDS says that it is highly reactive and air sensitive. How do you store it when a bottle is open?Label indicates that it should be stored under nitrogen, so what does this mean? How do you dispose of it? Can stock solution be made ahead of time and safely store in the fridge? Thanks ============================================================================ ====== So far as I know, all of these (organic) ruthenium compounds are quite unstable. That is why it is difficult to find someone who would ship for example, ruthenium tetroxide.
For staining, we have offered for some years a ruthenium tetroxide staining kit. It is described on URL www.2spi.com/catalog/chem/chem1c.shtml
The concept of the kit is that is allows you to make up tiny amounts of the tetroxide, in tiny amounts, which is the "active" species that does the staining. The ingredients that are used to produce the much more dangerous (actually potentially explosive tetroxide) are relatively inert and the amount of tetroxide that is produced is quite small, thereby minimizing any risk to the user.
If you think that there is some kind of specificity of the staining unique to ruthenium hexaammine trichloride, relative to the more straight-forward tetroxide, I (and probably others) would be very eager to hear more about it
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id JAA08440 for dist-Microscopy; Mon, 14 Jul 2003 09:13:30 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id JAA08435 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Mon, 14 Jul 2003 09:12:59 -0500 (CDT) Received: from email.med.yale.edu (email.med.yale.edu [130.132.232.16]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id JAA08428 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 14 Jul 2003 09:12:47 -0500 (CDT) Received: from yale.edu (yale130132006119.med.yale.edu [130.132.6.119]) by email.med.yale.edu (PMDF V6.1-1 #40995) with ESMTP id {0HI00JXHPPUWQ3-at-email.med.yale.edu} for Microscopy-at-MSA.Microscopy.Com; Mon, 14 Jul 2003 10:06:33 -0400 (EDT)
Hi all;
I am interested in quick freezing and freeze substitution of human Red Blood Cells (I am looking for high quality membrane architecture). I have the freezing all set up, but am looking for the freeze sub protocol. Could anyone send me one, or point me in the correct direction?
Thanx David ____________________
David Elliott Ph.D.
Yale University School of Medicine 333 Cedar Street PO Box 208022 New Haven, CT 06520-8022
Can you clarify: is Ruthenium tetroxide unstable when it is in solution, or only in the crystalline form? Thanks.
Marie
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Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
Regarding ruthenium tetroxide, I use the stain routinely for polymer microscopy and make it fresh as needed. An alternative (not better or worse, just different) to the periodate preparation of which Charles speaks was first described for use in microscopy by Montezinos. I have only slightly modified his procedure, using 10 % sodium hypochlorite instead of bleach. I refer you to G. M. Brown and J. H. Butler, "New method for the characterization of domain morphology of polymer blends using ruthenium tetroxide staining and low voltage scanning electron microscopy (LVSEM)", Polymer 38 (15), 3937 (1997). The appendix provides detailed descriptions for preparing and using the stain for low voltage SEM as well as TEM, and how to safety handle and dispose of the ruthenium tetroxide. Also, the Montezinos paper is referenced in this paper.
Feel free to contact me off-line with any questions.
Cheers,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Garber, Charles A." To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com} {cgarber-at-2spi.com} cc: Subject: Ruthenium staining
07/12/03 07:16 PM
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
==== We found a staining procedure for ruthenium hexaammine trichloride and we would like to try it. Has anybody used this chemical in their labs? I have questions regarding safety and handling of it. Is it easy to dissolve? MSDS says that it is highly reactive and air sensitive. How do you store it when a bottle is open?Label indicates that it should be stored under nitrogen, so what does this mean? How do you dispose of it? Can stock solution be made ahead of time and safely store in the fridge? Thanks ============================================================================
====== So far as I know, all of these (organic) ruthenium compounds are quite unstable. That is why it is difficult to find someone who would ship for example, ruthenium tetroxide.
For staining, we have offered for some years a ruthenium tetroxide staining kit. It is described on URL www.2spi.com/catalog/chem/chem1c.shtml
The concept of the kit is that is allows you to make up tiny amounts of the tetroxide, in tiny amounts, which is the "active" species that does the staining. The ingredients that are used to produce the much more dangerous
(actually potentially explosive tetroxide) are relatively inert and the amount of tetroxide that is produced is quite small, thereby minimizing any risk to the user.
If you think that there is some kind of specificity of the staining unique to ruthenium hexaammine trichloride, relative to the more straight-forward tetroxide, I (and probably others) would be very eager to hear more about it
Dear Sir; I worked about stucture analysis of single crystal with rigaku afc7s single crystal diffractometer in my master thesis.Now I have been studying shape memory alloys since 2000 in my doctorate thesis and responsible user from JEM3010 Transmission Electron Microscopy since 1999.I was heard "tem single crystal sample investigation to Darmstat university in germany" by my teacher in University of Ankara.I put on grid my single crystal sample and I observe.But In my opinion this method may be dangerous if sample fall in microscopy. Sincerely.
***************************************************** ***************************************************** ** Research Assistant ** ** PhD.Student ERDEM YASAR ** ** University of Kirikkale ** ** Department of Physics ** ** Electron Microscopy Laboratory ** ** 71450 KIRIKKALE/TURKEY ** ** erdem.yasar-at-physics.org ** ** tem_sem-at-hotmail.com ** ** http://turkuaz.kku.edu.tr/~yasar/erdem.html ** ***************************************************** *****************************************************
On Fri, 25 Apr 2003, darryl krueger wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } } From: darryl krueger {dkruege-at-clemson.edu} } } Subject: TEM single crystal sample } } } } I have a friend that is doing Diffraction, who is looking for a source for } } single crystals on TEM grids. If there is source someone knows of could } } you please let us know. TIA } } } } Darryl } } }
HELP!!! I have an old MT 5000 and a MT2B both in need of repair. Does anyone have a number for someone working on these microtomes? I am located in the south Texas area. Thanks in advance.
Lauren E. Chesnut Technical Director Electron Microscopy Lab UTHSCSA Dept. of Pathology MC-7750 7703 Floyd Curl Drive San Antonio, TX. 78229-3900 (210)567-4052 CHESNUTL-at-UTHSCSA.EDU
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Hi everybody. Ruthenium that I am using is a hexamine trichloride. I already dissolve it in cacodylate buffer and it almost immediately turns yellow upon addition of glutaraldehyde. Alter a few hours is dark brown. I am following procedure of Michael D. Buschmann et al "Ruthenium hexaammine trichloride chemography for aggrecan mapping in cartilage is a sensitive indicator for matrix degradation", The Journal of Histochemistry and Cytochemistry, vol 48(1): 81-88, 2000. The information I was able to retrieve form the Internet sites indicates that it is very reactive compound, degrades under air and is slightly radioactive hence my concerns. Thanks for your inputs. Dorota
we have to prepare a polymere (elastomere) which contains Nanoparticle (SiO2) in low concentration. The aim of the investigation is to get a nanoparitcle size distribution?
Hi All, It is a beast, isn't it? We have had somewhat consistent results with the new film by watching the temperature carefully and using a constant gentle bubbling with my nitrogen burst system (rather that the 2 second burst every 10 seconds that Kodak recommends). I am also careful to make sure that the film does not rest on the support bar that runs the length of the film racks, but is raised a bit to allow fluid flow. Of course I realize that I may just have a "good" box and all bets may be off with the next one, but since we've have nice results 4 times in a row, I thought it was worth passing it on. Lee -- Lee Cohen-Gould Electron & Optical Microscopy Facilities Weill Medical College of Cornell U. (212)746-6146 Rms A-105, LC-207
Werner, you omit the important point of likely particle size, but if it is nanoscale, let's assume it is somewhere in the 30-100nm range. I'm also assuming you plan get the distribution via low voltage FE-SEM. The simplest method by far is visit your local medical people at NMI and ask for the person who runs their diamond knife ultramicrotome (used for preparing ultrathin sections of tissue and the like for TEM) to prepare and 'face off' (ie put a flat on the end) a block of your material. Many polymers are soft enough to require cryosectioning, but that is standard for medical applications (and many biological ones as well, so they should have that capability. SInce you say the SiO2 is 'low concentration', wear of the diamond knife edge should not be a problem, which is something that the person will immediately be concerned about. I recommend a fairly high sectioning speed since you want to fracture your way through the particles, not pull them out of the polymer.
If you want thin TEM sections (by not having access to an FE-SEM), matters are trickier in general re picking up the section without having all the sectioned particles fall out, but then you would still have the holes as a guide. If the particles are {30nm in size, you've got an additional problem, as mixed phase materials tend to be difficult to section at thicknesses less than about 20-30nm, at least in the few soft matrix/hard particle systems that I've looked at. But it would be worth a try if you can find that microtome and a willing operator. I can't see mechanical polishing being very successful with your system, but if you have access to a focused ion beam (FIB) system, that could be tried as well.
Best of luck and let me know how it turns out.
Tom
Dr. Tom Malis Scientist Advisor Mineral Technology Branch Natural Resources Canada 555 Booth St., Ottawa, Ontario K1A 0G1 Tel.: (613) 995-7358 FAX: (613) 947-6606 malis-at-nrcan.gc.ca
-----Original Message----- } From: Werner Dreher [mailto:Dreher-at-nmi.de] Sent: Tuesday, July 15, 2003 8:11 AM To: Microscopy-at-sparc5.microscopy.com
Hi all,
we have to prepare a polymere (elastomere) which contains Nanoparticle (SiO2) in low concentration. The aim of the investigation is to get a nanoparitcle size distribution?
Kodak Eastman Motion Picture Film 5302 fine grain release positive film cat 166 7229.
At 09:20 15-07-2003 -0400, Eleana Sphicas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Postdoctoral Position Transmission Electron Microscopy Acadia University
The Center for Microstructural Analysis is seeking a postdoctoral research associate to perform transmission electron microscopy of ferromagnetic shape memory alloys. The work will involve collaboration with an interdisciplinary group (physicists, metallurgists and chemists) at Acadia University, Dalhousie University and Defence Research and Development Canada. Duties will include the operation of a Philips CM30 300 kV analytical STEM equipped with EDS and PEELS and specimen preparation using a Hitachi FIB 2000A. Occasional travel to a reactor facility to assist in the performance of neutron diffraction experiments may also be required.
The successful applicant will have a doctoral degree in Physics, Metallurgy or Materials Science with demonstrated expertise using TEM. Specific knowledge of high resolution imaging techniques and prior experience working with shape memory alloy systems would be considerable assets. Applicants must also possess good organizational and communication skills.
The position is funded for 18 months at $41000 CDN per annum, including salary and benefits. The start date is planned as soon as possible after September 1, 2003.
Review of applications will begin July 25, 2003. Interested applicants should submit a cover letter highlighting their qualifications, a CV including a list of publications, and contact information for three references to:
Dr. Craig Bennett Physics Department Acadia University Wolfville, NS B4P 2R5
On Sunday, July 13, 2003, at 12:03 PM, Erdem Yasar wrote:
} I worked about stucture analysis of single crystal with rigaku } afc7s } single crystal diffractometer in my master thesis.Now I have been } studying } shape memory alloys since 2000 in my doctorate thesis and responsible } user } from JEM3010 Transmission Electron Microscopy since 1999.I was heard } "tem } single crystal sample investigation to Darmstat university in germany" } by } my teacher in University of Ankara.I put on grid my single crystal } sample } and I observe.But In my opinion this method may be dangerous if sample } fall in microscopy. } Dear Erdem, Depending on the nature of your single-crystal specimen, there may be reliable methods either to attach it to a grid or to sandwich it between two grids (or halves of a folding grid). For a crystal that is only about 10nm thick and a few hundred nm wide--reasonable dimensions for a SAED structural study--there might be little problem if the sample drops, depending on where it ends up, which depends on the construction of your microscope. It is, of course, best if the specimen stays on the grid. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Werner Dreher wrote: =================================================== we have to prepare a polymere (elastomere) which contains Nanoparticle (SiO2) in low concentration. The aim of the investigation is to get a nanoparitcle size distribution? ======================================================= To do this reliably, and with the greatest efficiency, you will have to separate the isolate and then concentrate the nanoparticles from the polymer matrix and this is most easily done by putting the filled polymer into a plasma etcher using oxygen (which will produce an oxygen plasma, which will etch away the organics) leaving the inorganic nanoparticles behind.
We would recommend the SPI Supplies Plasma Prep II etcher for this kind of work, but there are other etchers on the market that should do the same thing. The Plasma Prep II etcher is described on URL http://www.2spi.com/catalog/instruments/etchers1.shtml
You can then pick up the nanoparticles on either a carbon supported TEM grid or if the grain size of the carbon is such that it interferes with the analysis of the SiO2 nanoparticles, you can consider using the SPI silicon nitride membrane window grids, see URL http://www.2spi.com/catalog/instruments/silicon-nitride.shtml since they are both structureless and featureless.
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 ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id HAA11573 for dist-Microscopy; Wed, 16 Jul 2003 07:24:05 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id HAA11569 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Wed, 16 Jul 2003 07:23:34 -0500 (CDT) Received: from mx0.stlawu.edu (mx0.stlawu.edu [199.0.76.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id HAA11562 for {microscopy-at-sparc5.microscopy.com} ; Wed, 16 Jul 2003 07:23:22 -0500 (CDT) Received: from 127.0.0.1 (localhost.localdomain [127.0.0.1]) by dummy.domain.name (Postfix) with SMTP id AB53968020 for {microscopy-at-sparc5.microscopy.com} ; Wed, 16 Jul 2003 09:02:39 -0400 (EDT) Received: from mx.stlawu.edu (mx.stlawu.edu [10.1.10.82]) by mx0.stlawu.edu (Postfix) with SMTP id 712D968BBE for {microscopy-at-sparc5.microscopy.com} ; Wed, 16 Jul 2003 09:02:38 -0400 (EDT) Received: from 127.0.0.1 (localhost [127.0.0.1]) by mx.stlawu.edu (Postfix on SuSE eMail Server 2.0) with SMTP id 3EEF2B6816A for {microscopy-at-sparc5.microscopy.com} ; Wed, 16 Jul 2003 07:46:03 -0400 (EDT) Received: from mail.stlawu.edu (mail.stlawu.edu [10.1.10.12]) by mx.stlawu.edu (Postfix on SuSE eMail Server 2.0) with ESMTP id 157E3B6816C for {microscopy-at-sparc5.microscopy.com} ; Wed, 16 Jul 2003 07:46:03 -0400 (EDT) Received: from stlawu.edu ([10.32.122.56]) by mail.stlawu.edu (Netscape Messaging Server 3.6) with ESMTP id AAA130A0; Wed, 16 Jul 2003 08:15:36 -0400 Message-ID: {3F154245.34E3A031-at-stlawu.edu}
listers
St. Lawrence University is seeking to fill a full-time, 12 month academic support staff position as a MICROSCOPY SPECIALIST. A masters degree or equivalent experience is required as is experience with confocal microscopy as well as electron microscopy (TEM and/or SEM). Mechanical and laboratory aptitude, computer experience, a desire to learn and teach new methodologies, and a positive work ethic is sought. The successful candidate will help oversee a developing interdisciplinary, multi-user microscopy/imagery center, will assist faculty and student researchers in advanced microscopy techniques, help teach microscopy methods and provide for the routine maintenance of the instrumentation infrastructure. The major instruments at the facility are on service contracts and the candidate will be expected to develop good working relationships with the professional service personnel. The facility is housed in the biology department and the successful candidate will also serve other science departments.
Applicants should send a letter of application, a current curriculum vitae (including references), a statement of any relevant research and teaching interests related to microscopy, and have three letters of recommendation forwarded to Dr. T. Budd, Biology Department, St. Lawrence University, Romoda Drive, Canton, NY 13617. The search committee will begin reviewing applications on July 21 and the search will remain open until filled.
-- Dr. T. Budd Chair of Biology St. Lawrence University Canton, NY 13617 Phone = 315-229-5640 Fax = 315-229-7429 E-mail = tbudd-at-stlawu.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, July 16, 2003 at 00:38:42 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin
Organization: 54
Education: 6-8th Grade Middle School
Location: Sofia Bulgaria
Question: How can I clean my immersion oil lenses after work with immersion oil ?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alessandro-at-polymer.kth.se) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, July 16, 2003 at 06:15:00 ---------------------------------------------------------------------------
Question: Dear Sir I would be grateful if you can provide some advice in order to carbon coat my replicas for TEM. I am using a Jeol Vacuum evaporator. I would like to know: -which kind of geometry should I use for the carbon rods? -how big should be the distance between the tips? -which current density should I use (25-30A as for evaporating metals?)? Thank you very much Regards Alessandro Matttozzi
I have the possibility of obtaining an Cambridge S90 that includes the factory installed Digital Image Store, that will allow Frame Averaged, Frame Integrated or Line Integrated video at tv rate.
Does anyone have any experience of connecting the tv output of the SEM to a PC TV card to allow the image to be saved on the computer?
Or do I need to spend more money in buying a digital imaging system.
I welcome any suggestions or recommendations
many thanks
Kevin
Electron Microscope unit School of Biological Sciences University of Aberdeen Aberdeen AB24 2TZ
Tel 01224-272847 Fax 01224-272396 ------------ Kevin Mackenzie k.s.mackenzie-at-abdn.ac.uk
Light-Microscopist There is a full-time opening for a core facility light/confocal microscopist available immediately. The encumbent must demonstrate total facility with current technology and sample preparations. They will be responsible for 1) providing direct hands-on access to light microscopy in a core facility setting, 2) overseeing daily facility operations, 3) consulting with users on experiment design, materials and methods, and instrument optimization and assist in data acquisition and analysis, 4) troubleshooting problems with microscopes or samples and 5) assuring instrument integrity. Must maintain a current understanding of the microscopy field as necessary to implement new technologies on campus. Applicants must have a strong desire to participate actively as a member of a team, possess excellent written and oral communication skills, be comfortable working independently, communicate effectively with service users to understand their microscopy needs, be facile with PC and Mac computer platforms and demonstrate effective troubleshooting skills. M.S. in biological sciences with five years research experience in molecular biology, immunohistochemistry and cytology is required. Previous work in a core facility is preferred.
The Jackson Laboratory is one of the world's foremost centers for mammalian genetics research. Located in Bar Harbor, Maine, the lab is adjacent to Acadia National Park. Mountains, ocean, forests, lakes, and trails are all within walking distance. If you are looking for a more natural environment, this could be the opportunity you've been searching for.
Interested applicants should send cover letter and resume to: jax/210 Human Resources, Box 27 The Jackson Laboratory 600 Main Street Bar Harbor, Maine 04609 Fax: (207) 288-6106 Email (preferred option): jobs-at-jax.org The Jackson Laboratory is an Affirmative Action/Equal Opportunity Employer.
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Dear readers I need some advice in order to optimize he carbon coating process of my replicas for TEM. In particular I need some tips about: -geometry of carbon rods, sharpening, distance between the tips, etc; -current density; -use of carbon ropes: how do they work and how has to be changed the set up I would be really grateful if You can help me. Regards
Alessandro Mattozzi Dept. of Fibre and Polymer Technology
Please note that the Core Facility Management session at M&M 2003 has been rescheduled. It originally was scheduled for Thursday August 7 from 1:00pm to 3:00pm. NEW TIME is Thursday August 7 from 8:00am to 10:00am in room 202B. Please change the time in your meeting schedules.
Topics for this year are:
How to Increase the Use of the Core Facility...with a Corresponding Increase in Revenue. Facilitator: Elaine Humphrey, U. of British Columbia
Defining the Roles of the Lab manager/Director and the Advisory Committee of a major Core Facility. Facilitator: Debby Sherman, Purdue University.
The goal of this session to to promote discussion of topics of interest to facility managers/directors. Therefore attendees are encouraged to bring information relative to the topics that they would like to share. In this case it may include examples of ways to advertise facility services (brochures, web sites, etc). Or perhaps you have a mission statement that helps clarify roles of staff and advisory committee members. Please contact me if you would like to share information and what sort of audio-visual equipment you would like to have available. Although it helps to have advanced notice to better organize the session, last minute inclusions are always welcome.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
Dear Colleagues, On behalf of the Organizing Committee, we have the pleasure to inform you that the 4th ASEAN Microscopy Conference and the 3rd Vietnam Conference on Electron Microscopy will be held during 5th - 6th January, 2004 in Hanoi. We would like to ask for your participation with an oral presentation and / or a poster session. The deadline for article submission will be on 30, August 2003. The submitted and approved articles will be put in the Proceedings of the Conference For registration and for Hotel reservation, please use the second announcement and call for paper that you have already received. Looking forward for your answer and we will be happy to welcome you in Hanoi.
Chairman of the Organizing Committee Prof. Nguyen Van Man M.D., D.M.Sc
If you need more informations, please contact at: Assoc. Prof Nguyen Kim Giao Electron Microscopy Unit National Institute of Hygiene and Epidemiology 1- Yersin Str - HaiBaTrung Distr - HaNoi-VietNam Tel: 84.4.9715434 Fax: 84.4.8210853 Email: emlad-at-hn.vnn.vn or emunihe-at-vol.vnn.vn
Omega Optical, the worldwide leading supplier of optical filters for fluorescence microscopy, is seeking to fill a position to sell our products. Interested applicants may apply by sending a resume to:
rgorham-at-omegafilters.com
Ruth Gorham Houle V.P. of Business Development Omega Optical, Inc. 210 Main St., PO Box 573 Brattleboro, Vermont 05301 ph:802-254-2690 X-127 t.f.ph:866-488-1064 fax:802-254-3937
Most EM's I know use PCI Quartz. The output of your EM will not be a standard frame rate of 30 frames/sec. [60 fields per sec.] but something less, so you simply can't put it into a video "frame grabber" card that only accepts NTSC [RS-170] video.
Oops, I just noticed you are in the UK! Video frame rates are different there, more scan lines per field but less fields/sec. Nonetheless, PCI Quartz sould do the job.
I have no interest in PCI Quartz other than as a user.
Hope this is helpful.
Peter Tomic Agere Systems Allentown, PA USA
-----Original Message----- } From: Kevin Mackenzie [mailto:k.s.mackenzie-at-abdn.ac.uk] Sent: Wednesday, July 16, 2003 9:00 AM To: Microscopy-at-sparc5.microscopy.com
I have the possibility of obtaining an Cambridge S90 that includes the factory installed Digital Image Store, that will allow Frame Averaged, Frame Integrated or Line Integrated video at tv rate.
Does anyone have any experience of connecting the tv output of the SEM to a PC TV card to allow the image to be saved on the computer?
Or do I need to spend more money in buying a digital imaging system.
I welcome any suggestions or recommendations
many thanks
Kevin
Electron Microscope unit School of Biological Sciences University of Aberdeen Aberdeen AB24 2TZ
Tel 01224-272847 Fax 01224-272396 ------------ Kevin Mackenzie k.s.mackenzie-at-abdn.ac.uk
The accepted method of cleaning oil from immersion lenses is with lens tissue paper wetted with xylene. The lens tissue paper should be available from any camera store.
I am trying to explain and compare the similarities and differences of ESEM equipment and VPSEM equipment in plain language to our Administrators. I know that the 2 types of equipment share some characteristics but others are unique. I also understand that the 2 types of equipment are unique in that individual companies produce one or the other, but not both.
I know that there have been some discussions about this which are archived, but with the pace that technology changes, I wanted to ask these questions again to be sure that I could provide the most up to date information possible to my Administrators.
Could you please help?
Thank you in advance.
Kind regards,
Paula.
Paula M. Allan-Wojtas Research Scientist - Food Microstructure / Chercheur scientique - microstructure des aliments Food Safety and Quality Team / Salubrité et qualité des aliments Agriculture and Agri-Food Canada /Agriculture et Agroalimentaire Canada Telephone / Téléphone: 902-679-5566 Facsimile / Télécopieur: 902-679-2311 32 Main Street / 32 rue Main Kentville, Nova Scotia / Kentville (Nouvelle-Écosse) B4N 1J5 allanwojtasp-at-agr.gc.ca
The Spouses Program offered as a separate social event at the M&M 2003 meeting in San Antonio has been canceled. The number of people registering for the event fell far short of the 35 person minimum we needed. Refunds can be arranged at the meeting in the registration area by contacting Monica Eggers at the registration desk, or Ev Osten or Dwight Erickson at the Local Arrangements Committee booth.
At the Local Arrangements Committee booth we can give you information on some of the venues that were part of the program and some of the things you can do on your own.
Ev Osten
Local Arrangements Committee Chair Microscopy & Microanalysis 2003 August 3 - 7, San Antonio
efosten-at-mmm.com 651-736-0104 fax: 651-733-0648
3M Company Corporate Analytical Technology Center 3M Center, 201-BE-16 St. Paul, MN 55144-1000 USA
Good News. Almost all of the lost master tapes from The MSA video collection have been recovered. So, some of the titles that have not been available for a while can now be provided in VHS or DVD format. The current catalog is available as a link on the MSA web page. Mastercard and VISA are now accepted. We also accept institutional purchase orders, check, money order, cash and precious gems that are appropriately mounted. As always the MSA Education Committee welcomes new contributions to the video collection and encourages all members to consider presenting tutorials at our annual meeting on new topics or on topics previously covered, that are in need of updating.
Greg Erdos MSA Video Wrangler
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295
Dear Alessandro, I also use a JEOL Evaporator. I use 5mm diameter carbon or graphite rods, one just flattened on the end and the other sharpened to a 1mm diameter tip, about 4mm long. These rods just touch, the sharpened point in the centre of the flat one. I stick the rods out of the metal holders about 10 mm each, with the springs on one about one-half stretched, so that the two stay in contact as the sharpened tip burns down. I pump to at least a 5 X 10-3 torr vacuum, then turn the heater current up to 35A, not too quickly. Leave it there and the current will increase by itself. I go until it reaches 45 to 50 A, but do not leave the heat on for more than 30 seconds. If the vacuum degrades above 10-4 torr, turn the current off, wait for the vacuum to improve, then turn up the current again. The best way to decide on how much to coat is by coating a polished brass disc at the same time, beside your samples. The brass goes from yellow to gold to orange to red and then has a sudden transition from red to blue at 25 nm thickness. For TEM replicas I would use about the orange or red thickness, but experience will help you to know what works best. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "by way of Ask-A-Microscopist" {alessandro-at-polymer.kth.se} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, July 16, 2003 5:37 AM
Dear Alessandro Mattozzi,
We can share our optimized conditions for making carbon.
The carbon rod which we used is about 1/4" in diameter. We sharp one end (about 1/4" length) of rod to 1/8" in diameter.
According to our experiences, the vacuum in the evaporating chamber is the most important parameter for making the high quality carbon films. The higher is better. Normally, we use 1.5 X 10 -6 torr.
The best current value is really depended on the vocuum which you use. Lower vocuume needs low current to get sufficient speed of growthing carbon film, higher vocuum needs higher current value. However, the carbon film made under low vocuum is easy to be oxidized and the film is very soft and easy to be broken.
One more tip for you is that, the speed for growthing carbon film is related with the current value. However, maxima current for growing a high qulity carbon film is limited by if you could see the spark away from the filament. The spark is large group of carbon moleculars which will mass up your carbon film.
We are a manufactory of making TEM grids for more than 5 years in United States. We provide the high quality TEM grids and the grids pre-coated with carbon films. You are welcome to try our product. For more information about product, please visit our website as below,
http://www.grid-tech.com/
shall you have any questions, I will be happy to answer.
with our best regards
Wendy Zhang ===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
} } Dear readers } I need some advice in order to optimize he carbon } coating process of my } replicas for TEM. In particular I need some tips } about: } -geometry of carbon rods, sharpening, distance } between the tips, etc; } -current density; } -use of carbon ropes: how do they work and how has } to be changed the set up } I would be really grateful if You can help me. } Regards } } Alessandro Mattozzi } Dept. of Fibre and Polymer Technology } }
===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
In many cases you can simply connect the output of the SEM to a video card. However, those signals need to follow the various video standards (NTSC, RS-170, PAL, etc.) which limits their resolution (for example: 640x480 for RS-170). Typically the signals are transmitted as composite signals on a coax cable.
If you want to digitize higher resolution modes, you need some special hardware (for an alternative to the Quartz system, see the ADDA II on our web site). These systems are more expensive, as they use special hardware.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Kevin Mackenzie [mailto:k.s.mackenzie-at-abdn.ac.uk] Sent: Wednesday, July 16, 2003 7:00 AM To: Microscopy-at-sparc5.microscopy.com
I have the possibility of obtaining an Cambridge S90 that includes the factory installed Digital Image Store, that will allow Frame Averaged, Frame Integrated or Line Integrated video at tv rate.
Does anyone have any experience of connecting the tv output of the SEM to a PC TV card to allow the image to be saved on the computer?
Or do I need to spend more money in buying a digital imaging system.
I welcome any suggestions or recommendations
many thanks
Kevin
Electron Microscope unit School of Biological Sciences University of Aberdeen Aberdeen AB24 2TZ
Tel 01224-272847 Fax 01224-272396 ------------ Kevin Mackenzie k.s.mackenzie-at-abdn.ac.uk
Stu- Xylene is pretty strong stuff. Won't it hurt the coatings? I just use 1) a Q-tip with optical lens cleaner and then lens tissue paper with lens cleaner. Is that okay? Rgds, Mike Shaw Roselle, NJ
} The accepted method of cleaning oil from immersion } lenses is with lens tissue paper wetted with xylene. } The lens tissue paper should be available from any } camera store. } } Stu Smalinskas } SKF USA } Plymouth, Michigan
Dear Wendy Zhang Yes, you right: better vacuum, better carbon film quality. But, using Electron Gun evaporation in combination with good vacuum delivered even better results. Thermal evaporation is not effective to produce mono atomic vapors. It's more like clouds of 10-15+ atom's clusters. Electron beam evaporation utilized bombardment of carbon by accelerated electrons. This way is more effective in formation nearly mono atomic carbon "clouds" and therefore better carbon film homogeneity/uniformity. This process is also may be controlled better than thermal evaporation. Finally, you may produce very thin, uniform and stable films. Personally, I am using 1.2-1.5 nm thick films for routine work in shadowing and negative staining. Best wishes, Sergey.
At 02:24 PM 7/17/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Things are changing rapidly. Five years ago only the ESEM could offer SE imaging and water on/in the specimen. Now many of the rivals can offer SE imaging in VPSEM. At a recent meeting one competitor claimed, in converstion, to be able to image water. I have not got round to asking for some images from them. I would check out the suppliers for current performance information.
Dave
On Thu, 17 Jul 2003 15:05:48 -0400 Paula Allan-Wojtas {AllanWojtasP-at-agr.gc.ca} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, all, } } I am trying to explain and compare the similarities and differences of } ESEM equipment and VPSEM equipment in plain language to our } Administrators. I know that the 2 types of equipment share some } characteristics but others are unique. I also understand that the 2 } types of equipment are unique in that individual companies produce one } or the other, but not both. } } I know that there have been some discussions about this which are } archived, but with the pace that technology changes, I wanted to ask } these questions again to be sure that I could provide the most up to } date information possible to my Administrators. } } Could you please help? } } Thank you in advance. } } Kind regards, } } Paula. } } } } Paula M. Allan-Wojtas } Research Scientist - Food Microstructure / Chercheur scientique - } microstructure des aliments } Food Safety and Quality Team / Salubrité et qualité des aliments } Agriculture and Agri-Food Canada /Agriculture et Agroalimentaire } Canada } Telephone / Téléphone: 902-679-5566 } Facsimile / Télécopieur: 902-679-2311 } 32 Main Street / 32 rue Main } Kentville, Nova Scotia / Kentville (Nouvelle-Écosse) } B4N 1J5 } allanwojtasp-at-agr.gc.ca } } } } } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
We'll soon be getting a Nikon SMZ 1000 stereoscopic zoom microscope with a beam splitter for a digital camera. This stereomicroscope will be used to help with the making and cleaning of TEM specimens. We'd like to attach a digital camera to catalog the specimen, but really don't need a high end digital camera. Just something to take a quick image of the sample to place in the notebook. So my questions are:
Does anyone know where I can find an adapter to attach a consumer grade digital camera (one bought at Target or Best Buy) to a Nikon SMZ 1000 stereoscopic zoom microscope?
Any recommendations on which consumer grade camera to get?
Thanks for your help everyone!
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
I enclose details of our latest microscopy report.
The report will include market segment size, growth rates, ten year projections, major players and competitive strategies for the microscopy market segment. The report will include an analysis of technology platforms, descriptions of companies in the field, and trends in technology and business impacting this market segment.
It will discuss technology platforms in various forms of microscopy including light, confocal, electron, and scanning probe. The report will examine products in development, perceived medical and market needs, the market outlook, economic considerations, and pricing for the microscopy market segment.
Each corporate profile includes the following elements:
History
Unique Company Strengths Corporate Research and Development Strategy Intellectual Property Assessment Corporate Collaborations Products and Developmental Pipeline Discovery-Stage Projects Summary Assessment of Market Prospect Direct Pipeline Competition Potential Improvements Over Existing Products Financial Details Summary
The information in this report is based upon interviews with sales and marketing professionals of companies in the biotechnology laboratory instrument market. Professionals at laboratories and dealers around the U.S. were queried, some several times, about their institution's products and marketing strategies as well as their overall thoughts about their industry segment.
Other sources of information for the report were trade association publications and meetings, product brochures and catalogs, and company literature. An examination was made of a number of company internet sites, and telephone interviews were conducted with a number of industry marketing executives. Where possible, an examination of the annual reports, 10k filings, and financial reports were used as the basis of the data reported.
Some of the information obtained for the report was taken from Biotechnology Associates databases and from the private data stores of the author. The information set forth in this study was obtained from sources that we believe to be reliable, but we do not guarantee the accuracy, adequacy or completeness of any information or the results obtained by the use of such information.
For a complete index of this report click on http://www.researchandmarkets.com/reports/5531
Report Index:
1. Executive Summary
2. Introduction
3. Light Microscopy 3.1 Products and Technologies 3.2 Market Analysis
4. Confocal Microscopy 4.1 Technologies and Products 4.2 Market Analysis
5. Electron Microscopy 5.1 Technologies and Products 5.2 Market Analysis
7. Semiconductor Processing Systems 7.1 Technologies and Products 7.2 Market Analysis
8. Automated Imaging Systems 8.1 Technologies and Products 8.2 Market Analysis
9. Summary and Conclusions
10. Company Profiles 10.1 Thermo Spectra Corporation 10.2 Applied Imaging Corporation 10.3 Bio-Rad Laboratories, Incorporated 10.4 Digital Instruments 10.5 Applied Precision 10.6 International Remote Imaging Systems (IRIS) 10.7 KLA-Tencor Instruments Corporation 10.8 Leica Incorporated 10.9 Autocyte/Neopath/Cytyc/Neuromedical Systems, Inc. 10.10 Nikon, Inc. 10.11 Olympus Optical Company Limited, Tokyo, Japan 10.12 Carl Zeiss, Inc.
11. Company Directory
List of Tables
Table 1 U.S. Sales of Light Microscopes 1996-2005 (in Dollars) Table 2 U.S. Sales of Light Microscopes 1996-2005 (in Units) Table 3 Breakdown of U.S. Sales of Light Microscopes by Price Range 1999 (in Dollars) Table 4 Breakdown of U.S. Sales of Light Microscopes by Price Range 1999 (in Units) Table 5 Breakdown of U.S. Sales of Light Microscopes by User Segment 1999 (in Dollars) Table 6 Breakdown of U.S. Sales of Light Microscopes by User Segment 1999 (in Units) Table 7 Breakdown of U.S. Sales of Light Microscopes by Manufacturer 1999 (in Dollars) Table 8 U.S. Sales of Confocal Microscopes 1996-2005 (in Dollars) Table 9 U.S. Sales of Confocal Microscopes 1996-2005 (in Units) Table 10 Breakdown of U.S. Sales of Confocal Microscopes by User Segment 1999 (in Dollars) Table 11 Breakdown of U.S. Sales of Confocal Microscopes by User Segment 1999 (in Units) Table 12 Breakdown of U.S. Sales of Confocal Microscopes by Manufacturer 1999 (in Dollars) Table 13 US. Sales of Electron Microscopes 1996-2005 (in Dollars) Table 14 U.S. Sales of Electron Microscopes 1996-2005 (in Units) Table 15 Breakdown of U.S. Sales of Electron Microscopes by Technology 1999 (in Dollars) Table 16 Breakdown of U.S. Sales of Electron Microscopes by Technology 1999 (in Units) Table 17 Breakdown of U.S. Sales of Electron Microscopes by User Segment 1999 (in Dollars) Table 18 Breakdown of U.S. Sales of Electron Microscopes by User Segment 1999 (in Units) Table 19 Breakdown of U.S. Sales of Electron Microscopes by Manufacturer 1999 (in Dollars) Table 20 U.S. Sales of Scanning Probe Microscopes 1996-2005 (in Dollars) Table 21 U.S. Sales of Scanning Probe Microscopes 1996-2005 (in Units) Table 22 U.S. Sales of Scanning Probe Microscopes by Type 1999 (in Dollars) Table 23 U.S. Sales of Scanning Probe Microscopes by Type 1999 (in Units) Table 24 U.S. Sales of Scanning Probe Microscopes by Application 1999 (in Dollars) Table 25 U.S. Sales of Scanning Probe Microscopes by Application 1999 (in Units) Table 26 U.S. Sales of Scanning Probe Microscopes by Manufacturer 1999 (in Dollars) Table 27 U.S. Sales of Semiconductor Processing Equipment 1996-2005 (in Dollars) Table 28 U.S. Sales of Semiconductor Processing Equipment 1996-2005 (in Units) Table 29 Breakdown of U.S. Sales of Semiconductor Processing Equipment by Manufacturer 1999 (in Dollars) Table 30 U.S. Sales of Automated Imaging Systems 1996-2005 (in Dollars) Table 31 U.S. Sales of Automated Imaging Systems 1996-2005 (in Units) Table 32 Breakdown of U.S. Sales of Automated Imaging Systems by Application 1999 (in Dollars) Table 33 Breakdown of U.S. Sales of Automated Imaging Systems by Application 1999 (in Units) Table 34 Breakdown of U.S. Sales of Automated Imaging Systems by Manufacturer 1999 (in Dollars) Table 35 U.S. Sales of Light, Confocal, Electron, And Scanning Probe Microscopes 1996-2005 (in Dollars) Table 36 U.S. Sales of Confocal, Electron And Scanning Probe Microscopes 1996-2005 (in Dollars) Table 37 U.S. Sales of Confocal, Electron And Scanning Probe Microscopes 1996-2005 (in Units) Table 38 1996 and 2005 Sales By Category Of Light, Confocal Electron, and Scanning Probe Microscopes (in Dollars) Table 39 1996 and 2005 Sales By Category of Confocal, Electron, And Scanning Probe Microscopes (in Dollars) Table 40 1996 and 2005 Sales By Category of Confocal, Electron And Scanning Probe Microscopes (in Units) Table 41 Breakdown of U.S. Sales of Light, Confocal, Electron, and Scanning Probe Microscopes by Manufacturer 1999 (in Dollars)
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The method we use is to use a solution of Windex (the original blue one) and water 1:1 and lens tissue. Moisten the lens tissue, DO NOT CRUMPLE it up, and gently sweep it across the front element of the lens. Repeat as needed. Never apply pressure to the lens. You should check with the manufacturer of your lens to see what they recommend, since different companies may use different coatings on their lenses and different coatings may need different handling (a lot of differences!). A build-up of old, dried oil may need more aggressive cleaning, but again, check with the manufacturer or distributor of your microscope. Lee -- Lee Cohen-Gould Electron & Optical Microscopy Facilities Weill Medical College of Cornell U. (212)746-6146 Rms A-105, LC-207
as far as 5302 is concerned, good luck. according to kodak canada it is no longer produced. i have not talked to kodak in new york, but your only available source may be supply houses which have stockpiled it.
Ev; I can certainly understand your disappointment at the lack of registration for the spouses program, but it doesn't appear that you have given much consideration to the people who purchased non-refundable airplane tickets to come to San Antonio, expecting to participate in the spouses program for which they registered.
John Mardinly Intel
-----Original Message----- } From: "efosten-at-mmm.com"-at-sparc5.microscopy.com [mailto:"efosten-at-mmm.com"-at-sparc5.microscopy.com] Sent: Thursday, July 17, 2003 1:06 PM To: Microscopy-at-sparc5.microscopy.com
To M&M 2003 attendees,
The Spouses Program offered as a separate social event at the M&M 2003 meeting in San Antonio has been canceled. The number of people registering for the event fell far short of the 35 person minimum we needed. Refunds can be arranged at the meeting in the registration area by contacting Monica Eggers at the registration desk, or Ev Osten or Dwight Erickson at the Local Arrangements Committee booth.
At the Local Arrangements Committee booth we can give you information on some of the venues that were part of the program and some of the things you can do on your own.
Ev Osten
Local Arrangements Committee Chair Microscopy & Microanalysis 2003 August 3 - 7, San Antonio
efosten-at-mmm.com 651-736-0104 fax: 651-733-0648
3M Company Corporate Analytical Technology Center 3M Center, 201-BE-16 St. Paul, MN 55144-1000 USA
Dear Paula, ESEM is a brand name of the FEI company and their term "Environmental SEM" refers to higher pressures. VPSEM is a generic term that includes ESEM and the other SEMs that use this form of imaging. Each manufacturer will give the range of pressures that they will operate at in their specifications. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Paula Allan-Wojtas" {AllanWojtasP-at-agr.gc.ca} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, July 17, 2003 12:05 PM
Hi, all,
I am trying to explain and compare the similarities and differences of ESEM equipment and VPSEM equipment in plain language to our Administrators. I know that the 2 types of equipment share some characteristics but others are unique. I also understand that the 2 types of equipment are unique in that individual companies produce one or the other, but not both.
I know that there have been some discussions about this which are archived, but with the pace that technology changes, I wanted to ask these questions again to be sure that I could provide the most up to date information possible to my Administrators.
Could you please help?
Thank you in advance.
Kind regards,
Paula.
Paula M. Allan-Wojtas Research Scientist - Food Microstructure / Chercheur scientique - microstructure des aliments Food Safety and Quality Team / Salubrité et qualité des aliments Agriculture and Agri-Food Canada /Agriculture et Agroalimentaire Canada Telephone / Téléphone: 902-679-5566 Facsimile / Télécopieur: 902-679-2311 32 Main Street / 32 rue Main Kentville, Nova Scotia / Kentville (Nouvelle-Écosse) B4N 1J5 allanwojtasp-at-agr.gc.ca
I've worked in many labs where xylene was set up as the solvent to clean immersion lenses, so I figure this solvent is most widely accepted. I know it smells strong, but I don't consider it a powerful solvent. I'm open to other ideas. Perhaps we should look at this the other way and ask which chemicals we should avoid. Harm may come to the optical coating and glue used to set the lenses in place if the wrong solvent is used. I believe we can cross off acetone from the accepted solvent list.
Stu- Xylene is pretty strong stuff. Won't it hurt the coatings? I just use 1) a Q-tip with optical lens cleaner and then lens tissue paper with lens cleaner.
Is that okay? Rgds, Mike Shaw Roselle, NJ
} The accepted method of cleaning oil from immersion } lenses is with lens tissue paper wetted with xylene.
} The lens tissue paper should be available from any } camera store. } } Stu Smalinskas } SKF USA } Plymouth, Michigan
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To all, The literature that accompanies each box of 4489 film, including the New Formulation still indicates processing "with nitrogen-burst agitation (1-second burst every 8 seconds)." Has there been an (un)official statement from Kodak suggesting otherwise?
Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Thanks, Stu- Will not use Acetone. Am also checking with manufacturer of my lenses for additoinal info. Rgds, Mike Shaw
In a message dated 7/18/2003 1:31:29 PM Eastern Standard Time, smalinskas-at-yahoo.com writes:
} Perhaps we should } look at this the other way and ask which chemicals we } should avoid. Harm may come to the optical coating } and glue used to set the lenses in place if the wrong } solvent is used. I believe we can cross off acetone } from the accepted solvent list.
William- I have a Canon A40, and I hook it up to a TV monitor for viewing and focusing. The nice thing about it is that you can buy a ready-made Canon bayonette adapter-to-52mm right from Canon at camera store. This pops onto the front of the camera, and it is a simple matter to hook up to a T-mount or any other ring from 52mm to whatever you need. You then have easy-on and easy-off of the camera from the microscope without having to unscrew the camera and remove cords... That's why I chose the Canon A-40. If you want more info- I have pictures as well as the set up on my website, so let me know if you are intersted. Rgds, Mike Shaw Roselle, NJ
In a message dated 7/18/2003 8:13:29 AM Eastern Standard Time, wgstratton-at-wisc.edu writes:
} Does anyone know where I can find an adapter to attach a consumer grade } digital camera (one bought at Target or Best Buy) to a } Nikon SMZ 1000 } stereoscopic zoom microscope? } } Any recommendations on which consumer grade camera to get? } } } Thanks for your help everyone! } } William Stratton
The Nikon Coolpix cameras have threaded lenses for various adapters that can be used to attach these consumer grade cameras to your microscope. Nikon and some aftermarket suppliers provide an adapter from the threads on the lens to a C-mount that should be on your micoscope. The adapter will also fit right into some eyepiece tubes for use on microscopes without a C-mount.
We have used the Coolpix 990 and 4500 models in this way. Obviously, the images are not of the same quality as the dedicated digital acquisition systems, but are pretty good quality for the cost. The cameras use a Compact Flash card for image storage, which is not as convenient as direct saves to disk but is relatively convenient with an inexpensive USB card reader. I am not so found of the direct USB attachment to the camera and Nikon softward for retrieving images.
Your Nikon rep should be able to provide information on the adapter, or do a quick search on google for "coolpix microscope adapter" to find information on Nikon and aftermarker alternatives.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
A company rep reads this mailing list and he will, I hope let you know that he has what is needed to connect the Coolpix to microscopes. We bought their generic adapter and the pieces needed to attach it to an inverted, a metallurgical and to a sophisticated trinocular. As it turns out, with just the generic, we can put it on our dissecting scope, and into the port on our hardness tester. We even used it on a trinocular scope in another lab. It is quite versatile.
Ron L
-----Original Message----- } From: Larry Hanke [mailto:hanke-at-mee-inc.com] Sent: Sunday, July 20, 2003 5:12 PM To: William Stratton Cc: Microscopy-at-sparc5.microscopy.com
The Nikon Coolpix cameras have threaded lenses for various adapters that
can be used to attach these consumer grade cameras to your microscope. Nikon and some aftermarket suppliers provide an adapter from the threads
on the lens to a C-mount that should be on your micoscope. The adapter will also fit right into some eyepiece tubes for use on microscopes without a C-mount.
We have used the Coolpix 990 and 4500 models in this way. Obviously, the
images are not of the same quality as the dedicated digital acquisition systems, but are pretty good quality for the cost. The cameras use a Compact Flash card for image storage, which is not as convenient as direct saves to disk but is relatively convenient with an inexpensive USB card reader. I am not so found of the direct USB attachment to the camera and Nikon softward for retrieving images.
Your Nikon rep should be able to provide information on the adapter, or do a quick search on google for "coolpix microscope adapter" to find information on Nikon and aftermarker alternatives.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
I'm having some trouble aligning the imaging system of an Hitachi H-7000. When it comes to changing the spot sizes from 1-7, and then moving it back to position, I find that even if I move just from 1 spot size to the next, the spot moves right off the screen, and it takes a very long period of time to find it. If I tried to directly move to spot 7 from say spot 5, when I finally do find spot 7 with condensor shift controls, I find that the beam is not perfectly round anymore, but half of the beam is obscured somehow, even though all of the apertures are out, and the specimen rod is out of the beam.
I tried for a long time working on this problem with no success. It seemed to take forever to find the beam after just changing from one spot size to the adjacent spot size. IT would just FLY off the screen, and it didnt' help to try "reset" button of the microscope or even just resetting the condensor shift controls.
I haven't used this H-7000 for quite some time, and now I'm only used to the JEOL, where we have a "bright tilt" button to turn on first. Is there a corresponding "bright tilt" button somewhere on the H-7000 that I have forgotten about?
On most TEMs (I don't know specifically about the H-7000) this alignment is done with the gun translation control. Most TEMs have 2 sets of deflectors between the gun and the sample. You are controlling only one set with the beam shifts. This set of deflectors is between the C2 lens and the sample. The other set of deflectors is between the anode and the C1 lens. These are the gun tilt and gun translate coils.
The gun translate is usually set by varying the C1 lens between a strong and weak setting. Think about the case where the beam (coming out of the gun) is entering the C1 lens off-axis.
Consider a strong C1 - the focal length is short and the cross-over will be close to the optic axis. So as a 1st approximation we consider it to be on the axis and center the beam with the normal beam translation knobs.
Now weaken the C1 lens - the focal length gets longer and the cross-over is proportionately farther away from the axis. Now use the gun translate (or shift) control to align the beam entering the C1 lens.
Repeat the process until the shift is minimal. Check your owners book to see how Hitachi recommends doing this alignment.
Cheers, Henk
At 09:30 AM 7/21/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Xiao.ming.wang-at-mail.mcgill.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, July 21, 2003 at 08:55:22 ---------------------------------------------------------------------------
Email: Xiao.ming.wang-at-mail.mcgill.ca Name: Xiaoming Wang
Organization: McGill Univesity
Education: Graduate College
Location: Montreal Canada
Question: Dear Sir/Madam,
I am looking for a software on trace analysis on TEM. Do you know any body who has the software or who is working on it?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jaideepp-at-rci.rutgers.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, July 21, 2003 at 10:07:26 ---------------------------------------------------------------------------
Question: Are there any softwares available to calculate the edge orientation polarograms and and/or anisotropy index from SEM images of oriented particles
In a message dated 7/21/03 2:09:28 PM, jaideepp-at-rci.rutgers.edu writes:
} Question: Are there any softwares available to calculate the edge orientation } polarograms and and/or anisotropy index from SEM images of oriented particles
That function is included in the Image Processing Tool Kit (www.ReindeerGraphics.com)
As previously announced, the spouses/companions program had to be canceled because we fell far short of the minimum number required by the tour company. As an alternative, I suggest that the people who are still interested in such activities get together at the Local Arrangements Committee booth in the registration area of the Convention Center at 8:30 on Monday morning, August 4. There you can meet the other people, and we (the LAC) can provide you with suggestions and information on things that an unstructured group can do.
Ev Osten
Local Arrangements Committee Chair Microscopy & Microanalysis 2003 August 3 - 7, San Antonio
efosten-at-mmm.com 651-736-0104 fax: 651-733-0648
3M Company Corporate Analytical Technology Center 3M Center, 201-BE-16 St. Paul, MN 55144-1000
Garry, The following is an alignment procedure we use for our Hitachi H-7000. You might try following it to see if it will help you. This method always works for us, so if you still have problems you might need to call service. Good luck. Mary Gail Engle
ALIGNMENT OF HITACHI H 7000
Center and Saturate Filament Turn acc voltage on- press 75, located above ready/on button Set magnification to 5000X Turn filament knob clockwise until screen is just illuminated. Condense beam with brightness knob to see wehnelt image Center wehnelt image with gun tilt and gun horizontal pods (lower left panel). Saturate filament by turning filament knob until image barely disappears. (Do not oversaturate) Spread beam with brightness knob
Center Condenser Aperture (two methods for this) Take objective aperture (center pod on column) out by moving lever to right. Condenser aperture (top pod on column) should be in #2 position. Expand beam with brightness knob to edge of focusing screen. Center beam with condenser aperture X/Y knobs Reverse brightness knob and repeat above (expand beam and center with X/Y knobs) If beam spot is elliptical correct with condenser stigmator knobs (lower right panel) until beam spot is circular.
OR:
Center Condenser Aperture Take objective aperture (center pod on column) out by moving lever to right. Condenser aperture (top pod on column) should be in #2 position. Press Index, press enter, press 11, press Index Move cursor to C3 lens using arrows Press C3/OBJ Modulation Button (upper right panel) Adjust X/Y knobs of the movable condenser aperture until beam spreads concentrically. Press C3/OBJ; Press Index If beam spot is elliptical correct with condenser stigmator knobs (lower right panel).
Center Beam Condense and center beam (with brightness knob) Set spot size to 2 (lower left panel). Center spot with brightness centering knobs. Set spot size to 7. Center spot with gun horizontal knobs (lower left panel). Repeat until beam is centered and doesn't move off center. Set spot size to 5. Center beam with brightness centering and spread beam Insert objective aperture (move lever to left)
FOR THE FOLLOWING PROCEDURES INSERT HOLEY GRID OR SAMPLE
Center Objective Aperture Make sure beam is on specimen and not a grid bar or empty space Condense beam with brightness knob. Press DIFF button. Set camera length to 0.4 with magnification knob. Adjust bright spot to very small size with diffraction spot knob Center aperture with X/Y knobs on the objective aperture pod (on the column). **DO NOT LEAVE IN DIFFRACTION MODE FOR MORE THAN 30 SECONDS** Press zoom button and spread beam with brightness knob
Align Imaging System Turn magnification to 20,000X and focus image with focus pods Turn on HV modulation (top right panel) Center beam with brightness centering pods. Adjust beam tilt knobs (lower right panel) until imaged moves in and out and not side to side. Turn off HV modulator button
Check Objective Aperture Astigmation Turn magnification up to 80,000. Press objective stigmatior reset switch (lower left panel) Look for a small hole. Under focus (counterclockwise) so white edge appears inside the hole evenly. If the white rim is not even around the hole, then use the objective stigmator X/Y knobs (lower left panel) to make it even around the entire hole
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
This is a common problem which arises due to alignment conflicts between the electron gun and the condenser system. The alignment procedures is set out below, it is a generic alignment for any TEM.
1. Large spot size centre the gun alignment shift (use some if the spot and halo moves out of alignment) 2. Small(er) spot size centre the illumination alignment shift. 3 Repeat for a constant centre.
Note 1 - most experienced operators simply align for the positions either side of the spot size they use most. Note 2 - to reduce this misalignment, train ALL operators that if they are looking at a filament image in order to align the gun, they must ONLY use gun tilt and shift, never use illumination shift as this causes the misalignment.
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, July 21, 2003 3:30 PM
Dear readers I am using TEM in order to look at my polymer samples. I replicated it with the double stage system. Do I need some kind of support films (Formvar-Carbon) on the grids or the carbon-metal replicas can be observed with a normal copper grid? I would be grateful if You can help me. Regards Alessandro Mattozzi
Many moons ago I was given the task of embedding and sectioning grey hair (human hair). I had a heck of a time getting the epoxy to infiltrate the cuticle, hence the sections tore and the images were less than stellar. The project ended before I could figure out what the best solution to the problem would be. Many moons later the problem has raised its ugly head and a solution must be found. The infiltration times have been expanded, different epoxies have been tried, and several different protocols have been enacted. The samples must not be subject to harsh chemicals because the integrity of the sample must not be compromised.
Any suggestions would help; I am being vague because the samples are not mine and you all know how companies get when it comes to their hair care products, just send some ideas.
thanks,
John Grazul TEM Facility Manager Cornell University Ithaca, NY 607 255 6421
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Thanks for the great responses to my question regarding attaching a digital camera to a Nikon SMZ 1000 stereomicroscope. To make sure this info is placed in the archives, I'm sending out this email with the responses I received. Hopefully this info can help some of you as well.
Thanks again,
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
The Nikon Coolpix cameras have threaded lenses for various adapters that can be used to attach these consumer grade cameras to your microscope. Nikon and some aftermarket suppliers provide an adapter from the threads on the lens to a C-mount that should be on your micoscope. The adapter will also fit right into some eyepiece tubes for use on microscopes without a C-mount.
We have used the Coolpix 990 and 4500 models in this way. Obviously, the images are not of the same quality as the dedicated digital acquisition systems, but are pretty good quality for the cost. The cameras use a Compact Flash card for image storage, which is not as convenient as direct saves to disk but is relatively convenient with an inexpensive USB card reader. I am not so found of the direct USB attachment to the camera and Nikon softward for retrieving images.
Your Nikon rep should be able to provide information on the adapter, or do a quick search on google for "coolpix microscope adapter" to find information on Nikon and aftermarker alternatives.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
*************************************************
Hello William,
Correct me if I am wrong, but I thought that in many cases the problem is much less one of resolution than it is one of available light and the sensitivity of the camera and the ability to view specimens at low light levels.
The so-called "high end" digital cameras have also dropped way down in price , so that there is not as much difference as previously between the "high end" vs. "consumer" cameras, but with the better camera, you also get better sensitivity.
The Pixera line of cameras is one of those so called "high end" cameras, some of which sell for a low price, which also includes a lot of useful software that does not come with a consumer camera. Now I am not exactly a disinterested thirty party since we now offer the Pixera line of cameras, see URL http://www.2spi.com/catalog/photo/pixera/pixera.html
But for lower available light levels, you might not get what you need with the best of consumer cameras, that is why I make this point to you. Note that I not posted this message through the listserver outof concern that someone might think it "too commercial".
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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William- I have a Canon A40, and I hook it up to a TV monitor for viewing and focusing. The nice thing about it is that you can buy a ready-made Canon bayonette adapter-to-52mm right from Canon at camera store. This pops onto the front of the camera, and it is a simple matter to hook up to a T-mount or any other ring from 52mm to whatever you need. You then have easy-on and easy-off of the camera from the microscope without having to unscrew the camera and remove cords... That's why I chose the Canon A-40. If you want more info- I have pictures as well as the set up on my website, so let me know if you are intersted. Rgds, Mike Shaw Roselle, NJ
see ig you can get a some sort of consumer nikon digital camera is my advise. They have less sexpensive modles. They have an incredible macro mode (2CM away). Optem sells and adaptor for about 135 there are many others. good luck.
I've run across some commercially made adapters for consumer cameras, but don't have the info at home. I'll try and dig it out and send it to you on Monday if I can find it (or post to the list).
As far as what grade camera to get, I'd offer the following thoughts:
It sounds like your use on the stereo won't require much, so just about anything over the $100 threshold should work for that.
You may want to consider something a little upscale from that for other uses. If you don't already have one, these things are great for documenting procedures, getting photo's of staff for the web or bulletin board, and just about anything where "a picture is worth a thousand words". If you haven't gotten into the habit already, it can happen quick. I also use mine to take pictures of my white board before cleaning it off. It sure beats manual transcription, and lets me get back to work faster.
For reference, at about 3 Gigapixels you can take a picture (assuming a decent lens system) and print it with a decent inkjet on photo paper and most people would have a hard time telling an 8 x 10 from a film based 8 x 10. (If you know what to look for, you can tell the difference.)
Just food for thought from a non-microscopist.
GO RED!
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc. Milwaukee, WI
These three pictures were taken with a SONY DSC-P31 held up against a small maginfying glass up against the eyepiece of a stereodissection 'scope from Fisher Scientific. I would recoemmend something better, but, as you can see, even a crappy setup can yield decent pics.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
Greetings William. We use a Nikon Coolpix (990 - I think the latest model is 999) with our Olympus stereomicroscope. Coupler options seem to be best for this series, and it takes good pictures for us. Optem International, www.OptemIntl.com, has lots of couplers and can help you choose the right components (coupler and C-mount). You might also check with your Nikon microscope rep; ours (Mager Scientific in Michigan) sells the Coolpix and adapters for customers who don't have 4-5k$ for their fancy digital camera options. I don't think you'll have problems finding what you need, but if you do just let me know and I will put you in touch with a rep at Mager.
Sincerely, Matt
Matthew Stephenson Analytical Associate Impact Analytical/MMI 1910 West Saint Andrews Road Midland, MI 48640 (989) 832-5555 X506 stephenson-at-impactanalytical.com
Alessandro metal-shadowed carbon replicas can be supported on normal copper grids without a support film. Chris
Dr. Chris Jeffree Inveresk Cottage 26, Carberry Road Inveresk Musselburgh Midlothian EH21 8PR Tel: +44 131 665 6062 FAX +44 131 653 6248 Mobile 07710 585 401 ----- Original Message ----- } From: "Alessandro Mattozzi" {alessandro-at-polymer.kth.se} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, July 22, 2003 3:25 PM
Good Day. On behalf of the Local Arrangements Committee of the 2003 Microscopy and Microanalysis Meeting in San Antonio, Texas, I would like to invite you (again) to the 2003 Golf Outing to be held Sunday, August 3 at the Pecan Valley Golf Club. Pecan Valley is a majestic golf course, located only six miles from downtown San Antonio and the Riverwalk. Pecan Valley was the site of the 50th Anniversary PGA Championship in 1968 when Julius Boros edged Arnold Palmer on the 18th hole and has also hosted three Texas Opens. It has been rated in Golf Digest¹s Top 50 Public Courses and #1 Public Golf Course in the State of Texas for 2002 (http://www.golftexas.com/pecan1.htm Rating: 74.5 - Slope: 136 - Yards: 7,071). The cost will be $70.00 and will include greens fees, cart, transportation to and from the Convention Center, driving range, lunch buffet and awards banquet. The bus will pick up people at three hotels: Marriott at 6:15, Hyatt at6:45, Hilton at 6:50. Leave the Hilton at 7:00 and arrive at the golf course at 7:15. Callaway club rental is available onsite at $35.00 per set.
People who want to play golf but don't go through online registration should get me a check for $70 made out to "Microscopy & Microanalysis" or pay onsite by check the M&M representative.
I would appreciate you contacting me by e-mail if you will be joining us for the golf outing. Please reply by Friday, July 25, 2003 (even if you have already registered) to Mark Sanders (msanders-at-cbs.umn.edu) with the following information:
Name:
Address:
Phone number:
E-mail:
Hotel in San Antonio:
Have you registered and paid: yes or no
Do you need rental clubs:
Do you have special dietary needs?
Do you want to play in a particular foursome?
Another addition to this year¹s outing is the inclusion of various types of sponsorships. We are also looking for additional raffle prizes and / or giveaways for the event. Any company or individual that becomes a sponsor will be promoted in the following ways: * On the hole of their choice (for hole sponsorships) * On the lunch placemats * In each ³goodie² bag that every golfer will receive at the course If you are interested in playing in the outing, please contact me directly (msanders-at-cbs.umn.edu). If you have a foursome, please let me know and include all four names. If you do not have a foursome, that is fine as well we can pair you into a foursome at the course. If you are interested in becoming a sponsor, please contact me directly and I can get further details to you.
I would also like to announce that there would be 2 ³free² slots for students (preferably). These will be in the name of a former member, golfer and a good friend of mine who passed away a few years ago, Joe Polak.
Contact Mark Sanders with any questions (msanders-at-cbs.umn.edu)
Thanks in advance for your consideration in this matter!
At Pecan Valley you can truly ³Play where Champions Have Left Their Footprints.²
My Best Regards,
Mark Sanders
MSA/MAS Golf Tournament at the Pecan Valley Golf Club
Alessandro Mattozzi wrote: ========================================================= I am using TEM in order to look at my polymer samples. I replicated it with the double stage system. Do I need some kind of support films (Formvar-Carbon) on the grids or the carbon-metal replicas can be observed with a normal copper grid? I would be grateful if You can help me. ========================================================== A lot will depend on the nature of the surface itself. Generally speaking, if you are talking about Pt/C replicas, we normally apply, after the Pt/C a carbon "backing" film, evaporated more or less vertically over the sample (without shadowing). That usually is enough to keep surface tension forces from pulling apart the replica film on the floating water surface when the plastic is dissolved. I have assumed you used polyacrylic acid (PAA) as the replicating polymer. A relatively flat surface results in an inherently more robust replica film and a rough (e.g. fracture) surface results in a replica film that is more likely to need some kind of backing film (because cracks tend to form out of any long straight "shadows").
You can of course always use a support film but any support film can take away some of the otherwise nice contrast in the replica film.
Of course, everyone has their own personal variation on this theme, and I have cited the one we normally use in our own laboratory for making replicas of polymer surfaces.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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I would like to use an IEEE 1394 or FireWire camera as a replacement for a PAL video camera on a microscope for regular image capture, but also for time-lapse and real-time recordings. The camera is to be used mainly for widefield microscopy (brightfield and fluorescence), but later on Nipkow disc based confocal microscopy is an option.
I am interested in finding a digital (IEEE 1394 or FireWire) color camera in combination with a FireWire interface (PCI) which can deliver frames at least at a rate of 25 fps. The size of the frames should be at least PAL video size (PAL = 760 x 576 pixels) or more if possible.
To observe polymer sample in TEM, you do need the grids pre-coated with carbon film.
The carbon-metal film is stable, but it could create a strong background of metals on your polymer images, that result the image resolution reducing. In contrast, formvar-carbon film or pure carbon film could provide a clean, weak and amorphous background.
On 200 mesh grid, many carbon films on the holes could be broken autmatically by the time going or by the huminity changing, which is because the holes size is too big to support the pure carbon film strongly. Normally, people use the formvar-carbon film on 200 mesh grids. However the formvar-carbon film has its own weakness, such as the formvar film is easy to be burn by electron beam under higher manification and the formvar could have the chemical reaction with the organic matter in your buffer.
The pure carbon film on 300 mesh and 400 mesh grids is much more stable than that on 200 mesh. I suggest you could try the 300 mesh or 400 mesh grids pre-coated with pure carbon film. I hope the hole size didn't block too much interest area under the magnification which you used.
Our company product any kinds of above carbon coated grids. If you are interested, you could get more information from our website,
http://www.grid-tech.com/
Shall you have more question, I will be happy to answer.
with my best regards,
Wendy Zhang ===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
} } Dear readers } I am using TEM in order to look at my polymer } samples. I replicated it with } the double stage system. Do I need some kind of } support films } (Formvar-Carbon) on the grids or the carbon-metal } replicas can be observed } with a normal copper grid? } I would be grateful if You can help me. } Regards } Alessandro Mattozzi } }
===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
This sounds like one more case of the proverbial "hair of the dog that bit you."
Seriously, in case you haven't already tried this: embed in Spurr or LR White. Both are very low viscosity and may offer a better infiltration. Alternatively, look at different infiltration solvents (i.e. ethanol vs acetone vs ...).
Good luck.
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
john grazul {grazul-at-ccmr.corne To: microscopy-at-sparc5.microscopy.com ll.edu} cc: Subject: embedding grey hair
07/22/03 04:01 PM
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All,
Many moons ago I was given the task of embedding and sectioning grey hair (human hair). I had a heck of a time getting the epoxy to infiltrate the cuticle, hence the sections tore and the images were less than stellar. The
project ended before I could figure out what the best solution to the problem would be. Many moons later the problem has raised its ugly head and
a solution must be found. The infiltration times have been expanded, different epoxies have been tried, and several different protocols have been enacted. The samples must not be subject to harsh chemicals because the integrity of the sample must not be compromised.
Any suggestions would help; I am being vague because the samples are not mine and you all know how companies get when it comes to their hair care products, just send some ideas.
thanks,
John Grazul TEM Facility Manager Cornell University Ithaca, NY 607 255 6421
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (manton-at-biol.uoa.gr) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, July 23, 2003 at 09:45:09 ---------------------------------------------------------------------------
Email: manton-at-biol.uoa.gr Name: Dr. Marianna Antonelou
Organization: Univ. Athens, Fac. Biology, Dpt. Cell Biology and Biophysics
Education: Graduate College
Location: Athens, Greece
Question: My goal is to do TEM-immunolabelling on human cells in culture using polyclonal and monoclonal primary antibodies, gold-labeled secondary ntibodies and the post-embedding method. What kind of acrylic embedding resin (compatible with agar) and what protocol (fixatives, time, buffers) would you suggest?
Probably not since Hepes is an organic buffer and KMnO4 is a strong oxidizer that would strongly react with it. Phosphate or an inorganic buffer would be better. Anyway, KMnO4 is such a "destroyer" of fine detail (good for membranes but not much else) that buffers are probably not very useful.
JB
} Hello, } Quick question, Is Hepes compatible with potassium permanganate for } yeast fix? } Mike D
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NO! I did this experiment recently and the mix formed a sludge rapidly and then over the course of hours it turned into a clear solution with a black precipitate. I didn't have sodium cacodylate around so after paraformaldehyde/glut fix, i rinsed in buffer, did 3 quick rinses in water and then fixed in KMnO4 in water and got great results with plant seeds. You risk some osmotic effects with the water rinse but I got away with it. if you are worried, stick with cacodylate (my least favorite buffer). good luck.
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} } } Dear Micronetters, } } } } } } Does anyone know what does the average Image Analysis technician makes } in } } } salary or hourly wage? } } } Is there an image analysis salary survey out there somewhere? } } } } } } I know this is a broad question, but there are so few IA technicians out } } } there. } } } } } } I perform DNA ploidy on breast carcinomas. } } } } } } Thanks for any advance info. } } } } } } Donald G. Awbrey, HT(ASCP) QIHC } } } Electron Microscopy / Image Analysis } } } 817-878-5647 } } } donaldawbrey-at-texashealth.org } } }
Oh! Thanks Nester......
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I don't think you'd beed to buffer the KMnO4 to use as a fixative.
The really good prep method for TEM of yeast that I have tried with pretty good results, uses 2% aqueous potassium permanganate (NOT buffered) as a 2nd fixative, after washing out all traces of the primary glutaraldehyde fix buffered with the PIPES buffer (also containing other ingredients).
The reference is: Robin Wright, Transmission Electron Microscopy of Yeast, Microscopoy Research and Technique 51:496-510 (2000).
Hope this helps,
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Bera
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I am using fresh aqueous 4% solution of KMnO4, 2 hour on ice with good results. For some unclear to me reason, permanganate fixation always better on yeasts than osmium. Sergey
At 08:10 AM 7/23/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I'm looking for some kind of mesh, or screen-like material that I can put on double sided tape such that small, hand-picked mineral grains can be dropped and nicely organized in the rectangular openings. After mounting, the minerals and grid will be covered in epoxy, and polished for WDS analyis.
I'm looking for 0.5-1mm -sized openings (that's millimeter, not micron). I've had a wire mesh company send me some samples but as the openings increase in size, so does the diameter of the wire. WIth the right sized holes the wire diameter is way too fat, and also the metal mesh doesn't stay flat on the tape, it curls up. I have a piece of no-see-um tent screen, which has an amazing, yet unacceptable weave under the microscope. The best I have so far is screen-door material, but this has 2mm squares. I've tried fabric shops, and the web. Any other ideas would be appreciated. Scott
************************************************ ....amphiboles do violence to history... T. Feininger, 2001. (taken out of context) **************************** Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Earth and Space Sciences ph.206-543-8393 Mail Stop 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
I just measured the magnification of the Zeiss 25X plan Neofluar N.A. 0.8 multi immersion objective and found that it has an apparent magnification of 26.88X. Independently, one of our technicians measured it and came out with 27X. (I was checking because I expected to measure 400 um and she had measured 370 um.)
Has anybody else noticed the discrepancy between the advertised value and the measured value? Is this one of those "known" things that we just didn't know?
The Zeiss 10, 40 and 100X objectives gave the expected results.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
} Many moons ago I was given the task of embedding and sectioning grey } hair (human hair). I had a heck of a time getting the epoxy to } infiltrate the cuticle, hence the sections tore and the images were } less than stellar. The project ended before I could figure out what } the best solution to the problem would be. Many moons later the } problem has raised its ugly head and a solution must be found. The } infiltration times have been expanded, different epoxies have been } tried, and several different protocols have been enacted. The } samples must not be subject to harsh chemicals because the integrity } of the sample must not be compromised. } } Any suggestions would help; I am being vague because the samples are } not mine and you all know how companies get when it comes to their } hair care products, just send some ideas. } } thanks, } } John Grazul } TEM Facility Manager } Cornell University } Ithaca, NY } 607 255 6421
John, I have a similar problem with the cuticle of drosophila pupae and have found that the best that I can do is to face the block and trim with a glass knife. Orient the cuticle perpendicular to the knife edge. The hair should be exposed at the bottom of the block face to avoid chatter in the sections. Most likely the hair will still break out of the section so a supported grid (formvar or collodion) will be needed to photograph the edges of the sample. Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
In a previous E-mail I sent information on the facility manager¹s session at M&M2003. The information is repeated below.
While arranging to get the Core Facility management session included in the annual meeting, it was suggested that we might consider forming a Focused Interest Group (FIG) for facility managers. The idea would be to have a group to act as a unit for inclusion in the annual meetings and to interact in other ways (web, newsletter, etc) to disseminate information of interest to this group and discuss common problems and concerns.
I hope to assemble a group of volunteers to discuss this idea of whether the core facility managers should petition the MSA Board to form a Focused Interest Group (MSA) within MSA. We would have to meet during the meetings, Tuesday or Wednesday with room and time to be arranged, to discuss this possibility and draft a request document. The summary of this discussion on forming a FIG and the draft document will be presented at the beginning of the Core Facility Management session for brief discussion and a vote.
Please contact me ASAP (dsherman-at-purdue.edu) if you would be willing to attend the initial meeting or would like to weigh in with your opinions. I will contact you either before the meeting or via the message board at the meeting with time and place. Feel free to comment on this possibility even though you may not be going to the meeting this year. All E-mails will be presented as part of the discussions. Thanks
Debby
============================== Original E-mail as follows:
Please note that the Core Facility Management session at M&M 2003 has been rescheduled. It originally was scheduled for Thursday August 7 from 1:00pm to 3:00pm. NEW TIME is Thursday August 7 from 8:00am to 10:00am in room 202B. Please change the time in your meeting schedules.
Topics for this year are:
How to Increase the Use of the Core Facility...with a Corresponding Increase in Revenue. Facilitator: Elaine Humphrey, U. of British Columbia
Defining the Roles of the Lab manager/Director and the Advisory Committee of a major Core Facility. Facilitator: Debby Sherman, Purdue University.
The goal of this session to to promote discussion of topics of interest to facility managers/directors. Therefore attendees are encouraged to bring information relative to the topics that they would like to share. In this case it may include examples of ways to advertise facility services (brochures, web sites, etc). Or perhaps you have a mission statement that helps clarify roles of staff and advisory committee members. Please contact me if you would like to share information and what sort of audio-visual equipment you would like to have available. Although it helps to have advanced notice to better organize the session, last minute inclusions are always welcome.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
The two main types of color cameras used in microscopy I know of are the ones with 3 separate CCDs for each of the three primary colors and the ones with a Bayer filter design. In the 3CCD camera the spatial sampling is the same for each of the three colors, but not in the Bayer filter camera design ? How do you deal with this difference when doing quantitative spatial measurements of colored samples in microscopy ?
What is the relative actual resolution seen with a 3CCD color camera and a camera with Bayer filter reconstruction? How to take into account the reduced sensitivity of the Bayer filter type for each color, compared with the 3CCD camera if any ? When are Bayer filter type cameras a valid alternative for a 3CCD camera in microscopy, besides the price ?
What about pixel shifts in a 3CCD camera due to misalignment of the color filters/splitter and the CCDs ?
Scott- You may find what you are looking for at the following site:
http://www.internetplastic.com/intromesh.html
They sell electroformed (flat) grid material with a wide range of pitches out of Ni, Cu, and Au. It is more expensive than what you have been using though. I don't know what thickness you need, this may be too thin, but they have all sorts of other grid materials as well. Good Luck.
Sincerely, Matthew Ervin, Ph.D. (301)394-0017 phone, (301)394-1559 fax MErvin-at-ARL.Army.mil
M/S: AMSRL-SE-RL US Army Research Laboratory 2800 Powder Mill Road Adelphi, MD 20783-1197
Disclaimer: The opinions and views expressed above are those of the author and do not necessarily represent those of the U.S. Army Research Laboratory or any other government agency
-----Original Message----- } From: S. Kuehner [mailto:kuehner-at-u.washington.edu] Sent: Wednesday, July 23, 2003 4:14 PM To: MSA
to the list-
I'm looking for some kind of mesh, or screen-like material that I can put on double sided tape such that small, hand-picked mineral grains can be dropped and nicely organized in the rectangular openings. After mounting, the minerals and grid will be covered in epoxy, and polished for WDS analyis.
I'm looking for 0.5-1mm -sized openings (that's millimeter, not micron). I've had a wire mesh company send me some samples but as the openings increase in size, so does the diameter of the wire. WIth the right sized holes the wire diameter is way too fat, and also the metal mesh doesn't stay flat on the tape, it curls up. I have a piece of no-see-um tent screen, which has an amazing, yet unacceptable weave under the microscope. The best I have so far is screen-door material, but this has 2mm squares. I've tried fabric shops, and the web. Any other ideas would be appreciated. Scott
************************************************ ....amphiboles do violence to history... T. Feininger, 2001. (taken out of context) **************************** Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Earth and Space Sciences ph.206-543-8393 Mail Stop 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
It is not unusual for objectives to vary by several percent. That is why,when measurements are critical and/or you are using video systems, we recommend actually calibrating the image using a stage micrometer and the full set of optics used for the experiment.
If you have earlier measurements, all is not lost. You can determine an "adjustment" ratio between the old measurements and your new, calibrated values.
Good hunting!
Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration. Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
At 04:21 PM 7/23/03 -0400, Michael Cammer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Contact the apps lab at Buehler (Waukegan, IL). They have a great little vacuum embedder which might help you to get better infiltration.
Good hunting, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration. Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
At 05:34 PM 7/23/03 -0400, Pat Connelly wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A product is available from SPI, and probably the other EM supply companies as well, that consists of a substrate (glass, conductive material, plastic, .. ???) upon which is imprinted a matrix of tacky elastomeric dots. Particles can be placed on the tacky dots and held in place for analysis, etc. In your case, you might consider using the tacky dots to hold the isolated particles in place during segregation and subsequent embedment.
I'm sure that my "tacky dot" terminology is wrong but I have actually seen the product at the 2002 M&M exhibition. It looks like it could work for your application.
Good luck,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"S. Kuehner" {kuehner-at-u.washing To: MSA {microscopy-at-sparc5.microscopy.com} ton.edu} cc: Subject: mineral mount grids
07/23/03 03:13 PM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
to the list-
I'm looking for some kind of mesh, or screen-like material that I can put on double sided tape such that small, hand-picked mineral grains can be dropped and nicely organized in the rectangular openings. After mounting, the minerals and grid will be covered in epoxy, and polished for WDS analyis.
I'm looking for 0.5-1mm -sized openings (that's millimeter, not micron). I've had a wire mesh company send me some samples but as the openings increase in size, so does the diameter of the wire. WIth the right sized holes the wire diameter is way too fat, and also the metal mesh doesn't stay flat on the tape, it curls up. I have a piece of no-see-um tent screen, which has an amazing, yet unacceptable weave under the microscope. The best I have so far is screen-door material, but this has 2mm squares. I've tried fabric shops, and the web. Any other ideas would be appreciated. Scott
************************************************ ....amphiboles do violence to history... T. Feininger, 2001. (taken out of context) **************************** Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Earth and Space Sciences ph.206-543-8393 Mail Stop 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
I'm not aware of any Zeiss specs for objective accuracy. There must certainly be some amount of allowed error. +/- 0% would be a really expensive objective!!
If you measured 26.88X, that would be about 7.5% error. If she measured 27X, that is 8% error. If you measured 400u and she got 370u, then the 30u difference is 7.5% error relative to your reading. So this jives with the mag readings. It seems that your method of checking magnification may not be as perfect as the objective. But if so, why do the other objectives check out OK? Perhaps there is an issue with two different pairs of eyes looking through the same oculars? Were the oculars tweaked for diopter correction?
Try a calibrated stage micrometer and see what you get. For critical measurement work, the micrometer is used for each objective. Then, that difference in mag is cranked into subsequent measurement readings.
gary g.
At 01:21 PM 7/23/2003, you wrote:
} I just measured the magnification of the Zeiss 25X plan Neofluar N.A. 0.8 } multi immersion objective and found that it has an apparent magnification } of 26.88X. Independently, one of our technicians measured it and came out } with 27X. (I was checking because I expected to measure 400 um and she } had measured 370 um.) } } Has anybody else noticed the discrepancy between the advertised value and } the measured value? Is this one of those "known" things that we just } didn't know? } } The Zeiss 10, 40 and 100X objectives gave the expected results. } } ____________________________________________________________________________ } Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. } Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 } (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ } }
Good Afternoon, We are starting some eye projects in my laboratory, and I have done very little with eyes except diagnostic biopsies, etc. Is there a good reference or tips on processing rat eyes for ultrastructural studies. Since there is interest in finding specific areas within the eye (therefore the need to know lateral, medial, rostral, and cranial zones of the eye), how usually is the eye cut (minced) for ultrastructural studies? Is perfusion superior to immersion for fixation? Thank you for any help you can give me. Stanley E. Hansen
I am going to buy an achromatic objective 20x n.a. 0.45 (spring loaded) from the Lambda Praha Ltd. and manufactured as DIN standard i.e. for 160 mm. tube length. I wonder if someone knows that kind of objective and he could tell me something about its quality.
If the measurement is done on a digital image acquired by a digital camera and not directly, this could cause the problem. The variation in the measurement could be caused by inaccuracy of spatial sampling. Calibrating a microscope equipped with a digital camera at "low" magnification with a lens with a relative high N.A. is prone to spatial sampling errors.
If the the Zeiss 25X plan Neofluar N.A. 0.8 objective is not matched to your camera, you get an error, caused by misalignment of the sample relative to the CCD-grid of the camera.
There is an excellent article on this issue by Prof. T. Young from the T.U.Delft in the Netherlands, scroll down a bit and have a look at the section on "Sampling Density":
} } I just measured the magnification of the Zeiss 25X plan Neofluar N.A. 0.8 } } multi immersion objective and found that it has an apparent magnification } } of 26.88X. Independently, one of our technicians measured it and came out } } with 27X. (I was checking because I expected to measure 400 um and she } } had measured 370 um.) } } } } Has anybody else noticed the discrepancy between the advertised value and } } the measured value? Is this one of those "known" things that we just } } didn't know? } } } } The Zeiss 10, 40 and 100X objectives gave the expected results. } } } } ____________________________________________________________________________ } } Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. } } Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 } } (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ } } } }
A variety of polymer or metal screens are available from Small Parts, Inc. www.smallparts.com. They sell plain weave polyester with 1mm openings and the open area is 57.5%. With Nylon the openings can be 2 mm with an open area of 53%. If you can tolerate metal then Buckbee-Mears Company, (612) 228-6400, makes metal masks by electropatterning. They have a free sample sheet that contains a flat (no weave) stainless steel screen with 1mm X 0.5 mm openings. They can also make custom parts.
Edison Analytical Laboratories, Inc. is an analytical services laboratory that has no commercial interest in either of the cited sources other than as a user of their products.
Dr. James Carnahan
Edison Analytical Laboratories, Inc. 301 Nott Street Schenectady, NY 12305 (518) 393-2112
In our lab we calibrated every combination of lenses and magnifications we use on our metallograph microscope. We did this with a calibrated stage micrometer and keep this calibration chart nearby for when we need to present accurate measurements.
Stu Smalinskas SKF USA Plymouth, Michigan
At 01:21 PM 7/23/2003, you wrote:
} I just measured the magnification of the Zeiss 25X plan Neofluar N.A. 0.8 } multi immersion objective and found that it has an apparent magnification } of 26.88X. Independently, one of our technicians measured it and came out } with 27X. (I was checking because I expected to measure 400 um and she } had measured 370 um.) } } Has anybody else noticed the discrepancy between the advertised value and } the measured value? Is this one of those "known" things that we just } didn't know? } } The Zeiss 10, 40 and 100X objectives gave the expected results. } } ____________________________________________________________________________ } Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. } Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 } (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ }
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Dear Fellow Listers, I am interested in purchasing a Cryo-Ultramicrotome for our user facility; we deal mainly with materials science applications. Would any vendors specializing in this type of product please contact me off-line with information about your products.
Also, if anyone out there has strong opinions, pros or cons, about different systems please contact me. Your input would be greatly appreciated.
Best regards, Ray Twesten
Ray D. Twesten, PhD. Center for Microanalysis of Materials Seitz Materials Research Laboratory 104 S. Goodwin Ave., Urbana, IL 61801 +1 217 244-6177 (Fax -2278)
} From:Dr.ALHAJI USMAN ALIEGO Nigerian National Pet.Corp. Lagos-Nigeria. Attn: President/Ceo.
STRICTLY CONFIDENTIAL BUSINESS PROPOSAL RE: TRANSFER OF US$58.5 MILLION (FIFTY EIGHT MILLION, FIVE HUNDRED THOUSAND US DOLLARS ONLY).
I know this email will reach you as a surprise,but need not to worry as we are using the only secured and confidential medium available to seek for foreign assistance/partnership in a business transaction which is of mutual benefit.
I am a member of the Government of Nigeria Contract Award and Monitoring Committee in the Nigerian National Petroleum Corporation(NNPC)Sometime ago,a contract was awarded to a foreign firm in NNPC by my Committee.This contract was over invoiced to the tune of US$58.5Million .U.S.Dollars. This was done deliberately,the over-invoicing was a deal by my committee to benefit from the project. We now want to transfer this money which is in a suspense Account with NNPC into any Overseas Account which we expect you to provide for us. SHARE: - For providing the account where we shall remit the money into,you will be entitled to 20% of the money,70% will be for me and my partners while 10% has been mapped out from the total sum to cover any upfront or out of pocket expenses that may be incurred by all parties during the course of this transfer,both locally and international expenses.In order for us to commence the transaction i would require the following: -
1.Your company's name,address,telephone and fax numbers.
2.Your bank name,address and account details.(May an account with zero balance)
The above information would be use to make formal applications as a matter of procedure for the release of the money and onward transfer to your account.It does not matter whether or not your company does contract projects of this nature described here. The assumption is that your company won the major contract and subcontracted it out to other companies.More often than not,big trading companies or firms of unrelated fields win major contracts and subcontracts to more specialized firms for execution of such contracts. We have strong reliable connections and contacts at the Central Bank Of Nigeria,as well as the Ministry of Finance and we have no doubt that all the money will be released and transferred if we get the necessary foreign partner to assist us in this deal.Therefore, when the business is successfully concluded we shall through our same connections withdraw all documents used from all the concerned Government Ministeries for 100% security.
We are ordinary Government workers and we will not want to miss this once in a lifetime opportunity to get rich. We want this money to be transferred to the overseas Accounts for us,before the present Government start Auditing all Government owned Parastatals accounts. Please contact me immediately through my email addresses whether or not you are interested in this deal.If you are not,it will enable me scout for another foreign partner to carry out this deal.But where you are interested, send the required documents aforementioned herein through my CONFIDENTIAL email address {usman009-at-freesurf.fr} as time is of the essence in this business. I wait in anticipation of your fullest co-operation.
Yours faithfully,
Dr.ALHAJI USMAN ALIEGO
PLEASE IF YOU ARE NOT INTERESTED ON THIS, KINDLY DESTROY THIS MESSAGE AND MENTAIN THE ADEQUATE SECRET FOR THE SAFETY OF ME AND MY COLEAGUES.
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
Scott Kuehner wrote and Gary M. Brown commented on the following request: ========================================================== I'm looking for some kind of mesh, or screen-like material that I can put on double sided tape such that small, hand-picked mineral grains can be dropped and nicely organized in the rectangular openings. After mounting, the minerals and grid will be covered in epoxy, and polished for WDS analysis.
I'm looking for 0.5-1mm -sized openings (that's millimeter, not micron). I've had a wire mesh company send me some samples but as the openings increase in size, so does the diameter of the wire. WIth the right sized holes the wire diameter is way too fat, and also the metal mesh doesn't stay flat on the tape, it curls up. I have a piece of no-see-um tent screen, which has an amazing, yet unacceptable weave under the microscope. The best I have so far is screen-door material, but this has 2mm squares. I've tried fabric shops, and the web. Any other ideas would be appreciated. ============================================================ Gary is right, the SPI Tacky Dot Slides (a proprietary product of SPI Supplies) would probably work, the product being described on URL http://www.2spi.com/catalog/new/tacky.shtml
The standard slide product with the largest dots is the one of 300 µm dots on 2,000 µm centers. So the dots might be a bit too small and the center to center spacing might be too small as well for this particular application... .unless one was to custom fabricate a slide with a larger center to center spacing (not a problem to do).
But the technique will work, and as evidence of that statement, see URL http://www.2spi.com/catalog/new/tackdot_array.html They were prepared exactly as proposed, except with the use of a Tacky Dot Slide instead of a screen mesh.
The cross-sectioned particles are all arranged in an orthogonal array, ready for automated analysis if one has stage automation on their system.
If you still wanted a mesh, you might want to see URL http://www.2spi.com/catalog/standards/stndcal6.shtml
A 30 mesh electroformed screen is available and it has a hole size of 785 µm or 0.785 mm which is in the range of your requirement. The "wire" is 61.5 µm , so I don't know if it will work for you, but that seems to be what you requested.
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
I am Dr.Ezenwa Chukwu. Chairman of the Tender Committee of the Nigerian National Petroleum Corporation (NNPC). My Committee is principally concerned with payment of all contract awarded from 1998 to date, in order of priority as regard capital projects of the NNPC. The information we gathered from the Foreign Office of the Nigeria Chambers of Commerce and Industries is so positive as to convince us that you would provide us with solution to a money transfer deal valued at Thirty One Million United States Dollars and subsequently a joint business venture. In the course of our duties as values, and project inspectors for the on-going liquefied Natural Gas (LNG) project, we have over-invoiced the value of some jobs done by foreign contractors for the NNPC to the tune of US$31M. As follows: - Computer optimization and Installation $16,000.000.00 Installation of 250,000.00 Monax Turbine$10,000.000.00 Turn Around Maintenance $5,000,000.00 Our aim of over-invoicing this payment is to divert the excess amount to a discrete account abroad. This fund is now floating in suspense account at the Central Bank of Nigeria (CBN). This is the fund my colleagues and I have decided to transfer into your account since we, as civil servants are not allowed to operate or own foreign account. The money will be shared as follows after transfer:30% for you (Account Owner) 60% for me and my colleagues10% to off-set both local and international expenses that Would be incurred in the course of this transaction. To be able to claim the funds, we will be purporting your company to be the original contractor / beneficiary of the funds so all procedures for international transfer shall be strictly followed, as we have worked out all modalities for a swift and riskfree transfer.
If this proposal satisfies you, please contact me Through ezenwachukwu009-at-go.com with the following important information. Bank Name / Address Account Name ,Account NumberTel/Fax Telex of Bank Personal Phone / Fax Numbers for easy communication. This transaction will last for 14 working days from the time we submit the required information, as all modalities concerning this transaction have been worked out and it is completely risk free.
Please be informed that this subject is classified sensitive. Therefore treat the transaction with utmost confidentiality and urgency.
Yours Faithfully. Dr.Ezenwa Chukwu URGENT REPLY:Please reply to: ezenwachukwu009-at-netzero.com
I am Dr.Ezenwa Chukwu. Chairman of the Tender Committee of the Nigerian National Petroleum Corporation (NNPC). My Committee is principally concerned with payment of all contract awarded from 1998 to date, in order of priority as regard capital projects of the NNPC. The information we gathered from the Foreign Office of the Nigeria Chambers of Commerce and Industries is so positive as to convince us that you would provide us with solution to a money transfer deal valued at Thirty One Million United States Dollars and subsequently a joint business venture. In the course of our duties as values, and project inspectors for the on-going liquefied Natural Gas (LNG) project, we have over-invoiced the value of some jobs done by foreign contractors for the NNPC to the tune of US$31M. As follows: - Computer optimization and Installation $16,000.000.00 Installation of 250,000.00 Monax Turbine$10,000.000.00 Turn Around Maintenance $5,000,000.00 Our aim of over-invoicing this payment is to divert the excess amount to a discrete account abroad. This fund is now floating in suspense account at the Central Bank of Nigeria (CBN). This is the fund my colleagues and I have decided to transfer into your account since we, as civil servants are not allowed to operate or own foreign account. The money will be shared as follows after transfer:30% for you (Account Owner) 60% for me and my colleagues10% to off-set both local and international expenses that Would be incurred in the course of this transaction. To be able to claim the funds, we will be purporting your company to be the original contractor / beneficiary of the funds so all procedures for international transfer shall be strictly followed, as we have worked out all modalities for a swift and riskfree transfer.
If this proposal satisfies you, please contact me Through ezenwachukwu009-at-go.com with the following important information. Bank Name / Address Account Name ,Account NumberTel/Fax Telex of Bank Personal Phone / Fax Numbers for easy communication. This transaction will last for 14 working days from the time we submit the required information, as all modalities concerning this transaction have been worked out and it is completely risk free.
Please be informed that this subject is classified sensitive. Therefore treat the transaction with utmost confidentiality and urgency.
Yours Faithfully. Dr.Ezenwa Chukwu URGENT REPLY:Please reply to: ezenwachukwu009-at-netzero.com
I mostly have to deal with digital microscopy in extreme low-light conditions (fluorescence microscopy) and the quality of the images is very important for the subsequent quantitative analysis.
I would like to know if the following formula for calculating the maximum Signal to Noise Ratio (SNR) for a CCD camera still holds (in general). Photon noise or photon shot noise, refers to the inherent natural variation of the incident photon flux and a such this form of noise limits the maximum SNR for a given CCD-element and the CCD camera ?
If we consider the capacity of one CCD element of a (Silicon-based) CCD, the capacity for photoelectrons "C" relates to the "maximum SNR" of a CCD camera:
SNRmax = 10 log10(C)
In general a CCD camera seems to have a photoelectron capacity per square micron of about 700 photoelectrons/µm^2. So, the maximum SNR of a CCD camera is defined by the "well" capacity of each individual CCD-element (pixel) and cannot be surpassed. So, there is no substitute for square microns per CCD-element to get a higher maximum SNR ? Increasing the SNR in this way however relates inversely to the spatial sampling ?
The total well capacity divided by the noise of the camera system, gives the dynamic range of the camera. The higher the dynamic range, the less noisy the camera becomes. So, there is a direct relation between individual CCD-element surface area and dynamic range ?
There is a lot more to be said on this in digital micrscopy, but I want to know wether this relation is (still) valid and that I did not miss some recent developments. What about the varying quantum efficiency depending on the wavelength of the incident photons ? What about coating CCD-cameras to improve the quantum efficiency in the shorter wavelengths or blocking near-infrared for which CCD's seem to be very sensitive in order to improve the SNR over the entire visible spectrum ?
We are using multiwell plates, mostly according to SBS standards on our automated microscope systems, ranging from 6 wells up to 1536 wells (and beyond for arrays if necessary). I am looking for something which makes life easier to calibrate the XY (and maybe Z) positioning of the sytem.
Are there plates or molds available which contain pinholes or other marks which make it easier to locate the center of the wells for SBS-standard plates (or others formats) ? We can position in XYZ with submicron precision, but XY-positioning in the sub-mm. range should do to start with. This would allow for an automated calibration procedure on the system.
Are there glass plates available in SBS-size format, which can be used for scanning arrays beyond 6144 format density ?
Dear List, We probably have a leak in the body of the Diffstak oil pump in our Philips SEM 515 microscope. Did anybody try to cure such a leak using a leak sealent? Is there any chance to succeed? Another question is on possibility of replacement of the pump (Edwards Diffstak PH 63 58 03, 230 V). Is it still produced or maybe is available as "second hand" item? Thank you for any suggestions,
Leszek Kepinski
Institute of Low Temperature and Structure Research, Polish Academy of Sciences, P.O.Box 1410, 50-950 Wroclaw, Poland kepinski-at-int.pan.wroc.pl
Has anyone been experiencing any section contamination recently that is extremely hard to tract down? Something resembling pepper, but is not due to staining artifacts? Especially in sections with an Epon or EmBed resin component?
??????
Curiously, Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! 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/
Just a reminder that we are hosting the 8th Annual Materials Microtomy Course in Tucson, Arizona October 28 - 31, 2003.
The course is filling up rapidly so if you wish to participate please drop me an email, or stop by our booth in San Antonio.
Full details, including a printable version of the course brochure, can be found at our website: www.rmcproducts.com
Dave Roberts Director - RMC EM Products Boeckeler Instruments, Inc 4650 South Butterfield Drive Tucson, AZ 85714 Tel: (520) 745-0001 Fax: (520) 745-0004 website: www.rmcproducts.com
A researcher on campus is working with photoreceptor cells, that have been dissociated from the retina. This lab would like to know if anyone on the list serve has a procedure for fixing the cells for light microscopy? They are also interested in a procedure for lectin binding? They would be grateful for any help. Thank you. Nancy Cherim University of New Hampshire
On Monday, July 28, 2003, at 02:21 AM, Peter Van Osta wrote:
} Hi, } } I mostly have to deal with digital microscopy in extreme low-light } conditions (fluorescence microscopy) and the quality of the images is } very important for the subsequent quantitative analysis. } } I would like to know if the following formula for calculating the } maximum Signal to Noise Ratio (SNR) for a CCD camera still holds (in } general). Photon noise or photon shot noise, refers to the inherent } natural variation of the incident photon flux and a such this form of } noise limits the maximum SNR for a given CCD-element and the CCD camera } ? } } If we consider the capacity of one CCD element of a (Silicon-based) } CCD, } the capacity for photoelectrons "C" relates to the "maximum SNR" of a } CCD camera: } } SNRmax = 10 log10(C) } } In general a CCD camera seems to have a photoelectron capacity per } square micron of about 700 photoelectrons/µm^2. So, the maximum SNR of } a } CCD camera is defined by the "well" capacity of each individual } CCD-element (pixel) and cannot be surpassed. So, there is no substitute } for square microns per CCD-element to get a higher maximum SNR ? } Increasing the SNR in this way however relates inversely to the spatial } sampling ? } } The total well capacity divided by the noise of the camera system, } gives } the dynamic range of the camera. The higher the dynamic range, the less } noisy the camera becomes. So, there is a direct relation between } individual CCD-element surface area and dynamic range ? } } There is a lot more to be said on this in digital micrscopy, but I want } to know wether this relation is (still) valid and that I did not miss } some recent developments. What about the varying quantum efficiency } depending on the wavelength of the incident photons ? What about } coating } CCD-cameras to improve the quantum efficiency in the shorter } wavelengths } or blocking near-infrared for which CCD's seem to be very sensitive in } order to improve the SNR over the entire visible spectrum ? } } Best regards, } } Peter Van Osta } Dear Peter, Please post the replies you get to the list. We have a similar problem, since we are doing low-dose EM. The questions asked in your post are the kinds of things we have been considering. We have also considered such problems as readout noise and whether the electronics gain can be optimized to give better S/N. We also need to get high spatial frequencies, so small pixel size and narrow point-spread function are important to us as well. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Hello. If you see pepper precipitate just over the tissue it might be osmium. Often when sample is not washed properly after fixation with glut osmium will precipitate. I do not think there is a cure for that when sample if already embedded. Dorota
Still looking for something to do Sunday morning in San Antonio? There is still space available for the 2003 golf outing at Pecan Valley Golf Club! On behalf of the Local Arrangements Committee of the 2003 Microscopy and Microanalysis Meeting in San Antonio, Texas, I would like to invite you (again) to the 2003 Golf Outing to be held Sunday, August 3 at the Pecan Valley Golf Club. Pecan Valley is a majestic golf course, located only six miles from downtown San Antonio and the Riverwalk. Pecan Valley was the site of the 50th Anniversary PGA Championship in 1968 when Julius Boros edged Arnold Palmer on the 18th hole and has also hosted three Texas Opens. It has been rated in Golf Digest¹s Top 50 Public Courses and #1 Public Golf Course in the State of Texas for 2002 (http://www.golftexas.com/pecan1.htm Rating: 74.5 - Slope: 136 - Yards: 7,071). The cost will be $70.00 and will include greens fees, cart, transportation to and from the Convention Center, driving range, lunch buffet and awards banquet. The bus will pick up people at three hotels: Marriott at 6:15, Hyatt at6:45, Hilton at 6:50. Leave the Hilton at 7:00 and arrive at the golf course at 7:15. Callaway club rental is available onsite at $35.00 per set.
People who want to play golf but don't go through online registration should get can pay at the registration booth on Saturday or pay onsite at the course by check to the M&M representative. The cost is $70 made out to "Microscopy & Microanalysis"
I would appreciate you contacting me by e-mail if you will be joining us for the golf outing. Please reply by Wed July 30, 2003 (even if you have already registered) to Mark Sanders (msanders-at-cbs.umn.edu) with the following information:
Name:
Address:
Phone number:
E-mail:
Hotel in San Antonio:
Have you registered and paid: yes or no
Do you need rental clubs:
Do you have special dietary needs?
Do you want to play in a particular foursome?
Shirt size: M L XL XXL
Another addition to this year¹s outing is the inclusion of various types of sponsorships. We are also looking for additional raffle prizes and / or giveaways for the event. Any company or individual that becomes a sponsor will be promoted in the following ways: * On the hole of their choice (for hole sponsorships) * On the lunch placemats * In each ³goodie² bag that every golfer will receive at the course If you are interested in playing in the outing, please contact me directly (msanders-at-cbs.umn.edu). If you have a foursome, please let me know and include all four names. If you do not have a foursome, that is fine as well we can pair you into a foursome at the course. If you are interested in becoming a sponsor, please contact me directly and I can get further details to you.
Contact Mark Sanders with any questions (msanders-at-cbs.umn.edu)
Thanks in advance for your consideration in this matter!
At Pecan Valley you can truly ³Play where Champions Have Left Their Footprints.²
My Best Regards,
Mark "keep your head down" Sanders
MSA/MAS Golf Tournament at the Pecan Valley Golf Club
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hoganbecky-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, July 28, 2003 at 20:07:39 ---------------------------------------------------------------------------
Question: I will be teaching gifted 5th graders in a new math/science program this year and need to know what type of microscope would be best for my program. Also, where can I buy a microscope for a reasonable price? Do colleges ever donate used microscopes to schools? I am working with a shoestring budget of $0.00 so I will have to make a personal purchase in order for my students to use this wonderful teaching tool. Thank you for asssisting me with this matter. I want to offer the best science program possible! Becky Hogan
} On Monday, July 28, 2003, at 02:21 AM, Peter Van Osta wrote: } } } Hi, } } } } I mostly have to deal with digital microscopy in extreme low-light } } conditions (fluorescence microscopy) and the quality of the images is } } very important for the subsequent quantitative analysis. } } } } I would like to know if the following formula for calculating the } } maximum Signal to Noise Ratio (SNR) for a CCD camera still holds (in } } general). Photon noise or photon shot noise, refers to the inherent } } natural variation of the incident photon flux and a such this form of } } noise limits the maximum SNR for a given CCD-element and the CCD camera } } ?
There are two excellent articles by J. M. Zuo in Ultramicroscopy,
which detail how to measure the gain, modulation transfer function, and detector quantum efficiency of a CCD imaging system. These articles deal with TEM, but I think the methods should be applicable to light microscopy as well.
For imaging in low signal conditions, I think the DQE is the most relevant parameter. It's defined as
SNR_OUT^2 DQE = ------------------ SNR_IN^2
The electronics that read the CCD chip and convert it to a digital image also add some noise, so you want to characterize the entire system, not just the chip.
Best wishes, Paul Voyles
Paul Voyles Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
The performance of a CCD camera depends on a lot of components - the characteristics of the detector on the chip (materials, dimensions, front or rear illumination) as well as the readout electronics (analog) and the subsequent digitization, all of which contribute noise (some additive, some multiplicative). It is a very complicated subject not easily reduced to a simple measurement, unfortunately. The best reference I have seen to the various sources of noise and other characteristics of performance is a very thick but comprehensive book: J. R. Janesick "Scientific Charge-Coupled Devices" SPIE Press, 2001 (isbn 0-8194-3698-4)
First try to determine the exact location of the leak. If it is in the steel/aluminum body it probably could be welded. Are you convinced it is not an O-ring? Torr seal will hold up to 120 degrees C. E-mail our evaporator manufacturing department directly at mb-at-laddresearch.com if you have any further questions and they can probably help.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Leszek Kêpiñski" {kepinski-at-int.pan.wroc.pl} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, July 28, 2003 10:01 AM
Randy,
We can't tell exactly the cause of your problem, but a number of years ago we experienced some problems with raw DDSA. We would think it could be dust particles, but the source could be anywhere. If it is Ladd LX112 Epon or our accelerators or hardeners please let us know and we'll investigate. There have been no other such reports.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, July 28, 2003 10:56 AM
Bill,
As I see it, the question should be answered differently for the light and electron microscopy cases. I have nothing to add to Peter Van Osta's comments on the light microscopy case. However, I think I can comment productively on the electron microscopy case. Note that it is beyond me to express the following without resorting to some mathematical formalism, so here goes:
The problem with a straightforward examination of pixel capacity and readout noise in the case of electron image capture is that the sequential image conversion (EM electron to light to CCD electron to DN) constitutes a stochastic chain of neighboring and hence non-independent detectors. The non-independence means that it is not possible to do a meaningful noise budget for the pixel in the CCD and it's mapped effective pixels at the scintillator and in the optical chain. As an example, early excitement over the possibility of measuring the gain of a CCD camera by measuring the shot noise in a uniform image met with the discovery that the noise was much less than could be accounted for by shot noise. The explanation was in the mixing of the light signal between neighboring pixels. Several attempts ensued to manage the mixing by means of a mixing factor. These attempts all failed due to the fact that noise and mixing occurred interspersed throughout the stochastic chain of image conversion and transfer - and were therefore inseparable.
It turns out that the non-independence of physical pixels can be dealt with through a conversion to Fourier space due to the convolution theorem. Pixel mixing in real space transforms to independent scaled channels in Fourier space and noise budgeting becomes possible once again. The formalism for this is stated (exactly as in Paul Voyles' email) but with an additional variable: spatial frequency.
After painful efforts to clarify these issues for electron microscopy it was discovered (by Hans DeRuijter) that an extensive literature existed already in the Medical Imaging field on exactly this concept (DQE (N, S) where S is the spatial frequency). Medical Imaging references can be found by searching on the names: Ian Cunningham and Rodney Shaw. Electron microscopy references can be found by searching on the names: Hans DeRuijter and Ruediger Meyer. Rudy has published a series of both theoretical and experimental analyses of electron cameras in terms of DQE in Ultramicroscopy.
In short, the SNR of an image (transformed into Fourier space) is the signal power spectrum divided by the noise power spectrum. The DQE is defined as the square of the output SNR over the input SNR. A simple regrouping of terms gives:
Which is fairly straightforward to measure since for low-contrast, low-dose imaging, the noise power spectrum "in" is just the Poisson noise in the incident beam, and since that noise distributes evenly in Fourier space (it is "white"), the NPSin = N. The NPSout is just the power spectrum of the output image provided the image is recalibrated in units of EM electrons before transformation. (The Fourier transform must be normalized to conserve sum of squares.) The MTF can be measured by the "tilted edge method" using a relatively clean beam-stop edge.
It can also be seen that the SNR out of a detector is just sqrt (DQE * specimen contrast * dose). Thus DQE can be seen as the "dose-efficiency" of a detector as a function of dose and spatial frequency. Twice the DQE means you need half the dose at any given spatial frequency of interest.
Hope this helped,
Paul
Paul Mooney
Program Manager, Imaging Product Development Gatan, Inc. 6678 Owens Dr. Pleasanton, CA 94588
tel: 925 224-7335 FAX: 925 463-0204
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Monday, July 28, 2003 4:40 PM To: microscopy-at-sparc5.microscopy.com
On Monday, July 28, 2003, at 02:21 AM, Peter Van Osta wrote:
} Hi, } } I mostly have to deal with digital microscopy in extreme low-light } conditions (fluorescence microscopy) and the quality of the images is } very important for the subsequent quantitative analysis. } } I would like to know if the following formula for calculating the } maximum Signal to Noise Ratio (SNR) for a CCD camera still holds (in } general). Photon noise or photon shot noise, refers to the inherent } natural variation of the incident photon flux and a such this form of } noise limits the maximum SNR for a given CCD-element and the CCD camera } ? } } If we consider the capacity of one CCD element of a (Silicon-based) } CCD, } the capacity for photoelectrons "C" relates to the "maximum SNR" of a } CCD camera: } } SNRmax = 10 log10(C) } } In general a CCD camera seems to have a photoelectron capacity per } square micron of about 700 photoelectrons/µm^2. So, the maximum SNR of } a } CCD camera is defined by the "well" capacity of each individual } CCD-element (pixel) and cannot be surpassed. So, there is no substitute } for square microns per CCD-element to get a higher maximum SNR ? } Increasing the SNR in this way however relates inversely to the spatial } sampling ? } } The total well capacity divided by the noise of the camera system, } gives } the dynamic range of the camera. The higher the dynamic range, the less } noisy the camera becomes. So, there is a direct relation between } individual CCD-element surface area and dynamic range ? } } There is a lot more to be said on this in digital micrscopy, but I want } to know wether this relation is (still) valid and that I did not miss } some recent developments. What about the varying quantum efficiency } depending on the wavelength of the incident photons ? What about } coating } CCD-cameras to improve the quantum efficiency in the shorter } wavelengths } or blocking near-infrared for which CCD's seem to be very sensitive in } order to improve the SNR over the entire visible spectrum ? } } Best regards, } } Peter Van Osta } Dear Peter, Please post the replies you get to the list. We have a similar problem, since we are doing low-dose EM. The questions asked in your post are the kinds of things we have been considering. We have also considered such problems as readout noise and whether the electronics gain can be optimized to give better S/N. We also need to get high spatial frequencies, so small pixel size and narrow point-spread function are important to us as well. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
I am looking for a new/used EM tissue processor. Any suggestions/recommendations/info are highly appreciated.
TIA,
GN
Gang (Greg) Ning Director, Electron Microscopy Facility Huck Institute for Life Sciences The Pennsylvania State University 1 South Frear Lab University Park, PA 16802 Phone: 814-863-0994 Fax: 814-863-1357 Email: gxn7-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Caroline Schooley, the project MICRO coordinator, gave me this advice when I turned to her with a similar question some time ago:
First, I went to the Project MICRO webpage http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html {http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html} .
Youâll find very helpful the section âBuying microscopesâ in the Microscopic Explorations part, as well as great projects ideas for the class.
For the microscope purchase, you can go to http://www.microscopeworld.com/
There are several options according to your needs and budget. I bought a few #185 monocular dissecting scopes for $75 each (what a deal!) for the class of second graders, both the kids and the teachers love them. The parents of the kids in the class decided to sponsor this, and contributed about $30 per family - that might be an option in a class with no funding for new equipment.
Good luck,
Alice.
Alice Dohnalkova Pacific Northwest National Laboratory Richland, WA 99352 (509) 372-0692
-----Original Message----- From: by way of Ask-A-Microscopist [mailto:hoganbecky-at-hotmail.com] Sent: Tue 29/07/2003 07:26 To: Microscopy-at-sparc5.microscopy.com Cc: Subject: Ask-A-Microscopist: teaching gifted 5th graders
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hoganbecky-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, July 28, 2003 at 20:07:39 ---------------------------------------------------------------------------
Question: I will be teaching gifted 5th graders in a new math/science program this year and need to know what type of microscope would be best for my program. Also, where can I buy a microscope for a reasonable price? Do colleges ever donate used microscopes to schools? I am working with a shoestring budget of $0.00 so I will have to make a personal purchase in order for my students to use this wonderful teaching tool. Thank you for asssisting me with this matter. I want to offer the best science program possible! Becky Hogan
The Department of Molecular, Cellular and Developmental Biology and the Neuroscience Research Institute of the University of California at Santa Barbara are sponsoring a Microscopy and Digital Imaging workshop on Sept. 15-19, 2003. Participants will get hands-on instruction and experience with advanced light, fluorescence, and confocal microscopy and imaging techniques that are essential tools for research. For further information and registration, please check our web site:
'Once again the MSA Education Committee is organizing Exhibitor Demonstrations and Tutorials on Tuesday, August 5 from 6:00 to 8:00pm in the Exhibit Hall. These mini-seminars or tutorial demonstrations are held in the booths of the participating companies after the Hall is closed to non-participants at 5:00 pm.
Signup sheets with titles and descriptions are at the MSA Education table in the MSA Mega Booth. When you sign up you will be issued a ticket, which you will need to renter the Hall after it is closed. You need to sign up no later than noon on Tuesday. The number of attendees is limited, so visit the MSA Education table soon, since the demonstrations get filled up quickly!
Here's a list of participating Exhibitors and titles:
Accurion Scientific Instruments "Atomic Force Microscopy meets Optical Microscopy"
Asylum Research "Simultaneous AFM and Fluorescence Imaging with the MFP-3D AFM"
Bitplane, Inc. "4D Particle Tracking"
Delong America "LVEM5: Applications and Operations"
EDAX "Stereo Imaging - how to collect and view images in 3D" "What you can do with an Orientation Imaging Microscopy EBSD System"
Emispec Systems, Inc. "A new software architecture for SPM imaging and spectroscopy; a joint development between Emispec System, Inc. and Molecular Imaging Corporation"
Extec Corp./A4I America "SIMPLICITYIntegrated Image Analysis commencing with Sample Preparation"
FEI Company "Overcoming Hurdles in High-resolution TEM Tomography"
Gatan, Inc. "The Gatan User Tutorial"
HACKER Instruments & Industries Inc. "Advances in Microwave Specimen Preparation for TEM"
Hitachi High Technologies America, Inc. "FE-SEM: SE/BSE Mixing- Pros and Cons"
Horiba, Inc. "Transmission X-Ray Analysis on your Desktop"
Imago Scientific Instruments Corporation "Introducing Imago's LEAPTM Atom Probe Microscope: innovative microscopy for rapid 3-D imaging and analysis at the atomic scale"
Microscopy/Microscopy Education, Inc. "Calibrating Fluorescence"
Molecular Imaging "Simultaneous Sample Topography and BioMolecular Force Recognition Mapping with a Versatile New Single Molecule Force Recognition Tool Kit"
Nanoptek Corporation "Photon Tunneling Microscopy: Sub-nanometer profiling in real time"
Nanotech-America "Breaking the bounds of AFM: New Hybrid Techniques"
Quantum Dot Corp "Lighting Up Cells and Tissues with Qdot Labels"
RMC-Boeckeler Instruments Inc. "Cryofixation & Freeze Substitution - A Complete Solution"
SensIR Technologies "The IlluminatIR - Combining light microscopy with infrared spectral analysis"
Soft Imaging System (SIS) "Laboratory Image Management"
South Bay Technology, Inc. "Ion Beam Sputtering: Applications to Electron Microscopy"
To All Those who are attending, or may be interested in attending, the Annual MSA "Microscopy & Microanalysis" meeting in San Antonio, TX next week (August 3-8), there will be:
A SPECIAL SESSION ON
LEGAL ISSUES
ASSOCIATED WITH MICROSCOPY AND RELATED TECHNOLOGIES
IN A
PUBLIC POLICY FORUM
"PATENT AND INTELLECTUAL PROPERTY ISSUES FOR MICROSCOPISTS"
Henry M. Schaffer Esq., MSA Counsel
A POTPOURRI OF PATENT TOPICS OF INTEREST TO MICROSCOPISTS INCLUDING:
* WHAT CAN AND CANNOT BE PATENTED
* CAN ANYONE USE YOUR PATENT - FOR FREE?
* WHAT ARE THE DIFFERENCES BETWEEN PATENTS AND COPYRIGHTS?
* HOW LONG DOES A PATENT LAST? CAN IT BE RENEWED?
* PATENTS AND STANDARDS - A RISKY MIX
} WEDNESDAY AUGUST 6, ROOM 206 A&B, 2.00 PM, CONVENTION CENTER, SAN ANTONIO
FOR MORE INFORMATION: CONTACT - www.msa.microscopy.com/MMHomePage.html
-- Peter Ingram Chair, MSA Public Policy Committee Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
Hi Everyone, I have a bunch of apertures for a Zeiss TEM (the TEM has been decommissioned). Most are from Ted Pella, Inc., cat #69020-30. I don't know if they are new or not, but they are all the same price (free). I will be in San Antonio for M&M, and if anyone who will also be in attendance wants them, drop me an email and I'll bring them along. Otherwise, they get pitched. Randy
Randy Nessler University of Iowa Central Microscopy Research Facility Phone 319-335-8142 Fax 319-384-4469
} Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (hoganbecky-at-hotmail.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, } July 28, 2003 at 20:07:39 } --------------------------------------------------------------------------- } } Email: hoganbecky-at-hotmail.com } Name: Becky Hogan } } Organization: Summerville Elementary } } Education: K-8 Grade Grammar School } } Location: Summerville, South Carolina } } Question: I will be teaching gifted 5th graders in a new } math/science program this year and need to know what type of } microscope would be best for my program. Also, where can I buy a } microscope for a reasonable price? Do colleges ever donate used } microscopes to schools? I am working with a shoestring budget of } $0.00 so I will have to make a personal purchase in order for my } students to use this wonderful teaching tool. } Thank you for asssisting me with this matter. I want to offer the } best science program possible! } Becky Hogan } } --------------------------------------------------------------------------- Becky -
In general, used college microscopes won't work well for you at the 5th grade level. Please look at the detailed advice on the Project MICRO website (URL below). If you have questions after reading that advice, I'll be happy to answer them directly. Thank YOU for your enthusiasm!
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
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jwirth4-at-juno.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, July 30, 2003 at 10:30:25 ---------------------------------------------------------------------------
Email: jwirth4-at-juno.com Name: Jim Wirth
Organization: Piedmont Community College
Education: Undergraduate College
Location: Charlottesville, VA 22903
Question: Hello, When using an inverted microscope, is it typical to focus on the bottom or top plane of the stage? Is it necessary to drop the specimen to the lower plane?
Is there a typical stage opening and insert dimension across the major manufacturers?
John Hoffpauir has sent me his comments. We believe his comments are quite plausible.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: JHoffpa464-at-aol.com To: sales-at-laddresearch.com Sent: Wednesday, July 30, 2003 1:40 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (asmus-at-centre.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, July 30, 2003 at 13:41:25 ---------------------------------------------------------------------------
Email: asmus-at-centre.edu Name: Steve Asmus
Organization: Centre College
Education: Graduate College
Location: Danville, Kentucky, US
Question: I'm comparing 2 fluorescence microscopes and would like feedback on how these scopes compare or any info on rankings of these scopes. I'm choosing between a Nikon E600 and an Olympus BX51. They have identical capabilites and similar prices. I am also interested in adding confocal capabilies eventually, and both appear to be upgradeable. Any advice from someone familiar with these scopes would be appreciated.
It is a very good and practical question! There are more than one source of the Cu characteristic X-rays your described. The TEM column is not "clean", since there are bremsstrahlung X-rays and uncollimated electrons, as a result of the electrons interacting with column components such as the diaphragms and polepieces. The bremsstrahlung X-rays and uncollimated electrons can strike the Cu grid and your specimen, producing Cu x-rays which are not distinguishable in your EDS spectra.
Some of the transmitted electrons are scattered at sufficiently high angles and strike other TEM column components such as the objective lens polepieces and objective diaphragm if it is not removed. They would also give rise to spurious X-rays. You can also check the probe size by directly imaging the probe, and check if there is any long tail. To get a feel of the background X-rays, you can place the beam through a hole in the sample, and collect a spectrum, which would show some peaks.
I hope this is somewhat helpful to you.
Kind Regards.
Wentao
__________________________________ Do you Yahoo!? The New Yahoo! Search - Faster. Easier. Bingo. http://search.yahoo.com
Dear Fellow Listers, There is quite a good book -Artifacts in Biological Electron Microscopy edit Richard Crang and Karen Klomparens (Plenum Press 1988) (dont know if still in print though) which has pictures of 'pepper' caused by embedding, fixation and staining as well as a host of other oddities. I have a few questions though as I was surprised to see such a question posted, Where do electron microscopists train and get experience these days, many labs seem to have one routine method and they stick to it? Is there a good web site that maybe images (descriptions just don't do it,) such as those found in the above book, could be posted to?
Cheers
Gillian Brown Histology Section, Asthma Biology RI CEDD http://ukdiscovery.gsk.com/histopathology/ ----- Forwarded by Gillian 2 Brown/PharmRD/GSK on 31-Jul-2003 09:44 -----
"Ladd Research" {ladres-at-worldnet.att.net}
30-Jul-2003 20:01 Please respond to "Ladd Research" {sales-at-laddresearch.com}
To: "Microscopy Listserver"
cc: Subject: Re: Contamination in resin sections---TEM
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John Hoffpauir has sent me his comments. We believe his comments are quite plausible.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: JHoffpa464-at-aol.com To: sales-at-laddresearch.com Sent: Wednesday, July 30, 2003 1:40 PM
Jim;
Is this inverted microscope for potted cross-sections? I generally use an inverted microscope for encapsulated cross-sections so that the polished face is perpendicular to the objective lens and the plane of focus would be that face or the top of the stage.
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: jwirth4-at-juno.com [mailto:jwirth4-at-juno.com] Sent: Wednesday, July 30, 2003 2:06 PM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jwirth4-at-juno.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, July 30, 2003 at 10:30:25 ---------------------------------------------------------------------------
Email: jwirth4-at-juno.com Name: Jim Wirth
Organization: Piedmont Community College
Education: Undergraduate College
Location: Charlottesville, VA 22903
Question: Hello, When using an inverted microscope, is it typical to focus on the bottom or top plane of the stage? Is it necessary to drop the specimen to the lower plane?
Is there a typical stage opening and insert dimension across the major manufacturers?
When I use TEM sections in SEM, I see Cu peaks also. I place grids on a carbon mounts, so backscattered electrons from this substrate generate the peaks very similar to ones we see with TEM.
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: Wentao Qin [mailto:wentao_qin-at-yahoo.com] } Sent: Thursday, July 31, 2003 1:08 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Cu peaks in STEM/EDS } } Hi Dr. Pan, } } It is a very good and practical question! There are } more than one source of the Cu characteristic X-rays } your described. The TEM column is not "clean", since } there are bremsstrahlung X-rays and uncollimated } electrons, as a result of the electrons interacting } with column components such as the diaphragms and } polepieces. The bremsstrahlung X-rays and } uncollimated electrons can strike the Cu grid and your } specimen, producing Cu x-rays which are not } distinguishable in your EDS spectra. } } Some of the transmitted electrons are scattered at } sufficiently high angles and strike other TEM column } components such as the objective lens polepieces and } objective diaphragm if it is not removed. They would } also give rise to spurious X-rays. You can also check } the probe size by directly imaging the probe, and } check if there is any long tail. To get a feel of the } background X-rays, you can place the beam through a } hole in the sample, and collect a spectrum, which } would show some peaks. } } I hope this is somewhat helpful to you. } } Kind Regards. } } Wentao } } __________________________________ } Do you Yahoo!? } The New Yahoo! Search - Faster. Easier. Bingo. } http://search.yahoo.com } }
"Biomedical Electron Microscopy" by Maunsbach and Afzelius (Academic Press, 1999) has lots of examples of artifacts. It is an excellent book. The Crang and Klomparens book mentioned below is also excellent. Glutaraldehyde in tissues can react with osmium to form artifacts, as John mentioned earlier. I did not know that it was buffer dependent.
} Dear Fellow Listers, } There is quite a good book -Artifacts in Biological Electron Microscopy } edit Richard Crang and Karen Klomparens (Plenum Press 1988) (dont know if } still in print though) which has pictures of 'pepper' caused by embedding, } fixation and staining as well as a host of other oddities. } I have a few questions though as I was surprised to see such a question } posted, Where do electron microscopists train and get experience these days, many labs seem to have one routine method and they stick to } it? Is there a good web site that maybe images (descriptions just don't } do it,) such as those found in the above book, could be posted to? } } Cheers } } Gillian Brown } Histology Section, Asthma Biology } RI CEDD } http://ukdiscovery.gsk.com/histopathology/ } ----- Forwarded by Gillian 2 Brown/PharmRD/GSK on 31-Jul-2003 09:44 ----- } } } "Ladd Research" {ladres-at-worldnet.att.net} } } 30-Jul-2003 20:01 } Please respond to "Ladd Research" {sales-at-laddresearch.com} } } } } } To: "Microscopy Listserver" } } cc: } Subject: Re: Contamination in resin sections---TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 **********************************************
Many thanks to all of you who responded to my first query on this topic. I was deliberately vague, hoping to get the widest range of ideas. We got some very good thoughts and we are trying all of them, in one form or another.
To be more specific, we are getting pepper in nearly every tissue we look at, but not uniformly everywhere in the tissue. I will email images to anyone requesting them. For example, in retinal tissue it most consistently shows up in the rods. Our standard processing involves 2% gluteraldehyed/2% paraformaldehyde in 0.1M cacodylate buffer, but we often use 0.17M cac buffer, Sorenson's phosphate buffer, and varying concentrations of fixatives, depending upon the project. Immunocytochemical processing is generally done in 4% paraformaldehyde/0.1% glut in cacodylate or phosphate buffers, usually with a small amount of CaCl2.
Here is what we have tried:
1) Redoing our water purification system and trying water from other labs (we're now on our 4th type of water, including purchased distilled water, reverse osmosis feed water into a Millipore Simplicity polishing system; two-tank-in-series deionized water system feeding into the Millipore; and house deionized water run through another lab's wall-mounted Millipore polishing system. No other lab using the latter water supply is reporting any problems; 2) Rewashing ALL of lab glassware; 3) Remixing ALL of our solutions, using fixatives from different lot numbers; 4) Remixing ALL of our resins; 5) Processing with and without secondary and tertiary fixatives (osmium and uranyl acetate)in all combinations; 6) Looking at both stained and unstained sections (Pb and UA); 7) Looking at different tissue types; 8) Embedding in different resins, including Epon/Araldite, Spurr's, pure Epon (actually Embed 812), LR White, and Araldite. 9) Cutting on different diamond knives, with different types of syringe-filtered water in the boats.
So far the cleanest sections have been in LR White, while the same tissue processed for Epon/Araldite showed contamination, but the use of several other resins seems to indicate that resin is probably not the issue. BUT, the LRW material was processed with a much lower percentage of gluteraldehyde in the fixative. But wouldn't the use of different lots of gluteraldehyde argue against this as the culprit?
We have been able to successfully remove or minimize the contamination by post-treating sections with 2% periodic acid, which also chews up our copper grids and adds its own yuck to the process. However, we are now cutting on gold and nickel grids to give us this option until we figure out what's going on here.
A genuine puzzler and a major nuisance. I guarantee that when we get this solved, I will post a detailed cure in hopes of saving anyone else this nightmare.
Thanks much for any further advice!
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! 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/
The "red-gren-red" indicator light for density adjust always seems to remain lit as red on the left side, making me believe that there is a problem with the light meter.
Changes in brightness, and exposure time don't seem to have any effect on the density readings.
Is there anyone else who has had problems with their light meter, and knows any simple remedies to this problem?
The camera does advance, and the beams seems to go on and off making an exposure for the time set by the exposure time, so it can take pictures. It's just that we seem to making the pictures in the dark.
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Dear Jim, Our inverted microscopes are meant to focus on the face of the sample that is face-down on the stage, so we focus on the top surface of the stage. These microscopes are used for large, reflected-light samples, because the design of the stage does not restrict the size of the sample. As for openings, our microscopes come with a variety of openings available. These are discs with an outside diameter of about two inches, that fit into an opening in the stage, and a selection of inside diameters, to accommodate different sized samples. The top of these discs is flat, while the bottom of them is concave, to allow the objectives to swing by. I hope this helps. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {jwirth4-at-juno.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, July 30, 2003 11:06 AM
Garry
The H7000 that I use has another exposure meter display inside the column just behind the screen. this consists of two red LEDs (ie no green middle. If the two come on together that is a correct exposure but increasing or decreasing the beam density on the screen should make one or the other come on. I apologise if you have checked this but if the column meter works then it sounds like it is the other meter. Also have you checked that no one has 'mucked about' with the meter calibration settings.
If everything else checks out then maybe it's just the link from the central screen that has broken - remember this is a current density rather than light meter.
I hope this helps.
Malcolm
Malcolm Haswell e.m. unit Chemispec School of Health, Natural and Social Sciences Fleming Building* University of Sunderland Tyne & Wear SR1 3SD UK tel no: 0191 515 2872 / 3468 e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca}
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Hi Randy Do you have the same problem when you use gold or nickel grids? Some time ago I worked on immunogold technique in tissue and I had problem with pepper/dirt film over the sections. I used nickel grids that were bought ages ago. I bought a new batch, cleaned it and sections were clean, no pepper/dirt film. Dorota
Hi all, I have 2 standard cubes I use (blue: excites at 490, emits at 515, for chlorophyll a, and green: excites at 545 and emits at 590 for phycoerythrin/cryptomonads), plus I have an additional cube for UV that I use for DAPI and calcafluor white. My problem is taxa that are from humic stained lakes. Does anyone have a better Excite/Emission range suggestion for these systems, especially for phycoerythin or cryptomonads? Pigments are measuring high, cells look healthy and are well colored, but fluoresce poorly. I think the pigments have shifted their absorption ranges, rather than the humics interfering (The preps I'm working with are very thin, with no overlying water). This has been troublesome for a while, but lately, I seem to be getting lots of samples from humic systems and I can ID and count much faster if I have good, active fluorescence. I'll post responses to the list if folks are interested. Thanks! Ann.
Ann St. Amand, Ph.D. President PhycoTech, Inc. 620 Broad St., Suite 100 St. Joseph, Michigan 49085 USA
I am having some trouble embedding arabidopsis seedling for TEM. I am using epon-araldite mixture and my tissue just dissolves away upon sectioning of my samples. I am assuming it is an embedding problem. I dehydrate my samples using an ETOH gradient then switch to 100% propylene oxide. I embed with a 2:1 (PO:epon-araldite) mixture for 12 hours then a 1:2 mixture for 12 hours then 100% epon-araldite at room temp followed by 24 hours at 30 degrees then 48 hours at 45 degrees. Should I change something in my protocol or is there another embedding medium that may work better for arabidopsis seedlings? Any help would be appreciated. Thankyou.
Becky- I gave a microscope talk to my daughter's 4th grade class. On a Friday, I gave the teacher 24 plastic petri dishes, one for each student and the teacher as well. Then I had my daughter collect them on Monday. The rule was that - 1) no liquids, and 2) each dish had to be scotch-taped closed, with only the student's name on the label. The kids were free to collect a small specimen of anything animal (not living), vegetable, or mineral. I then made slides of each item, and tried to guess what each was. Some were obvious. I came back with my microscope, and the slides, and made handouts for each child (and the teacher). The 24 page handout had a microphotograph of each slide and the student's name under the relative picture. The hand out also had a diagram of the microscope, and some brief information (4th grade level). I let the students come up one, by one to look through the microscope at this or that more interesting specimen. To save money on the booklet, I made two sets of color prints: One booklet contained all color photos (for the teacher's copy) and I Black and White photocopied the book (in photo-mode) for the other 22 student copies. I used gluestick to glue an original color print of each student's specimen in their respective booklet, and handed them out accordingly. It was great fun, and it worked out perfectly, and didn't take up too much of the teacher's time. Total was about an hour total class time. The students were very creative in their choices, and brought a full spectrum of things microscopic into the class! Rgds, Mike Shaw
Question: I will be teaching gifted 5th graders in a new math/science program this year and need to know what type of microscope would be best for my program. Also, where can I buy a microscope for a reasonable price? Do colleges ever donate used microscopes to schools? I am working with a shoestring budget of $0.00 so I will have to make a personal purchase in order for my students to use this wonderful teaching tool. Thank you for asssisting me with this matter. I want to offer the best science program possible! Becky Hogan } }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sconnell-at-liai.org) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, August 1, 2003 at 15:58:16 ---------------------------------------------------------------------------
Email: sconnell-at-liai.org Name: samuel connell
Organization: La Jolla Institute for Allergy and Immunology
Education: Graduate College
Location: San Diego, CA
Question: Iím heading into confocal microscopy calcium flux land for the first time. Iíve done a fair amount in flow cytometry using Indo-1. Iím interested in speaking with someone who has experience in doing this work with visible wavelength probes, preferably with the BioRad MRC 1024 system. Our system has the Kr/Ar mixed gas with the 488, 568, and 647 laser lines available. Weíre planning on looking at 512x512 XY images over time, potentially with scans every 5-10 seconds over 10-20 minutes or so. If someone would be so kind as to touch base with me, I would be truly indebted for the shared expertise. Thanks, -- Samuel Connell
We've done some work with Ca green. Basically, it gets brighter and dimmer depending on the free Ca. It works with blue excitation. Not particulary quantitative, but it gets the point across.
I know that Gatan makes various vacuum transfer holders for the TEM but are there any other suppliers?
Many thanks,
Ralph Haswell Research Scientist Shell International Chemicals B.V. Shell Research & Technology Centre, P.O. Box 38000, 1030 BN Amsterdam, The Netherlands
I really am wondering why nobody has raised this important question earlier. Yes, RGB color interpolation with Bayer filter cameras can become an problematic factor when it comes to quantitative spatial measurements of coloured samples. Especially when working with low power magnification where camera resolution is crucial.
Regarding Bayer filter cameras, you might want to consider the pixel shifting technology used in the ProgRes C14 digital microscopy camera from JENOPTIK Laser, Optik, Systeme GmbH.
The ProgRes C14 also uses a single CCD with Bayer filter design. The sensor can be moved by means of a piezoelectric shifting device as many other pixel shifting cameras use.
But there are different ways to shift the pixels to produce new information. Most pixel shifting camera use the sensor shifting only to gain optical resolution, i.e. scanning the image more accurately, which makes much sense when using high resolution optics. Thereby the sensor is shifted in fractions of the pixels' spacing, producing a slight overlap of the pixels. But: quantitative spatial measurements might become problematic due to the RGB color interpolation, where some spatial resolution is defintively lost.
In contrast to some other pixel shifting cameras, JENOPTIK's ProgRes C14 also shifts the sensor in full pixel steps, where the pixels overlap 100%. Doing this at least four times in x- and y- direction you sample the full RGB color information for each single pixel position. In the result you get a true color RGB image without interpolation, comparable to a RGB filter wheel camera. The file size (and thus nominal image resolution) stays the same. But precise quantitative spatial measurements of colored samples is now possible. We call this technique "Color-Co-Site-Sampling" and as far as I know only ZEISS's Axicam HRc and JENOPTIK's ProgRes C14 do use this technique in micro-photography.
You can combine this full pixel shifting increasing "color resolution" also with fractional pixel shifting increasing spatial resolution.
An image says more than a thousand words - so if you want some illustration for this technique please contact me.
The benefit of this technique compared to a filter wheel camera is, that you can use the camera also for moving specimens in single shot mode, where colors then are interpolated. And compared to a 3 CCD camera, the Bayer filer camera is more light sensitive, because incoming light does not need to be split up by a prism directing it onto the three single CCDs.
Of course, also in pixel shifting cameras pixel misalignment can occur - in principle. You need to keep in mind that the sensor shifting has do be done very precisely in the sub-µm range. To guarantee optimum alignment, the ProgRes C14 comes with a built-in calibration procedure that allows the user to calibrate the piezo scanner for best results.
I hope this information does add a new facette to your discussion about quantitative spatial measurements of coloured samples with Bayer filter cameras.
} -----Original Message----- } From: Peter Van Osta [mailto:pvosta-at-unionbio-eu.com] } Sent: Thursday, July 24, 2003 10:52 AM } To: MSA } Subject: Bayer filter versus 3CCD camera } ---------. } } } Hi, } } The two main types of color cameras used in microscopy I know } of are the } ones with 3 separate CCDs for each of the three primary colors and the } ones with a Bayer filter design. In the 3CCD camera the } spatial sampling } is the same for each of the three colors, but not in the Bayer filter } camera design ? How do you deal with this difference when doing } quantitative spatial measurements of colored samples in microscopy ? } } What is the relative actual resolution seen with a 3CCD color } camera and } a camera with Bayer filter reconstruction? How to take into } account the } reduced sensitivity of the Bayer filter type for each color, compared } with the 3CCD camera if any ? When are Bayer filter type } cameras a valid } alternative for a 3CCD camera in microscopy, besides the price ? } } What about pixel shifts in a 3CCD camera due to misalignment of the } color filters/splitter and the CCDs ? } } Best regards, } } Peter Van Osta } } Union Biometrica N.V./S.A. } European Scientific Operations (ESO) } Cipalstraat 3 } B-2440 Geel } Belgium } Tel.: +32 (0)14 570 619 } Fax.: +32 (0)14 570 621 } } http://www.unionbio.com/ } } http://ourworld.compuserve.com/homepages/pvosta/cvwww.htm }
Hi, My department is currently considering to buy a low voltage ion mill. The users I've spoken to up to date all agree concerning the good quality of the samples. However, the maintenance of many low energy ion
mills seems be a major problem, especially in the case of a large number
of users. We are also concerned about technical support from suppliers located overseas. Does anybody have experience with suppliers like Fischione and South Bay
Technology with respect to maintenance and technical support in Europe? Certainly, I am also interested in comments on the effectiveness of low energy ion mills in general.
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ON CONCLUSION OF THIS TRANSACTION,YOU WILL BE ENTITLED TO 25% OF THE TOTAL SUM AS GRATIFICATION.5% OF THE TOTAL SUM WILL BE USED TO REINBURSE EXPENSES THAT MIGHT ARISE FROM TELEPHONE BILLS AND OTHER EXPENSES DURING THE TRANSACTION,WHILE 70% WILL BE FOR ME AND MYPARTNERS HERE. PLEASE YOU HAVE BEEN ADVICED TO KEEP TOP SECRET AS WE ARE STILL IN SERVICE AND INTEND TO RETIRE
FROM SERVICE AFTER WE CONCLUDE THIS DEAL WITH YOU. I WILL BE MONITORING THE WHOLE SITUATION HERE IN THIS BANK UNTIL YOU CONFIRM THE MONEY IN YOUR ACCOUNT.WE THEN COME DOWN TO YOUR COUNTRY FOR SUBSEQUENT SHARING OF THE FUND ACCORDING TO THE PERCENTAGES PREVIOUSLY INDICATED AND FOR INVESTMENT IN ANY COUNTRY YOU MAY ADVICE US TOO. ALL OTHER NECESSARY INFORMATION WILL BE SENT TO YOU WHEN I HEAR FROM YOU. I SUGGEST YOU GET BACK TO ME AS SOON AS POSSIBLE,STATING YOUR WISH IN THIS DEAL.
YOURS FAITHFULLY, MR JACKSON BENSON. 00228-9118757
________________________________________________ Get your own "800" number Voicemail, fax, email, and a lot more http://www.ureach.com/reg/tag
Nestor Is there anything that can be done about the volume of SPAM this board sends to my inbox? I don't really want to, as this is a useful resource, but I am considering unsubscribing because of it. Cheers Ady
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I have started using a free spam filter called "Spam Assasin" or SAproxy. It was rated best by Consumer Reports. It is available from Blomba.com. For a free thing it is doing a fairly good job. I would guess that my spam has been cut by 70%. (now I feel alone and unwanted)
Greg
At 11:36 AM 8/6/2003 -0700, ady jenkinson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295
With all due respect and thanks to the manager of this extremely valuable list server, would it be possible to put a filter on the e-mail sent out? It should be possible for the list server to except mail ONLY from registered users. This list has become my number one source of spam (I am not interested in all of these people who wish to give me lots of money).
Thank you
David ____________________
David Elliott Ph.D.
Yale University School of Medicine Campus Address: TAC S160 333 Cedar Street PO Box 208022 New Haven, CT 06520-8022
Hmmm. I get maybe one spam a day from the list. Nothing compared to what is out there.
geoff
ady jenkinson wrote:
} Nestor } Is there anything that can be done about the volume of } SPAM this board sends to my inbox? } I don't really want to, as this is a useful resource, } but I am considering unsubscribing because of it. } Cheers } Ady } } __________________________________ } Do you Yahoo!? } Yahoo! SiteBuilder - Free, easy-to-use web site design software } http://sitebuilder.yahoo.com } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
The Listserver has a spam filter on it, and while admittedly a few messages get through it is a very small fraction of the "attempts".
Can you please check and insure that the spam is really coming from the Listserver and is not being "faked". This is becoming a common tactic among spammers. They find a real address and subsitute it for the address in the FROM field to make it appear that mail is coming from a legit address.
I saw 2 simultaneous spams that got through the filter yesterday (and they are now blocked) but nothing else for the week.
Just for the record in the last 48 hours there were 162 spam messages blocked by the filter. This does not count the 2 that got through.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hmmm. I get maybe one spam a day from the list. Nothing compared to what } is out there. } } geoff } } ady jenkinson wrote: } } } Nestor } } Is there anything that can be done about the volume of } } SPAM this board sends to my inbox? } } I don't really want to, as this is a useful resource, } } but I am considering unsubscribing because of it. } } Cheers } } Ady } } } } __________________________________ } } Do you Yahoo!? } } Yahoo! SiteBuilder - Free, easy-to-use web site design software } } http://sitebuilder.yahoo.com } } } } } } } }
I wonder if anyone knows where we could find highly crystalline fine graphite powder with a platelet size in the range of 50-100nm (smaller is still OK). I have tried to grind an old AFM HOPG crystal and lost all my optimism after 2 hours...
You've been doing a great job on keeping the filter "tuned", keep up the good work.
Fellow Listers:
To complain about one or two spam messages here and there seems unrealistic. As a system administrator, I can say that the volume of spam, and the various tactics (and sophistication of them) that are used by the spammers, is constantly increasing.
The miniscule volume of spam on this list, and high signal to noise ratio, is a testament to Nestor's vigilance against this plague.
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
Colleagues
The Listserver has a spam filter on it, and while admittedly a few messages get through it is a very small fraction of the "attempts".
Can you please check and insure that the spam is really coming from the Listserver and is not being "faked". This is becoming a common tactic among spammers. They find a real address and subsitute it for the address in the FROM field to make it appear that mail is coming from a legit address.
I saw 2 simultaneous spams that got through the filter yesterday (and they are now blocked) but nothing else for the week.
Just for the record in the last 48 hours there were 162 spam messages blocked by the filter. This does not count the 2 that got through.
Dear Sir, I am MALIK ALLI from Iraq.I decided to write you due my situation now.There is an information I would like you to keep very confidential.
There is sum amount of money my boss deposited in Europe, out side our country,and followiong the battle at Basra by the American Millitary forces,he was killed .It is only me that knows these because I'm his second,I will not be {BR} able to give you the full details that led to that urgly incidents during the War. Presently I am staying in another country,just to save my life,with my three children. The money in question, is Twentyfive million five hundred thousand united state dollars (40.500,000.00Milllion) U.S.Dollars. What I would want you to do, is to assits me to recouver the money as it can only be identified only by calling the pin no to the company concerned, There is no risk in this transaction. I will use the money for an investiment in your country for the future of my children. If you are intrested, and can maintain the very confidentiality of this transaction,you e-mail me on email;bashiralli-at-123.com immediately for more clearification,and also note that I am a refugees now since the americans've taken over our country and as a soldier,I'll not come back now becuase of the the situation in Iraq.I can speak little English,so dont mind my English. Thank you very much.
MALIK ALLI
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
Dear Sir, I am MALIK ALLI from Iraq.I decided to write you due my situation now.There is an information I would like you to keep very confidential.
There is sum amount of money my boss deposited in Europe, out side our country,and followiong the battle at Basra by the American Millitary forces,he was killed .It is only me that knows these because I'm his second,I will not be {BR} able to give you the full details that led to that urgly incidents during the War. Presently I am staying in another country,just to save my life,with my three children. The money in question, is Twentyfive million five hundred thousand united state dollars (40.500,000.00Milllion) U.S.Dollars. What I would want you to do, is to assits me to recouver the money as it can only be identified only by calling the pin no to the company concerned, There is no risk in this transaction. I will use the money for an investiment in your country for the future of my children. If you are intrested, and can maintain the very confidentiality of this transaction,you e-mail me on email;bashiralli-at-123.com immediately for more clearification,and also note that I am a refugees now since the americans've taken over our country and as a soldier,I'll not come back now becuase of the the situation in Iraq.I can speak little English,so dont mind my English. Thank you very much.
MALIK ALLI
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
Dear Sir, I am MALIK ALLI from Iraq.I decided to write you due my situation now.There is an information I would like you to keep very confidential.
There is sum amount of money my boss deposited in Europe, out side our country,and followiong the battle at Basra by the American Millitary forces,he was killed .It is only me that knows these because I'm his second,I will not be {BR} able to give you the full details that led to that urgly incidents during the War. Presently I am staying in another country,just to save my life,with my three children. The money in question, is Twentyfive million five hundred thousand united state dollars (40.500,000.00Milllion) U.S.Dollars. What I would want you to do, is to assits me to recouver the money as it can only be identified only by calling the pin no to the company concerned, There is no risk in this transaction. I will use the money for an investiment in your country for the future of my children. If you are intrested, and can maintain the very confidentiality of this transaction,you e-mail me on email;bashiralli-at-123.com immediately for more clearification,and also note that I am a refugees now since the americans've taken over our country and as a soldier,I'll not come back now becuase of the the situation in Iraq.I can speak little English,so dont mind my English. Thank you very much.
MALIK ALLI
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
Dear Sir, I am MALIK ALLI from Iraq.I decided to write you due my situation now.There is an information I would like you to keep very confidential.
There is sum amount of money my boss deposited in Europe, out side our country,and followiong the battle at Basra by the American Millitary forces,he was killed .It is only me that knows these because I'm his second,I will not be {BR} able to give you the full details that led to that urgly incidents during the War. Presently I am staying in another country,just to save my life,with my three children. The money in question, is Twentyfive million five hundred thousand united state dollars (40.500,000.00Milllion) U.S.Dollars. What I would want you to do, is to assits me to recouver the money as it can only be identified only by calling the pin no to the company concerned, There is no risk in this transaction. I will use the money for an investiment in your country for the future of my children. If you are intrested, and can maintain the very confidentiality of this transaction,you e-mail me on email;bashiralli-at-123.com immediately for more clearification,and also note that I am a refugees now since the americans've taken over our country and as a soldier,I'll not come back now becuase of the the situation in Iraq.I can speak little English,so dont mind my English. Thank you very much.
MALIK ALLI
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
I was a little surprised by the recent requests for spam filtering, and the threats to unsubscribe. Then I realized that they must be newer members. Those of us that have been around for a while, know the excellent job you do at keeping unwanted traffic to a minimum.
Here at work, the spam filtering had gotten pretty good, but the spammers are always working hard at getting in. Of late, there has been an increase in the spam, but I know the filtering will be fixed to reduce it again.
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Just for example, I just received FOUR spam messages that contain the Listserver header (visible above these lines), but the "from" and "to" addresses were both "malikalli-at-123.com". This is a clear indication that they did NOT come from the Listserver.
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Could anybody suggest references I could use to learn more about analyzing adhesives? Nobody in our group has a lot of experience/knowledge about adhesives and that is something I've been asked to rectify. In particular I'm talking about the adhesives that would be used to adhere a label onto a plastic material. When there are problems with the adhesion (for example bubbles under the label) we want to be able to determine if the adhesive has failed, if it isn't uniformly distributed, or if we should be looking to the package for answers. Ideally we would like to build some in-house expertise in the evaluation of adhesives, the distinction between different ones, and develop a feel for what adhesives are ideal for which materials. So, if there are any books, journal articles, courses, etc. that anyone thinks would be beneficial I would appreciate any information you can send my way.
Does anyone have experience using Nanoplast (FB101) Water Soluble Melamine Resin (from Agar Scientific)? I would really appreciate any hints on how to get rid of bubbles generated during drying/polymerisation and how to stop the tissue floating up towards the top of the embedding mold.
Does anyone know the refractive index of the polymerised resin?
Thanks Hilary
-- Hilary Holloway Technical Manager Biomedical Imaging and Research Unit Division of Anatomy with Radiology Faculty of Medical & Health Science University of Auckland, Post Bag 92109 Auckland, New Zealand
*Subject: Re: Administrivia: Can you do anything about SPAM Nestor? *To: Microscopy-at-sparc5.microscopy.com *From: "Darrell Miles" {milesd-at-US.ibm.com} *Date sent: Thu, 7 Aug 2003 12:38:53 -0400
*------------------------------------------------------------------------ *The Microscopy ListServer -- Sponsor: The Microscopy Society of America
That's right. I have received them too.
Regards,
Witold Zielinski
* * *Just for example, I just received FOUR spam messages *that contain the Listserver header (visible above these *lines), but the "from" and "to" addresses were both *"malikalli-at-123.com". This is a clear indication that they *did NOT come from the Listserver. * *Darrell * *(me, and me alone) * * * * :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) ;)
Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
I'm getting more messages about spam than actual spam now.
Let's congratulate Nestor once more for excellent de-spamming and leave the spam thread alone now.
Regards
Pete -- Peter Bond Scientific Officer Plymouth Electron Microscopy Centre University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel: 44 (0)1752 233092 pbond-at-plymouth.ac.uk
The workshop will consist of four consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of microscopy. The course instructors include Jan Hinsch of Leica, Inc., Dennis O'Leary of Micro-Optical Methods, Mary McCann of McCann Imaging, John Reffner of SensIR Technologies, Inc. and N.Y.M.S. Instructor Don O'Leary.
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Here are a few observations on Nanoplast (FB101) resin that we made here a few years ago:
1. First of all, the amount of catalyst, B-52, called for in the tech sheet that came with the kit gave blocks that were way too hard to cut in our experience, like glass. The instructions were as follows:
Soft block: 10.0g MME 7002(the resin), 0.15 g B-52 (catalyst) Medium: 10.0g 0.20 g hard: 10.0g 0.25 g
So we greatly reduced the amount to 25% of the amounts called for and then got blocks that were easy to cut.
2. As for bubbles during polymerization, not sure about that, don't recall having that problem. Are you sure they were not formed during too vigorous mixing of resin and catalyst? Also, Nanoplast instructions warn not to use silicone moulds: "Silicone moulds are not recommended because air bubbles form during the period of drying". As the resin is 30% water, there is a drying period of 2 days at 40C, then boost temperature to 60C for final plymerization. I don't understand why silicone moulds should cause resin to bubble, but check your moulds and try poly-whatever plastic ones, like BEEM capsules.
3. Tech sheet gives refractive index of 1.47 for the "solution", but not any data for cured blocks.
4. For more info, check this reference for details about using Nanoplast, especially for the drying steps during embedding prior to heat curing:
Weyda, Frantisek. Rapid Communication: Notes on the use of Nanoplast FB 101 for transmission electron microscopy. 1990. Journal of Electron Microscxopy Technique 16:356-357.
The above paper also discusses use of Nanoplast for immunolabeling.
I don't use this resin anymore, it was for special application that a client was following, but may be worth a second look - one of these days.
I'm also sending you a short enclosure off-line with some other tech tips we compiled here on Nanoplast, its a summary of some discussion on this List and off-line that took place in 1998.
Hope this helps!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Bera
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers } } Does anyone have experience using Nanoplast (FB101) Water Soluble } Melamine Resin (from Agar Scientific)? } I would really appreciate any hints on how to get rid of bubbles } generated during drying/polymerisation and how to stop the tissue } floating up towards the top of the embedding mold. } } Does anyone know the refractive index of the polymerised resin? } } Thanks } Hilary } } -- } Hilary Holloway } Technical Manager } Biomedical Imaging and Research Unit } Division of Anatomy with Radiology } Faculty of Medical & Health Science } University of Auckland, Post Bag 92109 } Auckland, New Zealand } } Tel : +64 9 373 7599 Ext.: 86761 } Fax: +64 9 373 7484
At 12:36 PM 08/08/2003 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Sunday, August 10, 2003 at 14:24:08 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin Andreev
Organization: 54 "st.Ivan Rilski"
Education: 6-8th Grade Middle School
Location: Sofia Bulgaria
Question: What solvent should I use to disolve the paraffin in the embedding of the speciment?
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by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id TAA02081 for dist-Microscopy; Sun, 10 Aug 2003 19:00:06 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id SAA02078 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Sun, 10 Aug 2003 18:59:35 -0500 (CDT) Received: from col-msxproto1.col.missouri.edu (col-msxproto1.col.missouri.edu [128.206.3.151]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id SAA02071 for {Microscopy-at-msa.microscopy.com} ; Sun, 10 Aug 2003 18:59:23 -0500 (CDT) Received: from phillips-dell.missouri.edu ([128.206.80.38]) by col-msxproto1.col.missouri.edu over TLS secured channel with Microsoft SMTPSVC(5.0.2195.6673); Sun, 10 Aug 2003 18:58:15 -0500 Message-Id: {5.1.1.5.2.20030810185425.029ad350-at-pop.missouri.edu} X-Sender: phillipst-at-pop.missouri.edu X-Mailer: QUALCOMM Windows Eudora Version 5.1.1
Our JEOL 1200 has started giving us lots of trouble with holding the correct stigmation. We set it and everything is fine and 15 minutes later, it is way out of stig. The factory service guys found nothing wrong - naturally the intermitment problem refused to materialize when the service rep was here. We have been doing mostly immuno EM lately so we are using nickel grids - is there any chance this could be having an effect? Your thoughts on this maddening problem would be appreciated. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
We are relative novices on the SEM and are seeking advice on ways to minimize our charging problems. We would appreciate any tips from the mavens out there.
We are looking at biological specimens (tissue explants) coated either with platinum or carbon (details below). The carbon coating protocol is for when we are trying to image at colloidal gold labeling on the surface using the BSE detector. The charging is much worse on the carbon coated specimens compared to the platinum coated ones. I don't have a feel for how much of this problem is "normal" and one has to live with it like so much else in EM! But if anyone out there has some ideas or tricks to reduce charging, we are willing to try it. I would also be interested in anyone's nomination for a great reference book on SEM techniques. Thanks. Tom
Platinum coating protocol:
Tissue (2x2x2 mm) fixed in parformaldehyde/glutaraldehyde (or 2% PF) for several hours. Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. Tissue dehydrated in EtOH series. Tissue critical point dried. Tissue mounted on stub with conductive tape. Next, silver painted the edges of the specimen and allowed to dry. Sputter coated the specimen with platinum at 10 mA for 70 sec. Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications ranging from 1-20k and sometimes up to 50k. Specimens stored in a desiccation chamber. *Re-coating for another minute fixes the problem some of the time.
Carbon coating protocol:
Tissue (from 1x1x1 to 3x3x3 mm) fixed in 2% PF for several hours. Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. Tissue dehydrated in EtOH series. Tissue critical point dried. Tissue mounted on stub with conductive tape. Next, silver painted the edges of the specimen and allowed to dry. Sputter coated the specimen with high purity carbon for 3 sec (~ 20 nm coating?). Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications ranging from 1-20k using both BSE and SE to image. Specimens stored in a desiccation chamber.
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
With silver paint being used, the sample should be well earthed to the support. Presumably, the support is well earthed/clamped to the sample stage in the microscope? If so, then the sample must be inherently the problem - Is the sample undercut or spongey? We have sometimes re-coated from three or four directions by laying the sample on its side or tilted at 45 degrees or thereabouts. This can help.
Keith Ryan Marine biological Association of the UK & University of Plymouth, UK _____________________________
} from: Tom Phillips {phillipst-at-missouri.edu} } date: Mon, 11 Aug 2003 01:17:56 } to: Microscopy-at-sparc5.microscopy.com } subject: Re: SEM - charging problems with bio samples } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are relative novices on the SEM and are seeking advice on ways to } minimize our charging problems. We would appreciate any tips from the } mavens out there. } } We are looking at biological specimens (tissue explants) coated either with } platinum or carbon (details below). The carbon coating protocol is for } when we are trying to image at colloidal gold labeling on the surface using } the BSE detector. The charging is much worse on the carbon coated } specimens compared to the platinum coated ones. I don't have a feel for } how much of this problem is "normal" and one has to live with it like so } much else in EM! But if anyone out there has some ideas or tricks to } reduce charging, we are willing to try it. I would also be interested in } anyone's nomination for a great reference book on SEM techniques. Thanks. Tom } } Platinum coating protocol: } } Tissue (2x2x2 mm) fixed in parformaldehyde/glutaraldehyde (or 2% PF) for } several hours. } Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. } Tissue dehydrated in EtOH series. } Tissue critical point dried. } Tissue mounted on stub with conductive tape. } Next, silver painted the edges of the specimen and allowed to dry. } Sputter coated the specimen with platinum at 10 mA for 70 sec. } Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications ranging from } 1-20k and sometimes up to 50k. } Specimens stored in a desiccation chamber. } *Re-coating for another minute fixes the problem some of the time. } } Carbon coating protocol: } } Tissue (from 1x1x1 to 3x3x3 mm) fixed in 2% PF for several hours. } Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. } Tissue dehydrated in EtOH series. } Tissue critical point dried. } Tissue mounted on stub with conductive tape. } Next, silver painted the edges of the specimen and allowed to dry. } Sputter coated the specimen with high purity carbon for 3 sec (~ 20 nm } coating?). } Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications ranging from } 1-20k using both BSE and SE to image. } Specimens stored in a desiccation chamber. } } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } }
Nickel is one of the few metals other than iron that is ferromagnetic. Try running the same imaging protocol without nickel grids to verify that this is your problem.
Another thought is that you can check the grids for residual magnetism by suspending a piece of metal - such as a pin or paper clip - on a thread and see if the grids "stick" to the pin. If they are deemed to be magnetic, they may be salvaged by running through a degausser.
Stu Smalinskas Metallurgist SKF NATC 46815 Port Street Plymouth, Michigan 48170-6060 (734) 414-6862
Our JEOL 1200 has started giving us lots of trouble with holding the correct stigmation. We set it and everything is fine and 15 minutes later, it is way out of stig. The factory service guys found nothing wrong - naturally the intermitment problem refused to materialize when the service rep was here. We have been doing mostly immuno EM lately so we are using nickel grids - is there any chance this could be having an effect? Your thoughts on this maddening problem would be appreciated. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 11, 2003 at 01:42:01 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin Andreev
Organization: 54 "st.Ivan Rilski"
Education: 6-8th Grade Middle School
Location: Sofia Bulgaria
Question: Can I use acetone for disolving paraffin in paraffin embedding.What else should I use exept xylene/toluene.Can I use benzine.Or some other domestic solvent or some which is easy to find at the stores?
There was another massive spam attack this weekend. I have updated the filters, but I'm leerie of the fact that this may catch additional valid postings.
If you receive a "rejected mail messages" please remember to follow the directions you receive so that I can resolve the issues and get you back into the system.
I got a ton of replies on my problems with stigmation drift in our JEOL TEM and the consensus seems to be that my Nickel grids are causing the trouble. I have to use Ni grids since copper causes problems in the immunocytochemical post embedding staining protocols that I must use. So the big question is how to demagnetize the grids. I am thinking about buying a small unit used to demagnitize recording heads on tape recorders (e.g. the Han D Mag by RB Annis http://www.usrecordingmedia.com/handmagdebyr.html). Has anybody done this type of thing? Thanks again, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Organic solvents and solid compounds differ in a property called "polarity". Those with similar polarity tend to dissolve in each other, while those with large differences do not. Since paraffin has a very low polarity, the most effective solvents for it are ones with low polarity. Examples of low polarity solvents are petroleum distillates (like the benzine that you mention) or materials such as naphtha, ligroin, white spirit and mineral spirits. For something that is readily available and evaporates easily you could try the liquid used as fuel for cigarette lighters. Acetone has a relatively high polarity and therefore is not very effective in dissolving paraffin.
The liquids sold to remove spots from clothing might work but these can contain chlorinated compounds that are more dangerous to health and they may leave residues that do not evaporate easily.
You should be aware that different countries and languages may use the terms "naphtha" and "benzine" to designate slightly different materials refined from petroleum. But all should be effective for dissolving paraffin. Be careful, however, to note the difference between "benzine" and "benzene". Benzene is a specific chemical compound with very serious health hazards - more serious than those of benzine.
You should also avoid xylene and toluene because they have greater health risks than the materials mentioned above which work just as well. Regardless of the material used, use ventilation and avoid breathing the vapor to protect your health and those around you.
John
by way of Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 11, 2003 at 01:42:01 } --------------------------------------------------------------------------- } } Email: sswaffe-at-abv.bg } Name: Veselin Andreev } } Organization: 54 "st.Ivan Rilski" } } Education: 6-8th Grade Middle School } } Location: Sofia Bulgaria } } Question: Can I use acetone for disolving paraffin in paraffin embedding.What else should I use exept xylene/toluene.Can I use benzine.Or some other domestic solvent or some which is easy to find at the stores? } } ---------------------------------------------------------------------------
Gold is the standard that most people use for sputter coating so you must be aware that platinum takes twice as long to put down. You do not say what type of sputter coater you use (important as there are about five different styles of coater needing different techniques) but if it is a coater graduated in kV I would say that 70 seconds at 10mA was much too short and with too low a current. I would expect 20mA for 1.5 to 3 minutes absolute minimum with 5 cms target to specimen distance. Secondly if your materials have complex/rough structures you will need to coat at at least two different angles. Coat for say 2 minutes with 45 degs tilt in one direction and then 2 minutes with 4 degs tilt in the other.
Carbon coating is much less efficient than sputtering, so unless you spin or tilt the specimen during coating one would expect the results to be far worse.
You are using a very low kV and an extremely low emission current (unless you have a FEG which I guess you do?) which should be ideal, so I would also ask which Hitachi machine you are using? If you have a double detector system use the lower detector as this relies less on SE, reducing the level of visible charge. Upper detectors are almost pure SE detectors so are prone to showing charge.
What type of conducting tape do you use the only god tapes are double sided carbon, any other style and in my experience you are asking for trouble.
Please come back for more info if the above does not move you out of trouble.
Regards
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "Tom Phillips" {phillipst-at-missouri.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, August 11, 2003 1:17 AM
Is there any chance that you have a lost nickel grid stuck inside the microscope? The grids are magnetic, but typically don't change the stigmation by much. However, you should see if the loss of astigmatism (I am assuming Objective lens) changes only when you move the sample. This will be more of a problem at higher mags. You should also put a sample on a copper grid and see if you have the same problem. Try correcting the image and not moving the sample for a long time to see if it changes. Do this with the two types of grids.
If it is related to the motion of the sample on the Ni grid, then it is grid related. You should use one set of stig correctors when non-Ni grids are used and the other set for when the Ni ones are used. You can use the computer readout values of the non-Ni stig correctors to set the ones used for the Ni grids.
In general with magnetic samples, the key is to correct the stigmation after every time you move the sample and to minimize the amount of magnetic material that you have (the case with a Ni grid). You should also take hysteresis of the lenses into account when changing them. Always come in from the same direction when you change a lens or alignment setting. For example, overfocus and come down in objective lens strength for focusing or go through the crossover for the condenser and condense the beam from the over strength value.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Sunday, August 10, 2003 7:58 PM To: Microscopy-at-sparc5.microscopy.com
Our JEOL 1200 has started giving us lots of trouble with holding the correct stigmation. We set it and everything is fine and 15 minutes later, it is way out of stig. The factory service guys found nothing wrong - naturally the intermitment problem refused to materialize when the service rep was here. We have been doing mostly immuno EM lately so we are using nickel grids - is there any chance this could be having an effect? Your thoughts on this maddening problem would be appreciated. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
A degausser won't work to solve the problem. Once they are put in the strong magnetic field of the objective lens, they are magnetized again. By being aware of the magnetic hysteresis of the material and the lens, you can minimize the effect of the ferromagnetism of the grid by always having it at the same value by magnetizing it the same way.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Kestutis Smalinskas [mailto:smalinskas-at-yahoo.com] Sent: Monday, August 11, 2003 8:44 AM To: microscopy-at-sparc5.microscopy.com
Nickel is one of the few metals other than iron that is ferromagnetic. Try running the same imaging protocol without nickel grids to verify that this is your problem.
Another thought is that you can check the grids for residual magnetism by suspending a piece of metal - such as a pin or paper clip - on a thread and see if the grids "stick" to the pin. If they are deemed to be magnetic, they may be salvaged by running through a degausser.
Stu Smalinskas Metallurgist SKF NATC 46815 Port Street Plymouth, Michigan 48170-6060 (734) 414-6862
Our JEOL 1200 has started giving us lots of trouble with holding the correct stigmation. We set it and everything is fine and 15 minutes later, it is way out of stig. The factory service guys found nothing wrong - naturally the intermitment problem refused to materialize when the service rep was here. We have been doing mostly immuno EM lately so we are using nickel grids - is there any chance this could be having an effect? Your thoughts on this maddening problem would be appreciated. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
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i apologize if this too basic and for the double post some of you will receive.
can you fix gfp fusion proteins and obtain a fluorescent emission? i would think the answer is no, as most, if not all fixation procedures affect the conformational state/shape of a protein. and i think gfp needs its shape to be excited/emit.
if the answer is yes, can someone direct me to a reference?
thank you all in advance.
best, gary
Gary S. Laevsky, Ph.D. Research Associate The Scripps Research Institute 10550 N. Torrey Pines Road/IMM-24 La Jolla, CA 92037 (858) 784-9372
Apparently, my second reply on this topic did not go through.
A demagnetizer will work outside the microscope, but once inside the strong magnetic field of the objective lens, the sample will be saturated again. Incidentally, one of the nice things about magnetic samples at a university is that students learn to correct astigmatism because they have to do it all the time. I worked with Ni emitters in graduate school. You just have to assume that every time that you take an image, you must stigmate it first.
If you want a demagnetizer for the lab, you don't have to spend a lot of money on a unit. All they are is a solenoid that you slowly insert the piece and slowly withdraw it along the axis. What you are doing is putting it in a strong field and then slowly decreasing that field as you slowly pull it out. You can use a soldering gun for this that has a "loop" the heats the tip. Craftsman makes them and so does Wen. You can also find solenoid coils that will work. Even a transformer can be used. I've used a Sears Craftsman soldering gun to demagnetize tweezers effectively for years. The one that I have has a low heat and a high heat that changes the current in the loop.
There are also other grid materials. Have you looked at gold, Moly, Ta, carbon, diamond, stainless steel, aluminum, or Be grids? Look in all of the EM supply companies'' catalogs because I don't think that any one has all of these. All of the above are non-magnetic materials. (The stainless steel should be austenitic stainless steel, 300 series alloys.)
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Monday, August 11, 2003 12:06 PM To: Microscopy-at-sparc5.microscopy.com
I got a ton of replies on my problems with stigmation drift in our JEOL TEM and the consensus seems to be that my Nickel grids are causing the trouble. I have to use Ni grids since copper causes problems in the immunocytochemical post embedding staining protocols that I must use. So the big question is how to demagnetize the grids. I am thinking about buying a small unit used to demagnitize recording heads on tape recorders (e.g. the Han D Mag by RB Annis http://www.usrecordingmedia.com/handmagdebyr.html). Has anybody done this type of thing? Thanks again, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Tom Personally, I never had stigmation problems with Ni grids on my JEM1200EX. I would more suspect sample itself. It's easy to check: just see astigmatism without any grids in the grid holder. When I have problems with astigmatism, I always start from the empty holder to be sure, it's not fingerprints of my lovely customers. If, it's not a case, then I'll check objective aperture and see does astigmatism changed when you change the aperture hole size. The worse scenario if something from the sample dropped into the column and stick to the pole-piece... As for your grids, I would suggest to use gold plated copper grids specifically designed for immuno EM. It's about $25/100. Major EM suppliers do have it. Good luck, Sergey.
At 09:06 AM 8/11/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
On Sunday, August 10, 2003, at 04:58 PM, Tom Phillips wrote:
} We have been doing mostly immuno EM lately so we are using nickel } grids - is there any chance this could be having an effect?
Dear List, I'd also like to know what the effect of using Ni (or other magnetic) grids is; that is, are they OK for lower mag/lower res, but not for higher mag/res? TIA. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
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Hilary Holloway wrote: ============================================================== Does anyone have experience using Nanoplast (FB101) Water Soluble Melamine Resin (from Agar Scientific)? I would really appreciate any hints on how to get rid of bubbles generated during drying/polymerisation and how to stop the tissue floating up towards the top of the embedding mold.
Does anyone know the refractive index of the polymerised resin? =============================================================== Refractive index and extensive other information about Nanoplast FB 101 can be found on the SPI Supplies website page http://www.2spi.com/catalog/chem/nanopl.shtml
The bubble problem is discussed and is usually related to the use of silicone embedding molds, which is not recommended.
The use of Nanoplast is not what I would call "extensive" but there is a definite number of researchers who find that they can obtain results with this resin system that are not obtainable with any other resin system. We maintain a list of publications on URL http://www.2spi.com/catalog/chem/fb101.html If you have suggestions for publications we have missed, I would welcome any suggestions for publication additions.
Disclaimer: SPI Supplies has been a long time worldwide distributor of the Nanoplast FB 101 product and would naturally like to see more people using it.......
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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Thomas E. Phillips wrote: ============================================================================ ======== We are relative novices on the SEM and are seeking advice on ways to minimize our charging problems. We would appreciate any tips from the mavens out there.
We are looking at biological specimens (tissue explants) coated either with platinum or carbon (details below). The carbon coating protocol is for when we are trying to image at colloidal gold labeling on the surface using the BSE detector. The charging is much worse on the carbon coated specimens compared to the platinum coated ones. I don't have a feel for how much of this problem is "normal" and one has to live with it like so much else in EM! But if anyone out there has some ideas or tricks to reduce charging, we are willing to try it. {snip} ============================================================================ ====== For cell surface tagging, a relatively new option is the use of osmium metal coating, as demonstrated on URL http://www.2spi.com/catalog/osmium-plasma-coater-demonstration.html
I know this defies most conventional wisdom, but the secret is that the coating is **so** thin (e.g. 2 nm), that it results in the needed conductivity but without disrupting the BSE signal from the colloidal gold probes. And one does not give up the topographical information, as they would with carbon coating. The instrument that puts on this kind of coating is described on URL http://www.2spi.com/catalog/osmi-coat.html
Disclaimer: SPI Supplies distributes the osmium plasma coater so we would have a vested in seeing more people using the technique and purchasing this kind of equipment.
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id HAA19432 for dist-Microscopy; Tue, 12 Aug 2003 07:30:09 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id HAA19429 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Tue, 12 Aug 2003 07:29:38 -0500 (CDT) Received: from gw-nl1.akzonobel.com ([193.173.44.2]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id HAA19421 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 12 Aug 2003 07:29:25 -0500 (CDT) Received: from vscan02.xah.intra (vscan02.xah.intra [10.2.224.141]) by mailgate01.xah.intra (Postfix) with ESMTP id 0D1C1549BE for {Microscopy-at-sparc5.microscopy.com} ; Tue, 12 Aug 2003 14:09:23 +0200 (MET DST) Received: from vscan02.xah.intra (localhost [127.0.0.1]) by localhost.xah.intra (Postfix) with ESMTP id B9EA717620 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 12 Aug 2003 14:09:22 +0200 (MEST) Received: from ahmnaw.d10.intra (ahmnaw.ahm.intra [10.2.0.206]) by vscan02.xah.intra (Postfix) with ESMTP id 83CBD17616 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 12 Aug 2003 14:09:22 +0200 (MEST) Received: from CHMNAE.d20.intra ([10.12.0.62]) by ahmnaw.d10.intra with Microsoft SMTPSVC(5.0.2195.5329); Tue, 12 Aug 2003 14:09:22 +0200 Received: from ASDN24.d20.intra ([10.6.17.24]) by CHMNAE.d20.intra with Microsoft SMTPSVC(5.0.2195.5329); Tue, 12 Aug 2003 14:09:20 +0200 content-class: urn:content-classes:message MIME-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Disposition-Notification-To: "Klijn, F. (Frits)" {Frits.Klijn-at-AkzonobelCatalysts.com} X-MimeOLE: Produced By Microsoft Exchange V6.0.5762.3
Try petroleum ether. This is available in different boiling point ranges, eg 60-80°C might be very useful for your purpose. This is available from several suppliers (like Baker, Merck).
Frits Klijn
-----Original Message----- } From: John Twilley [mailto:jtwilley-at-sprynet.com] Sent: Monday, August 11, 2003 20:01 To: by way of Ask-A-Microscopist Cc: Microscopy-at-sparc5.microscopy.com
Vaselin,
Organic solvents and solid compounds differ in a property called "polarity". Those with similar polarity tend to dissolve in each other, while those with large differences do not. Since paraffin has a very low polarity, the most effective solvents for it are ones with low polarity. Examples of low polarity solvents are petroleum distillates (like the benzine that you mention) or materials such as naphtha, ligroin, white spirit and mineral spirits. For something that is readily available and evaporates easily you could try the liquid used as fuel for cigarette lighters. Acetone has a relatively high polarity and therefore is not very effective in dissolving paraffin.
The liquids sold to remove spots from clothing might work but these can contain chlorinated compounds that are more dangerous to health and they may leave residues that do not evaporate easily.
You should be aware that different countries and languages may use the terms "naphtha" and "benzine" to designate slightly different materials refined from petroleum. But all should be effective for dissolving paraffin. Be careful, however, to note the difference between "benzine" and "benzene". Benzene is a specific chemical compound with very serious health hazards - more serious than those of benzine.
You should also avoid xylene and toluene because they have greater health risks than the materials mentioned above which work just as well. Regardless of the material used, use ventilation and avoid breathing the vapor to protect your health and those around you.
John
by way of Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 11, 2003 at 01:42:01 } --------------------------------------------------------------------------- } } Email: sswaffe-at-abv.bg } Name: Veselin Andreev } } Organization: 54 "st.Ivan Rilski" } } Education: 6-8th Grade Middle School } } Location: Sofia Bulgaria } } Question: Can I use acetone for disolving paraffin in paraffin embedding.What else should I use exept xylene/toluene.Can I use benzine.Or some other domestic solvent or some which is easy to find at the stores? } } ---------------------------------------------------------------------------
Try this from Cobehn Systems; http://www.cobehn.com/page2.htm
Peter Tomic Agere Systems Allentown, PA USA
-----Original Message----- } From: sswaffe-at-abv.bg [mailto:sswaffe-at-abv.bg] Sent: Monday, August 11, 2003 8:53 AM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 11, 2003 at 01:42:01 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin Andreev
Organization: 54 "st.Ivan Rilski"
Education: 6-8th Grade Middle School
Location: Sofia Bulgaria
Question: Can I use acetone for disolving paraffin in paraffin embedding.What else should I use exept xylene/toluene.Can I use benzine.Or some other domestic solvent or some which is easy to find at the stores?
I think it is better not to use conductive tape since anyway you are using silver paint (I prefer to use graphite paint). Next step is to coat for one additional minute at different tilt angle - for specimens with rough surface it could be helpful. Also you can try to rise voltage to 15-25 kV to check if you can see gold with BSE on platinum coated specimens.
I have never used pure platinum for coating, but with two coatings with gold/palladium (15 ma, 30 sec. each) I usually get good results.
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: Phillips, Thomas E. } Sent: Sunday, August 10, 2003 7:18 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM - charging problems with bio samples } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } We are relative novices on the SEM and are seeking advice on ways to } minimize our charging problems. We would appreciate any tips } from the } mavens out there. } } We are looking at biological specimens (tissue explants) } coated either with } platinum or carbon (details below). The carbon coating } protocol is for } when we are trying to image at colloidal gold labeling on the } surface using } the BSE detector. The charging is much worse on the carbon coated } specimens compared to the platinum coated ones. I don't have } a feel for } how much of this problem is "normal" and one has to live } with it like so } much else in EM! But if anyone out there has some ideas or tricks to } reduce charging, we are willing to try it. I would also be } interested in } anyone's nomination for a great reference book on SEM } techniques. Thanks. Tom } } Platinum coating protocol: } } Tissue (2x2x2 mm) fixed in parformaldehyde/glutaraldehyde (or } 2% PF) for } several hours. } Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. } Tissue dehydrated in EtOH series. } Tissue critical point dried. } Tissue mounted on stub with conductive tape. } Next, silver painted the edges of the specimen and allowed to dry. } Sputter coated the specimen with platinum at 10 mA for 70 sec. } Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications } ranging from } 1-20k and sometimes up to 50k. } Specimens stored in a desiccation chamber. } *Re-coating for another minute fixes the problem some of the time. } } Carbon coating protocol: } } Tissue (from 1x1x1 to 3x3x3 mm) fixed in 2% PF for several hours. } Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. } Tissue dehydrated in EtOH series. } Tissue critical point dried. } Tissue mounted on stub with conductive tape. } Next, silver painted the edges of the specimen and allowed to dry. } Sputter coated the specimen with high purity carbon for 3 sec } (~ 20 nm } coating?). } Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications } ranging from } 1-20k using both BSE and SE to image. } Specimens stored in a desiccation chamber. } } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } } }
Are you doing silver enhancement? If not, did you try gold-gilded copper grids? That's what I use. Reasonably sturdy, don't do funny things when approached with tweezers, not to mention astigmatism in the scope, and they are not too expensive. Gold coating seems to prevent any contact of Cu with the PBS.
I did have what seemed to be Ni-induced astigmatism a few times, but it was always one particular grid, and the next Ni grid would be OK.
Vlad -- ------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
I am going to make up a batch of Formvar in Ethylene Dichloride and would like some suggestions, please.
* Having not heard back from my safety people yet--how are you safely & properly disposing of the waste? * What is the best/safest way to clean my glassware and stir bar afterwards? * What seems to be the best dilution? I had read to use between 0.1 % and 0.3%, but could have sworn we used 0.75% in a lab years ago.
Thanks in advance for any helpful advice!
Donna R. Clarkson
Northrop Grumman Information Technology for U S Army Medical Research Detachment at Brooks City-Base Phone (210) 536-1416 FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil
Nestor; Great job on the SPAM filtering. My Intel inbox was clogged with 627 emails after 4 days at MSA, and it was 98% SPAM. Any chance you could submit your resume to the Intel IT department? They certainly could use your expertise!
John Mardinly Phone: 408-765-2346 Pager: 877-277-1182
-----Original Message----- } From: zaluzec-at-sparc5.microscopy.com [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Monday, August 11, 2003 6:12 AM To: Microscopy-at-sparc5.microscopy.com
Colleagues...
There was another massive spam attack this weekend. I have updated the filters, but I'm leerie of the fact that this may catch additional valid postings.
If you receive a "rejected mail messages" please remember to follow the directions you receive so that I can resolve the issues and get you back into the system.
I appreciate all the advice on nickel grids potentially causing me the problems with stigmation. I agree this is a likely culprit but that doesn't explain why the problem is intermittent - some times I experience no problems at all. I thought perhaps some batches of grids were magnetized and some weren't but the consensus seems to be that the EM itself will make them magnetic so that eliminates that possibility. I am thinking about switching to gold grids but the cost is not insignificant. I need to use formvar/carbon coated grids to support my very unstable acrylic resin sections of plant material for the rigorous immunolabeling protocol that I have to follow. Since i would like to avoid making my own films, my choice of vendors is limited. It looks to me like switching to gold will cost 40 or 50 cents more a grid. That would be several hundred dollars a year for me so I would like to avoid it if possible. Clearly Ni grids can't always be problematic or nobody would buy them. Thanks for all the comments on this issue. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Wendy Clark, Librarian Tennessee Valley Authority Research Library Muscle Shoals, AL 35661 256-386-2872 whclark-at-tva.gov
Has contacted me and informed me that the TVA has closed its microscopy facility at her location and has a long list of relevant journals and books that will be discarded if no one wants them. I suggested that she give me the list and I would post it on the listserver.
The list is below. Sorry about the length. If you want anything on the list, PLEASE contact Wendy.
Ron Anderson, Editor Microscopy Today
JOURNALS AVAILABLE FOR DONATION:
Agricultural ammonia news, 1954 - 1967 Agricultural and biological chemistry, 1966 - 1991 (Tokyo) Agricultural history, 1978 - 1989 Agricultural nitrogen news, 1967 - 1970 Ambio, 1976 - 1984 American laboratory, 1969 - 1990 Applied engineering in agriculture, 1985 - 1994 Archiv fur Acker- und Pflanzenbau und bodenkunde/Deutsche Demokratische Republik, Deutsche Akademie der Landwirtschftswissenschaften zu Berlin, 1972 - 1987 Archives of environmental contamination and toxicology, 1994 - 1997 ASTM standardization news, 1973 - 1984 Bacteriological reviews, 1961 - 1975 Bibliography of agriculture, Section A, agricultural economics and rural sociology, 1942 - 1943 Bibliography of agriculture, Section B, agricultural engineering, 1942 - 1943 Bibliography of agriculture, section C, entomology, 1942 - 1943 Bibliography of agriculture, section D, plant science, 1942 - 1943 Bibliography of agriculture, section E, forestry, 1942 - 1943 Bibliography of agriculture, section F, food processing and distribution, 1943 Bibliography of agriculture/U.S. Department of Agriculture, Library, 1943 - 1975 Biochemistry, 1962 - 1974 Biochemistry and cell biology, 1986 - 1987 Biological abstracts, 1945 - 1969 Biological conservation, 1968 - 1981 Biotechnology letters, 1982 - 1983; 1996 - 1998 The Botanical review, 1964 - 1988 Bulletin of environmental contamination and toxicology, 1994 - 1997 Bulletin of the Academy of Sciences of the USSR, Division of Chemical Science, 1952 - 1991 Canadian chemical processing, 1952 - 1984 Canadian chemistry and process industries, 1944 - 1951 Canadian journal of botany, 1996 - 1997 Canadian journal of chemical engineering, 1967 - 1993 Canadian journal of forest research, 1971 - 1981 Canadian journal of plant science, 1969 - 1995 Canadian journal of research, Sec. B, 1942 - 1943 Canadian journal of research, Sec. B, Chemical sciences, 1944 - 1950 Chemical and engineering news, 1942 - 2002 Chemical engineering, 1947 - 2002 Chemical engineering progress, 1947 - 1995 Chemical engineering science, 1966 - 1982 Chemical market reporter, 1996 - 1999 Chemical marketing reporter, 1984 - 1996 Chemical processing, 1960 - 1986 Chemistry and industry, 1936 - 1988 Chemistry in Britain, 1965 - 1995 Control engineering, 1960 - 1988; 1992 Corrosion, 1954 - 1995 Corrosion science, 1966 - 1974 Dissertation abstracts international, A, the humanities and social sciences, 1969 - 1994 Ecology, 1971 - 1997 Energy progress, 1981 - 1988 Engineering news-record, 1980 - 1986 ENR, 1987 - 2002 Environmental affairs, 1971 - 1978 Environmental pollution, 1973 - 1979 Environmental pollution, Series A, Ecological and biological, 1980 - 1982 The Environmental professional: the official journal of the National Association of Environmental Professionals, 1988 - 1995 Environmental progress, 1982 - 1994 Forest science, 1955 - 1995 Growth and change, 1970 - 1987 Hazardous waste consultant, 1984 - 1995 Inorganic and nuclear chemistry letters, 1969 - 1981 Instrumentation science & technology, 1994 - 1997 Instrumentation technology: the journal of the Instrumentation Society of America, 1963 - 1978 InTech, 1979 - 1992 International chemical engineering, 1961 - 1994 International journal of chemical kinetics, 1969 - 1989 International journal of sulfur chemistry, 1973 - 1976 International journal of sulfur chemistry, A, Original, experimental, and theoretical studies, 1971 - 1972 International journal of sulfur chemistry, B, quarterly reports on sulfur chemistry, 1971 - 1972 International journal of sulfur chemistry, C, Mechanisms of reactions of sulfur compounds, 1971 - 1972 Journal of analytical atomic spectrometry, 1986 - 1995 Journal of applied spectroscopy, 1965 - 1972 Journal of chemical engineering of Japan, 1968 - 1982 The Journal of chemical thermodynamics, 1969 - 1993 Journal of inorganic and nuclear chemistry, 1955 - 1981 Journal of regional science, 1958 - 1988 Journal of research of the National Bureau of Standards, 1977 - 1988 Journal of research of the National Bureau of Standards, 1934 - 1959 Journal of research of the National Bureau of Standards, B, Mathematical sciences, 1968 - 1977 Journal of research of the National Bureau of Standards, B, mathematics and mathematical physics, 1959 - 1967 Journal of research of the National Bureau of Standards, C, engineering and instrumentation, 1959 - 1972 Journal of research of the National Bureau of Standards, Section A, Physics and chemistry, 1959 - 1977 Journal of research of the National Institute of Standards and Technology, 1989 - 1995 Land and water: the magazine of natural resource management and restoration, 1994 - 1998 Langmuir: the ACS journal of surfaces and colloids, 1985 - 1995 Mendeleev chemistry journal 1983 - 1989 Nuclear instruments & methods, 1967 - 1981 Nuclear instruments & methods in physics research, 1981 - 1983 Nuclear instruments & methods in physics research, Section A, accelerators, spectrometers, detectors, and associated equipment, 1984 - 1988 Nuclear instruments & methods in physics research, Section B, Beam interactions with materials and atoms, 1984 - 1989 Pesticide science, 1992 - 1995 Phosphorus and sulfur and the related elements, 1976 - 1988 Phosphorus and the heavier group Va elements, 1971 Phosphorus and the related group V elements, 1972 - 1975 Phosphorus, sulfur, and silicon and the related elements, 1989 - 1991 Pollution engineering, 1969 - 1995 Proceedings of the Academy of Sciences of the USSR, 1965 - 1990 Proceedings of the Analytical Division of the Chemical Society, 1975 - 1979 Proceedings of the Chemical Society, 1957 - 1964 The Progressive fish-culturist, 1947 - 1968, 1992 - 1994 The quarterly journal of economics, 1964 - 1987 Quarterly reports on sulfur chemistry, 1966 - 1970 Quarterly reviews - chemical society, 1960 - 1971 Record of chemical progress, 1939 - 1971 Recueil des travaux chimiques des Pays-Bas, 1920 - 1991 Recueil: journal of the Royal Netherlands Chemical Society, 1980 - 1991 Remote sensing of environment, 1969 - 1981 The Science of the total environment, 1994 - 1995 Sensors and actuators, B, Chemical, 1992 - 1997 Separation and purification methods, 1972 - 1983 Separation science, 1966 - 1977 Soviet Materials Science, 1965 - 1981 Soviet Physics, Crystallography, 1969 - 1982 Soviet Plant Physiology, 1962 - 1982 Soviet Progress in Chemistry, 1984 - 1991 Standardization news, 1985 - 1992 Studies in applied mathematics, 1969 - 1987 Successful farming, 1960 - 1995 Synthesis and reactivity in inorganic and metal-organic chemistry, 1974 - 1988 Synthesis in inorganic and metal-organic chemistry, 1971 - 1973 Tree physiology, 1986 - 1994 Tree planters' notes, 1967 - 1994 Weed Science, 1996 - 1998 Weed technology: a journal of the weed science society of America, 1996 -
BOOK SETS
Annual Review of Plant Physiology v.15, 1964 - V48, 1987 Handbuch der Pflanzenphysiologie (Encyclopedia of Plant Physiology) 18v. SpringerVerlag, 1967 (German-English) Plant Physiology v.1-5. ed. F.C. Steward. Academic, 1960 Annual Review of Phytopathlogy v.4, 1966-v.31,1993 Annual Review of Microbiology v.14, 1960-v.47, 1993 Annual Review of Entomology v.1,1956-v.37,1992 Annual Review of Biochemistry v.30,1961-v.55,1986 Industrial Waste Conference. Purdue University. 2nd,1946-48th,1993 Proceedings American Society for Horticultual Science v.36,1938; v.53,1949 - v.93,1968 Proceedings Annual Convention, Association of Land Grant Colleges & Universities. 47, 1933 - 80th, 1966. Florida State Horticultural Society Proceedings v.75,1962 - v.100, 1987 Proceedings Agricultural Research Institute, 9th, 1960 - 32nd, 1983 Transactions of the International Congress of Soil Science 1st,1927; 2nd, 1930; 3rd., 1956; 7th, 1960; 8th, 1964; 9th, 1968; 10th, 1974; 11th, 1978; 12th., 1982; 13th, 1986. Proceedings Soil Science Society of Florida, v.1, 1939 - v.53, 1994
} Donna R. Clarkson } Northrop Grumman Information Technology } for U S Army Medical Research Detachment } at Brooks City-Base } Phone (210) 536-1416 } FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil
} I am going to make up a batch of Formvar in Ethylene Dichloride and } would like some suggestions, please. } } * Having not heard back from my safety people yet--how are you safely & properly disposing of the waste?
Work in a fume hood as the fumes are considered carcinogenic. Keep all wastes in a container (vented to hood and away from flames) for disposal as a volatile chemical.
} * What is the best/safest way to clean my glassware and stir bar } afterwards?
Rinse in ethylene dichloride several times and put the washes in the disposal container, above.
} * What seems to be the best dilution? I had read to use between 0.1 % } and 0.3%, but could have sworn we used 0.75% in a lab years ago. } I have used 0.25% Formvar or Butvar in ethylene dichloride. My recommendation is that you purchase the solution already made up (all EM Supply houses carry it). That way you have less waste and the product is guaranteed.
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 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
I have been trying to label V5 postive neurons of 4.0% paraform. fixed brain sections with a polyclonal V5 antibody. These neurons are also gfp (green fluores. protein) labelled so we know that the vector has delivered the V5 sequence. The gfp is not fused to the V5 coding sequence.
My question is does the gfp fluores. inhibit any labeling with the avidin-biotin immunocytochemistry procedure to labelling with the V5 antibody (like steric hindrance which is sometimes a problem with gold immunoEM labelling)? So far we have tried 3 times to label with no success. We make sure we have gfp positive cells in the brain tissue slices before we start any immunocytochemistry procedure.
I am using DAB (diaminobenzidine) as a chromagen which would be silver enhanced and then the 70 micron brain tissue slab would be post fixed in 2.0% glut, osmicated and embedded in Epon.
Any help would be very much appreciated!
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642
GFP may not interfere with biotin-avidin reaction. The constant of association for biotin-avidin complex is about 10^12M^-1, which is pretty high. 4% formaldehyde is sounds to me too much. It may kill antigenic determinants if you are using primary antibodies. From another hand (opposite to the previous), V5 may be easily washed out if it's small peptide (have no idea what it is). I would suggest to do immuno-staining at LM level first to be sure that your procedure works, then switch to EM. Brain, also contains a lot of lipids, so you may need to use detergents to open cells up. Good luck, Sergey.
At 12:56 PM 8/12/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I have recently switched to Formvar in Diethylene Chloride as a base for making carbon coated grids, because I was told that I would get bettter continuous films, the problem I am having is that I cannot get rid of the forvar after coating. Does anyone out there have a method for making C coated grids that works everytime?? I am getting to the end of my patience and have wasted an awful lot of grids over the last few months.
Thanks
Steve Parry -- Steve Parry Centre for Microscopy & Microanalysis, (M010) The University of Western Australia, 35 Sterling Highway, Crawley, WA 6009, Australia.
Does anybbody have some experiences in the application of fluorescents in combination with silica/alumina based catalysts? What type of fluorescents? Etc.
I am quite new in this field so all information is welcome.
Regards,
Frits Klijn Akzo Nobel Catalysts b.v. Dept. SMA - R&D1 P.O.Box 37650 1030 BE Amsterdam Netherlands
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I am currently using a CP-Research AFM/STM unit from ThermoMicroscopes/Veeco. I would like to find someone who has made improvements in the machine for electronic noise reduction in the STM mode. I notice that when the tip is retracted, a leakage current of hundreds of picoamps can be measured. While scanning, I can only stabilize the image using three nanoamps or more.
thank you
Prof. Dr. Frederico Cunha Physics Department Universidade Federal de Sergipe Brazil
Hello: We are getting rid of a Philips 400T TEM. It was in working order as of last year. It has been listed on Govt' surplus for a few months with no takers. It is now being offered to Universities, and other similar organization. You will have to pay for packing and shipping. We are located in Maryland. Drop me an email if interested, Charlie Murphy murphyc-at-ba.ars.usda.gov
Charles Murphy USDA, ARS, Electron Microscopy Unit Bldg. 177B, BARC East Beltsville, MD, 20705 (301) 504-8046 (301) 504-8923 fax murphyc-at-ba.ars.usda.gov
On Tuesday, August 12, 2003, at 09:10 AM, Clarkson Donna R Contr USAMRD wrote:
} * Having not heard back from my safety people yet--how are you safely } & properly disposing of the waste? } Dear Donna, I second what John Bozzola said, and check the local regulations for proper disposal of Cl-containing hydrocarbons. In NY they were separated from other HCs, but in CA they are not. In both cases, the safety offices took care of the disposal. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Steve, When I want just carbon as a substrate I will coat freshly cleaved mica with carbon in a metal evaporator. This allows me to dictate the carbon thickness and I have no trouble floating off grid size squares(prescored)in a drop of water. Picking up the carbon with glow discharged grids (flat side) is a snap. Make sure everything is clean. This should give you thin clean substrates. Mike D.
Steve Parry wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have recently switched to Formvar in Diethylene Chloride as a base } for making carbon coated grids, because I was told that I would get } bettter continuous films, the problem I am having is that I cannot } get rid of the forvar after coating. } Does anyone out there have a method for making C coated grids that } works everytime?? I am getting to the end of my patience and have } wasted an awful lot of grids over the last few months. } } Thanks } } Steve Parry } -- } Steve Parry } Centre for Microscopy & Microanalysis, (M010) } The University of Western Australia, } 35 Sterling Highway, Crawley, } WA 6009, Australia. } } Fax: +61-8-9380-1087 } Email: sparry-at-cmm.uwa.edu.au } Phone: +61-8-9380-8057 [24 hour voicemail attached]
Based in West London, the Clinical Sciences Centre (CSC) is an Institute of the Medical Research Council and an academic division of the Faculty of Medicine, Imperial College, London, UK. The CSC has an international reputation and its interdisciplinary research and training programmes in cell biology, genomics, and biological and clinical imaging, address key questions underpinning human health and disease.
The imaging of molecules within cells, organs and whole organisms is at the forefront of scientific developments in the life sciences. The CSC is a leading research institute with groups developing new avenues in the areas of confocal laser scanning microscopy (CLSM), live-cell imaging (CCD imaging and spinning-disk confocal), transmission electron microscopy (TEM), scanning ion conductance microscopy, position emission tomography (PET), and nuclear magnetic resonance (NMR).
We are seeking a highly motivated individual to be responsible for this state-of-the-art microscope core facility at the CSC, to provide full support for confocal, live-cell imaging, and other fluorescence microscope users within the institute. Duties will include direct technical support, training and supervision of users, assuring instrument integrity, maintaining the facility web site. Direct involvement in research project using fluorescence microscopy will be expected (further information about research at the CSC can be found at www.csc.mrc.ac.uk ). The candidate will have to maintain a current understanding of the microscopy field as necessary to implement new technologies in the department.
The successful candidate should have a science degree in a relevant subject, with 5 years research experience or a PhD in a relevant subject, and must demonstrate a good understanding of current fluorescence imaging technology and sample preparation. Experience in confocal, live-cell and digital imaging (particularly Leica and Perkin-Elmer confocal microscopes and/or Deltavision microscope) are highly desirable. Applicants at this level will be appointed at MRC Pay Band 4. Please quote Ref: MRA/RA/D.
We also actively encourage applications from candidates with less experience but a minimum of 3 years of research experience or imaging facility management who will run the facility and assist in research projects. Applicants should have a science degree in a relevant subject and will be appointed at MRC Pay Band 5. Please quote Ref: MRA/RAS/D.
All applicants must have a strong wish to participate actively in team work, possess excellent written and oral communication skills, be comfortable working independently, communicate effectively with service users to understand their microscopy needs, be at ease with PC computer platforms and demonstrate effective troubleshooting skills.
Further details on how to apply are available from www.csc.mrc.ac.uk or email recruit-at-csc.mrc.ac.uk, quoting the relevant reference above. Informal enquiries and further information about the position to Drs Ana Pombo (tel: 020 8383 8232; ana.pombo-at-csc.mrc.ac.uk) or Alex Sardini (tel: 020 8383 8270, a.sardini-at-csc.mrc.ac.uk).
The closing date for applications is 20 September 2003. Interviews will take place 22 September-15 October 2003.
Ana Pombo, D.Phil. Group Head Nuclear Organisation Group MRC Clinical Sciences Centre Hammersmith Hospital Campus Du Cane Road London W12 0NN UK
I would like to add a few cents to the Mike Delannoy posting. Instead picking carbon on individual grid in the drop of water, I am doing as a following:
- you need some "flask" diameter about 90 mm and 40-50 mm high with deionized ultrapure water filled up to about 5 mm lower than edge (flask itself should be clean as well). You also may use any suitable in size glass container. The idea is to have relatively large flat area of water surface. - you need fluorescent lamp to illuminate the water surface in the way it happening when you illuminate water surface in the ultratome knife boat. So, you should see the reflection of the bulb on the surface. Playing with the angle ant location, you need to find the orientation, which gives you well illuminated large area on the water surface. - depends, how many grids you want to made, cut the piece of mica with carbon of correspondent size. Important: all FOUR sides of your mica piece should be fresh cut. - slowly under 45 deg angle move mica (carbon is up) into the water, so carbon will float on the surface. It should be visible if illumination is set right. - put grids on the floating carbon. DO NOT touch carbon by tweezers. Try to drop grids on the carbon from a few mm distance. - cut the piece of Parafilm slightly bigger that carbon. Using two tweezers, very carefully put Parafilm on the carbon (with grids). Hold one side of the Parafilm sheet with tweezer; with one smooth movement move Parafilm (with carbon and grids) under the water and then out of water. - place Parafilm sheet with attached grids (up) on the piece of filter paper and dry at 60oC for about 10 min. -Grids are ready. - Personally, I prefer to use fresh carbon for every experiment, so I made about 6 grids at the time and them make more if needed. If you are using extremely thick carbon film, you may not need good illumination. - I don't see any point to glow discharge grids itself. If glow discharge "works" on the naked grids, it simply meant that grids are greasy and that contamination has been ionized by glow discharge. Clean metal grids will not hold charge at all (electro-conductivity, you know). Covering grids with very diluted plastic solution (like parlodium 0.01-0.005%) may help to attach carbon to the grids. Just put grids on the filter paper and put a drop of plastic on it, let it dry. More effective way to mount carbon film on the grids is to use "holey" carbon coated grids. Carbon sticks nicely to carbon.
Sergey
At 10:24 AM 8/13/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
On Sun, Aug 10, 2003 at 07:17:56PM -0500, Tom Phillips wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are relative novices on the SEM and are seeking advice on ways to } minimize our charging problems. We would appreciate any tips from the } mavens out there. } } We are looking at biological specimens (tissue explants) coated either with } platinum or carbon (details below). The carbon coating protocol is for } when we are trying to image at colloidal gold labeling on the surface using } the BSE detector. The charging is much worse on the carbon coated } specimens compared to the platinum coated ones. I don't have a feel for } how much of this problem is "normal" and one has to live with it like so } much else in EM! But if anyone out there has some ideas or tricks to } reduce charging, we are willing to try it. I would also be interested in } anyone's nomination for a great reference book on SEM techniques. Thanks. } Tom
I only know a little bit about colloidal gold labeling but I would think if you're using a BSE detector to look for the gold then I don't think you'd want to coat the sample with gold or platinum (unless the gold particles are significant topographic features and you're using the BSE detector in 'topo' mode) -- that pretty much leaves you with carbon coating -- more prone to charging but still usable.
Here are some general SEM techniques you can try to reduce charging for 'problem' samples.
* Use smaller spot size * Use smaller aperature * Use a LOWER magnification if feasable * Increase scan rate
All these basically reduce the rate at which electrons hit a given area of your sample.
If you're still having problems you may have to coat with more carbon.
I hope this helps. Good luck!
-- James Firmiss ------ firmiss-at-granicus.if.org ------ "Let Sanity Prevail"
Dear Steve, The method I use to make carbon-coated grids uses collodion in amyl acetate. A drop or two is put onto the surface of a large dish of distilled water. This dries to a circle about three to four inches in diameter on the surface of the water. I drop copper grids onto this surface, preferably before it dries too much, so the collodion is still a bit sticky. After I have about thirty grids arranged in a neat rectangle on the collodion film, I drop a filter paper on top and bring the edges of the film over the edges of the filter paper. As soon as the paper is wet, I lift it out by one edge and lay it down, grids up, on more filter paper to dry. This may take some practice to get the filter paper up out of the water with the grids still stuck on. When the paper is dry, I cut out the rectangle of paper with the grids on it with scissors and carbon coat it (grids side up) in the evaporator. The grids are then removed from the paper and put in a Jaffe washer (collodion/carbon side up) filled with chloroform for 48 hours. (A Jaffe washer is a petri plate with a stack of four glass slides and a stack of filter paper over the glass slides. The bottom of the petri plate is filled with solvent and the lid of the petri plate put on. Then, another, larger dish is put over it so the solvent doesn't evaporate too quickly. The material put on the top of the filter paper stays wet with solvent and the solvent washes the material). This will dissolve the collodion and bring the carbon film down to stick it to the copper grid. These films are conductive, continuous and strong enough for routine work at 200kV. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Steve Parry" {sparry-at-cmm.uwa.edu.au} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, August 12, 2003 8:18 PM
James/Tom Yes, it is certainly my experience that gold palladium sputter coating quickly kills the BSE signal from 5 and 10nm colloidal gold. Carbon provides the best signal from small gold, but a thin sputtered chromium coating is also very good, and has the advantage of providing more fine topographical detail than can be obtained from carbon-coated specimens. I have used the Denton head in my Gatan Alto to do this, and have also had excellent results from specimens sputtered for us by Emitech.
Unfortunately, the techniques James suggests for reducing charging, while effective for reducing charging, may be just exactly what you want to avoid in BSE imaging of gold. Signal and signal/noise is often a problem, so one needs more beam current rather than less, and long record times, and it is therefore important that the coating and mounting technique is perfect. I often use the Analysis mode with the normal aperture 2/3 and and a relatively large spot size 5 on my Hitachi 4700, with the emission current increased to 15 or even 20µA, and find I need to be working at higher mags (say 20-100k) to get the best images.
Things to consider when optimizing carbon coating and specimen mounting are:
*Use rotary-tilt coating technique to get even coating all over. Static coating will leave a 3-d specimen with large uncoated shadow areas *Make sure the coating really is thick enough. A piece of white paper or masking tape placed near the specimens should be mid grey *Ensure the coating is in electrical continuity with the stub. If there is the slightest gap under the tissue block then paint it up with carbon dag. *If the specimens are large cubes (} 1mm) make sure the sides are adequately coated with carbon, and if in doubt dag them *Paint dag lines to the metal of the stub. Don't rely entirely on the conductivity of sticky carbon tabs, which is often not good enough.
Chris
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Facility
} ----- Original Message ----- } From: "firmiss" {firmiss-at-granicus.if.org} } To: "MSA ListServer" {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, August 13, 2003 11:42 PM } Subject: Re: SEM - charging problems with bio samples } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ---. } } } } } } On Sun, Aug 10, 2003 at 07:17:56PM -0500, Tom Phillips wrote: } } } } -------------------------------------------------------------------- } ---- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -------------------------------------------------------------------- } ---. } } } } } } } } } We are relative novices on the SEM and are seeking advice on ways } to } } } minimize our charging problems. We would appreciate any tips from } the } } } mavens out there. } } } } } } We are looking at biological specimens (tissue explants) coated } either with } } } platinum or carbon (details below). The carbon coating protocol } is for } } } when we are trying to image at colloidal gold labeling on the } surface using } } } the BSE detector. The charging is much worse on the carbon coated } } } specimens compared to the platinum coated ones. I don't have a } feel for } } } how much of this problem is "normal" and one has to live with it } like so } } } much else in EM! But if anyone out there has some ideas or tricks } to } } } reduce charging, we are willing to try it. I would also be } interested in } } } anyone's nomination for a great reference book on SEM techniques. } Thanks. } } } Tom } } } } I only know a little bit about colloidal gold labeling but I would } think } } if you're using a BSE detector to look for the gold then I don't } think } } you'd want to coat the sample with gold or platinum (unless the gold } } particles are significant topographic features and you're using the } } BSE detector in 'topo' mode) -- that pretty much leaves you with } carbon } } coating -- more prone to charging but still usable. } } } } Here are some general SEM techniques you can try to reduce charging } for } } 'problem' samples. } } } } * Use smaller spot size } } * Use smaller aperature } } * Use a LOWER magnification if feasable } } * Increase scan rate } } } } All these basically reduce the rate at which electrons hit a given } area } } of your sample. } } } } If you're still having problems you may have to coat with more } carbon. } } } } I hope this helps. Good luck! } } } } -- } } James Firmiss ------ firmiss-at-granicus.if.org ------ } } "Let Sanity Prevail" } } }
I second Mike Delannoy's suggestion. Mica should give you the smoothest continuous carbon films. In my experience one problem is that the C-films can wrinkle terribly if floated off too soon from the mica. This can be simply avoided by letting the film first "mature" overnight on the mica. Good luck,
Jim
-----Original Message----- } From: Steve Parry [mailto:sparry-at-cmm.uwa.edu.au] Sent: Wednesday, August 13, 2003 5:19 AM To: Microscopy Listserver
I have recently switched to Formvar in Diethylene Chloride as a base for making carbon coated grids, because I was told that I would get bettter continuous films, the problem I am having is that I cannot get rid of the forvar after coating. Does anyone out there have a method for making C coated grids that works everytime?? I am getting to the end of my patience and have wasted an awful lot of grids over the last few months.
Thanks
Steve Parry -- Steve Parry Centre for Microscopy & Microanalysis, (M010) The University of Western Australia, 35 Sterling Highway, Crawley, WA 6009, Australia.
We have an EM autoradiography project to carry out. It is way too long since I did any of this so I need some advice about exposure times. If anyone is current on AR please contact me offline if you can help. It was great to see everyone again in San Antonio - well done MSA for another great meeting.
Regards
Chris
Christopher J. Gilpin Ph.D. Assistant Professor Director Molecular and Cellular Imaging Facility K1.246 Department of Cell Biology University of Texas Southwestern Medical Center 5323 Harry Hines Boulevard Dallas, Texas 75390-9039 +1 214 648 2827 Phone +1 214 648 6408 Fax christopher.gilpin-at-utsouthwestern.edu
Does anybody know a quick recipe for "grid glue" made out of scotch tape (or other sticky stuff) and some solvent for glueing films to TEM grids? Thanks in advance. -Karl
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
I am attempting to do a negative stain (which I know how to do) on a sample. the problem I'm having is that according to the protocol I have to glow discharge the grid before I use it.
I don't have access to a vacuum evaporator with a glow discharge unit. So what's a tech to do?
Any suggestions as to how to fake a glow are greatly appreciated.
Please help me to glimmer, glimmer. ;-)
Unable to glow,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We have had a lot of success using ion beam sputter deposited Cr thin films for colloidal gold labeling of biological samples. Due to the nature of ion beam sputtered films we can precisely control the deposited thickness to less than 10 angstroms and still provide a uniform layer of Cr for charge reduction. The gold label on the sample provide high contrast in backscatter mode and the deposited Cr does not interfere with the signal.
If you would like additional information on this type of equipment or would like to see some examples I would be happy to provide this to you off-list. I hope this helps!
Best Regards,
Shane Roberts Applications Engineering Manager South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 www.southbaytech.com roberts-at-southbaytech.com
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Thursday, August 14, 2003 12:15 AM To: microscopy-at-sparc5.microscopy.com
James/Tom Yes, it is certainly my experience that gold palladium sputter coating quickly kills the BSE signal from 5 and 10nm colloidal gold. Carbon provides the best signal from small gold, but a thin sputtered chromium coating is also very good, and has the advantage of providing more fine topographical detail than can be obtained from carbon-coated specimens. I have used the Denton head in my Gatan Alto to do this, and have also had excellent results from specimens sputtered for us by Emitech.
Unfortunately, the techniques James suggests for reducing charging, while effective for reducing charging, may be just exactly what you want to avoid in BSE imaging of gold. Signal and signal/noise is often a problem, so one needs more beam current rather than less, and long record times, and it is therefore important that the coating and mounting technique is perfect. I often use the Analysis mode with the normal aperture 2/3 and and a relatively large spot size 5 on my Hitachi 4700, with the emission current increased to 15 or even 20µA, and find I need to be working at higher mags (say 20-100k) to get the best images.
Things to consider when optimizing carbon coating and specimen mounting are:
*Use rotary-tilt coating technique to get even coating all over. Static coating will leave a 3-d specimen with large uncoated shadow areas *Make sure the coating really is thick enough. A piece of white paper or masking tape placed near the specimens should be mid grey *Ensure the coating is in electrical continuity with the stub. If there is the slightest gap under the tissue block then paint it up with carbon dag. *If the specimens are large cubes (} 1mm) make sure the sides are adequately coated with carbon, and if in doubt dag them *Paint dag lines to the metal of the stub. Don't rely entirely on the conductivity of sticky carbon tabs, which is often not good enough.
Chris
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Facility
} ----- Original Message ----- } From: "firmiss" {firmiss-at-granicus.if.org} } To: "MSA ListServer" {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, August 13, 2003 11:42 PM } Subject: Re: SEM - charging problems with bio samples } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ---. } } } } } } On Sun, Aug 10, 2003 at 07:17:56PM -0500, Tom Phillips wrote: } } } } -------------------------------------------------------------------- } ---- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -------------------------------------------------------------------- } ---. } } } } } } } } } We are relative novices on the SEM and are seeking advice on ways } to } } } minimize our charging problems. We would appreciate any tips from } the } } } mavens out there. } } } } } } We are looking at biological specimens (tissue explants) coated } either with } } } platinum or carbon (details below). The carbon coating protocol } is for } } } when we are trying to image at colloidal gold labeling on the } surface using } } } the BSE detector. The charging is much worse on the carbon coated } } } specimens compared to the platinum coated ones. I don't have a } feel for } } } how much of this problem is "normal" and one has to live with it } like so } } } much else in EM! But if anyone out there has some ideas or tricks } to } } } reduce charging, we are willing to try it. I would also be } interested in } } } anyone's nomination for a great reference book on SEM techniques. } Thanks. } } } Tom } } } } I only know a little bit about colloidal gold labeling but I would } think } } if you're using a BSE detector to look for the gold then I don't } think } } you'd want to coat the sample with gold or platinum (unless the gold } } particles are significant topographic features and you're using the } } BSE detector in 'topo' mode) -- that pretty much leaves you with } carbon } } coating -- more prone to charging but still usable. } } } } Here are some general SEM techniques you can try to reduce charging } for } } 'problem' samples. } } } } * Use smaller spot size } } * Use smaller aperature } } * Use a LOWER magnification if feasable } } * Increase scan rate } } } } All these basically reduce the rate at which electrons hit a given } area } } of your sample. } } } } If you're still having problems you may have to coat with more } carbon. } } } } I hope this helps. Good luck! } } } } -- } } James Firmiss ------ firmiss-at-granicus.if.org ------ } } "Let Sanity Prevail" } } }
We've been looking at the Nikon and Olympus TIRF systems.
Question 1: Is the 100X 1.65 N.A. objective really necessary?
Question 2: I would love to have an informal discussion off list with anybody who has experience with one or both of these systems. If I may speak with you, please contact me off list by email and we could write or set up a time that I could call you at your convenience.
Thanks! ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
Light/ Confocal Microscopist: There is a full-time opening for a core facility light/confocal microscopist available immediately. The encumbent must demonstrate total facility with current technology and sample preparations. They will be responsible for 1) providing direct hands-on access to light microscopy in a core facility setting, 2) overseeing daily facility operations, 3) consulting with users on experiment design, materials and methods, and instrument optimization and assist in data acquisition and analysis, 4) troubleshooting problems with microscopes or samples and 5) assuring instrument integrity. Must maintain a current understanding of the microscopy field as necessary to implement new technologies on campus. Applicants must have a strong desire to participate actively as a member of a team, possess excellent written and oral communication skills, be comfortable working independently, communicate effectively with service users to understand their microscopy needs, be facile with PC and Mac computer platforms and demonstrate effective troubleshooting skills. M.S. in biological sciences with five years research experience in molecular biology, immunohistochemistry and cytology is required. Previous work in a core facility is preferred.
The Jackson Laboratory is one of the world's foremost centers for mammalian genetics research. Located in Bar Harbor, Maine, the lab is adjacent to Acadia National Park. Mountains, ocean, forests, lakes, and trails are all within walking distance. If you are looking for a more natural environment, this could be the opportunity you've been searching for.
Interested applicants should send cover letter and resume to: jax/210 Human Resources, Box 27 The Jackson Laboratory 600 Main Street Bar Harbor, Maine 04609 Fax: (207) 288-6106 Email (preferred option): jobs-at-jax.org The Jackson Laboratory is an Affirmative Action/Equal Opportunity Employer.
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Paula Sicurello wrote: ================================================================== I am attempting to do a negative stain (which I know how to do) on a sample. the problem I'm having is that according to the protocol I have to glow discharge the grid before I use it.
I don't have access to a vacuum evaporator with a glow discharge unit. So what's a tech to do?
Any suggestions as to how to fake a glow are greatly appreciated.
Please help me to glimmer, glimmer. ;-)
Unable to glow, =================================================================== If this is the kind of treatment one would apply to carbon coated grids to make them more hydrophilic, we can produce the same effect in the SPI Plasma Prep II Plasma Etcher but with an "air" plasma instead of an oxygen plasma (using oxygen would change your sample, actually it would probably etch away and effectively destroy your sample). The Plasma Prep II unit is shown on URL http://www.2spi.com/catalog/instruments/etchers1.shtml
We would be happy to do a "demo" of the technique for you if you sent us your grids in a grid box. Contact me off-line and I will give you shipping instructions. When this technique is used to make carbon coated grids hydrophilic, the effect seems to last something like 90 days.
Disclaimer: SPI Supplies is the manufacturer of the Plasma Prep II plasma etcher/cleaner/asher so we would have a vested interest in seeing more of them sold.....
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 ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id PAA16630 for dist-Microscopy; Thu, 14 Aug 2003 15:24:37 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id PAA16626 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Thu, 14 Aug 2003 15:24:06 -0500 (CDT) Received: from web40206.mail.yahoo.com (web40206.mail.yahoo.com [66.218.78.67]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id PAA16618 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 14 Aug 2003 15:23:53 -0500 (CDT) Message-ID: {20030814202245.40055.qmail-at-web40206.mail.yahoo.com} Received: from [137.131.240.13] by web40206.mail.yahoo.com via HTTP; Thu, 14 Aug 2003 13:22:45 PDT
Dear Steve and other microscopist,
The Formvar on your Carbon film is hard to be cleaned completely. The rest of Formvar could cause several problems, such as charging, poor stigmatism, drifting of specimen and even chemical reaction with your sample in EM.
I suggest you to try the pure carbon film coated grid, therefore you don't need to clean the Formvar on your grids. This carbon film didn't touch with any plastics while it was made. The pure carbon film is floated on the surface of water and sit on the cleaned grids.
We do produce this kind of grid. For more information, please check our website at
http://www.grid-tech.com/
Good luck.
Wendy ===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
--- Steve Parry {sparry-at-cmm.uwa.edu.au} wrote: } } } I have recently switched to Formvar in Diethylene } Chloride as a base } for making carbon coated grids, because I was told } that I would get } bettter continuous films, the problem I am having is } that I cannot } get rid of the forvar after coating. } Does anyone out there have a method for making C } coated grids that } works everytime?? I am getting to the end of my } patience and have } wasted an awful lot of grids over the last few } months. } } Thanks } } Steve Parry } -- } Steve Parry } Centre for Microscopy & Microanalysis, } (M010) } The University of Western Australia, } 35 Sterling Highway, Crawley, } WA 6009, Australia. } } Fax: +61-8-9380-1087 } Email: sparry-at-cmm.uwa.edu.au } Phone: +61-8-9380-8057 [24 hour voicemail } attached] } }
===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
Karl Garsha {garsha-at-itg.uiuc.edu} ASKED THE FOLLOWING QUESTION:
} Does anybody know a quick recipe for "grid glue" made out of scotch } tape (or other sticky stuff) and some solvent for glueing films to } TEM grids? Thanks in advance. } -Karl } RESPONSE:
GRID GLUE can be prepared by dissolving the adhesive from 2 inches of Scotch Brand transparent tape in 10 ml of ethylene dichloride. Please be sure to use the transparent tape (old style) rather than the Magik Tape since the adhesives are apparently different. I believe that chloroform can also be used as a solvent.
It might also be posible (and I invite microscopy vendors to chime in) to use the commercially prepared Grid-Stick Glue (EMS sells it, for example) but it will have to be diluted.... I have not tried this, however.
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 Email: bozzola-at-siu.edu ##############################################################
We are getting rid of a functioning Philips 505 SEM. Anyone with a 500 series scope that might like parts let me know ASAP. SEI,BEI,CL,W-hairpin (boxes of filaments), LAB6 (sorry no filaments - got IG pumps), pneumatic valves, bit and pieces, etc. etc.. . .
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
There are several possibilities here, but none of them are as good as the real thing.
1. Put your grids in the chamber of a sputter coater but cover the grids with a "tent" of filter paper. The tent should completely cover over the grids so that few of the metal particles will strike it but the argon ions will. Alternatively, you could turn the slide containing the grids upside down so that they face away from the target. Just be sure to leave some space underneath, so the ions can contact the grids. Activate the sputtering process so that you see the plasma for 40-50 sec. You might have to experiment here so that you minimize the presence of metal. If you have a carbon sputterer, I believe that would be the best.
2. You might try a dozen hits of the grids with one of the electrostatic guns (used by photographers). This sometimes does work.
3. Finally, and don't ask why this works, just leaving the grids for several days inside of a refrigerator will cause them to become hydrophilic. I am guessing that the trapped organics (odors, fumes, etc.) are depositing onto the active surface of the carbon. This is what happens when you place sodium bicarb in the refrig to trap odors.
Let us know which work best for you.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
A couple of times I just dissolved cellotape in chloroform - as much as could be squeezed into a small 10 ml sample bottle. Saturated solution? Not scientific but it helped when needed!
Keith Ryan Marine Biological Association of the UK & University of Plymouth, UK
----- Original Message ----- } From: "Karl Garsha" {garsha-at-itg.uiuc.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, August 14, 2003 4:19 PM
Hello, Just another question about our Electroscan E3 ESEM. Lately, we've noticed that when you start up the main console power, the right screen remains grey and doesn't load the microscope software such as the vacuum/gun/image menus. The left screen has a scale bar on it, but nothing works. I have to power down and then power up, sometimes 3x for it to boot up correctly.
Anyone have any advice where I should look for problems? Thanks for the help.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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 make a general purpose glue by stuffing several feet of Scotch Double Stick tape (other types would probably work as well) into a 100ml bottle then adding about 50 ml of heptane. Shake the bottle several times a day and in a few days you'll have a nice solution that will leave a sticky residue when it dries. This adhesive may not be the best to use in an electron microscope since it may outgas and contaminate your system--do some tests. Back in the 1970's there was a paper given at the MSA (EMSA) meeting describing a solution of Neoprene W in chloroform. It worked well with minimal outgassing but the only source of the Neoprene was DuPont. I have a few pieces buried somewhere but hopefully someone else has a beter source.
On Thu, 14 Aug 2003 10:19:19 -0500 Karl Garsha {garsha-at-itg.uiuc.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America
Larry Ackerman Advanced Imaging Lab/Dr. John Sedat/Dr. David Agard Dept. of Biochemistry & Biophysics University of California San Francisco 600-16th Street, Box 2140 San Francisco, CA 94143
This might be due to a bad connection / malfunctioning of a SCSI-card inserted in your computer to connect the computer with the microscope. If one connection fails, ik can interfere other connections too. Try to see how the pc starts up when you disconnect the microscope from it. Then start up again after connecting one by one.
Another possibility is, if you have SCSI-hardware, there are conflicts with the startup-sequence. In other words, SCSI-hardware should start up in a particular order (one has to start up BEFORE/AFTER another). If you just added a new piece of hardware, the numbering might be conflicting.
Sven Terclavers
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We sometimes have the same problem with our E-3 scope. In our case it occurs because the disk drive does not begin to read the disk. In that case the yellow light on the disk drive will not light up when you start the microscope.
We have developed a special trick for our E-3 scope, where we give the disk a slight pressure to the left at the end of the insertion into the drive. When the disk has been inserted that way, it can usually be read by the drive.
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: 14. august 2003 23:56 To: Microscopy-at-sparc5.microscopy.com
Hello, Just another question about our Electroscan E3 ESEM. Lately, we've noticed that when you start up the main console power, the right screen remains grey and doesn't load the microscope software such as the vacuum/gun/image menus. The left screen has a scale bar on it, but nothing works. I have to power down and then power up, sometimes 3x for it to boot up correctly.
Anyone have any advice where I should look for problems? Thanks for the help.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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 use double sided tape in chloroform (a ball of tape that is just covered by the chloroform). Shake the bottle for 20 sec. and remove what is left of the tape (you want the glue off of the tape, not the plastic). Not too scientific, but it works. David
On Thursday, August 14, 2003, at 11:19 AM, Karl Garsha wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Does anybody know a quick recipe for "grid glue" made out of scotch } tape (or other sticky stuff) and some solvent for glueing films to TEM } grids? Thanks in advance. } -Karl } } -- } Karl Garsha } Light Microscopy Specialist } Imaging Technology Group } Beckman Institute for Advanced Science and Technology } University of Illinois at Urbana-Champaign } 405 North Mathews Avenue } Urbana, IL 61801 } Office: B650J } Phone: 217.244.6292 } Fax: 217.244.6219 } Mobile: 217.390.1874 } www.itg.uiuc.edu } } } } ____________________
David Elliott Ph.D.
Yale University School of Medicine Campus Address: TAC S160 333 Cedar Street PO Box 208022 New Haven, CT 06520-8022
Re method 3. placing grids in a fridge, do you put them in on filter paper in a petri dish with the lid on, for instance?
Dave
On Thu, 14 Aug 2003 15:45:26 -0500 "John J. Bozzola" {bozzola-at-siu.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Paula, } } There are several possibilities here, but none of them are as good as } the real thing. } } 1. Put your grids in the chamber of a sputter coater but cover the } grids with a "tent" of filter paper. The tent should completely cover } over the grids so that few of the metal particles will strike it but } the argon ions will. Alternatively, you could turn the slide } containing the grids upside down so that they face away from the } target. Just be sure to leave some space underneath, so the ions can } contact the grids. Activate the sputtering process so that you see } the plasma for 40-50 sec. You might have to experiment here so that } you minimize the presence of metal. If you have a carbon sputterer, I } believe that would be the best. } } 2. You might try a dozen hits of the grids with one of the } electrostatic guns (used by photographers). This sometimes does work. } } 3. Finally, and don't ask why this works, just leaving the grids for } several days inside of a refrigerator will cause them to become } hydrophilic. I am guessing that the trapped organics (odors, fumes, } etc.) are depositing onto the active surface of the carbon. This is } what happens when you place sodium bicarb in the refrig to trap odors. } } Let us know which work best for you. } } JB } } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/~image/ } ############################################################## } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
We have used "grid pens" with glue with great success for our immunocytochemistry work. Initially, we had put up with LR White and Unicryl sections looking like a ragged piece of kleenex in the TEM. Then we moved to formvar/carbon grids, but had severe problems with "stickiness" causing high background labeling, apparently due to both antibody binding and mechanical trapping of the gold conjugate, not to mention various other folding and layering artifacts. Finally, after a suggestion from someone on this list, we tried the pens. Now our sections remain largely intact, the background problem is under control, and we are much happier campers. We do not dilute the glue in the pens, but just put one dab on grids on filter paper and let them dry a couple minutes before picking up our sections.
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Thursday, August 14, 2003 3:29 PM To: Microscopy-at-sparc5.microscopy.com
Karl Garsha {garsha-at-itg.uiuc.edu} ASKED THE FOLLOWING QUESTION:
} Does anybody know a quick recipe for "grid glue" made out of scotch } tape (or other sticky stuff) and some solvent for glueing films to } TEM grids? Thanks in advance. } -Karl } RESPONSE:
GRID GLUE can be prepared by dissolving the adhesive from 2 inches of Scotch Brand transparent tape in 10 ml of ethylene dichloride. Please be sure to use the transparent tape (old style) rather than the Magik Tape since the adhesives are apparently different. I believe that chloroform can also be used as a solvent.
It might also be posible (and I invite microscopy vendors to chime in) to use the commercially prepared Grid-Stick Glue (EMS sells it, for example) but it will have to be diluted.... I have not tried this, however.
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 Email: bozzola-at-siu.edu ##############################################################
first, have you tried a standard negative stain preparation? a lot of the time there is enough protein present in the sample to overcome the hydrophobicity. just leave the sample on the grid for about a minute.
second, try an airfuge, EM-90 preparation. the combination of centrifugal force for 1/2 hr plus protein present in the preparation usually overcomes the need for glow discharge.
third, try pre-treating the grids with either poly-L-lysine or Alcian blue. use a 1% solution, float the grids on the solution for 1-2 minutes, wick off the excess, then let dry
one of these should work
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
Michael, The 1.65 NA objective is not absolutely necessary, but as you aware TIRF needs higher NA (NA} n). But you can obtain fairly good images with 1.45 NA lenses available now with Nikon and Zeiss. The 1.65 NA available only with Olympus as of now and you may need to pay a higher price for that. I had an introductory practical course experience only with Olympus system with both 1.45 and 1.65 lenses. If you need any other information please let me know by email.
Shiv
Mayandi Sivaguru, Ph.D., Associate Director Molecular Cytology Core Facility Molecular Biology Program 2, Tucker Hall University of Missouri Columbia, MO 65211-7400 sivagurum-at-missouri.edu
Voice: 1-573-882-4895 Fax: 1-573-882-0123
www.biotech.missouri.edu/mcc/
At 12:42 PM 8/14/03 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It came to my mind, that exposure of the grids to the strong UV (real UV) light do the trick also. Effect will vary depends from the UV source and intensity. As I remember, grids are good for about 40 min after all. Personally, I prefer to use poly-lysine treatment. In my hands it works nearly the same as glow discharge. Even better, because it does not made surface rough. Sergey
At 01:45 PM 8/14/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, August 13, 2003 at 02:56:55 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin Andreev
Organization: 54 "st.Ivan Rilski"
Education: 6-8th Grade Middle School
Location: Sofia Bulgaria
Question: How can I prepare permanent slides for examination under microscope?What materials should I use for preserving the speciment for a very long time?
Veselin- There are commercial preparations you can buy, but you can also use clear nail polish if you do not have access to these cements. But first- you must remove the water from your specimen, by soaking in alcohol. First water plus alcohol, then gradually reducing the water content. Making permanent slides takes a different process depending upon what the specimen is. What are you mounting on the slide? Rgds, Mike Shaw } } Email: sswaffe-at-abv.bg } Name: Veselin Andreev } } Organization: 54 "st.Ivan Rilski" } } Education: 6-8th Grade Middle School } } Location: Sofia Bulgaria } } Question: How can I prepare permanent slides for examination under microscope?What materials should I use for preserving the speciment for a very long time? } } ------------------------------------------------------------ } ---------------
Richard I use Electron Microscopy, 2nd edition by John Bozzola and Lonnie Russell for my graduate TEM course. It is an excellent book and my students really enjoy it, especially the survey of biological ultrastructure at the back.
My favorite is "A beginners handbook in Biological Transmission Electron Microscopy", by Brenda S. Weakley, Chruchill Livingstone is the publisher. My second edition is 1981, there may be a more recent edition. "Biological Techinques in Electron Microscopy" by Clinton Dawes is also good but my copy is so old (1971) it may be out of print.
Geoff
Richard Gardiner wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
You can easily use the method described by Mike to make about 50 pure carbon grids at a time (much cheaper than buying if you already have the equipment).
You place about 50 grids on a square piece of filter paper lying on a little "table" made of wire mesh under the water surface. To get the grids under water make them wet first and push them through the surface vertically. Then you lower the water surface by removing water from underneath (for example with a syringe or a plastic tube with a valve) to place the carbon film (floated off the mica) on top of the grids. (Of course the carbon film has to be in one piece in this case.) Push the carbon into position with something blunt (a paddle ? :)
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em _______________________________________
} } Steve, } } When I want just carbon as a substrate I will } } coat freshly cleaved mica with carbon in a metal } } evaporator. This allows me to dictate the carbon } } thickness and I have no trouble floating off grid } } size squares(prescored)in a drop of water. Picking up the } } carbon with glow discharged grids (flat side) } } is a snap. Make sure everything is clean. } } This should give you thin clean substrates. } } Mike D.
I've used the Bozzola and Russell textbook entitled "Electron Microscopy: Principles and Techniques for Electron Microscopy" since the first edition was published in 1992. A second edition was published in 1999. It was especially suited to my needs as I taught both an SEM and a TEM course each year - primarily to undergraduate students. The students were able to use the same text for both courses. The authors provide a good general background on theory, and an excellent treatment of procedures and techniques for both types of microscopy. Although there may be other texts out there that are suitable as well, I highly recommend this one as a text to teach from.
Doug Bray
Richard Gardiner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello: } } I was wondering what text books people are using to teach either } Undergraduate or Graduate courses in Biological TEM. } } Richard } Gardiner
Bozzola, John J., Russell, Lonie D. - Electron Microscopy - Principles and Techniques for Biologists.
Alice Dohnalkova Environmental Microbiology Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 tel. (509) 372-0692 office (509) 376-3654 TEM lab fax (509) 376-1321
} -----Original Message----- } From: Richard Gardiner [SMTP:rbgardiner-at-rogers.com] } Sent: Saturday, August 16, 2003 12:09 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: EM Textbooks Used } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello: } } I was wondering what text books people are using to teach either } Undergraduate or Graduate courses in Biological TEM. } } Richard } Gardiner } }
Hi, We use PTA to stain thin sections of human skin for collagen. I make up a fresh solution of 1% PTA in dH20. Mix thoroughly and filter twice through a #1 Whatman. Leave the pH acid. Don't adjust it toward neutral (it won't stain as well). Stain sections 30 mimutes. staining sequence should be: 1) PTA 2) UA 3) PB
Robert Underwood Dermatology Research Center University of Washington
On Mon, 18 Aug 2003, Dusevich, Vladimir wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need a protocol for staining collagen thin sections with } phosphotungstic acid. } } Thanks a lot, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } }
With many of you looking for extra money for microscopy labs and the recent convergence of microscopy and spectroscopy, I thought this small grant program might be of interest.
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ANNOUNCEMENT
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5. Previous awardee colleges are ineligible for the PCMNCG program for a three-year period following receipt of the grant (awardee colleges from 2001, 2002 and 2003 are not eligible for the 2004 program).
6. Grant applications and proposals must be received no later than December 1, 2003. Faculty members may obtain application forms and additional information from:
Rita M. Windisch, Ph.D. The Pittsburgh Conference PCMNCG 300 Penn Center Blvd., Suite 332 Pittsburgh, PA 15235-5503 Telephone (412) 825-3220, Ext. 204 FAX (412) 825-3224 E-mail: windisch-at-pittcon.org
or from the Pittsburgh Conference Website: www.pittcon.org Announcement of the awards will be made by February 2004. Schools chosen will join the list of over 200 institutions honored since the inception of this program in 1974.
I think Dawes is still available from Ladd Research. But I prefer Bozzola and Russell for a text, and Maunsbach and Afzelius "Biomedical Electron Micropscopy" for lots of EMs comparing fixatives, buffers, embedding media, and so forth.
Phil
} My favorite is "A beginners handbook in Biological Transmission } Electron Microscopy", by Brenda S. Weakley, Chruchill Livingstone } is the publisher. My second edition is 1981, there may be a more } recent edition. "Biological Techinques in Electron Microscopy" by } Clinton Dawes is also good but my copy is so old (1971) it may be } out of print. } } Geoff } } Richard Gardiner wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Phillip Your way to mount carbon on the grids has one very serious disadvantage: when you mount carbon this way, you will use "air-side" of carbon as a working surface. This side is usually more grainy and dirty. The whole idea of using carbon from mica is to use "mica-side" of carbon as a working surface. The idea here is that the carbon face toward mica is more uniform and perfectly clean (it was not exposured to the air). In order to use "good" side of carbon, you have to put grids on floated carbon and then pick the grids with Parafilm for instance. Similarly, you need to do the same for plastic film created on water - working side is "water-side", not air. Only one exception I do know is Formvar film created on the glass. Glass is not such clean and perfect as mica or water surface, therefore, you need to use "air-side". Then your technique will work perfectly. I also use your technique to mount holey film on the grids. Best wishes, Sergey.
At 04:07 PM 8/18/03 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ssamuelsson-at-eyetk.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, August 18, 2003 at 14:09:26 ---------------------------------------------------------------------------
Email: ssamuelsson-at-eyetk.com Name: Steve Samuelsson
Organization: Eyetech Pharmaceuticals
Title-Subject: Whole Mounts
Question: Regarding growing endothelial cells on nickel grids for whole mount TEM analysis: our luck is inconsistent and we now need to dig further for the reason. 200 and 300 mesh nickel grids are coated with ~0.5% formvar in ethylene dichloride (we also tried chloroform) and picked up on a 13mm cover slip. These are allowed to dry and age a day or longer. Prior to plating, the slips are UV sterilized 10-15', incubated in 1% gelatin ~45', washed 1x with PBS, culture medium added followed by cells. Our issue is incomplete (or downright lousy) growth of cells on the grids. Cells are robust and growing fine on the bottom of the 24 well plate so we suspect something with the grids. My first thought was that we needed carbon coating of the grids but have some trouble with this explanation. Second thought was that the solvent was still coming out of the films and poisoning the cells; thus the 'aging' issue with the cover slips. Any suggestions?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hafeez189-at-yahoo.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 18, 2003 at 14:38:56 ---------------------------------------------------------------------------
Email: hafeez189-at-yahoo.com Name: Hafeez Anwar
Organization: University of Agriculture, Faisalabad, Pakistan
Education: Graduate College
Location: Pakistan
Question: hi, i am working on optical polarization with hot stage and i am studying the morphology of polymer blends. my question is that why the colors of the sample change when we rotate the analyzer ?(kindly ,explain in detail)
A user has the following question. Please reply off line to her at: DCOLLISO-at-glcc.com (Donna Collison)
Does anyone have experience in staining 'toughened nylon' for backscattered compositional analysis. Toughened nylon as we are describing it is nylon having butadiene rubber phases within the nylon matrix. We need to determine were an antimony component is within the formulation. So far we have placed fractured pieces of this material directly in osmium tetroxide for a 24 hour period and was not able to detect any contrast. Any suggestions and help would be greatly appreciated.
Many thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
I need a little advice on safely examining plants infected with Salmonella sp. I have a user who wants to use cryofixation of melon and tomato plants that have been inoculated with the Salmonella sp. bacterium. She would like to bring the inoculated plants into the EM facility, dissect a small bit of the plant surface and then do a conventional cryo prep and examine in the SEM. I am not familiar with the safest method of examining pathogens in the SEM. Can the bacteria survive cryo fixation? What is the best method of cleaning the scope after use? What about the instruments used for dissection? As you can see, I have a lot of questions and before we proceed with this project I need to satisfy myself that we are doing this correctly.
Thanks for you help.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://microscopy.mcb.ucdavis.edu raharris-at-ucdavis.edu
On Mon, 18 Aug 2003 17:59:25 -0500 by way of Ask-A-Microscopist {ssamuelsson-at-eyetk.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ssamuelsson-at-eyetk.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, August 18, 2003 at 14:09:26 } --------------------------------------------------------------------------- } } Email: ssamuelsson-at-eyetk.com } Name: Steve Samuelsson } } Organization: Eyetech Pharmaceuticals } } Title-Subject: Whole Mounts } } Question: Regarding growing endothelial cells on nickel grids for whole mount TEM analysis: our luck is inconsistent and we now need to dig further for the reason. 200 and 300 mesh nickel grids are coated with ~0.5% formvar in ethylene dichloride (we also tried chloroform) and picked up on a 13mm cover slip. These are allowed to dry and age a day or longer. Prior to plating, the slips are UV sterilized 10-15', incubated in 1% gelatin ~45', washed 1x with PBS, culture medium added followed by cells. Our issue is incomplete (or downright lousy) growth of cells on the grids. Cells are robust and growing fine on the bottom of the 24 well plate so we suspect something with the grids. My first thought was that we needed carbon coating of the grids but have some trouble with this explanation. Second thought was that the solvent was still coming out of the films and poisoning the cells; thus the 'aging' issue with the cover slips. Any suggestions? } } } --------------------------------------------------------------------------- } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
My own experience has been that only gold or titanium grids give consistent results with this procedure. Both copper and nickel tend to be quite toxic.
Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals Ridgefield, CT
-- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Could the nickel be toxic? Why not try gold? } } Dave } } } On Mon, 18 Aug 2003 17:59:25 -0500 by way of } Ask-A-Microscopist {ssamuelsson-at-eyetk.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted } by (ssamuelsson-at-eyetk.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, } August 18, 2003 at 14:09:26 } } --------------------------------------------------------------------------- } } } } Email: ssamuelsson-at-eyetk.com } } Name: Steve Samuelsson } } } } Organization: Eyetech Pharmaceuticals } } } } Title-Subject: Whole Mounts } } } } Question: Regarding growing endothelial cells on nickel grids for whole mount } TEM analysis: our luck is inconsistent and we now need to dig further for the } reason. 200 and 300 mesh nickel grids are coated with ~0.5% formvar in ethylene } dichloride (we also tried chloroform) and picked up on a 13mm cover slip. These } are allowed to dry and age a day or longer. Prior to plating, the slips are UV } sterilized 10-15', incubated in 1% gelatin ~45', washed 1x with PBS, culture } medium added followed by cells. Our issue is incomplete (or downright lousy) } growth of cells on the grids. Cells are robust and growing fine on the bottom } of the 24 well plate so we suspect something with the grids. My first thought } was that we needed carbon coating of the grids but have some trouble with this } explanation. Second thought was that the solvent was still coming out of the } films and poisoning the cells; thus the 'aging' issue with the cover slips. Any } suggestions? } } } } } } --------------------------------------------------------------------------- } } } } } } } } This incoming email to UWE has been independently scanned for viruses and any } virus detected has been removed using McAfee anti-virus software } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } }
Suppliers are, in my experience, reluctant to comment on the problem potential hazards of non chemically fixed microorganisms in cryoSEM orlow vacuum/ESEM.
I have seen a paper showing that some bacteria (which species?) survive cryoSEM sessions. In theory they could get pumped into your rotary pump and out of the room (comments anyone?).
A colleague looked for presence of bacteria in the chamber after ESEM and found little, I think. (I will e-mail her for comment).
Here if a colleague can show the bacteria are harmless we would consider looking at them unfixed. If they are pathogenic I insist they are fixed first.
Your colleague should advise on cleaning tools. One of our users cleaned forceps in 90% ethanol. You can wipe over the stage and chamber but what about the column?
I hope you have started an interesting debate on a topic we mull over here periodically.
Dave
On Mon, 18 Aug 2003 16:52:52 -0700 "Rick A. Harris" {raharris-at-ucdavis.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need a little advice on safely examining plants infected with Salmonella } sp. I have a user who wants to use cryofixation of melon and tomato plants } that have been inoculated with the Salmonella sp. bacterium. She would like } to bring the inoculated plants into the EM facility, dissect a small bit of } the plant surface and then do a conventional cryo prep and examine in the } SEM. I am not familiar with the safest method of examining pathogens in the } SEM. Can the bacteria survive cryo fixation? What is the best method of } cleaning the scope after use? What about the instruments used for } dissection? As you can see, I have a lot of questions and before we proceed } with this project I need to satisfy myself that we are doing this correctly. } } Thanks for you help. } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } http://microscopy.mcb.ucdavis.edu } raharris-at-ucdavis.edu } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I highly recommend Bozzola & Russell's Electron Microscopy 2nd ed. I use it for my graduate level EM course.
Cheers,
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://confocal.nmsu.edu/eml
On Sat, 16 Aug 2003, Richard Gardiner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello: } } I was wondering what text books people are using to teach either } Undergraduate or Graduate courses in Biological TEM. } } Richard } Gardiner } } }
I have a lot of trouble making too Epon/Araldite in this weather of high humidity. Although our hospital is air conditioned, it still seems to leave a lot of humidity in the air [read: it's almost useless], and it seems that we always having a lot of trouble infiltrating and getting the thin sections to stay together in this sort of weather. Our thin sections break either in the boat when we are cutting, or rapidly under the beam of the microscope.
Is there any way of modifying the formula of our plastic to compensate for this problem?
This is the formula that we presently use to make our plastic:
On Monday, August 18, 2003, at 04:52 PM, Rick A. Harris wrote:
} I need a little advice on safely examining plants infected with } Salmonella } sp. I have a user who wants to use cryofixation of melon and tomato } plants } that have been inoculated with the Salmonella sp. bacterium. She } would like } to bring the inoculated plants into the EM facility, dissect a small } bit of } the plant surface and then do a conventional cryo prep and examine in } the } SEM. I am not familiar with the safest method of examining pathogens } in the } SEM. Can the bacteria survive cryo fixation? What is the best method } of } cleaning the scope after use? What about the instruments used for } dissection? As you can see, I have a lot of questions and before we } proceed } with this project I need to satisfy myself that we are doing this } correctly. } Dear Rick, We do some cryo-TEM on (mild) pathogens, and our safety-office-approved protocol is to do the preparation--growth and freezing--in a biosafety hood. We are allowed to transfer the grids, which are kept under LN2, to the cryostage as with any other specimen, then the grids are dropped into bleach after examination. You should assume that Salmonella will survive cryofixation, and treat your specimens, tools, stubs, etc. as potential sources of infection. Check with your safety office for the best sterilization method for Salmonella, but bleach or detergent will usually disrupt bacteria. I'm not at all certain what would be best for the inside of the SEM, but, if your specimens remain frozen for the duration of their time inside the scope, you may be able to avoid putting bleach or detergent into your instrument. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
On Monday, August 18, 2003, at 04:00 PM, by way of Ask-A-Microscopist wrote:
} Question: hi, i am working on optical polarization with hot stage and } i am studying the morphology of polymer blends. } my question is that why the colors of the sample change when we rotate } the analyzer ?(kindly ,explain in detail) } Dear Hafeez, Your sample causes a rotation of the polarization vector of the light passing through it. The amount of the rotation is proportional to the thickness of the sample, and it will vary with the wavelength of the light. The phenomenon is called "optical rotary dispersion" (if I remember my optics correctly), and the details should be available in a physics text. Consider a simple case. If you are looking through a slab of material with uniform thickness, the light entering the sample will be polarized, say, at 12 o'clock. As it passes through the slab, the polarization of each wavelength is rotated to a different extent, say, red to 5 o'clock, green to 6 o'clock, and blue to 7 o'clock. When the analyzer is oriented at 5 o'clock, the sample will appear red, etc. If the sample thickness varies, the apparent color at a given analyzer position will vary with thickness, and, as the analyzer orientation changes, the colors will shift. If your specimen is wedge-shaped, you should see the colors in the same order as in a rainbow--assuming that the dispersion is monotonic with wavelength, as it usually is--and the colors will move pretty much in unison along the slope of the wedge as the analyzer is rotated. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Epon is somewhat miscible with water, a colleague regularly goes from 95% to an Epon:alcohol mix, so humidity may not be the problem. First I would try all fresh plastics, Epon, Araldite and especially DMP-30 which has a short shelf life. You could also try longer times (or smaller pieces of tissue) and agitation to be sure infiltration is not the problem. It could be that your plastics have gotten 'tired' just as the weather has become more humid?
Geoff
Garry Burgess wrote:
} I have a lot of trouble making too Epon/Araldite in this weather of high } humidity. Although our hospital is air conditioned, it still seems to leave } a lot of humidity in the air [read: it's almost useless], and it seems that } we always having a lot of trouble infiltrating and getting the thin sections } to stay together in this sort of weather. Our thin sections break either in } the boat when we are cutting, or rapidly under the beam of the microscope. } } Is there any way of modifying the formula of our plastic to compensate for } this problem? } } This is the formula that we presently use to make our plastic: } } Araldite: 65 grams } Epon [JEMBED 812 Epon relacement]: 81 grams } DDSA: 232 grams } DMP-30: 7.2 grams. } } } Garry Burgess } Charge Technologist } Department of Pathology - EM Laboratory } Health Sciences Centre } Winnipeg } } }
-- -- ********************************************** 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 **********************************************
One thing I've found that at least helps is to pre-cook the embedding molds to sort of dry them out. Also, store the resin components under vacuum. Don't know about changing your recipe....
Good luck!
Tamara
On Tue, 19 Aug 2003, Garry Burgess wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have a lot of trouble making too Epon/Araldite in this weather of high } humidity. Although our hospital is air conditioned, it still seems to leave } a lot of humidity in the air [read: it's almost useless], and it seems that } we always having a lot of trouble infiltrating and getting the thin sections } to stay together in this sort of weather. Our thin sections break either in } the boat when we are cutting, or rapidly under the beam of the microscope. } } Is there any way of modifying the formula of our plastic to compensate for } this problem? } } This is the formula that we presently use to make our plastic: } } Araldite: 65 grams } Epon [JEMBED 812 Epon relacement]: 81 grams } DDSA: 232 grams } DMP-30: 7.2 grams. } } } Garry Burgess } Charge Technologist } Department of Pathology - EM Laboratory } Health Sciences Centre } Winnipeg } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
Now that the safety nazis have forced us to wear face shields, smocks and gloves for handling liquid nitrogen, they want us to wear safety glasses while doing wedge polishing, which entails frequent looking through a microscope to judge the progress. The concern is that some polish slurry 'could' splash into the eyes of a technician. Of course the safety glasses really get in the way when looking through a microscope, and our technicians are really not happy about this. I would like to hear as many opinions as possible on whether there is significant likelyhood of such a splash, and if polishing slurry, like Siton or Glanzox really poses a hazard to the eyes of a person using it. Thanks.
This is a simple matter of your company doing risk assessment and covering their butts to prevent law suits and higher insurance premiums. I don't think it matters if it is silicon polish, rock chips off a jack hammer or any other flying material, a company/institution wants to lessen liability and increase safety. By telling you and your techs that you MUST wear these items, they have less liability if someone does get something in their eye.
Example A: Occupational safety does not say to wear the eye shields and Technician A gets abrasive slurry compound in his/her eye. Depending on how that compound affects the eye, he/she is now getting paid comp/injury leave and possibly preparing a lawsuit against the company for not providing a safe work place while OSHA is looking for answers and handing out fines as well.
Example B: Occupational safety says to wear the eye shields and Technician A does not because its an "inconvenience". Technician A gets compound in his/her eye. The company can now say, "Sorry but you were negligent in following proper protocol." The company's management and insurance company can rest easy knowing that any lawsuit has little to no validity due to the negligence of the employee.
I doubt there is much you can do about a perceived safety threat. If you can state that productivity is being hampered due to the implementation of these new safety protocols, management will probably listen to your complaint. What do the directions and MSDSs say for these materials? Do the manufacturers suggest wearing eye shields? I hear Oakley safety spec glasses are expensive but comfortable.
Probably not what you wanted to hear but, for the most part, it's the hard truth. Whether these materials pose a real threat or not, occupational safety views it as a risk and you have to follow the rules. Allowing anyone you supervise to take their glasses off would put your position in jeopardy. Just something to think about.
Chris Zurenko Univ. of Michigan KHRI/Otopathology
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Now that the safety nazis have forced us to wear face shields, smocks } and gloves for handling liquid nitrogen, they want us to wear safety } glasses while doing wedge polishing, which entails frequent looking } through a microscope to judge the progress. The concern is that some } polish slurry 'could' splash into the eyes of a technician. Of course } the safety glasses really get in the way when looking through a } microscope, and our technicians are really not happy about this. I would } like to hear as many opinions as possible on whether there is } significant likelyhood of such a splash, and if polishing slurry, like } Siton or Glanzox really poses a hazard to the eyes of a person using it. } Thanks. } } John Mardinly } Intel
I follow your logic, but it is logic I will resist to the day I retire (which may just have come a little closer!). Possible outcomes for this line of thinking are that many legitimate research activities will effectively be denied us, and more worryingly that this approach to safety is a way for comapnies to deny their responsibilities to employees for their safety. Would you want to work for an employer whose rules effectively abolished any accident insurance. Just something to think about. Chris
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Facility
----- Original Message ----- } From: "Christopher S. Zurenko" {czurenko-at-umich.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, August 20, 2003 3:14 AM
In high humidity Michigan, I routinely used an Epon-Araldite receipe from Palevitz and Newcombe (JCB 45(2):383, 1970) that used 40ml DDSA, 20ml Epon (or replacement), 20ml Araldite 6005 and 1.44ml DMP-30. I used a glass syringe for dispensing the DMP-30, a glass rod to stir the resin, de-gassed the resin before use and between resin changes, and kept all components under vacuum. When my resin started behaving oddly I tossed all components and started with fresh.
Good luck!
Angela Welford, EMT Dept of Pathology University of New Mexico Albuquerque, NM, 87131 505-272-1445
The formula should not be changed. We would suggest storing the chemicals in a desiccator cabinet with a desiccant such as silica gel..
JD Arnott
Disclaimer: Ladd Research sells the chemicals and supplies mentioned in this e-mail.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, August 19, 2003 12:25 PM
Garry: I have been using an Epon/Araldite mixture here in Houston for at least the past 15 years with no problems. I doubt that you are any more hot and humid than I am here. I do not measure mine by weight but by volume. I use the following formula: 12 ml of Epon 812 (any substitute 812) 10 ml Araldite 502 24 ml DDSA 1 ml DMP 30
I do all of my measurements using a disposable plastic syringe. I remove the plunger and pour in an amount just over what I need (be sure to keep the tip cover on or it will flow out while you are pouring). After filling the syringe with slightly more than the desired amount, I carefully replace the plunger and invert the syringe, remove the cap and expel the air. Then I discard the small amount over what I need and then expel the measured portion into a disposable 50 ml polypropylene centrifuge tube. Brand does not matter as long as it has a sealable top (I am currently using Falcon Bluemax 2098). I use the same syringe for all of the first 3 components. I measure the DMP30 by placing the tip of a 2 or 3 ml syringe in the bottle and drawing up the desired amount into the syringe. After all the components have been place in the centrifuge tube, I mix by inversion on a rotator for at least 15 minutes. I know many people say you shouldn't mix by inversion because of the air bubbles but I find this is not a problem if I let the mix set for 15 to 20 minutes after mixing. My embedding protocol is as follows 50% ethanol for 15 minutes 70% ethanol for 15 minutes 95% ethanol for 15 minutes 100% ethanol for 15 minutes X2 100% propylene oxide for 15 minutes X2 1:1 Epon/araldite:propylene oxide for 1 hour Pure Epon/Araldite for 1 hour Place one piece of tissue on fresh plastic in a beem capsule and let it slowly settle to the bottom of the capsule. After 3 to 4 hours place in oven at 80o C overnight to polymerize. If you need more info you can email me at msteglic-at-mdanderson.org.
Mannie Steglich Tech Dir E M Lab U T M D Anderson Cancer Center Houston, TX
Steve, Karen, who was Dr. Keith Porter's Research Specialist, was in my department several years back. They were studing fish scale cells growing on gold grids that were coated but I do not recall with which plastic. The grids were carbon coated indirectly (a shield was placed between the rods and the grids). I do know that the grids were glow discharged while still on the coverslip which was used to pick up the coated grids. A thin strip of aluminum was used instead of the usual copper wire for the glow. The grids were UV sterilized and I believe they were put directly into the culture dishes with no washing/rinsing. They did use 400 kV. to see more than just the edge of the cells. I believe that this work was published before Dr. Porter passed away. Pat
Patricia Stranen Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
} Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (ssamuelsson-at-eyetk.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html } on Monday, August 18, 2003 } --------------------------------------------------------------------------- } Email: ssamuelsson-at-eyetk.com } Name: Steve Samuelsson } Organization: Eyetech Pharmaceuticals } Title-Subject: Whole Mounts } Question: Regarding growing endothelial cells on nickel grids for } whole mount TEM analysis: our luck is inconsistent and we now need } to dig further for the reason. 200 and 300 mesh nickel grids are } coated with ~0.5% formvar in ethylene dichloride (we also tried } chloroform) and picked up on a 13mm cover slip. These are allowed } to dry and age a day or longer. Prior to plating, the slips are UV } sterilized 10-15', incubated in 1% gelatin ~45', washed 1x with PBS, } culture medium added followed by cells. Our issue is incomplete (or } downright lousy) growth of cells on the grids. Cells are robust and } growing fine on the bottom of the 24 well plate so we suspect } something with the grids. My first thought was that we needed } carbon coating of the grids but have some trouble with this } explanation. Second thought was that the solvent was still coming } out of the films and poisoning the cells; thus the 'aging' issue } with the cover slips. Any suggestions? } ---------------------------------------------------------------------------
This has happened at our institution also, but for short-sighted folk like me who have to wear glasses on the microscope anyway, "they" made a real effort to get reasonably comfortable and not too large fully enclosed safety glasses with prescription lenses - must have cost a packet. They do have to worry about insurance, etc, but here, at least, although it's an annoyance, it's not too hard to follow the regulations.
} } Now that the safety nazis have forced us to wear face shields, smocks and } gloves for handling liquid nitrogen, they want us to wear safety glasses } while doing wedge polishing, which entails frequent looking through a } microscope to judge the progress. The concern is that some polish slurry } 'could' splash into the eyes of a technician. Of course the safety glasses } really get in the way when looking through a microscope, and our } technicians are really not happy about this. I would like to hear as many } opinions as possible on whether there is significant likelyhood of such a } splash, and if polishing slurry, like Siton or Glanzox really poses a } hazard to the eyes of a person using it. Thanks. } } John Mardinly } Intel
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
Years ago I grew crustacean neurosecretory cells on Formvar-coated gold grids, fixed, postfixed, dehydrated, and critical point dried them (we had a grid holder for our Tousimis CPD) for 400 kV TEM. They were pretty happy cells, and it worked well.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Being in the US university system for ten years now (wow, time is just flying) I was and is amused how this system handles safety issue. Students and most technical stuff have very little idea how to perform safe in the laboratory environment. Safety issue is limited to some bureaucratic tasks not related to the real safety (mostly keeping paperwork in a good shape and accurately fill out chemical waste disposal tags). So, in such situation, I think, it's superviser's personal responsibility to provide safely environment to the workers and train them accordingly. I think, the health issue is much, much more important than any personal or corporative ambitious. If it's SAFER to use protective glasses, it's your responsibility to ensure that your workers do use glasses. It's really sad, but I agree at some degree with Chris Zurenko, that safety issue sometime reflects not real carry about worker's health but attempt to avoid legal responsibility (which, in my point of view, nothing to do with real safety). Sergey
P.S. By the way: in my EM work, my glasses, I wear to correct my vision, save my eyes in uncounted number of times. I, also, used to remove them every time I am using light microscope or binoculars. I don't see big problem removing them if necessary. But: yes, it decreases my productivity. Ok, let say, I need about 5 sec. to remove them and put back. Let say I do it 40 times a day: 5 secx40=200 sec/3.3 min per day.
At 12:36 AM 8/20/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I'm not saying that I follow my own logic 100%. There are tasks that I perform in the lab everyday that I probably "should" be wearing more protective gear, but I don't, and I'm sure we all do. With regard to the original post, I'm not sure that I would want to wear a full smock for pouring liquid nitrogen either, but gloves and eye protection are a good idea.
The point I'm trying to make is that in a day and age of fast food chains getting sued because their fatty food makes people overweight and cigarette companies handing out huge settlements because people now have cancer, etc. (after doctors have been stating for YEARS that fatty food and cigarettes are not good for you), companies can not be too careful. That is the case here. The company implementing the eye guards does not want someone to get abrasive compound in their eyes. They are attempting to limit their risk and liability for someone being injured, very simple. You would like to think that the company has acted on more of a humanitarian agenda to provide for and protect its employees (and it may be a small piece of the pie) but I think it's more to protect the company.
I don't see how legitimate research would be denied due to the implementation of safety rules. There should be rules to limit risk, but you can never 100% eliminate it. Legitimate research grinds on everyday (following internal, local, state and federal safety mandates) and I don't see that ending anytime soon.
Back to the original post - if using this eyewear is creating a major problem for the type of research being conducted, talk with management and the safety people to attempt to implement a mutually beneficial setup where an eye splash could mostly be eliminated. Maybe some sort of Lexan guards or shielding rather than eyewear. As far as the specific question about the abrasive getting in the eye, I don't know but I don't think that matters (though I can't imagine any sort of abrasive in the eye would ever be a good or tolerable thing). It probably wouldn't matter if it's abrasive, oil, sewage, tap water or sterile saline that possesses a risk of getting slashed in the eye. Your safety people have identified a safety risk and have moved to protect the employees and company. Whether you or anyone else agrees with me or not, it is now the responsibility of techs to wear the glasses and supervisors to make sure they are worn, whether anyone is happy about it or not.
Cheers
-------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648
Garry: Please check to storing conditions of your components, the DDSA and less the DMP 30 are very hidroscopyc. The last one can be replaced with BDMA with very good results at ultramicrotomy time. Have a good luck!
Francisco Freire FRMS Head Electron Microscopy Service University of Las Palmas de Gran Canaria Canary Islands Spain
In my labs it is mandatory to wear safety glasses. Risk assessments are carried out on all microscope usage and, if deemed safe, can be given an exemption sticker (signed off by management) so the user can remove specs. That RA would cover what activities are occurring adjacent and whether the operative has wet hands/gloves as any contaminants could be transferred to hair/skin and indeed eyes during removal of the glasses.
Where the 'environment' remains unsafe we are now moving to replacing the eyepieces with those suitable for spectacle wearers (safety or not). Non spectacle users in Histopath labs here are also having them fitted for doing their 'safe' microscopy as they prefer the ergonomics of them.
So change the microscopy set up and every-one should be safe and happy.
Gillian Brown Histology Section, Asthma Biology RI CEDD, GSK.
After a massive (or at least sobig) virus attack I'm back online.
That's a very interesting point Sergej makes.
We work with thin (1 layer) protein crystals and large individual protein molecules (chaperones etc.).
For crystals the flatness of the carbon surface is obviously important but for single proteins I've always thought it shouldn't make any difference. Initially we use negative stain to look at the quality of a specimen. To get higher resolution data we use cryo-specimens on holey carbon so that is a different story. Back to the negative stain: I've been wondering about two possible effects when staining: 1. When you stain a specimen on the rough side the stain might fill all the valleys of the carbon surface leading to large background noise and making it difficult to see the particles (some grids seem to have a lot of background). 2. When you stain a specimen on the smooth side (which is facing the grid) the stain might get trapped between the grid bars leading to an unnecessarily thick layer of stain which again increases the noise.
Are either of the two real problems?
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em _______________________________________
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, August 18, 2003 10:06 PM
The attachments with .pif extensions
This is the virus you may have. http://securityresponse.symantec.com/avcenter/venc/data/w32.sobig.f-at-mm.html
Note that the virus is spoofing e-mail addresses. so your system could be clean.
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
first, i am not really sure what the position of chris jeffree is, i found his response a bit confusing. sergey's comments considering time are to the point, and rosemary describes the situation i have experienced quite accurately. from my view, after thirty years i still have my eyes. last time i checked, they were a bit important for my work.
in a previous persona (before i grew up and went to graduate school) i had the privilege of negotiating collective agreements on behalf of an independent union which represents university non-academic staff. at the university of manitoba this means about 2000 administrative, clerical, computer, library, security and technical employees. the technical staff were, and still are about 40% of the population. for the record, i was chief negotiator, not member of the team, and in one of those cases we were on strike for 7 weeks. hopefully this sets it clear with chris zurenko that i was not a management stooge. the issue of safety supplies and equipment was always important, even in the dark ages of 1975. in fact, historically most improvements in working conditions and staff safety are the result of labour organizations fighting for them. five day work weeks, vacations, and child labour laws were not the result of an insurance company telling managers they had to make the changes. they were the result of the workers fighting for better and safer working conditions. in our case, we identified problems long before any insurer or government agency ever did. what is most important is that the university of manitoba, at least, never argued the importance of the issues. usually, our only problem was figuring out how we could ensure payment for the needed materials in individual grant funded positions.
having worked in the university research system for over 30 years i have seen most different approaches. to make it clear, i do not think universities are any more concerned with safety than any employer. have the 'safety nazis' made it more difficult, yes. but at least the safety issues are being dealt with in many areas, not by a few benevolent employers. no insurance payment will compensate for the use of a hand, hearing, life, or in most of our cases, an eye. if you have a problem with implimentation do what rosemary did, discuss the methods, rationally. do it as if you were defending your thesis again. sell the problem and get a cure. but let's listen to sergey and not complain about the few seconds needed to take off glasses or shields.
finally, to make it clear, i learned my attitudes from my manufacturer father, who taught me that the most important resource he had was his workers. i am not a left wing pinko, i am very middle of the road. and perhaps a little to the right of centre on economic issues
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
Can anyone recommend a microscopy method/technique to visualize and/or possibly quantify the amount of free plasticizer vs the amount of bound plasticizer in cast PVC skins? We suspect a ratio difference of free vs bound plasticizer when comparing PVC raw materials vs cast PVC skins which are exhibiting adhesive failure between the PVC skin and foam. We think there may be a higher amount than normal, of free plasticizer migrating to the PVC skin surface.
Fred A. Hayes Polymer Microscopy Collins & Aikman IntelliMold Div 4651 Platt Lane Ann Arbor, MI 48108 734-477-7029 office 734-477-9214 fax
Only for windows. Mac users never have the problem. :) I received about 30..opened some..no problem. :)
on 8/21/03 7:24 AM, "atcsem-at-earthlink.net"-at-sparc5.microscopy.com at "atcsem-at-earthlink.net"-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } The attachments with .pif extensions } } This is the virus you may have. } http://securityresponse.symantec.com/avcenter/venc/data/w32.sobig.f-at-mm.html } } Note that the virus is spoofing e-mail addresses. so your system could be } clean. } } Pavel Lozovyy } ATC SEM Lab } Phone: 216-692-6637 } E-mail: atclabs-at-sbcglobal.net } web: www.atclabs.com } } }
In Europe (I am not sure of the USA) there is a general system of controlling exposure to hazards which involves in order of preference: 1.Prevent exposure (eg remove the hazardous process or substitute hazardous material with safer one). 2.Control process by design/engineering (eg shielding, fume hood). 3.Use procedural controls (eg less people involved, good hygiene, reduce quantities). 4.and only finally use PPE (personal protective equipment) such as safety glasses.
If the first three can be applied successfully then there should be no need for PPE such as safety glasses. An obvious point may be to separate the preparation procedure from the microscopy because if there is a risk of splashing the eyes then surely there is a risk of splashing the optics of microscopes.
It should also be borne in mind that if an accident (at least in the UK) results in an injury and the employer didn't take reasonable precautions and enforce them then that employer could be deemed to be negligent. Health and safety issues are often covered by criminal law so the employer or their representative could go to prison (doesn't often happen - but can) and any damages would not normally be insurable so the employer would have to pay and not the insurer. It gives a different slant on the "safety nazis" .
I am still a union health and safety officer and so my sympathies are with the staff rather than employers but I do recognize how difficult a job employers can sometimes have.
I'm sorry for extending an already lengthy debate but the principles are extremely important and I have tried to be brief.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: "paul r hazelton, PhD" {paul_hazelton-at-umanitoba.ca}
Sergey! You say that you can't use the bar side of the grids. Why is that so clear? A thicker layer of stain might not be a disadvantage as long as it is even.
(I'm assuming specimens that are much smaller than the mesh)
Philip
} Phillip - you right: on the rough surface negative stain fills the } imperfections of the surface and therefore increase background noise. It } also makes stain's layer thicker (holding more stain then necessary). All } these things decrease the resolution of your images. You could not use, of } coarse, the opposite side of the grid with bars. So, you need to put flat } side of the carbon up: mount your grids ON the floating carbon. If you are } trying to do high resolution EM on individual proteins, you definitely need } to pay attention to the quality of the carbon: this is a whole point to } use "mica-side" of the carbon, because mica surface is atomically perfect } (it's a single crystal), so the carbon surface will be nearly perfect as } well. The quality of mica is important too. Have a good day, Sergey. } } } } } } } } After a massive (or at least sobig) virus attack I'm back online. } } } } That's a very interesting point Sergej makes. } } } } We work with thin (1 layer) protein crystals and large individual protein } } molecules (chaperones etc.). } } } } For crystals the flatness of the carbon surface is obviously important } } but for single proteins I've always thought it shouldn't make any } } difference. } } Initially we use negative stain to look at the quality of a specimen. } } To get higher resolution data we use cryo-specimens on holey carbon so } } that is a different story. } } Back to the negative stain: I've been wondering about two possible effects } } when staining: } } 1. When you stain a specimen on the rough side the stain might fill all the } } valleys } } of the carbon surface leading to large background noise and making it } } difficult } } to see the particles (some grids seem to have a lot of background). } } 2. When you stain a specimen on the smooth side (which is facing the grid) } } the stain } } might get trapped between the grid bars leading to an unnecessarily thick } } layer of } } stain which again increases the noise. } } } } Are either of the two real problems? } } } } } } } } } } Phillip } } } Your way to mount carbon on the grids has one very serious disadvantage: } } } } } when you mount carbon this way, you will use "air-side" of carbon as a } } } working surface. This side is usually more grainy and dirty. The whole } } } idea of using carbon from mica is to use "mica-side" of carbon as a } } working } } } surface. The idea here is that the carbon face toward mica is more } } uniform } } } and perfectly clean (it was not exposured to the air). In order to use } } } "good" side of carbon, you have to put grids on floated carbon and then } } } pick the grids with Parafilm for instance. Similarly, you need to do the } } } same for plastic film created on water - working side is "water-side", not } } } air. Only one exception I do know is Formvar film created on the } } } glass. Glass is not such clean and perfect as mica or water surface, } } } therefore, you need to use "air-side". Then your technique will work } } } perfectly. I also use your technique to mount holey film on the grids. } } } Best wishes, Sergey. } } } } } } } } } } } } } } } } } } You can easily use the method described by Mike to make } } } } about 50 pure carbon grids at a time (much cheaper than } } } } buying if you already have the equipment). } } } } } } } } You place about 50 grids on a square piece of filter paper } } } } lying on a little "table" made of wire mesh under the water surface. } } } } To get the grids under water make them wet first and push them } } } } through the surface vertically. } } } } Then you lower the water surface by removing water from underneath } } } } (for example with a syringe or a plastic tube with a valve) to place } } } } the carbon film (floated off the mica) on top of the grids. } } } } (Of course the carbon film has to be in one piece in this case.) } } } } Push the carbon into position with something blunt (a paddle ? :) } } } } } } } } Philip Koeck
} } } } } } Steve, } } } } } } When I want just carbon as a substrate I will } } } } } } coat freshly cleaved mica with carbon in a metal } } } } } } evaporator. This allows me to dictate the carbon } } } } } } thickness and I have no trouble floating off grid } } } } } } size squares(prescored)in a drop of water. Picking up the } } } } } } carbon with glow discharged grids (flat side) } } } } } } is a snap. Make sure everything is clean. } } } } } } This should give you thin clean substrates. } } } } } } Mike D. } } } } } } }
You mentioned a good point with regard to chemistry and microscopy, wet gloves. Many people do etching, polishing, and wet chemistry with iterations between a hood/sink and a microscope. Some also feel that they are saving time/money by not removing their gloves whilst adjusting the focus, stage position and worst of all the eyepieces. I can say from experience how this caused me great pain and an emergency hospital visit because someone had their gloved hands in nitric acid and decided to adjust the inter-ocular distance on a microscope that I was alternately using.
My rule was that no gloved hand may touch a microscope whether it's been in a hood, sink or whatever. This way there can be no mistake unless some fool likes to work with bare skin, and there is no protecting oneself from the super-idiot. The only remedy for them is banishment to a desk job where the limit of injury is paper cuts or burns from hot coffee.
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: "gillian.2.brown-at-gsk.com"-at-sparc5.microscopy.com [mailto:"gillian.2.brown-at-gsk.com"-at-sparc5.microscopy.com] Sent: Thursday, August 21, 2003 5:46 AM To: microscopy-at-sparc5.microscopy.com
In my labs it is mandatory to wear safety glasses. Risk assessments are carried out on all microscope usage and, if deemed safe, can be given an exemption sticker (signed off by management) so the user can remove specs. That RA would cover what activities are occurring adjacent and whether the operative has wet hands/gloves as any contaminants could be transferred to hair/skin and indeed eyes during removal of the glasses.
Where the 'environment' remains unsafe we are now moving to replacing the eyepieces with those suitable for spectacle wearers (safety or not). Non spectacle users in Histopath labs here are also having them fitted for doing their 'safe' microscopy as they prefer the ergonomics of them.
So change the microscopy set up and every-one should be safe and happy.
Gillian Brown Histology Section, Asthma Biology RI CEDD, GSK.
your points are very good. i don't know that i have seen them written down here, but they fit exactly the process we use here. there is heavy emphasis on point three. procedure controls. if a person is only going to do something only 1x or 2x, we do it for them. this does not matter if it is dealing with pathogens, radiation, osmium, or other chemical or biologicals.
for what it is worth, i have been trying to eliminate all use of radioactives from my work - not because of the necessary controls and protection, but because of the paper work, that unmentioned administrative point, #5.
thanks for the points, i will forward them to my former union.
I see you've already had a detailed explanation (which I've lost - accidentally deleted because of something odd in my software - never mind, I can pick it up again once it's in the archive), however, I would like to share with you some experience of observing polymer spherulites under the polarizing microscope.
Normally, one observes spherulites between crossed polars, and if the birefringence is strong, you get a four lobed pattern with colours in the sequence you also observe with the quartz wedge. However, if you rotate the analyser to be parallel with the polarizer, you see different colours, in a two lobe pattern. These colours are the complementary ones to those observed between crossed polars. Going from no birefringence to just over the 1st order, one has:
Crossed - Parallel
Black - White (zero order) Grey - not so bright Yellow - Dark Blue Purple - Green (1st order) Blue - Reddish Orange
I have often looked at polymers (including blends) under the polarizing optical microscope. I do not find that rotating the analyser gives extra information, it simply changes the colour of the presentation. However, rotating the specimen between crossed polars, and watching the colours brighten and darken (though they remain the same colour) does give information about the orientation of the molecules.
Hope this is of use.
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +----------------------------------------+ Phone:+44 (0) 118 9318572 Fax: +44 (0) 118 9750203 University internal extension 7867 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +----------------------------------------+
My company is in the semiconductor industry. We current have an AMRAY 2030 C for performing defect review on 8-inch wafers with the ability to read defect files from wafer inspection tools. Our AMRAY is on its last legs.
Are there lab SEM available that can inspect 8-inch wafers edge to edge with the ability to read defect files?
The semiconductor SEM's are expensive and restrictive for r&d work. They are design for production purposes and many SEM controls are removed or automated that limit the tools usefulness.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yezer-at-barak-online.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, August 23, 2003 at 10:16:49 ---------------------------------------------------------------------------
Email: yezer-at-barak-online.net Name: Ezer Yosi
Title-Subject: MListserver:
Question: I am looking for a fast glue which is compatible with UHV (for TEM sample). Can you kindly recomend on one of loctite system for this application.
Ezer Yossef wrote: ===================================================== Question: I am looking for a fast glue which is compatible with UHV (for TEM sample). Can you kindly recomend on one of loctite system for this application. --------------------------------------------------------------------------- The M-Bond 610 system seems to be the most widely used glue for this application, and it can be purchased from SPI Supplies (or some of the other major supply houses for EM consumables) and is described on URL http://www.2spi.com/catalog/spec_prep/glue.shtml
I do hear reports of some who claim to get comparable results with some of the "5 minute" epoxy systems, such as is shown on URL http://www.2spi.com/catalog/spec_prep/5minglue.shtml
The latter is obviously much cheaper but most people seem to prefer M-Bond 610 (or possibly other alternatives that might be comparable to M-Bond 610).
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One more thing add under your message. The Mo grid with very smooth hole will provide the most flat carbon film without cryo-wrink. For more details, please cheak Pacific Grid-Tech website.
http://www.grid-tech.com/
regards
Wendy
} -----------------------------------------------------------------------. } } } After a massive (or at least sobig) virus attack I'm } back online. } } That's a very interesting point Sergej makes. } } We work with thin (1 layer) protein crystals and } large individual protein } molecules (chaperones etc.). } } For crystals the flatness of the carbon surface is } obviously important } but for single proteins I've always thought it } shouldn't make any } difference. } Initially we use negative stain to look at the } quality of a specimen. } To get higher resolution data we use cryo-specimens } on holey carbon so } that is a different story. } Back to the negative stain: I've been wondering } about two possible effects } when staining: } 1. When you stain a specimen on the rough side the } stain might fill all the } valleys } of the carbon surface leading to large background } noise and making it } difficult } to see the particles (some grids seem to have a lot } of background). } 2. When you stain a specimen on the smooth side } (which is facing the grid) } the stain } might get trapped between the grid bars leading to } an unnecessarily thick } layer of } stain which again increases the noise. } } Are either of the two real problems? } } } Philip Koeck } Svdertvrns Hvgskola and } Karolinska Institutet } Dept. of Bioscience at Novum } S-14157 Huddinge } Sweden } phone: +46-8-6089186 } fax: +46-8-6089290 } http://www.biosci.ki.se/em } _______________________________________ } } } ----- Original Message ----- } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {microscopy-at-sparc5.microscopy.com} } Sent: Monday, August 18, 2003 10:06 PM } Subject: Re: Carbon Support Films for TEM } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Phillip } } Your way to mount carbon on the grids has one very } serious disadvantage: } } } when you mount carbon this way, you will use } "air-side" of carbon as a } } working surface. This side is usually more grainy } and dirty. The whole } } idea of using carbon from mica is to use } "mica-side" of carbon as a } working } } surface. The idea here is that the carbon face } toward mica is more } uniform } } and perfectly clean (it was not exposured to the } air). In order to use } } "good" side of carbon, you have to put grids on } floated carbon and then } } pick the grids with Parafilm for instance. } Similarly, you need to do the } } same for plastic film created on water - working } side is "water-side", not } } air. Only one exception I do know is Formvar film } created on the } } glass. Glass is not such clean and perfect as } mica or water surface, } } therefore, you need to use "air-side". Then your } technique will work } } perfectly. I also use your technique to mount } holey film on the grids. } } Best wishes, Sergey. } } } } } } At 04:07 PM 8/18/03 +0200, you wrote: } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } You can easily use the method described by Mike } to make } } } about 50 pure carbon grids at a time (much } cheaper than } } } buying if you already have the equipment). } } } } } } You place about 50 grids on a square piece of } filter paper } } } lying on a little "table" made of wire mesh under } the water surface. } } } To get the grids under water make them wet first } and push them } } } through the surface vertically. } } } Then you lower the water surface by removing } water from underneath } } } (for example with a syringe or a plastic tube } with a valve) to place } } } the carbon film (floated off the mica) on top of } the grids. } } } (Of course the carbon film has to be in one piece } in this case.) } } } Push the carbon into position with something } blunt (a paddle ? :) } } } } } } Philip Koeck } } } Svdertvrns Hvgskola and } } } Karolinska Institutet } } } Dept. of Bioscience at Novum } } } S-14157 Huddinge } } } Sweden } } } phone: +46-8-6089186 } } } fax: +46-8-6089290 } } } http://www.biosci.ki.se/em } } } _______________________________________ } } } } } } } } Steve, } } } } } When I want just carbon as a substrate I } will } } } } } coat freshly cleaved mica with carbon in a } metal } } } } } evaporator. This allows me to dictate the } carbon } } } } } thickness and I have no trouble floating off } grid } } } } } size squares(prescored)in a drop of water. } Picking up the } } } } } carbon with glow discharged grids (flat side) } } } } } is a snap. Make sure everything is clean. } } } } } This should give you thin clean substrates. } } } } } Mike D. } } } } } }
===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
First off, let me comment about UHV and TEM. UHV is generally considered to be in the vacuum range where the pressure is less than 1E-9 Torr. Although the microscope manufacturers would like you to believe that your sample is setting in an UHV environment, chances are it isn't. The dedicated STEM systems may come close. The reason for this is that the pressure is typically measured using the ion pump or a gauge located close to the pump flange. Your sample sits at the end of a conductance limited tube that connects to your pump. In vacuum systems, the throughput (quantity of gas per unit time), Q (Torr l/sec) = S * P, where S is the pump speed (l/sec) and P is pressure (Torr). S * P at the pump is equal to S(eff)*P(sample). S(eff) is the effective pump speed at your sample and is related to both the conductance and pump speed. However, if your diameter of the tubing is small and the length is long or has bends in it, the effective pump speed will be conductance limited and is approximately the same as the conductance. The net effect is that the effective pump speed at your sample is always less than the pump speed and as a result, the true pressure at your sample will be higher than the reported pressure. With all that said, if you cool an anti-contamination device in the neighborhood of your sample, that acts as a cryopump and will lower the pressure at your sample.
For high vacuum systems, epoxy based materials will work fine. However, you should make sure that the mix is right so that everything cross links well and you don't have any of either the hardener or the resin "left over". You should also minimize the amount that you use. Epoxies that have worked well in TEM sample prep are M-bond 610 (as Chuck Garber said), Gatan's G-1 which is equivalent to Epoxy Technologies' Epo-Tek 353ND. A silver filled conductive epoxy that works well is Epoxy Technologies' H-22. Instead of the 5 or 10 min epoxies, I would recommend the 90 min working time, super strength types that require an overnight cure.
If your interested in true, UHV compatible "glues", you are in trouble. There are two sealers that people use for this purpose in UHV systems. One is a product called VacSeal(R) and comes in a brush-on bottle form or a spray application. This stuff works very well when it is room temperature dried, but works even better with a 200C cure. I doubt that it would work well for TEM sample prep. Another material that is used is a two part putty-like material that used to be available from Perkin Elmer. Sorry, I can't remember the name. I would check with vacuum component manufacturers for the name. You might also check with Kurt Lesker. People have also used the water based "Aquadawg" or "Aquadog" to tack things down in UHV systems, but it has to be heated to drive off the water.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
We are in the market for an EBSD system. (I believe all the relevant vendors already know this.) This is a very new area for us and we have basically no experience in this area (to be very blunt). We are dealing with technical specs, vendor information, and demos. And as good and as hands- on as demos maybe they can not give you the true picture of working with a system for months or years or multiple users. What we are looking for is information from USERS of EBSD systems (Researchers, facility managers, etc.) - we are looking for comments, opinions, pro & cons, experiences of any EBSD systems.
Please send your comments directly to me so you maybe as candid as possible.
Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
I'd suggest you investigate both Raman confocal and FT-IR microscopy.
The FT-IR is probably easier to do, if you can get cross sections. Otherwise, Raman confocal can go down layer by layer. I don't think that there is any limitation for either system in terms of the chemical fingerprint they can provide to you. The bound v. unbound may have a better signature in Raman, however.
A number of companies, including BioRad and Nicolet, provide FT-IR microscopes. SensIR (Danbury, CT) provides the IlluminatIR - a unique microscopy accessory which turns a research microscope into an FT-IR microprobe without disabling the microscope.
Renishaw and Jobin-Yvon (JY) both provide Raman confocal.
All of the companies have apps labs which can run a sample or two for you. I'd suggest you visit microscopy.info for contact information. I'm going to be on vacation most of this week, but if you get stuck, drop me an email and I'll send you more specific contact information.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Optimizing Light Microscopy is still available through MME. For information on ordering single or small quantities, visit our website. For discounts on classroom-sized, call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& At 11:19 AM 8/21/03 -0400, Fred.Hayes-at-colaik.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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A position is available in the Materials Science and Engineering Department at Carnegie Mellon University (http://neon.mems.cmu.edu/newfront.shtml) for a full time staff member with experience in transmission electron microscopy. The position will focus on analysis of ceramic thin films having applications in the areas of electronics, magnetics, hard coatings, and catalysis. A significant portion will focus on nano-layered thin films for use as hard coatings. Tasks will include sample preparation (of plan-view and cross-sectional samples), electron microscopy (including high-resolution), and analysis of structural details (such as crystal, interfacial, grain, and superlattice structures). The position could be filled either at the research scientist or research associate (postdoctoral) level. Salary will be commensurate with experience. Carnegie Mellon University is an equal opportunity employer.
For Details and Inquiries Please Contact Paul Salvador Assistant Professor Department of Materials Science and Engineering Wean Hall 3301 Carnegie Mellon University Pittsburgh, PA 15206 TEL: 412-268-2702, FAX: 412-268-7596 paulsalvador-at-cmu.edu
To all those people out there who use the Gatan PIPS -
I thought this was common knowledge, but maybe not. Instead of polishing the sputtered deposits off the viewing window after every hour or so of milling time, just use a grade 0, 19 mm diameter glass cover slip. It sits nicely in the recess under the window - the only precaution you have to take is to make sure it doesn't slide to partially cover the O-ring. They are cheap and can be considered disposable, but if you are really tight and have a big jar of KOH in the cupboard (like me) you can clean the deposits off by boiling in caustic solution for a while and re-use them.
I just hate to think of all those cumulative hours spent by users around the world polishing glass when there is a simple alternative!
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I would like to try poly-L-lysine for sticking cells to coverslips etc for dehydration and SEM. My S**** catalogue offers about 15 variants. Can anyone advise me on which type to use?
Dave
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I'm looking for any feedback on fixation of eyes (small and large animal). More specifically: a fixative that would work well for Histology and EM. Any info. on: * Glutaraldehyde * Karnovsky's Fixative * McDowell - Trump's Fixative or any other fixative would be greatly appreciated.
Thanks in advance,
Pedro
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The 2-part epoxy is called Torr-seal, and is is very good. --John Mardinly
material that is used is a two part putty-like material that used to be available from Perkin Elmer. Sorry, I can't remember the name. I would check with vacuum component manufacturers for the name. You might also check with Kurt Lesker. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
David There is some trick with Poly-Lysine solution. I would suggest to use some from EM major suppliers. Personally I am using 1% solution from TedPella. I don't think there is a difference between different brands. In England you may contact Agar, very good EM supplier. Good luck, Sergey.
At 07:11 AM 8/26/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Dear Listers, I am looking for a lab that can perform TEM analysis of high impact polystyrene. I intend to obtain an extended support for next few months in order to analyze 30-35 samples. The analysis involves specimen preparation (cryomicrotomy, staining), imaging at 2-3 reasonable magnifications and calculation of rubber volume fraction/size of each sample. I will appreciate the help of the list members in this regard. The interested labs on the list server may want to contact me of line for further discussions regarding the project.
Best regards.
Toseef Ahmed, Ph.D. Electron Microscopy and Thermal Labs, Analytical Section SABIC R&T, P.O. Box 42503, Riyadh 11551, Saudi Arabia Tel: (966-1) 265-3333 Ext.6003, 6104 Fax: (966-1) 265-1101 E-mail: toseef-at-sabic.com,
My original post was ambiguous. I am looking for ways to stick fixed cells to coverslips to avoid repeated centrifugation during dehydration for SEM.
Dave
On Tue, 26 Aug 2003 15:11:46 +0100 (GMT Daylight Time) "Patton, David" {David.Patton-at-uwe.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would like to try poly-L-lysine for sticking cells to } coverslips etc for dehydration and SEM. My S**** catalogue } offers about 15 variants. Can anyone advise me on which } type to use? } } Dave } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Torr-Seal is available from many of the supply houses including Ladd. See http://www.laddresearch.com/General_Catalog/Chapter_11/Miscellaneous_Vacuum_ Accessori/miscellaneous_vacuum_accessori.html for our selection. MSDS is also available on line for those seeking more information.
Debra Sicard Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Mardinly, John" {john.mardinly-at-intel.com} To: "Walck, Scott D." {walck-at-ppg.com} ; "Microscopy (E-mail)" {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, August 26, 2003 1:12 PM
Hi all,
We are a University microscopy recharge center and will be installing a single beam FIB within the next 6 months. I've got to establish a reasonable initial hourly use rate without the benefit of experience with a FIB costs and use patterns. Will you university FIB users out there share some of your experience, wisdom and rates? Off line will be best I think.
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (maloneyb-at-fiu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, August 25, 2003 at 14:36:09 ---------------------------------------------------------------------------
Email: maloneyb-at-fiu.edu Name: Barb
Organization: FIU
Title-Subject: Phillips 200 TEM
Question: Does anyone have the specs on this particular model? Need ASAP Thanks Barb
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (info-at-klughammer.de) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, August 27, 2003 at 03:42:53 ---------------------------------------------------------------------------
does anybody know a EPI fluorescence microscope manufacturer which offers quite good quality at an affordable price. They will be used for university beginner groups.
This is way off the microscopy topic, but unfortunately, it is relevant to our daily activities of late. I am trying to contact someone probably associated with this list to inform them that they are infected with the SoBig virus and don't appear to know it yet.
Like many of us, I have been receiving dozens of copies of the SoBig virus of late. Being a curious type and being the IT guy for the computers in our labs, I have been poking into some of the messages to see where they came from. Even though the Return Path: and From: fields are usually spoofed, I found that many were coming from a couple of sources. I have included the relevant information below.
Since many of the alleged senders were frequent posters to this list I am guessing that the real sender may be a member of this list. So if BILLPC on SYMET.NET or PRIVAT on T-DIALIN.NET means something to one of you out there, please see about getting your PC disinfected.
Thanks.
Warren
Received: from PRIVAT (pD9E7724F.dip.t-dialin.net [217.231.114.79]) by despam-3.iastate.edu (8.12.4/8.12.4) with ESMTP id h7QCNLKG007344 for {wesaia-at-iastate.edu} ; Tue, 26 Aug 2003 07:23:22 -0500 } From: {martin.voecks-at-web.de}
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------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
We do have the capibiilty. What kind of arrangement would you propose to have the work done?
**************************************** Chaoying Ni, PhD The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
On Wed, 27 Aug 2003, AHMED, TOSEEF wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } I am looking for a lab that can perform TEM analysis of high impact polystyrene. I intend to obtain an extended support for next few months in order to analyze 30-35 samples. The analysis involves specimen preparation (cryomicrotomy, staining), imaging at 2-3 reasonable magnifications and calculation of rubber volume fraction/size of each sample. I will appreciate the help of the list members in this regard. The interested labs on the list server may want to contact me of line for further discussions regarding the project. } } Best regards. } } } Toseef Ahmed, Ph.D. } Electron Microscopy and Thermal Labs, Analytical Section } SABIC R&T, P.O. Box 42503, Riyadh 11551, Saudi Arabia } Tel: (966-1) 265-3333 Ext.6003, 6104 } Fax: (966-1) 265-1101 } E-mail: toseef-at-sabic.com, } }
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
I've found good success sticking live bacteria cells to the coverslips FOR fixing and dehydrating... Prep (slides or coverslips) Clean slides in Ethanol with a small amount of dilute HCl Rinse in Distilled water Soak for 30 minutes in your Poly-L solution Let air dry (can be stored desiccated for a long time) The Solution that works well and as is published in Steven E. Ruzin's Plant Microtechnique and Microscopy, calls for 100µg-1mg/ml and Tris 10mM, pH 8.0
I've had success with a "thats about enough" diluting the stock Poly-L with Distilled water (one or two mls per 100-200mls of dH20).
This has worked well for general histochemistry (wax sections) as well as liquid suspended bacterial preparations for the SEM. Samples survive fixation, washing, dehydration, and even CPD with significant coverage where the Poly-L is.
We now get our Poly-L-Lysine from the big F chemical/supply company (the S chemical company also has it) the 1% solution is what we get.
HTH
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] } Sent: Wednesday, August 27, 2003 4:20 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Poly-L-lysine: clarification } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } My original post was ambiguous. I am looking for ways to } stick fixed cells to coverslips to avoid repeated } centrifugation during dehydration for SEM. } } Dave } } } On Tue, 26 Aug 2003 15:11:46 +0100 (GMT Daylight Time) } "Patton, David" {David.Patton-at-uwe.ac.uk} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I would like to try poly-L-lysine for sticking cells to } } coverslips etc for dehydration and SEM. My S**** catalogue } } offers about 15 variants. Can anyone advise me on which } } type to use? } } } } Dave } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } } } This incoming email to UWE has been independently scanned for viruses } and any virus detected has been removed using McAfee anti-virus software } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England"
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pkrsko-at-stevens-tech.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, August 27, 2003 at 12:10:52 ---------------------------------------------------------------------------
Email: pkrsko-at-stevens-tech.edu Name: Peter Krsko
Organization: Stevens Inst Tech
Education: Graduate College
Location: Hoboken, NJ
Question: Hello,
I would like to find out if it's possible to get phosphorus used on TEM screens in solution. I am using simple electron beam mask and I want to highlight the apperture opening with this paint.
Your help is greatly appreciated.
Thank you.
Peter Krsko
P.S: Here are some images for you: http://attila.stevens-tech.edu/~pkrsko/billboart/
Pedro, Karnowsky's is the best. works both for LM and EM. Shashi CCMB Hyderabad INDIA
I'm looking for any feedback on fixation of eyes (small and large animal). More specifically: a fixative that would work well for Histology and EM. Any info. on: * Glutaraldehyde * Karnovsky's Fixative * McDowell - Trump's Fixative or any other fixative would be greatly appreciated.
Thanks in advance,
Pedro
__________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com
Toseef Ahmed, Ph.D. wrote: ======================================================= I am looking for a lab that can perform TEM analysis of high impact polystyrene. I intend to obtain an extended support for next few months in order to analyze 30-35 samples. The analysis involves specimen preparation (cryomicrotomy, staining), imaging at 2-3 reasonable magnifications and calculation of rubber volume fraction/size of each sample. I will appreciate the help of the list members in this regard. The interested labs on the list server may want to contact me of line for further discussions regarding the project. ======================================================= There are a number of independent analytical laboratories in the USA and elsewhere set up to do this type of TEM work for clients. We maintain a listing of some of these laboratories on our website under our "Hot Services" section, see URL http://www.2spi.com/hot-service7.html
Structure Probe, Inc. has been conducting this type of characterization work on high impact polystyrene (HIPS) since the inception of our business in 1970, both cryo and at RT. We are also accredited by A2LA to the standard of ISO Guide 17025. We are not a university, but are a commercial business. Providing a high standard of service to our clients is our #1 priority.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. 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: www.2spi.com ############################ ==================================================
Paula Sounds like you have the wrong bulb. An enlarger bulb is opal (milky-white) so the filament should not be visible, and the label should be printed on the base close to the cap, where it won't interfere with image formation . Chris
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "Paula Sicurello" {patpxs-at-gwumc.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, August 27, 2003 8:21 PM
Hi,
I am currently testing our new microscopy based reader on which I would like to test a variety of samples. One of the samples I am looking for is a plate which contains 6144 samples of cells or a tissue array within the boundaries of a SBS-standard plate. Arrays or plates with 1536 well or samples/spots on them would be useful too, if they fit into the SBS-standard plate boundaries. The arrays should be in a evenly spaced rectangular pattern. The carrier should be either glass or plastic.
Does anyone have such samples available which I could use for testing either in brightfield mode or with fluorescence or a combination of both ? Preferably fixed material which can be reused several times for testing.
The list of Local Arrangements Committee members shown in the EXPO and Proceedings for the 2003 Microscopy & Microanalysis meeting was based on a preliminary list I submitted two months before the meeting for those publications' deadlines. By the time of the meeting some of the people on the list were unable to participate, and others not on the list attended M&M 2003 and served on the LAC. To give the LAC members proper recognition for the tremendous job they did, here are their names:
} From Texas:
Pamela Neill
Jodi Roepsch
Robert Champaign
Becky Holdford
Ann Burke
Josephine Taylor
Ann Rushing
Susan Robbins
Sandra Westmoreland
Robert Droleskey
} From Oklahoma:
Mary Sheldon
Elizabeth Duncan
Rae Harrington
Amy Sheldon
} From Minnesota:
Dwight Erickson, LAC Treasurer
Thanks to all!
Ev Osten
Local Arrangements Committee Chair Microscopy & Microanalysis 2003 August 3 - 7, San Antonio
efosten-at-mmm.com 651-736-0104 fax: 651-733-0648
3M Company Corporate Analytical Technology Center 3M Center, 201-BE-16 St. Paul, MN 55144-1000
It sounds like you have the point-source bulb in there. The correct bulb for that Durst is a large frosted bulb, the number of which I can no longer remember. If your enlarger is set up for point source, you will need to do a point-source alignment of the system and use the proper condensers to get rid of the filament image. I no longer remember how to do that, either, except that the lens must be used at its largest f/stop. The point source lamp plugs into a rheostat to control the brightness, instead of using regular f-stops, so if you switch to regular condenser lighting, you will need to plug the lamp into a regular outlet. If you later switch back to point-source you MUST remember to switch back to the proper power source or the bulb will explode like a little grenade and scare the *^%*%* out of you. Speaking from experience here. :-)
Lots of luck, Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Paula Sicurello [mailto:patpxs-at-gwumc.edu] Sent: Wednesday, August 27, 2003 2:21 PM To: microscopy-at-sparc5.microscopy.com
Hi Listers,
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Barb, I think that I have everything that you would need. Please reply with what you need in particular. I am currently using the Philips 200 and have complete sets of the manuals and I think all the O-rings that you could possible need too. Pat Connelly Research Specialist Dept. of Biology Univ. of Pennsylvania Philadelphia, PA 19104-6018 215-898-7145
Yet another reason to go with Macintosh! Not a problem on our end..
Carol Jean Hirt, VP Materials Research Laboratories, Inc. 800-424-1776 www.mrllab.com
on 8/27/03 12:22 PM, Warren E Straszheim at wesaia-at-iastate.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This is way off the microscopy topic, but unfortunately, it is relevant to } our daily activities of late. } I am trying to contact someone probably associated with this list to inform } them that they are infected with the SoBig virus and don't appear to know } it yet. } } Like many of us, I have been receiving dozens of copies of the SoBig virus } of late. Being a curious type and being the IT guy for the computers in our } labs, I have been poking into some of the messages to see where they came } from. Even though the Return Path: and From: fields are usually spoofed, I } found that many were coming from a couple of sources. I have included the } relevant information below. } } Since many of the alleged senders were frequent posters to this list I am } guessing that the real sender may be a member of this list. So if BILLPC on } SYMET.NET or PRIVAT on T-DIALIN.NET means something to one of you out } there, please see about getting your PC disinfected. } } Thanks. } } Warren } } Received: from PRIVAT (pD9E7724F.dip.t-dialin.net [217.231.114.79]) } by despam-3.iastate.edu (8.12.4/8.12.4) with ESMTP id h7QCNLKG007344 } for {wesaia-at-iastate.edu} ; Tue, 26 Aug 2003 07:23:22 -0500 } } From: {martin.voecks-at-web.de} } } } Received: from BILLPC (dsl1-216-90-9-53.symet.net [216.90.9.53]) } by despam-2.iastate.edu (8.12.4/8.12.4) with ESMTP id h7REWjbX018529 } for {wesaia-at-iastate.edu} ; Wed, 27 Aug 2003 09:32:46 -0500 } Message-Id: {200308271432.h7REWjbX018529-at-despam-2.iastate.edu} } } From: {gary-at-gaugler.com} } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 46 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } } } }
You may wish to consider an alternative. The use of glass slides coated with 2% aminopropyltriethoxysilane in dry acetone is now commonly used to make sections adhere better. You dip once in 2% APTS for 10-30 sec, rinse in acetone and then rinse in distilled water. It works far better than poly-L-lysine or chrome-gel for this purpose. But it was originally described by Gottlieb & Glaser (1976 Biochem Biophys Res Comm 63:815-821) in conjunction with glutaraldehyde to get single cells to adhere to glass better. You might want to check this paper out. good luck. tom
At 09:20 AM 8/27/2003 +0100, you wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
That is one cofiguration and should work OK. I have also used 240/200 and a 105 mm lens. If all you are getting is the filament image, then you need to adjust the bellows and refocus on the grains in your film. From the bulb description, it sounds like you are using a regular enlarging bulb and not a point source. You can verify this by checking if the enlarger is plugged directly into the timer or is there a variable reostat inbetween that allows you to adjust the light intensity. If you need further help in aligment, email me directly.
Mannie Steglich Tech Dir E M Lab U T M D Anderson Cancer Center Houston TX msteglic-at-mdanderson.org ---------------------- Forwarded by Mannie Steglich/MDACC on 08/28/2003 11:16 AM ---------------------------
Microscopy-request-at-sparc5.microscopy.com on 08/27/2003 02:21:13 PM
To: microscopy-at-sparc5.microscopy.com cc:
Hi Listers,
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Or if you can't live without Windows - just get rid of outlook express on every windows machine you have. Replace it with something better like Eudora that doesn't have so many security flaws.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } Carol Jean Hirt, VP } Materials Research Laboratories, Inc. } 800-424-1776 } www.mrllab.com } } on 8/27/03 12:22 PM, Warren E Straszheim at wesaia-at-iastate.edu wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } This is way off the microscopy topic, but unfortunately, it is relevant to } } our daily activities of late. } } I am trying to contact someone probably associated with this list to inform } } them that they are infected with the SoBig virus and don't appear to know } } it yet. } } } } Like many of us, I have been receiving dozens of copies of the SoBig virus } } of late. Being a curious type and being the IT guy for the computers in our } } labs, I have been poking into some of the messages to see where they came } } from. Even though the Return Path: and From: fields are usually spoofed, I } } found that many were coming from a couple of sources. I have included the } } relevant information below. } } } } Since many of the alleged senders were frequent posters to this list I am } } guessing that the real sender may be a member of this list. So if BILLPC on } } SYMET.NET or PRIVAT on T-DIALIN.NET means something to one of you out } } there, please see about getting your PC disinfected. } } } } Thanks. } } } } Warren } } } } Received: from PRIVAT (pD9E7724F.dip.t-dialin.net [217.231.114.79]) } } by despam-3.iastate.edu (8.12.4/8.12.4) with ESMTP id h7QCNLKG007344 } } for {wesaia-at-iastate.edu} ; Tue, 26 Aug 2003 07:23:22 -0500 } } } From: {martin.voecks-at-web.de} } } } } } } Received: from BILLPC (dsl1-216-90-9-53.symet.net [216.90.9.53]) } } by despam-2.iastate.edu (8.12.4/8.12.4) with ESMTP id h7REWjbX018529 } } for {wesaia-at-iastate.edu} ; Wed, 27 Aug 2003 09:32:46 -0500 } } Message-Id: {200308271432.h7REWjbX018529-at-despam-2.iastate.edu} } } } From: {gary-at-gaugler.com} } } } } ------------------------------------------- } } No files should be attached to this message } } ------------------------------------------- } } Warren E. Straszheim, Ph.D. } } Materials Analysis and Research Lab } } Iowa State University } } 46 Town Engineering } } Ames IA, 50011-3232 } } } } Ph: 515-294-8187 } } FAX: 515-294-4563 } } } } E-Mail: wesaia-at-iastate.edu } } Web: www.marl.iastate.edu } } } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } } Computer applications and networking } } } } } } } } } }
Going with Linux or Mac OS will probably ensure that you don't get infected by a virus, but it won't spare you from receiving all the fool traffic that these viruses have generated. These viruses are equal opportunity pains in the butt in that regard.
But I agree with you, Windows is a high-profile target for virus writers. Those of us using it cannot afford to let our guard down and say "it won't happen to me". We should all have safeguards in place and make sure that virus definitions are up to date if we are going to connect to the net.
Then there is also the often forgotten caution, don't open unexpected attachments. I don't care how cute the clip or website is alleged to be, I don't care to risk my computer over that. And if someone is sending me a document, I want to have a clear indication that it is something worthwhile and and authentic and safe. Messages supposedly from colleagues that say simply "Here is the new file - check it out" don't do much to certify that it is a file that I should trust.
BTW, Gordon, one of the virus copies was _allegedly_ from you.
You all be careful out there. Warren
At 12:20 PM 8/28/2003 -0700, you wrote:
} Or better yet, go with Linux... } } Or if you can't live without Windows - just get rid of outlook express on } every windows machine you have. Replace it with something better like } Eudora that doesn't have so many security flaws. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } Carol Jean Hirt, VP } } Materials Research Laboratories, Inc. } } 800-424-1776 } } www.mrllab.com
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Paula, The Durst 45-S should have the two condensor lens configurations printed out on the cover, front panel, of the enlarger head. Some will be for 4x5, 120, and 35mm. Unfortunately I never did much work for 35mm since we had an easy to use Omega with the frosted bulb. A point-source bulb, which is what you have, will give you better images but may also exacerbate problems you may have on the film, such as dust and scratches, which is why many people opt for the frosted set-up, especially for 35mm. Good luck John
-- John Perrino Cell Sciences Imaging Facility Beckman B001 Stanford University Stanford, CA 94305 650-723-3462
Chris, It's not true, halogen bulbs in relatively modern enlargers are transparent. Halogen bulbs delivered much more "light" and it's spectral composition is better for color photography, which, actually, unimportant for B&W. The important thing is that those bulbs considered as a "pointed light source", so the image is sharper and more contrasty.
In my old Devere enlarger, I am using the following combinations:
Condenser: top lens (mm) 180 180 120 bottom 180 120 100 Objective lens------------} 150-105 105-75 60-50 mm
The bulb is transparent, but I am not quite sure it's halogen.
Unfortunately, I know nothing about Durst enlargers.
Sergey
At 12:01 AM 8/28/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Virus from me...? I doubt it. Send me the full headers and I'll check it out. My email is run through pine on a SunOS computer.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Thu, 28 Aug 2003, Warren E Straszheim wrote:
} Going with Linux or Mac OS will probably ensure that you don't get infected } by a virus, but it won't spare you from receiving all the fool traffic that } these viruses have generated. These viruses are equal opportunity pains in } the butt in that regard. } } But I agree with you, Windows is a high-profile target for virus writers. } Those of us using it cannot afford to let our guard down and say "it won't } happen to me". We should all have safeguards in place and make sure that } virus definitions are up to date if we are going to connect to the net. } } Then there is also the often forgotten caution, don't open unexpected } attachments. I don't care how cute the clip or website is alleged to be, I } don't care to risk my computer over that. And if someone is sending me a } document, I want to have a clear indication that it is something worthwhile } and and authentic and safe. Messages supposedly from colleagues that say } simply "Here is the new file - check it out" don't do much to certify that } it is a file that I should trust. } } BTW, Gordon, one of the virus copies was _allegedly_ from you. } } You all be careful out there. } Warren } } At 12:20 PM 8/28/2003 -0700, you wrote: } } } Or better yet, go with Linux... } } } } Or if you can't live without Windows - just get rid of outlook express on } } every windows machine you have. Replace it with something better like } } Eudora that doesn't have so many security flaws. } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } } Gordon Ante Vrdoljak Electron Microscope Lab } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } phone (510) 642-2085 Berkeley CA 94720-3330 } } fax (510) 643-6207 cell (510) 290-6793 } } } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } } } Carol Jean Hirt, VP } } } Materials Research Laboratories, Inc. } } } 800-424-1776 } } } www.mrllab.com } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 46 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } }
I've been asked to see what the EM difference is between rat eye lens epithelium in culture and in situ. So I'd like to fix the epithelium while it is still in the eye. I'm worried that if I do that it then won't be possible to get the epithelium off the lens without damaging it; I also seem to remember that lens becomes brittle and difficult to handle when fixed. I'd like to keep life simple, so it may be necessary to take the epithelium off the lens before fixing. Does anyone have experience with this?
Perhaps this is a good time to engage PDF as safe attachments(?). It would seem so.
The Mac is not particularly affected since it constitutes a small percentage of PC sales and total installed base. However, it is not immune. It is just that nobody wastes time on a minuscule platform. If it is hyped-up, of course that may change.
gary g.
At 01:30 PM 8/28/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The main thing with fixing eyes is to make sure that the fix can get in - so slit across the cornea and take out the lens (assuming you don't want either of these). For EM, glut is OK, but because an eye is big, the fast penetration/fixation of paraform combined with the thorough action of glut is better - so Karnovsky's. If you need to do immuno, neither of these is suitable; PLP (paraform/lysine/periodate) is good or just paraform. As for McD-Trump - I've never heard of it!
Diana
At 12:19 PM -0400 26/8/03, Louro, Pedro wrote: } I'm looking for any feedback on fixation of eyes (small and large animal). } More specifically: a fixative that would work well for Histology and EM. } Any info. on: } * Glutaraldehyde } * Karnovsky's Fixative } * McDowell - Trump's Fixative } or any other fixative would be greatly appreciated. } } Thanks in advance, } } Pedro
--
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
My linux server has not noticed, is there a new virus ?
but my windows box cannot find time to shut out the viri, the popups, the the cookies and the adware, spyware, and other stuff. Could it be because some of the browsers were written to accomodate, to force the user to put up with these things? Browsers simply read the file that are transmitted from the server to the client. If the browsers ignored the adware, the pop ups and stuff, there would be few people sending them out, because no one could read them.
What is needed is concerted organized effort by those of us in the Internet community to find the people that entered this virus into the system.
Interesting thought, back in 1947, 48, and 49 there was a similar problem with people on the radio, everybody kept trying to out maneuver the next guy so that no one could get anything on their personal radio receivers.
It turned out that the big stations were heavily financed and they wanted the little guys (competition) off the air. So they thought, we will keep doing this until we can get the government to license the airways, in that manner we can control the media called radio by forcing them to get licensed, and by limiting the number of licenses, and eliminate our competition, since the little guys do not have money, manpower, or knowledge about how to lobby the government.
Now this is not a result we want to happen with the internet. Government should not be involved in the email system. So let's start finding the persons that did this So Big Virus. Lets enlist everyone in the net to help back track it. until we find its source. That should not be too big of a job. it only happened last week.
I am betting the guys responsible for it, would suprise us all?
Any ideas how we could back trace this? one idea is two create two threads on this list: 1. to identify So Big by its entry date and source amoung members 2. the other to report any progress in backtracking to the origin that is known or reported by anyone anywhere through out the world as one or more of us becomes aware of it.
sterling
On Thu, 28 Aug 2003, Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Going with Linux or Mac OS will probably ensure that you don't get infected } by a virus, but it won't spare you from receiving all the fool traffic that } these viruses have generated. These viruses are equal opportunity pains in } the butt in that regard. } } But I agree with you, Windows is a high-profile target for virus writers. } Those of us using it cannot afford to let our guard down and say "it won't } happen to me". We should all have safeguards in place and make sure that } virus definitions are up to date if we are going to connect to the net. } } Then there is also the often forgotten caution, don't open unexpected } attachments. I don't care how cute the clip or website is alleged to be, I } don't care to risk my computer over that. And if someone is sending me a } document, I want to have a clear indication that it is something worthwhile } and and authentic and safe. Messages supposedly from colleagues that say } simply "Here is the new file - check it out" don't do much to certify that } it is a file that I should trust. } } BTW, Gordon, one of the virus copies was _allegedly_ from you. } } You all be careful out there. } Warren } } At 12:20 PM 8/28/2003 -0700, you wrote: } } } Or better yet, go with Linux... } } } } Or if you can't live without Windows - just get rid of outlook express on } } every windows machine you have. Replace it with something better like } } Eudora that doesn't have so many security flaws. } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } } Gordon Ante Vrdoljak Electron Microscope Lab } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } phone (510) 642-2085 Berkeley CA 94720-3330 } } fax (510) 643-6207 cell (510) 290-6793 } } } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } } } Carol Jean Hirt, VP } } } Materials Research Laboratories, Inc. } } } 800-424-1776 } } } www.mrllab.com } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 46 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } } }
Does anyone know what has become of DiaTech in Knoxville Tenn.? They were makers of fine diamond knives for ultramicrotomy and must have folded or been sold on to another company--any news of them? Many thanks, Lee F. Brunckhorst leeb-at-uow.edu.au (Wollongong University Materials Engineering, Australia)
I'll have to go with Gary on this one, at the risk of starting a flame war. My Timex Sinclair and Atari ST are completely immune to viruses for the same reason...
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Hi, Could those of you doing EBSD in an SEM kindly share your sample prep method with me. I welcome all feedback, but would particularly like to here from those working with geological samples. Is anyone using an Ion Mill? If so, what is the maximum size of the 'thinned area'?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
Mannie is correct in suggesting to adjust the bellow to focus the grain on your negative. Specifically, you should bring the 50mm lens closer to the condensers. You were at a wrong focal plane when you see filament. It does not matter what bulb you use in terms of focusing.
Ann Fook Yang
} } } {"msteglic-at-mdanderson.org"-at-sparc5.microscopy.com} 08/28/03 12:22PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
That is one cofiguration and should work OK. I have also used 240/200 and a 105 mm lens. If all you are getting is the filament image, then you need to adjust the bellows and refocus on the grains in your film. From the bulb description, it sounds like you are using a regular enlarging bulb and not a point source. You can verify this by checking if the enlarger is plugged directly into the timer or is there a variable reostat inbetween that allows you to adjust the light intensity. If you need further help in aligment, email me directly.
Mannie Steglich Tech Dir E M Lab U T M D Anderson Cancer Center Houston TX msteglic-at-mdanderson.org ---------------------- Forwarded by Mannie Steglich/MDACC on 08/28/2003 11:16 AM ---------------------------
Microscopy-request-at-sparc5.microscopy.com on 08/27/2003 02:21:13 PM
To: microscopy-at-sparc5.microscopy.com cc:
Hi Listers,
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Mannie is correct in suggesting to adjust the bellow to focus the grain on your negative. Specifically, you should bring the 50mm lens closer to the condensers. You were at a wrong focal plane when you see filament. It does not matter what bulb you use in terms of focusing.
Ann Fook Yang
} } } {"msteglic-at-mdanderson.org"-at-sparc5.microscopy.com} 08/28/03 12:22PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
That is one cofiguration and should work OK. I have also used 240/200 and a 105 mm lens. If all you are getting is the filament image, then you need to adjust the bellows and refocus on the grains in your film. From the bulb description, it sounds like you are using a regular enlarging bulb and not a point source. You can verify this by checking if the enlarger is plugged directly into the timer or is there a variable reostat inbetween that allows you to adjust the light intensity. If you need further help in aligment, email me directly.
Mannie Steglich Tech Dir E M Lab U T M D Anderson Cancer Center Houston TX msteglic-at-mdanderson.org ---------------------- Forwarded by Mannie Steglich/MDACC on 08/28/2003 11:16 AM ---------------------------
Microscopy-request-at-sparc5.microscopy.com on 08/27/2003 02:21:13 PM
To: microscopy-at-sparc5.microscopy.com cc:
Hi Listers,
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
I am glad you mentioned PDF, i have noticed that when I load some PDf's that all of a sudden my computer starts sending things outbound lots and lots of bits and I cannot stop it unless I shut down and reboot. anyone else notice that with PDF? Can someone say what it is that is happening?
sterling
On Thu, 28 Aug 2003, Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Perhaps this is a good time to engage PDF } as safe attachments(?). It would seem so. } } The Mac is not particularly affected since it } constitutes a small percentage of PC sales } and total installed base. However, it is not immune. } It is just that nobody wastes time on a minuscule } platform. If it is hyped-up, of course that } may change. } } gary g. } } } At 01:30 PM 8/28/2003, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Going with Linux or Mac OS will probably ensure that you don't get } } infected by a virus, but it won't spare you from receiving all the fool } } traffic that these viruses have generated. These viruses are equal } } opportunity pains in the butt in that regard. } } } } But I agree with you, Windows is a high-profile target for virus writers. } } Those of us using it cannot afford to let our guard down and say "it won't } } happen to me". We should all have safeguards in place and make sure that } } virus definitions are up to date if we are going to connect to the net. } } } } Then there is also the often forgotten caution, don't open unexpected } } attachments. I don't care how cute the clip or website is alleged to be, I } } don't care to risk my computer over that. And if someone is sending me a } } document, I want to have a clear indication that it is something } } worthwhile and and authentic and safe. Messages supposedly from colleagues } } that say simply "Here is the new file - check it out" don't do much to } } certify that it is a file that I should trust. } } } } BTW, Gordon, one of the virus copies was _allegedly_ from you. } } } } You all be careful out there. } } Warren } } } } At 12:20 PM 8/28/2003 -0700, you wrote: } } } } } Or better yet, go with Linux... } } } } } } Or if you can't live without Windows - just get rid of outlook express on } } } every windows machine you have. Replace it with something better like } } } Eudora that doesn't have so many security flaws. } } } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } } } Gordon Ante Vrdoljak Electron Microscope Lab } } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } } phone (510) 642-2085 Berkeley CA 94720-3330 } } } fax (510) 643-6207 cell (510) 290-6793 } } } } } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } } } } } Carol Jean Hirt, VP } } } } Materials Research Laboratories, Inc. } } } } 800-424-1776 } } } } www.mrllab.com } } } } ------------------------------------------- } } No files should be attached to this message } } ------------------------------------------- } } Warren E. Straszheim, Ph.D. } } Materials Analysis and Research Lab } } Iowa State University } } 46 Town Engineering } } Ames IA, 50011-3232 } } } } Ph: 515-294-8187 } } FAX: 515-294-4563 } } } } E-Mail: wesaia-at-iastate.edu } } Web: www.marl.iastate.edu } } } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } } Computer applications and networking } } } }
} Perhaps this is a good time to engage PDF } as safe attachments(?). It would seem so. } } The Mac is not particularly affected since it } constitutes a small percentage of PC sales } and total installed base. However, it is not immune. } It is just that nobody wastes time on a minuscule } platform. If it is hyped-up, of course that } may change. } } gary g.
Actually also not a good idea, there are such things as a PDF virus.
The biggest problem with Windows (any flavor) is Outlook Express. As installed, it auto executes any Attachment. This makes it a sucker punch for the common cracker. If Windows users turned that "feature" off or use a different email app (like Eudora) then virus propagation would drop by many factors. This is how the SoBig virus infects and propagates.
Another problem is Windows (any flavor), by default, has many network ports open that have no business being open thus exposing the machine to numerous buffer overflow cracks. This is how the MSBlaster virus infects and propagates.
As for the MacOS not being affected because the size of it's market. A small gain of truth but mostly FUD. Market share of linux is also small but that does not keep the virus and cracks away. Also statistically, even though the MacOS is smaller, one should see a few virus problems over the years. What do I see, zero, nothing. I don't even run virus protection software on the MacOS any more. Stopped doing that many years ago.
As a professional programmer fluent in apps and drivers in both the WinOS and MacOS world, the truth is there are fundamental differences in the way OS9 works from Windows that make 95% of the virus cracks impossible to implement. Can you design a virus to infect MacOS9, yes but you are going to have to work really hard at it. Script kiddies and hack virus programmers need not apply.
OSX is also different but OSX is an industrial strength OS that is installed in a safe manner (for example, there are no network ports open unless you open them. Note that does not mean you can't surf the net, it means that no one can surf you). Also Apple takes a defensive approach, they assume that there are evil people out there that want to mess with your computer. Microsoft does not design Windows with security in mind, they keep saying they are but as long as those extraneous network ports are open by default, I know they are not.
As a case in point, 4pi runs an powerpc linux server for web, ftp and mail services. We always get probed and I mean seriously probed, but they can't get in because 1) it's a powerpc box and intel linux cracks don't work. and 2) it's locked down with regard to security. We have all flavors of OSs here. Macs (both OS9 and OSX) and linux boxes are permitted email access. Windows boxes are not. Too much risk with cracks. Last time I saw a virus problem on a MacOS box was back in the pre-powerpc days and that was an programming experiment in self infection.
For more years, than I can count, non-MacOS users have been declaring victory and Apple with the MacOS dead because of its size. Apple is still around, Macs are still around and with OSX, the market size is even growing. OSX is pretty neat I must say.
And last, you will not keep out a professional cracker out of your computer in the same way you will not keep a professional thief out of your house. Professional are a different breed, they will get in. That's way they are called professionals. Thankfully, most all virus and crack writers are not real professionals. I can recommend several books that will put the fear of god in anyone that thinks the internet is a benign place. It's a real war zone and the sooner we accept that fact and convince companies such as Microsoft to default the OS in a secure mode, then we will start to see a reduction rather than a increase in cracks.
Microsoft resorted to linux to protect its servers. I found it funny. http://www.internetweek.com/story/showArticle.jhtml?articleID=13100775
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Thu, 28 Aug 2003, Gordon Vrololjak wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Or better yet, go with Linux... } } Or if you can't live without Windows - just get rid of outlook express on } every windows machine you have. Replace it with something better like } Eudora that doesn't have so many security flaws. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } Carol Jean Hirt, VP } } Materials Research Laboratories, Inc. } } 800-424-1776 } } www.mrllab.com } } } } on 8/27/03 12:22 PM, Warren E Straszheim at wesaia-at-iastate.edu wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } This is way off the microscopy topic, but unfortunately, it is relevant to } } } our daily activities of late. } } } I am trying to contact someone probably associated with this list to inform } } } them that they are infected with the SoBig virus and don't appear to know } } } it yet. } } } } } } Like many of us, I have been receiving dozens of copies of the SoBig virus } } } of late. Being a curious type and being the IT guy for the computers in our } } } labs, I have been poking into some of the messages to see where they came } } } from. Even though the Return Path: and From: fields are usually spoofed, I } } } found that many were coming from a couple of sources. I have included the } } } relevant information below. } } } } } } Since many of the alleged senders were frequent posters to this list I am } } } guessing that the real sender may be a member of this list. So if BILLPC on } } } SYMET.NET or PRIVAT on T-DIALIN.NET means something to one of you out } } } there, please see about getting your PC disinfected. } } } } } } Thanks. } } } } } } Warren } } } } } } Received: from PRIVAT (pD9E7724F.dip.t-dialin.net [217.231.114.79]) } } } by despam-3.iastate.edu (8.12.4/8.12.4) with ESMTP id h7QCNLKG007344 } } } for {wesaia-at-iastate.edu} ; Tue, 26 Aug 2003 07:23:22 -0500 } } } } From: {martin.voecks-at-web.de} } } } } } } } } } Received: from BILLPC (dsl1-216-90-9-53.symet.net [216.90.9.53]) } } } by despam-2.iastate.edu (8.12.4/8.12.4) with ESMTP id h7REWjbX018529 } } } for {wesaia-at-iastate.edu} ; Wed, 27 Aug 2003 09:32:46 -0500 } } } Message-Id: {200308271432.h7REWjbX018529-at-despam-2.iastate.edu} } } } } From: {gary-at-gaugler.com} } } } } } } ------------------------------------------- } } } No files should be attached to this message } } } ------------------------------------------- } } } Warren E. Straszheim, Ph.D. } } } Materials Analysis and Research Lab } } } Iowa State University } } } 46 Town Engineering } } } Ames IA, 50011-3232 } } } } } } Ph: 515-294-8187 } } } FAX: 515-294-4563 } } } } } } E-Mail: wesaia-at-iastate.edu } } } Web: www.marl.iastate.edu } } } } } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } } } Computer applications and networking } } } } } } } } } } } } } } } } }
What you describe sounds like you are mounting the 50mm lens below the lens mounting turret in the same manner as you mount the 135mm and 150mm lenses for larger format negatives.
The 50mm lens must be recessed into the bellows. There is a tube shaped mounting adapter that fits into the lens mounting turret.
Frank **************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
No, the virus was not from you. I said "_allegedly_" from you. You can check out the headers below. The culprit was BILLPC, whoever that may be. Your name (and many other listers including myself and Debby Sherman, etc) have been forged as the sources. I just wanted you to know you were included in the party. {G}
More viruses are hiding or forging the identity of the true sender. I don't assume anymore that the alleged source is the real source but instead examine the other headers before I try to alert someone to an infection on their end. I have been scolded as the source of a virus on several such occasions. In all cases, so far, I have not been the real source.
Warren
At 03:05 PM 8/28/2003 -0700, Gordon V. wrote: } Virus from me...? } I doubt it. Send me the full headers and I'll check it out. My email is } run through pine on a SunOS computer.
Received: from BILLPC (dsl1-216-90-9-53.symet.net [216.90.9.53]) by despam-2.iastate.edu (8.12.4/8.12.4) with ESMTP id h7RKJFbX022026 for {wesaia-at-iastate.edu} ; Wed, 27 Aug 2003 15:19:17 -0500 Message-Id: {200308272019.h7RKJFbX022026-at-despam-2.iastate.edu} } From: {gvrdolja-at-nature.berkeley.edu} To: {wesaia-at-iastate.edu}
I suppose it could be one of a couple things with PDF files.
First, I think PDF files may now allow for some code, either macros or embedded files, that could execute on your computer when you view the file. I would need a real PDF expert to comment on the current capabilities. I know MS Word document files were immune from infection until Word 97, IIRC. About that time, MS started supporting macros as valid parts of Word documents. It wasn't long before hackers started exploiting the "feature". Even before that, the Concept virus came out using a document template (.DOT) named as a document (.DOC) showing that documents could now be hosts for viruses. Maybe the same thing is happening to PDF files.
Second, it could be that PDF is not the real file extension. I know I have seen attachments with names such as "details.txt.vbs". The file could show up as "details.txt" in the mail window or an Explorer window if the system was set to hide known file extensions. Unfortunately, the default Windows installation hides extensions, ala Mac, and allows hackers to use this trick to fool users into thinking the file is just a benign text or PDF file. But when the file is executed, Windows uses the true, hidden extension to determine what to do with the file.
Either way, I come back to the conclusion that attachments should be received only cautiously. I appreciate Scott Davilla's comments that the Internet is a very dangerous place. If we knew just how dangerous, we might be more careful.
Warren
At 02:23 AM 8/29/2003 -0500, you wrote:
} I am glad you mentioned PDF, i have noticed that when I load some PDf's } that all of a sudden my computer starts sending things outbound lots and } lots of bits and I cannot stop it unless I shut down and reboot. anyone } else notice that with PDF? } Can someone say what it is that is happening? } } sterling
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Hello all, I am looking into getting a nebulizer to deposit proteins in glycerol as 25-150um diameter droplets onto cleaved sheets of mica that will later be rotary shadowed. I have heard that the bulb-driven nebulizers do not give enough condensed force of spray to create fine droplets. Does anyone have experience with this method and can suggest protocols and a nebulizer to purchase? John -- John Perrino Cell Sciences Imaging Facility Beckman B001 Stanford University Stanford, CA 94305 650-723-3462
Can anyone recommend a source for an antibody against rabbit-GFP to use for TEM immunocytochemistry? We are trying to localize a protein with a GFP construct in arabidopsis root tissue. We have tried one antibody but labeling is poor and not well localized. We want to make sure that it is not the antibody. The protein is an unknown and the research group has not yet been able to develop a clean antibody against it.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
Apparently they may have found him http://www.reuters.com/newsArticle.jhtml;jsessionid=1MBT2YBXGTM1QCRBAELCFEY?type=topNews&storyID=3361073
On Thu, 28 Aug 2003 sstouden-at-thelinks.com wrote:
} } My linux server has not noticed, is there a new virus ? } } but my windows box cannot find time to shut out the viri, the popups, the } the cookies and the adware, spyware, and other stuff. Could it be because } some of the browsers were written to accomodate, to force the user to put } up with these things? Browsers simply read the file that are transmitted } from the server to the client. If the browsers ignored the adware, the } pop ups and stuff, there would be few people sending them out, because no } one could read them. } } What is needed is concerted organized effort by those of us in the } Internet community to find the people that entered this virus into the } system. } } Interesting thought, back in 1947, 48, and 49 there was a similar problem } with people on the radio, everybody kept trying to out maneuver the next } guy so that no one could get anything on their personal radio receivers. } } It turned out that the big stations were heavily financed and they wanted } the little guys (competition) off the air. So they thought, we will keep } doing this until we can get the government to license the airways, in } that manner we can control the media called radio by forcing them to } get licensed, and by limiting the number of licenses, and eliminate our } competition, since the little guys do not have money, manpower, or } knowledge about how to lobby the government. } } Now this is not a result we want to happen with the internet. Government } should not be involved in the email system. So let's start finding the } persons that did this So Big Virus. Lets enlist everyone in the net to } help back track it. until we find its source. That should not be too big } of a job. it only happened last week. } } } I am betting the guys responsible for it, would suprise us all? } } Any ideas how we could back trace this? } one idea is two create two threads on this list: } 1. to identify So Big by its entry date and source amoung members } 2. the other to report any progress in backtracking to the origin } that is known or reported by anyone anywhere through out the } world as one or more of us becomes aware of it. } } sterling } } } On Thu, 28 Aug 2003, Warren E Straszheim wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Going with Linux or Mac OS will probably ensure that you don't get infected } } by a virus, but it won't spare you from receiving all the fool traffic that } } these viruses have generated. These viruses are equal opportunity pains in } } the butt in that regard. } } } } But I agree with you, Windows is a high-profile target for virus writers. } } Those of us using it cannot afford to let our guard down and say "it won't } } happen to me". We should all have safeguards in place and make sure that } } virus definitions are up to date if we are going to connect to the net. } } } } Then there is also the often forgotten caution, don't open unexpected } } attachments. I don't care how cute the clip or website is alleged to be, I } } don't care to risk my computer over that. And if someone is sending me a } } document, I want to have a clear indication that it is something worthwhile } } and and authentic and safe. Messages supposedly from colleagues that say } } simply "Here is the new file - check it out" don't do much to certify that } } it is a file that I should trust. } } } } BTW, Gordon, one of the virus copies was _allegedly_ from you. } } } } You all be careful out there. } } Warren } } } } At 12:20 PM 8/28/2003 -0700, you wrote: } } } } } Or better yet, go with Linux... } } } } } } Or if you can't live without Windows - just get rid of outlook express on } } } every windows machine you have. Replace it with something better like } } } Eudora that doesn't have so many security flaws. } } } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } } } Gordon Ante Vrdoljak Electron Microscope Lab } } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } } phone (510) 642-2085 Berkeley CA 94720-3330 } } } fax (510) 643-6207 cell (510) 290-6793 } } } } } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } } } } } Carol Jean Hirt, VP } } } } Materials Research Laboratories, Inc. } } } } 800-424-1776 } } } } www.mrllab.com } } } } ------------------------------------------- } } No files should be attached to this message } } ------------------------------------------- } } Warren E. Straszheim, Ph.D. } } Materials Analysis and Research Lab } } Iowa State University } } 46 Town Engineering } } Ames IA, 50011-3232 } } } } Ph: 515-294-8187 } } FAX: 515-294-4563 } } } } E-Mail: wesaia-at-iastate.edu } } Web: www.marl.iastate.edu } } } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } } Computer applications and networking } } } } } } } }
As the recipient of an excellent grant, I am contemplating a purchase of a digital camera for my High School classroom, to be used often on microscopes of various kinds---with 23mm and 30mm id eyepiece tubes, one of them a trinocular scope. These are not "up-to-date" scopes. Be that as it may. I would like to request comments from list members, who in the past have given lots of such advice. I apologize for covering familiar territory.
I note that many microscopists have mentioned Coolpix's on this list. I am interested in probably either a Coolpix or a Canon 1D EOS digital camera.
Although I would appreciate comments from users of the Canon, I fear that the 1:1.6 cmos photodetector size to 35mm film size would render it difficult to focus the EOS with a straightforward adaptor such as the Diagnostic Instruments adaptors for 35mm cameras---which works QUITE well with a Canon film EOS. THis was my plan, but, again, I am pretty discouraged by potential focusing problems.
I have ruled out the Olympus E20N, after some experience and alot of vignetting.
The Coolpix 5000 looks interesting. It does much of what I need, including monitoring the live specimen on a TV, Computer, or video projector. I like some of the features of the 5700, including better telephoto range for shore bird studies. I have a couple of questions I would refer to those with Coolpix experience:
How well does the 5700 work on a scope? Does the 5700 identically to the 5000 out of the box, with the Nikon adaptor kit? Are these cameras significantly better than the 4500? Are macro kits available to enable focus to within 1/8"? (I intend to use it as an "aquarium microscope" for example to study the community at the sediment/water column interface, for which the Videoflex camera works admirably well.) Can one take a shot easily an within ordinary lag values while monitoring in real time? Nikon has a relay lens (is that a projection lens?) for this purpose, that will work on Coolpix 5000; does it work ok on the 5700?
Another question: is the Nikon D-100 worth the extra cost, and does it incorporate the video monitoring Coolpix features? Is it useable on a scope, and how?
Thank you for any comments or suggestions. I apologize for asking so many questions at once...
Alan Davis Marianas High School Saipan
I wish to ask whether microscopists feel it
-- adavis-at-saipan.com 1-670-322-6580 Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI
I have steadily endeavored to keep my mind free, so as to give up any hypothesis, however much beloved -- and I cannot resist forming one on every subject -- as soon as facts are shown to be opposed to it. -- Charles Darwin (1809-1882)
The right to search for truth implies also a duty; one must not conceal any part of what one has recognized to be true. -- Albert Einstein
As we enjoy great advantages from the inventions of others we should be glad of an opportunity to serve others by any invention of ours, and this we should do freely and generously. -- Benjamin Franklin
Debby- Usually, GFP is fluorescent or luminesces on it own part, and it isn't necessary to localize it with an antibody.
Here, there seems to be some confusion about the subject protein. GFP is not a rabbit protein. If the protein of interest is being localized with a rabbit antibody, that makes sense. But what is the reagent with the GFP? Carol Heckman (Bowling Green State University)
} Date: Fri, 29 Aug 2003 17:28:42 -0500 } Subject: GFP antibody } From: Debby Sherman {dsherman-at-purdue.edu} } To: "message to: MSA list" {microscopy-at-sparc5.microscopy.com} } X-Virus-Scanned: by amavisd-new on Purdue Mailhub } X-Sig: 11b694fff7e42b4e8f56e22d68ad0d62 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Carol GFP is fluorescent, but there are many uses for antibodies against it nevertheless, including immunoblotting, immunoprecipitation and detection of the GFP-labelled protein by immunogold labelling for electron microscopy in the absence of antibodies against the target protein itself.
What is presumably being requested is a recommendation for a reliable anti-GFP.
Chris
----- Original Message ----- } From: "Carol Heckman" {heckman-at-bgnet.bgsu.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Sunday, August 31, 2003 12:19 AM
Carol,
You are correct in that GFP is often inserted into the code of a particular protein so that it is replicated as the protein in replicated. Then the fluorescent properties of the GFP molecule can be used to localize the particular target protein using fluorescent or confocal microscopy. However, this fluorescence is not useful if you want to do higher resolution localization of the protein. For that you need TEM immunocytochemical procedures utilizing a marker such as colloidal gold.
It is very helpful to identify GFP in living tissue and then take that same tissue, prepare it for TEM ICC and then localize the protein using an antibody against the GFP. If that antibody is made in rabbits than you need an anti-rabbit colloidal gold IgG probe.
What I wanted was suggestions on where to get the antibody against GFP that was been successfully used for TEM ICC. I would like to try a second antibody to make sure my localization, or lack of such, is not due to the antibody we have. Often an antibody will work well in Westerns, such as the one we have, and not work as well under ICC conditions. This is just like how one rabbit will produce what looks like a great antibody and another rabbit will produce one that does not appear to be as strong but actually will work better. Don't ask me why,...it just happens.
I did lead you astray in that I said rabbit -GFP in my original post. I am sorry for that in that It should have read rabbit antibody against GFP. We always have lots of anti-rabbit colloidal gold tagged IgG around so prefer rabbit antibodies. However, we can easily get IgG from other sources so that should not be a limit if there is a GFP antibody out there from another organism that works better than the rabbit ones.
Debby
On 8/30/03 6:19 PM, "Carol Heckman" {heckman-at-bgnet.bgsu.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Debby- } Usually, GFP is fluorescent or luminesces on it own part, and it } isn't necessary to localize it with an antibody. } } Here, there seems to be some confusion about the subject protein. } GFP is not a rabbit protein. If the protein of interest is being } localized with a rabbit antibody, that makes sense. But what is the } reagent with the GFP? } Carol Heckman } (Bowling Green State University) } } } Date: Fri, 29 Aug 2003 17:28:42 -0500 } } Subject: GFP antibody } } From: Debby Sherman {dsherman-at-purdue.edu} } } To: "message to: MSA list" {microscopy-at-sparc5.microscopy.com} } } X-Virus-Scanned: by amavisd-new on Purdue Mailhub } } X-Sig: 11b694fff7e42b4e8f56e22d68ad0d62 } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi, } } } } Can anyone recommend a source for an antibody against rabbit-GFP to use } } for TEM immunocytochemistry? We are trying to localize a protein with a GFP } } construct in arabidopsis root tissue. We have tried one antibody but } } labeling is poor and not well localized. We want to make sure that it is } } not the antibody. The protein is an unknown and the research group has not } } yet been able to develop a clean antibody against it. } } } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-052 Whistler Building } } 170 S. University Street } } West Lafayette, IN 47907 } } }
Apologies for the late reply. I don't want to overdo this topic, but few peeps have addressed one of the original questions: is there much danger to getting abrasive sloshed in your eye?
While SiC/water splashes might not be much of a chemical irritant, aqueous suspensions of metal oxide particles are typically not pH-neutral, e.g., alumina suspensions--} acidic, silica suspensions--} basic. So, if you work with those types of polishing suspensions, an eye splash seems likely to be more painful than just the irritation associated with having gritty stuff in your eye.
I work in a lab where I wear safety glasses and frequently go back and forth between the polisher and the optical microscope. It's a bit of a pain to take the glasses off and on, but you get used to it, and the slight inconvenience is worth preventing a splash in the eye.
Eve
-- Eve Donnelly Experimental Biomechanics Lab Sibley School of Mechanical & Aerospace Engineering Cornell University 130 Upson Hall Ithaca, NY 14853 tel. 607.255.3582 fax. 607.255.1222
I have two suggestions: 1. Cryo sections of the same should help 2. Application of alcohol helps to loosen the epithelium which can then be gently peeled off. This techique is used to debride the corneal epithelium during surgeries. Shashi CCMB, Hyderabad INDIA
Hi all,
I've been asked to see what the EM difference is between rat eye lens epithelium in culture and in situ. So I'd like to fix the epithelium while it is still in the eye. I'm worried that if I do that it then won't be possible to get the epithelium off the lens without damaging it; I also seem to remember that lens becomes brittle and difficult to handle when fixed. I'd like to keep life simple, so it may be necessary to take the epithelium off the lens before fixing. Does anyone have experience with this?
Thanks,
Diana
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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Temporary position (contractor status) offering 40 hrs/week with limited benefits
Job Description: Transmission Electron Microscopist / Analyst in Process Characterization Lab supporting the Advanced Technology Development Facility (ATDF) as well as both internal and external projects at International SEMATECH (ISMT) in Austin, Texas. The TEM group at ISMT provides materials characterization in support of research and process development projects for future integrated circuit manufacturing. Our research projects tend to focus on materials issues several years ahead of current semiconductor industry manufacturing trends, we therefore often study materials systems that offer new and exciting characterization problems and often collaborate with leading researchers at national labs and universities around the world, however we also have a significant workload involving routine characterization and process monitoring for systems that are already well understood.
Required: M.S. or Ph.D. in Materials Science, Solid-State Physics or Chemistry with strong analytical TEM experience.
For more details please contact: Brendan Foran -at- Brendan_Foran-at-SEMATECH.org
International SEMATECH, headquartered in Austin, Texas, is a global consortium of leading semiconductor manufacturers that represent about half the world's semiconductor production. International SEMATECH engages in cooperative, precompetitive efforts to improve semiconductor manufacturing technology through the support of our members. To learn more about International SEMATECH, visit our website at www.sematech.org.
} not related to the real safety (mostly keeping paperwork in a good shape } and accurately fill out chemical waste disposal tags). So, in such } situation, I think, it's superviser's personal responsibility to provide } safely environment to the workers and train them accordingly.
Look, we don't want to have this devolve into a session on what is safe and what isn't, but this is a very simplistic view. Paperwork may be essential for real safety of all handlers of material down the chain of disposal. If a supervisor has a limited budget, he/she is going to scrimp on safety.
Policies are in place to protect workers who historically have been mistreated. Yes, blanket policies do not apply in all situations. The bureaucracy simply needs some flexibility for individually documented exemptions.
} P.S. By the way: in my EM work, my glasses, I wear to correct my vision, } save my eyes in uncounted number of times. I, also, used to remove them } every time I am using light microscope or binoculars. I don't see big } problem removing them if necessary. But: yes, it decreases my } productivity. Ok, let say, I need about 5 sec. to remove them and put } back. Let say I do it 40 times a day: 5 secx40=200 sec/3.3 min per day.
But you're wearing gloves and your gloves shouldn't be near your unshielded eyes each time you have to handle the googles. So you really should take off your gloves before removing your goggles each time.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
Dear Listeners, I am making ferrite nanoparticles doped with Co and/or Mn in the size range of 20-100nm. I have looked at them with TEM and made up size distributions from the pictures. However, I would now like to check my distribution curves with a particle sizer. Does anyone now if this is possible? I suspect it to be difficult since my particles are magnetic. If it is possible, does anyone know what type of medium I can disperse my particles in? So far I have tried to alter the pH but it doesn't seem to be enough and the particles still agglomerate before the the measurement is over. Thanks .
Yours sincerely Richard
PhD student, Swedish Defense Research Agency Department of Polymer Technology Royal Institute of Technology Teknikringen 56 SE-100 44 Stockholm SWEDEN phone: +46-8-790 76 37
--------------------------------------------------------- Legal Notice: This electronic mail and its attachments are intended solely for the person(s) to whom they are addressed and contain information which is confidential or otherwise protected from disclosure, except for the purpose they are intended to. Dissemination, distribution, or reproduction by anyone other than their intended recipients is prohibited and may be illegal. If you are not an intended recipient, please immediately inform the sender and send him/her back the present e-mail and its attachments and destroy any copies which may be in your possession.
Our webppage on connecting a Coolpix to a microscope will answer a lot of your questions: http://www.mvia.com/Coolpix/clpxadpt.htm
Also, please see my answers below...
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Alan Davis wrote: } } The Coolpix 5000 looks interesting. It does much of what I need, } including monitoring the live specimen on a TV, Computer, or video } projector. I like some of the features of the 5700, including better } telephoto range for shore bird studies. I have a couple of questions I } would refer to those with Coolpix experience: } } How well does the 5700 work on a scope?
Very poorly - you'll need to use the digital zoom mode on the 5700. See our FAQ section: http://www.mvia.com/Coolpix/clpxadpt.htm#FAQ
} Does the 5700 identically to the 5000 out of the box, with the Nikon } adaptor kit?
NO! You'll need to use the digital zoom mode on the 5700 as opposed to the optical zoom mode on the 5000, which will result in very poor images.
} Are these cameras significantly better than the 4500?
The 5000 has a higher resolution, but does not have a swivel body as the 4500 model does, which make it a little less ergonomical. However, the 500 will give you better images.
} Are macro kits available to enable focus to within 1/8"? (I intend } to use it as an "aquarium microscope" for example to study the community } at the sediment/water column interface, for which the Videoflex camera } works admirably well.)
There are a couple of lenses available from Nikon for various wide angle and fisheye applications.
} Can one take a shot easily an within ordinary lag values while } monitoring in real time?
Yes, but the live image will pause when you capture your image.
} Nikon has a relay lens (is that a projection lens?) for this } purpose, that will work on Coolpix 5000; does it work ok on the 5700? } } Another question: is the Nikon D-100 worth the extra cost, and does it } incorporate the video monitoring Coolpix features? Is it useable on a } scope, and how? } } Thank you for any comments or suggestions. I apologize for asking so } many questions at once... } } Alan Davis } Marianas High School } Saipan } } I wish to ask whether microscopists feel it } } -- } adavis-at-saipan.com 1-670-322-6580 } Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI } } I have steadily endeavored to keep my mind free, so as to give up any } hypothesis, however much beloved -- and I cannot resist forming one } on every subject -- as soon as facts are shown to be opposed to it. } -- Charles Darwin (1809-1882) } } The right to search for truth implies also a duty; one must not } conceal any part of what one has recognized to be true. } -- Albert Einstein } } As we enjoy great advantages from the inventions of others we should } be glad of an opportunity to serve others by any invention of } ours, and this we should do freely and generously. } -- Benjamin Franklin } }
Here is our answer to the many responses concerning the previous thread on Torr Seal. Torr Seal is an extremely high quality epoxy resin manufactured under a private label agreement as Torr Seal. Since there are non-critical applications which do not require such a high quality epoxy we, like others do carry a product known as a Torr Seal equivalent. We do let all our users know that when they select an 'equivalent they are paying less, but naturally there is a sacrifice in quality.
John Arnott
Disclaimer: Ladd Research sells the products mentioned in this e-mail.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
Dear Listeners, I am making ferrite nanoparticles doped with Co and/or Mn in the size range of 20-100nm. I have looked at them with TEM and made up size distributions from the pictures. However, I would now like to check my distribution curves with a particle sizer. Does anyone now if this is possible? I suspect it to be difficult since my particles are magnetic. If it is possible, does anyone know what type of medium I can disperse my particles in? So far I have tried to alter the pH but it doesn't seem to be enough and the particles still agglomerate before the the measurement is over. Thanks .
Yours sincerely Richard
PhD student, Swedish Defense Research Agency Department of Polymer Technology Royal Institute of Technology Teknikringen 56 SE-100 44 Stockholm SWEDEN phone: +46-8-790 76 37
--------------------------------------------------------- Legal Notice: This electronic mail and its attachments are intended solely for the person(s) to whom they are addressed and contain information which is confidential or otherwise protected from disclosure, except for the purpose they are intended to. Dissemination, distribution, or reproduction by anyone other than their intended recipients is prohibited and may be illegal. If you are not an intended recipient, please immediately inform the sender and send him/her back the present e-mail and its attachments and destroy any copies which may be in your possession.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Jlindstrom-at-Hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, September 2, 2003 at 17:19:35 ---------------------------------------------------------------------------
Email: Jlindstrom-at-Hotmail.com Name: james lindstrom
I would appreciate off-list feedback from users of JEOL 6700F tools.
I'm looking at a system with turbo pump, large specimen exchange interlock, solid state BSE vs. Robinson BSE, beam stability, accommodation to EDAX Genesis EDS and overall usability and support. Location is Northern CA.
Off-list please. All are confidential. Trying to make a purchase decision between this and comparable Hitachi. Specimens are microchips in cross section, top down, metallurgical coupons of IC manufacturing tools, legacy support for Amray 12mm diameter 3.1mm pin specimen stubs. Also, there is a need for working with rather large specimens. Like blocks of NIST standards (100mm diameter, 50mm high).
Native digital capture resolution is low. Are there other options that work? Is anyone using Soft Imaging ADDA? 4Kx4K pixels minimum is needed.
} From: "Michael Cammer" {cammer-at-aecom.yu.edu} : : } not related to the real safety (mostly keeping paperwork in a good shape : } and accurately fill out chemical waste disposal tags). So, in such : } situation, I think, it's superviser's personal responsibility to provide : } safely environment to the workers and train them accordingly. : : Look, we don't want to have this devolve into a session on what is safe and : what isn't, but this is a very simplistic view. Paperwork may be essential : for real safety of all handlers of material down the chain of disposal. If : a supervisor has a limited budget, he/she is going to scrimp on safety. : : Policies are in place to protect workers who historically have been : mistreated. Yes, blanket policies do not apply in all situations. The : bureaucracy simply needs some flexibility for individually documented : exemptions. : : } P.S. By the way: in my EM work, my glasses, I wear to correct my vision, : } save my eyes in uncounted number of times. I, also, used to remove them : } every time I am using light microscope or binoculars. I don't see big : } problem removing them if necessary. But: yes, it decreases my : } productivity. Ok, let say, I need about 5 sec. to remove them and put : } back. Let say I do it 40 times a day: 5 secx40=200 sec/3.3 min per day. : : But you're wearing gloves and your gloves shouldn't be near your unshielded : eyes each time you have to handle the googles. So you really should take : off your gloves before removing your goggles each time. : Face shields will solve the glove problem. If the curve of plastic face shields interferes with your work a welders helmet with plain glass or you prescription lenses could be used. You would need some clearance behind the microscope but it would speed things up and not run the risk of getting something in your eye when you were removing goggles. With prescription lens in the helmet it should speed up work as you don't have to remove your glasses. It also solves the problem of scratched face shields.
Here is an example http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&item=2551210520&category=633 The part below the lens should be cut away as you don't need protection from the burning rays of the welders arc and the dark lens replaced with a clear one. Then just the push of your thumb will stand the helmet straight up out of the way of a microscope.
A welders helmet should be sturdy enough to satisfy any safety officer and quicker to tip up than removing goggles. Some surgery on the hood would make it lighter and reduce the clearance need behind the microscope with out compromising eye safety.
There are also some goggles that have a flip up lens available at welders supply houses as well. I don't know if they would clear a microscope. Here is an example http://cgi.ebay.com/ebaymotors/ws/eBayISAPI.dll?ViewItem&category=35000&item=2429373537 There are other designs that might be more suitable with the lens swinging 180 degrees. A call to your local Air Gas or other welding supply dealer will let you know what is available.
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
Dear Colleagues, We have following trouble with Zephyr ZEM 1000CW watercooling unit: When our CM12 is switched on from STAND BY, overload protector switches off the pump motor of ZEM 1000CW from time to time. If the reset button of overload protector is pressed down, the motor starts and all is OK. But after several days or weeks this occurs again. Please, can you give us some hints or suggestions. Many thanks in advance. Oldrich Benada
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-241062399 Fax: +420-241062347 WEB: http://www.biomed.cas.cz/mbu/lem113/lem.htm
Hello, I am searching for a manual for Reichert FC4 cryoultramicrotome. Is there anybody who could help me with that? Best regards,
Dr Dobroslawa Budka Materials Research Group iThemba LABS PO Box 722 Somerset West 7129 South Africa fax: +27-21-8433543 phone: +27-21-8431161 mobile phone: 0722656091
We are looking for volunteers to perform beta testing of some code to do HREM image processing and Direct Methods. The code is a gnu-like package (i.e. free) which should compile on any unix or Mac computer fairly simply using a conventional "./configure ; make ; make install" strategy. If you know what I am talking about and are interested, please email me (not the whole list). Thanks.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm2-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
I just about wrote "Urgent Assistance Needed" as the subject line, but then realized that would probably not make it past several dozen spam filters. I'm in my usual position (stuck between a rock and a deadline). I'm looking for a copy (original reprint or the journal issue) of:
MANTON I. & VON STOSCH H.A. 1966. Observations on the fine structure of the male gamete of the marine centric diatom Lithodesmium undulatum. Journal of the Royal Microscopical Society 85: 119-134.
I need to scan several figures from the paper, and our interlibrary loans can only get a xerox copy. Can anybody help? I'd gladly pay courier charges (both ways if you need the journal/paper back). Alternatively, if someone is willing, they could scan the figures from their end, but it would most likely be all of the plates in the paper, as I'm not sure right now which ones I need.
If you help, I'll make several offerings on your behalf to the gods of microscopy, requesting long filament life, long strings of buttery smooth silver-gray thin sections, and awesome powers against the Dark Forces of Bureaucracy. I'll also put you on the list to receive the coveted large format scan of the Philips 200 column cross-section poster (discussed earlier in this forum) which I should have constructed and cleaned up Real Soon Now.
Please contact me off-list. Thanks,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I'm trying to stain liver sections embedded in paraffin and fixed by metacarn fixative. My first antibody is monoclonal. The problem is that I only have a positive signal at the periphery of the section and nothing in the middle. Does someone have any clue why this could happen? Thanks in advance.
Prof. Dr. Francisco Javier Hernandez Blazquez Universidade de São Paulo Fac. de Medicina Veterinária e Zootecnia Departamento de Cirurgia - Setor de Anatomia Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
Thanks for the quick reply, Chris (and others who may be in the pipe),
Even my good luck seems to have a bad side! Not 5 minutes after posting this request, I found a copy of this paper, literally under my nose. Embarassing to say the least! I'll be quiet for at least a week....
Cheers,
Jim
Christopher F. Blanford wrote:
} James - } } Unfortunately we can't take any copies of the journal out of the } library. But as the RMS is just up the street from me, do you think } you could just give them a call? Either see if they have a copy they } could send you, or if you're still in a bind, see if they'd loan me a } copy for a day and I could scan in the pages you want. The number: +44 } 1865 248 768. } } Good luck. } } Chris } } On Wednesday, September 3, 2003, at 01:40 pm, James M. Ehrman wrote: } } } Hi all, } } } } I just about wrote "Urgent Assistance Needed" as the subject line, } } but then realized that would probably not make it past several dozen } } spam } } filters. I'm in my usual position (stuck between a rock and a } } deadline). I'm looking } } for a copy (original reprint or the journal issue) of: } } } } MANTON I. & VON STOSCH H.A. 1966. Observations on the fine structure } } of the male gamete of the marine centric diatom Lithodesmium } } undulatum. Journal of } } the Royal Microscopical Society 85: 119-134. } } } } I need to scan several figures from the paper, and our interlibrary } } loans can only } } get a xerox copy. Can anybody help? I'd gladly pay courier charges } } (both ways } } if you need the journal/paper back). Alternatively, if someone is } } willing, they could } } scan the figures from their end, but it would most likely be all of } } the plates in the paper, } } as I'm not sure right now which ones I need. } } } } If you help, I'll make several offerings on your behalf to the gods } } of microscopy, requesting } } long filament life, long strings of buttery smooth silver-gray thin } } sections, and awesome } } powers against the Dark Forces of Bureaucracy. I'll also put you on } } the list to receive } } the coveted large format scan of the Philips 200 column cross-section } } poster (discussed } } earlier in this forum) which I should have constructed and cleaned up } } Real Soon Now. } } } } Please contact me off-list. Thanks, } } } } Jim } } } } -- } } } } James M. Ehrman } } Digital Microscopy Facility } } Mount Allison University } } Sackville, NB E4L 1G7 } } CANADA } } } } phone: 506-364-2519 } } fax: 506-364-2505 } } email: jehrman-at-mta.ca } } www: http://www.mta.ca/~jehrman } } } } } } } } } }
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (taylor-at-bio.fsu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 3, 2003 at 11:21:33 ---------------------------------------------------------------------------
Email: taylor-at-bio.fsu.edu Name: Kenneth Taylor
Organization: Florida State University
Title-Subject: Fixation and Embedding of bacteria
Question: Hi,
I am looking for a protocol for fixing, embedding and sectioning bacteria, in particular E. coli. Good membrane preservation would be important for this study.
Does anyone have a protocol that they would be willing to share?
OK...thanks to all of the respondents about FESEMs. I did receive some good feedback.
I am trying to make a budget wedge for FY04 and need to know a basic idea of benefits of what I have selected as candidate systems. That said, I also need to put in funding wedges to cover the venue of appropriate SEMs.
JEOL 6700F Hitachi S-4800 Hitachi S-5200
Other options are welcome too.
I would appreciate knowing personal experiences based on differing sizes of specimens and different types of analysis. Budget time comes along one time, and is perversely ignorant of systems. Caveat emptor.
It may be time to upgrade. But to what? Cold FE with accommodation of my EDAX Cryospec is necessary. Robinson BSE is also a big plus.
Hello, could you please let me know who the suppliers of HISTORESIN are. It is a glycol methacrylate usful for light microscopy. It used to be marketed originally as LKB HISTORESIN. Thank you, Dobrusia
Dr Dobroslawa Budka Materials Research Group iThemba LABS PO Box 722 Somerset West 7129 South Africa fax: +27-21-8433543 phone: +27-21-8431161 mobile phone: 0722656091
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (reckert-at-nwlabs1896.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 3, 2003 at 21:24:36 ---------------------------------------------------------------------------
Question: We are interested in purchasing a CamScan CS44 SEM (appr. 10 years old) and I was wondering if anybody could share their experience (weak and strong points) about the scope. Thank you.
Well, BILLPC has stopped sending me copies of a virus, but I still get copies from PRIVAT (pD9E76707.dip.t-dialin.net [217.231.103.7], and MINIHOTEL (modemcable027.214-131-66.nowhere.mc.videotron.ca [66.131.214.27]
Some of the spoofed addresses/domains on the messages are ones I recognize from this list. So if any of you recognize these folks or their domains, would you please tell them to check out their machines. Thanks.
Hello Historesin is now produced and sold by Leica Microsystems Nussloch GmbH. Their address is Heidelberg Strabe 17-19 D-69226 Nussloch/Heidelberg Tel. 0 62 24/143-0 You may look at their homepage at http://www.histo-solutions.com/website/sc_hbu.nsf
Good luck
Prof. Dr. Francisco Javier Hernandez Blazquez University of São Paulo Fac. Veterinary Medicine and Animal Sciences Departament of Surgery - Sector of Anatomy Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) - Brasil Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
----- Original Message ----- } From: "dobrusia budka" {dobrusiabudka-at-tlabs.ac.za} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, September 04, 2003 8:26 AM
Hi:
Help!!! I'm presently having problems with photography of nerve tissue. My problem is that the tissue appears shredded, and it has multiple holes and splits along the tissue but not the plastic. Making the tissue impossible to photo. This problem can only be seen when we are to photograph in the microscope. 1 micron light slides look fine. The only thing we have change is the type of fixative. We went from glutaraldehyde to para formaldehyde, but nothing have change from my protocol. We think is an infiltration problem, because the tissue is big to start and humidity have been a problem in the past in other types of tissue, and because the holes and splits are only in the tissue. We are looking for processing protocols for Sural Nerve tissue in order to compare with the one we have. If anyone can help it will be greatly appreciated.
Thanks
Omayra Velez MS Electron Microscopy Specialist Pathology Department Weill Cornell Medical College 1300 York Ave NY,NY, 10021 212-746-6437 omv2001-at-med.cornell.edu
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Does anyone know anything about the ISI WB-6 that is currently being offered on eBay? A friend of mine was wondering about the specifics concerning the condition and usability of the instrument.
Omayra It looks like fixation/impregnation problem. Paraformaldehyde itself is not enough for EM. I know nothing about your particular sample, in general, we are using 4% paraformaldehyde+ 1% GA overnight (+4oC). Nervous tissue also contains a lot of lipids, so dehydratation should be slow: actually, all steps should be longer than usual. I do find that Spurr resin is good for brain tissue: it penetrates better than Epon or Araldite. Good luck, Sergey.
At 02:04 PM 9/4/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Historesin as such is now discontinued but lives on as a series of resins produced by Heraeus Kulzer in Germany - Technovit 7100 (Historesin), Technovit 8100 (Historesin Plus) and Technovit 9100 NEW (Histodur).
These resins are all identical to the Historesin products. TAAB Laboratories Equipment Ltd is the Distributor for these products in the UK, but if anyone has a problem finding the distributor in their country, I may be able to help,
Best regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, England Tel ++44 (0)118 981 7775 Fax ++44 (0)118 981 7881 e-mail sales-at-taab.co.uk www.taab.co.uk
I have problems with my EBSD measurements. The image of the phosphorscreen is distorted at the bottom of the screen while substracting the backgroung. instead of pure cirlces at the bottom you see more or less straight lines cutting the circles or the circles themselfs are distorted. When substracting at very low magnifiocations this problem doesnŽt occure. When you look at the RAW picture the area at the bottom shows a dark line. Please I need help. It is urgent
Regards Dirk Kirch -- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
I have problems with my EBSD measurements. The image of the phosphorscreen is distorted at the bottom of the screen while substracting the backgroung. instead of pure cirlces at the bottom you see more or less straight lines cutting the circles or the circles themselfs are distorted. When substracting at very low magnifiocations this problem doesnŽt occure. When you look at the RAW picture the area at the bottom shows a dark line. Please I need help. It is urgent
Regards Dirk Kirch
-- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
-- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
We are thinking to buy a liquid helium cooled double tilt TEM holder. Has anyone used this type of holder? Can you please tell me your experience with it? I would appreciate very much if you could tell me how well it works, and what problems it might have.
I would like to ask more specifically about the liquid helium cooling TEM holder made by USA side of the Gatan company. The UK made holder will be too expensive for us, so anyone has experience with the US made holder?
Hi to all: Thanks for the responce on the nerve problem. Some of you have asked how big the tissue samples are and the type of resin I'm using. We are using, EMBED 812 with DMP-30 as catalyst. The tissue size is about 5mm x 2mm x 3mm. Big for the most part. I already tryed to make the blocks smaller, but the nero-pathologist wanted the pieces this big. We have a 20 hour protocol from buffer to 100% epon. I'll try some of the sudjestions, lenthtening the protocol. Although we think we are also having a humidity issue as well, due to construction and AC problems.
Thanks again
Omayra Velez Electron Microscopy Specialist Pathology Department Weill Cornell Medical College 1300 York Ave NY,NY. 10021 212-746-6437 omv2001-at-med.cornell.edu
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Here is the table of contents for the September/October 2003 issue of Microscopy Today.
New Subscriptions via http://www.microscopy-today.com only, please New subscriptions will close on Tuesday 9 September for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting. Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA. MSA members anywhere have free subscriptions. Non-MSA, non-North American subscriptions have been reduced from $80 or $110US to $35US. Additional details in the magazine or on our web site.
Table of Contents:
Carmichael Print Your Own Organs! Kelly and Gribb Atom Probes LEAP Ahead Breger Microscopy At The Ends Of The Earth Barkan, et al. A Si Multi-Cathode Detector For Microanalysis Applications Sehgal, Karim, Stafford, & Fasolka Techniques for Combinatorial and High- Throughput Microscopy Part 1: Gradient Spec. Fabrication for Polymer Thin Film Research Millette, Boltin, Few, & Turner Microscopical Studies of World Trade Center Disaster Dust Particles Vrdoljak Prep. Of Soil Samples For Light And TEM Radzikowska A New Look At Cast Iron Microstructure Lifshin & Gauvin Precision and Detection Limits for EDS Analysis in the SEM Sedgewick Adding Color To Grayscale Images Downing Support Films with Uniform Hole Size Mills Cleaning a Cold Cathode Gauge Tube Yang and Kalab Zigzag Edges in SEM Micrographs Schooley Microscopy CD-ROMs For Children: A Bibliography
Does anyone have a package or know of a package that can calculate changes in sample thickness from thickness fringes observed in conventional TEM? I am imageing defect free single crystals on the order of 100nm. I need to know how the crystals thickness profile changes.
I have a student who wants to image magnetic particles with SEM. The particles are in the 400nm to 1µm size range. I had planned to stick them to double stick copper tape prior to imaging. However, I am concerned about their adversely affecting the microscope. Are there any suggestions as to mounting the particles, whether short working distance may magnify potential problems if some particles break loose and get into the lens. Etc?
If there is a concern then I guess it would be possible to mount on sticky carbon tape, overlay the sample with a formvar film, and then use backscattering to image the particles. However, is this necessary?
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
If one wants to do pure standards-based quantitative EDS analysis, protocol says that conditions for the standards must be the same as for the specimen. Well, if one is using a thermal FE SEM, then the beam current is very likely to be the same at any time. How about for a cold FE gun?
From what I can surmise, the cold guns vary much more than thermal FE guns, to the point that intervention is necessary. Does this mean that every quant EDS measurement has to have a precursor Faraday cup reading of beam current? Conversely, does this mean that this is not required when using a thermal FE gun?
The EDAX system supports a Beam Current Factor so that calculations are made based on as-read current when making the quant analysis. This seems like a lot of hassle. Is the difference between standardless and standards-based EDS so huge that the hassle is warranted? Or, is this nit picking?
Is it possible to simply degauss the samples before putting them in the SEM?
There is a border, I believe, between too much degaussing and too little. Looking at hard drive tracks is a case in point. Depending on your ultimate resolution, the working distance may facilitate your specimens by keeping them away from the final lens. 1um specimens should not require short WD.
Your BSE ought to be a separate issue depending on what type it is and what KV it works at.
gary g.
At 03:14 PM 9/5/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have routinely examined (gamma-)ferric oxide particles in the SEM by dusting the particles onto double-sided sticky carbon tape. Or you can paint a small dab of carbon paint onto your stub and dust the particles onto the paint while it is still wet. Then tap the side of the stub firmly on a hard surface to dislodge any loose particles. If you are really paranoid you can use a low-pressure gas stream (like a "Micro Duster") to make sure there are no loose particles present. However, the more aggressively you remove looser particles, the less you are likely to see larger particles that may exist in the specimen. This may be important if you are trying to do particle size distribution measurements.
Particles like Fe2O3 are weakly magnetic, so there is not much danger of them being sucked up into your lenses by the magnetic field.
You will probably want to keep the working distance fairly short ( {10mm) to achieve acceptable image quality.
Hope this helps, Ken Gaugler
Debby Sherman wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } } I have a student who wants to image magnetic particles with SEM. The } particles are in the 400nm to 1µm size range. I had planned to stick them } to double stick copper tape prior to imaging. However, I am concerned about } their adversely affecting the microscope. Are there any suggestions as to } mounting the particles, whether short working distance may magnify potential } problems if some particles break loose and get into the lens. Etc? } } If there is a concern then I guess it would be possible to mount on } sticky carbon tape, overlay the sample with a formvar film, and then use } backscattering to image the particles. However, is this necessary? } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } } }
Wouldn't it also be dependent on the type of SEM used (i.e. HRSEM)?
If the final lens (pole piece) is not energized for through-the-lens detection, the sticky Cu tape should hold them down. Disperse the particles then blow off the weakly held particles. Now lower a ferro magnetic material over the samples and see if any pop off. If particles do "pop" off then use formvar and a high kV. The magnetism of the particles may play a role in their physical construct so degauss with caution. I believe imaging aberrations will most likely be the issue over that of tool contamination. I have successfully imaged toner particles in a relatively low resolution SEM. It should be noted that this tool has been classified as a "dirty" tool. The toner particle were even dispersed on the "new" C sticky tabs. The ones that are rock hard with barely any adhesive.
Kristopher
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Friday, September 05, 2003 7:28 PM To: Debby Sherman Cc: MSA listserver
Is it possible to simply degauss the samples before putting them in the SEM?
There is a border, I believe, between too much degaussing and too little. Looking at hard drive tracks is a case in point. Depending on your ultimate resolution, the working distance may facilitate your specimens by keeping them away from the final lens. 1um specimens should not require short WD.
Your BSE ought to be a separate issue depending on what type it is and what KV it works at.
gary g.
At 03:14 PM 9/5/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You really don't need a package. You can do this by hand using basic TEM techniques.
I think that your best way to do this would be to set up a good 2-beam condition and use convergent beam electron diffraction. You use the distances between the symmetrical fringes (Kossel-Mollenstedt fringes) in the known reflection, calculate the deviation from exact Bragg condition for each fringe, and plot the results assuming that the first fringe that you see is the 1st, 2nd, or 3rd. You plot s(i)^2/n(i)^2 vs 1/n(i)^2, where n(i) is the index of the particular fringe. You try different starting values for n(1) starting from 1, then 2, then 3. You usually don't have to go higher than 3. When you get a straight line, the slope of the straight line is -1/(extdist)^2 and the intercept of the line is 1/t^2. This comes from the following relationship,
s(i)^2/n(i)^2 + 1/(extdist)^2/n(i)^2 = 1/t^2.
This technique is demonstrated in a number of TEM texts. The procedure is written in the Williams and Carter book.
You could apply this at two points on the sample along the wedge and get the angle.
You could also do this from the results of the two-beam approximation model. If you are at 2-beam Bright Field condition and you do not have bend contours present, then the first dark fringe that occurs in the wedge will occur at a thickness of 0.5 * (extdist(g)) and the second at 1.5 * (extdist(g)), (and so forth.) From the lateral separation distance between the fringes and the value of the extinction distance for that reflection, you can calculate the wedge angle. This is also written up in the Williams and Carter book and others as well.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: thurston e herricks [mailto:thurst0n-at-u.washington.edu] Sent: Friday, September 05, 2003 3:37 PM To: Microscopy-at-sparc5.microscopy.com
Hello all,
Does anyone have a package or know of a package that can calculate changes in sample thickness from thickness fringes observed in conventional TEM? I am imageing defect free single crystals on the order of 100nm. I need to know how the crystals thickness profile changes.
Take care
Thurston Herricks
From MicroscopyL-request-at-MSA.Microscopy.Com Sun Sep 7 20:14:53 2003 Return-Path: {"zaluzec-at-microscopy.com"} Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by microscopy.com (8.12.8/8.12.8) with SMTP id h881EDiI004403 for "MListServerFilteredEmail_5-at-microscopy.com"; Sun, 7 Sep 2003 20:14:33 -0500 Received: from [206.69.208.25] (mac25.zaluzec.com [206.69.208.25]) by microscopy.com (8.12.8/8.12.8) with ESMTP id h881ECiJ004400 for {microscopy-at-microscopy.com} ; Sun, 7 Sep 2003 20:14:12 -0500 Mime-Version: 1.0 Message-Id: {p05200f11bb8186881db1-at-[206.69.208.25]}
Colleagues:
The Microscopy Listserver Archives are now updated through August 2003.
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-MSA.Microscopy.Com Mon Sep 8 13:12:34 2003 Return-Path: {"taylor-at-bio.fsu.edu"} Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by microscopy.com (8.12.8/8.12.8) with SMTP id h88IBlo6011676 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 8 Sep 2003 13:12:14 -0500 Received: from [130.202.232.233] (mac21.zaluzec.com [206.69.208.21]) by microscopy.com (8.12.8/8.12.8) with ESMTP id h88IBUo7011672 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 8 Sep 2003 13:11:30 -0500 Mime-Version: 1.0 X-Sender: (Unverified) Message-Id: {p05111a0abb8274a145b5-at-[130.202.232.233]}
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Email: taylor-at-bio.fsu.edu Name: Kenneth Taylor
Organization: Florida State University
Title-Subject: Fixation and Embedding of bacteria
Question: Hi,
I am looking for a protocol for fixing, embedding and sectioning bacteria, in particular E. coli. Good membrane preservation would be important for this study.
Does anyone have a protocol that they would be willing to share?
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Email: Jlindstrom-at-Hotmail.com Name: james lindstrom
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Email: icastro-at-usiminas.co.br Name: Ivan de Castro
Organization: Usiminas
Education: Graduate College
Location: Brazil
Question: I'm having some difficulty on metallographic examination of Hot Dip Galvanized steels. Then I will shall be pleased if you could help me giving some information about papers that discrebe the better technique to metallographic prepation and etching of HDG steels. Thanking you Yours sincerely
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Email: Jlindstrom-at-Hotmail.com Name: james lindstrom
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Sorry Colleagues
I've run into a serious problem, which is taking the Listserver off-line for a few days.
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Email: maloneyb-at-fiu.edu Name: Barb
Organization: FIU
Title-Subject: Receipe for cultured cells on filter for SEM
Question: Dear Group - what is the newest and most successful receipe for prep for SEM on cultured cells on a filter? Thanks Barb
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (babbini-at-units.it) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 10, 2003 at 05:57:53 ---------------------------------------------------------------------------
Email: babbini-at-units.it Name: Lorenza Babbini
Organization: Dept. of Biology - University of Trieste - Italy
Title-Subject: MListserver:
Question: Hi all! I have to do my sections of decalcified coralline algae and I would like to try to use Histocryl resin. Is there anyone that ever used it for sectioning plants? Suggestions and comments are welcome! Lorenza
From MicroscopyL-request-at-microscopy.com Wed Sep 10 16:45:31 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8AKjJ9v011223 for {L1-at-ns.microscopy.com} ; Wed, 10 Sep 2003 16:45:19 -0400 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8AKjJEv011218 for L1; Wed, 10 Sep 2003 15:45:19 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8AKic9v011185 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 10 Sep 2003 16:44:58 -0400 Received: from smtp5.doit.wisc.edu (heimdall.doit.wisc.edu [144.92.197.159]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8AKiH9v011179 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 10 Sep 2003 16:44:28 -0400 Received: from conversion-daemon.smtp5.doit.wisc.edu by smtp5.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) id {0HL000G01M28JX-at-smtp5.doit.wisc.edu} for Microscopy-at-sparc5.microscopy.com; Wed, 10 Sep 2003 15:44:08 -0500 (CDT) Received: from [10.25.50.161] (ansci.wisc.edu [144.92.132.175]) by smtp5.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) with ESMTP id {0HL0005DMMXJ19-at-smtp5.doit.wisc.edu} for Microscopy-at-sparc5.microscopy.com; Wed, 10 Sep 2003 15:44:08 -0500 (CDT)
Micromavens,
Does anyone have for sale a used but still properly functioning Metaltek filter wheel, Model # 5240? The wheel itself is all we'd need, but the controller could be useful as well. Or possibly a Uniblitz shutter model # V505S1Z0? The shutter in our Metaltek has blitzed its last, and I am suspicious of the wheel itself as well. Apologies to those who get this message twice from both the microscopy and the confocal lists.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-microscopy.com Thu Sep 11 11:34:38 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8BFYS9v018775 for {L1-at-ns.microscopy.com} ; Thu, 11 Sep 2003 11:34:28 -0400 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8BFYSdJ018770 for L1; Thu, 11 Sep 2003 10:34:28 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8BFXl9v018739 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 11 Sep 2003 11:34:08 -0400 Received: from nowell.armstrong.com (nowell.armstrong.com [204.74.20.231]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8BFXU9v018735 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 11 Sep 2003 11:33:42 -0400 Received: from lccmx003.americas.armstrong.com (lccmx003.americas.armstrong.com [10.37.66.76]) by nowell.armstrong.com (8.12.9/8.12.9) with ESMTP id h8BFX1bl011513 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 11 Sep 2003 11:33:01 -0400
Calling all Old-Timers!
I have a 1950's vintage AO Spencer microtome with the porro prism, binocular head. In the past, we've had a stereomicroscope with the same head and the same problem, which is prisms breaking loose from the adhesive mounting. I know that it's next to impossible to try to achieve freehand alignment when re-cementing, and am told that there's a jig for this purpose. Not having the jig, and imagining that those who would posess one and the know-how to use it are superannuated, if still living, I'm turning to the group at large for any helpful leads in getting this instrument repaired. Or, does anyone have the heads with intact prisms that you would be willing to sell? Or, am I better off just getting another, used microtome altogether?
Much obliged,
Dave Stadden Research Scientist Testing & Analysis Lab Armstrong World Industries, Inc.
717-396-5109 717-396-5865 (fax)
From MicroscopyL-request-at-microscopy.com Thu Sep 11 13:36:53 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8BHak9v019815 for {L1-at-ns.microscopy.com} ; Thu, 11 Sep 2003 13:36:46 -0400 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8BHak9n019811 for L1; Thu, 11 Sep 2003 12:36:46 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8BHa59v019782 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 11 Sep 2003 13:36:25 -0400 Received: from mailhub-3.iastate.edu (mailhub-3.iastate.edu [129.186.140.13]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8BHZH9v019772 for {Microscopy-at-msa.microscopy.com} ; Thu, 11 Sep 2003 13:35:53 -0400 Received: from mailout-1.iastate.edu (mailout-1.iastate.edu [129.186.140.1]) by mailhub-3.iastate.edu (8.9.3p2/8.9.3) with SMTP id MAA01165 for {Microscopy-at-msa.microscopy.com} ; Thu, 11 Sep 2003 12:35:03 -0500 Received: from pdp2.marl.iastate.edu(129.186.227.2) by mailout-1.iastate.edu via csmap id 9586; Thu, 11 Sep 2003 17:32:52 +0000 (UTC) Message-Id: {5.2.0.9.2.20030911123618.01b71008-at-wesaia.mail.iastate.edu} X-Sender: wesaia-at-wesaia.mail.iastate.edu X-Mailer: QUALCOMM Windows Eudora Version 5.2.0.9
I really do have some worthwhile information to offer, but the short answer may be, it all depends.
The importance and frequency of beam current measurements will depend on the stability of your beam and your tolerance for error. If your beam current only fluctuates by a percent or so over a few minutes, you might be able to ignore it. The uncertainty in EDS results is going to be a larger error than the relative error from beam drift. However, if you experience 5% drift in 5 minutes, I would definitely recommend frequent checks of the beam current to correct for the drift. I say this not having worked with FE guns, so I don't know how much drift to expect. I definitely like to have current measurements often enough so that there is no more than a 5% change in beam current between measurements. More critical measurements would necessitate even more frequent checks.
The type of quant analysis will also make a difference. If you are monitoring all elements in the sample you can normalize your results which should cancel out the effects of beam drift. At moderate count rates all elements should be affected similarly, i.e., only the intensity should drop. The change should not affect the matrix correction, if the software is written correctly. You could check your software by forcing a major change in beam current via the condenser lens to see if the results are the same.
However, if you are not directly quantifying all elements present, e.g., you are analyzing an element by difference, then an accurate measure of beam current will be important. Whatever loss (or gain) in current you encounter will be attributed to the element determined by difference. If that element is not critical (e.g., C in a polymer), then you only need be concerned about the loss (or gain) of concentration in proportion to the current drift. But if you are trying to determine a small component (e.g., 5% Li by difference) it will be profoundly influenced by the beam drift.
The difference in standardless analysis and analysis with your own standards will depend on your system and its software. Some systems use built-in standards for their "standardless" analysis. Those built-in standards are probably collected on another scope in another lab under different conditions, but they can produce some pretty good results when corrected for current scope conditions.
If your technique is good, you might be able to improve on the "standardless" results by collecting your own standards. For instance, our internal standards were collected at the highest resolution mode on our analyzer while we typically do our x-ray work at a faster mode to improve statistics. Therefore, we can improve our deconvolution and thus our results by collecting our own standard profiles. You just need to be sure that your techniques are as good or better than those of the application engineers for your EDS system. I know it took me a while to relearn the lesson that the carbon coat on the standards and unknowns must be identical. The coat can cut the intensities by a few percent.
My personal experience is that when mixing line series (K, L, and M), the use of my own standards usually leads to a substantial improvement in accuracy. For analyses within the line series, I find that I cannot necessarily improve the results.
So like I said, "it depends" whether the close monitoring of beam current will be important or not.
HTH, Warren
At 04:45 PM 9/5/2003 -0700, you wrote:
} Greetings to EDS experts: } } If one wants to do pure standards-based } quantitative EDS analysis, protocol says } that conditions for the standards must } be the same as for the specimen. Well, } if one is using a thermal FE SEM, then } the beam current is very likely to be } the same at any time. How about for a } cold FE gun? } } From what I can surmise, the cold guns } vary much more than thermal FE guns, to } the point that intervention is necessary. } Does this mean that every quant EDS } measurement has to have a precursor } Faraday cup reading of beam current? } Conversely, does this mean that this is } not required when using a thermal FE gun? } } The EDAX system supports a Beam Current } Factor so that calculations are made based } on as-read current when making the quant } analysis. This seems like a lot of hassle. } Is the difference between standardless and } standards-based EDS so huge that the hassle } is warranted? Or, is this nit picking? } } gary g.
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Scanning electron microscopy, x-ray analysis, and image analysis of materials Computer applications and networking
From MicroscopyL-request-at-microscopy.com Thu Sep 11 15:59:05 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8BJwi9v020916 for {L1-at-ns.microscopy.com} ; Thu, 11 Sep 2003 15:58:44 -0400 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8BJwiwo020911 for L1; Thu, 11 Sep 2003 14:58:44 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8BJvq9v020867 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 11 Sep 2003 15:58:13 -0400 Received: from lakemtao06.cox.net (lakemtao06.cox.net [68.1.17.115]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8BJvJ9v020861 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 11 Sep 2003 15:57:30 -0400 Received: from home ([68.12.42.17]) by lakemtao06.cox.net (InterMail vM.5.01.06.05 201-253-122-130-105-20030824) with SMTP id {20030911195708.VPZD4662.lakemtao06.cox.net-at-home} ; Thu, 11 Sep 2003 15:57:08 -0400 Message-ID: {01b001c3789e$e3c4fe90$112a0c44-at-home}
Arthur Rosenfelder roseoptics-at-aol.com has the tools to rebuild these scopes or can probably swap out on that is already reconditioned.
Rosenfelder Optics, Specializing in Microscope Service and Repair. Maintaining research and instructional optics since 1980, P.O. Box 164, Bellvue, Colorado 80512, (970) 217-3368 RoseOptics-at-aol.com Art often has microscopes as well as parts and repair for sale email him for you needs. Art also remounts and realigns prisms in AO Spenser Cycloptic stereo microscopes.
Gordon Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. ----- Original Message ----- } From: "David R Stadden" {DRStadden-at-armstrong.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, September 11, 2003 10:33 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (halbers-at-doacs.state.fl.us) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, September 11, 2003 at 14:55:45 ---------------------------------------------------------------------------
Email: halbers-at-doacs.state.fl.us Name: Susan Halbert
Organization: Division of Plant Industry, Florida Department of Agriculture
Education: Graduate College
Location: Gainesville, FL USA
Question: I have a late '70s vintage Zeiss universal microscope with a built-in camera. Some time ago, the camera blew a fuse. We replaced the fuse, and it blew again, indicating an electrical malfunction. We have not been able to find anyone that can fix it. Does anyone have any suggestions?
From MicroscopyL-request-at-microscopy.com Fri Sep 12 07:24:17 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8CBO29v027269 for {L1-at-ns.microscopy.com} ; Fri, 12 Sep 2003 07:24:02 -0400 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8CBO21b027265 for L1; Fri, 12 Sep 2003 06:24:02 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8CBNG9v027230 for "MListServerFilteredEmail_5-at-microscopy.com"; Fri, 12 Sep 2003 07:23:42 -0400 Received: from mail1.hrz.uni-bielefeld.de (mail1.hrz.uni-bielefeld.de [129.70.4.88]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8CBMo9v027220 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 12 Sep 2003 07:23:06 -0400 Received: from conversion-daemon.mail1.hrz.uni-bielefeld.de by mail1.hrz.uni-bielefeld.de (iPlanet Messaging Server 5.2 HotFix 1.15 (built Apr 28 2003)) id {0HL300301LMQ6G-at-mail1.hrz.uni-bielefeld.de} for Microscopy-at-MSA.Microscopy.Com; Fri, 12 Sep 2003 13:22:41 +0200 (MEST) Received: from UNI-BIELEFELD.DE ([129.70.82.150]) by mail1.hrz.uni-bielefeld.de (iPlanet Messaging Server 5.2 HotFix 1.15 (built Apr 28 2003)) with ESMTPPA id {0HL3009O6M9STR-at-mail1.hrz.uni-bielefeld.de} for Microscopy-at-MSA.Microscopy.Com; Fri, 12 Sep 2003 13:22:40 +0200 (MEST)
Dear All, I would like to post the following question: Is there a sensitive method to measure the intracellular pH by a staining procedure of tissue sections (pancreas tissue)? Thanks for any suggestions! Daniel
-- (-)-(-) --------------------------- \"/ --- Dr. Daniel Eberhard =V= Developmental Biology & Molecular Pathology University of Bielefeld D 33501 Bielefeld/Germany
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Daniel,
By the time you fix the tissue, embed in paraffin (or other medium) and section, then the living intracellular pH is longsince gone. However, some work has been done on living cells in culture, using special fluorescent dyes that change their color or density at different pHs. I don't have any references handy, but you could probably find some by searching PubMed or Google.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Friday, September 12, 2003 1:22 PM +0200 DANIEL EBERHARD {daniel.eberhard-at-uni-bielefeld.de} wrote: } } Dear All, } I would like to post the following question: } Is there a sensitive method to measure the intracellular pH by a staining } procedure of tissue sections (pancreas tissue)? Thanks for any } suggestions! Daniel } } -- } (-)-(-) } --------------------------- \"/ --- } Dr. Daniel Eberhard =V= } Developmental Biology & Molecular Pathology University of Bielefeld } D 33501 Bielefeld/Germany } } FAX: xx49(0)521-106-5654 (-)-(-) } --------------------------- \"/ --- } =V= } }
From MicroscopyL-request-at-microscopy.com Fri Sep 12 13:52:28 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8CHq29v030221 for {L1-at-ns.microscopy.com} ; Fri, 12 Sep 2003 13:52:02 -0400 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8CHq2Nh030216 for L1; Fri, 12 Sep 2003 12:52:02 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8CHp29v030176 for "MListServerFilteredEmail_5-at-microscopy.com"; Fri, 12 Sep 2003 13:51:25 -0400 Received: from zeus.itg.uiuc.edu (zeus.itg.uiuc.edu [130.126.126.162]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8CHoP9v030169 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 12 Sep 2003 13:50:36 -0400 Received: from itg.uiuc.edu (rollins.itg.uiuc.edu [130.126.126.232]) by zeus.itg.uiuc.edu (8.11.6/8.11.6) with ESMTP id h8CHoFb00803; Fri, 12 Sep 2003 12:50:15 -0500 Message-ID: {3F620759.5030908-at-itg.uiuc.edu}
Hello Daniel, There are a number of fluorescent dyes which can be used to determine intracellular pH, the most useful variants exhibit shifting of the emmission spectra with changes in pH. You can check Molecular Probes' website for more information: http://www.probes.com/servlets/directory?id1=3&id2=76 -Karl G.
DANIEL EBERHARD wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } } Dear All, } I would like to post the following question: } Is there a sensitive method to measure the intracellular pH by a } staining procedure of tissue sections (pancreas tissue)? } Thanks for any suggestions! Daniel }
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
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I would be grateful for any input from the listserve for the following request I recently received. Rosemary
"I am working on a crime scene investigation outreach activity for 4th-7th graders. I have the activities prepared already, but I would like to include more hard science involving fiber analysis and fingerprinting. I am familiar with several types of microscopy, but I was wondering if you could help me with the explanations of their application to forensic science. We are also hoping to be able to put images on the outreach website and the final paper version so that students could see, for instance, the differences between types of fibers and have an idea of how a match is made."
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hgabrisc-at-uno.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, September 12, 2003 at 14:02:21 ---------------------------------------------------------------------------
Email: hgabrisc-at-uno.edu Name: Heike Gabrisch
Organization: University of New Orleans
Title-Subject: MListserver: negative scanner
Question: Can anybody recommend a good scanner for TEM negatives?
From MicroscopyL-request-at-microscopy.com Fri Sep 12 23:17:28 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8D3HD9v001629 for {L1-at-ns.microscopy.com} ; Fri, 12 Sep 2003 23:17:13 -0400 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8D3HDar001624 for L1; Fri, 12 Sep 2003 22:17:13 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8D3GR9v001564 for "MListServerFilteredEmail_5-at-microscopy.com"; Fri, 12 Sep 2003 23:16:52 -0400 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8D3GBA0001558 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 12 Sep 2003 23:16:12 -0400 Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) Message-Id: {p05111a02bb883a3ce727-at-[206.69.208.21]}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Gilweiss-at-worldnet.att.net) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, September 12, 2003 at 19:44:55 ---------------------------------------------------------------------------
Question: What is the practical use of circular polarization in microscopy? What physical phenomena initiate it? I have not found a good working source of info on the subject.
From MicroscopyL-request-at-microscopy.com Fri Sep 12 23:17:27 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8D3HC9v001598 for {L1-at-ns.microscopy.com} ; Fri, 12 Sep 2003 23:17:12 -0400 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8D3HCJ8001593 for L1; Fri, 12 Sep 2003 22:17:12 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8D3GQ9v001563 for "MListServerFilteredEmail_5-at-microscopy.com"; Fri, 12 Sep 2003 23:16:52 -0400 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8D3GB9w001558 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 12 Sep 2003 23:16:11 -0400 Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) Message-Id: {p05111a01bb883a1ee009-at-[206.69.208.21]}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Gilweiss-at-worldnet.att.net) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, September 12, 2003 at 19:44:55 ---------------------------------------------------------------------------
Question: What is the practical use of circular polarization in microscopy? What physical phenomena initiate it? I have not found a good working source of info on the subject.
From MicroscopyL-request-at-microscopy.com Mon Sep 15 08:31:38 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FDV94n006995 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 08:31:09 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FDV9m4006990 for L1; Mon, 15 Sep 2003 08:31:09 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FDUE4n006960 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 08:30:34 -0500 Received: from mail.med.cornell.edu (mail.med.cornell.edu [140.251.3.3]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FDTh4n006951 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 15 Sep 2003 08:29:53 -0500 Received: from [140.251.145.131] by mail.med.cornell.edu (Netscape Messaging Server 3.6) with ESMTP id AAA3B6AA; Mon, 15 Sep 2003 09:29:29 -0400 Mime-Version: 1.0 X-Sender: lcgould-at-pop.med.cornell.edu Message-Id: {p05100301bb8b6ea0f4d4-at-[140.251.145.131]} In-Reply-To: {p05111a00bb88391fa442-at-[206.69.208.21]} References: {p05111a00bb88391fa442-at-[206.69.208.21]}
There are scanners available over a wide price range. I bought one a few years ago, on a tight budget, and I'm happy with it. Its an Epson Expression Pro 1600. I came with a trans-illuminator for transparencies and runs with Photoshop. I use 400 dpi for "work print" type images, and a higher dpi for publication. 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
From MicroscopyL-request-at-microscopy.com Mon Sep 15 08:33:46 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FDXZ4n007045 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 08:33:35 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FDXZ2N007040 for L1; Mon, 15 Sep 2003 08:33:35 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FDWt4n007012 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 08:33:15 -0500 Received: from smtp0.libero.it (smtp0.libero.it [193.70.192.33]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FDWd4n007009 for {microscopy-at-MSA.microscopy.com} ; Mon, 15 Sep 2003 08:32:50 -0500 Received: from libero.it (193.70.192.91) by smtp0.libero.it (7.0.020-DD01) id 3F6031C1000647FA for microscopy-at-MSA.microscopy.com; Mon, 15 Sep 2003 15:32:29 +0200
Hi all,
IŽd like to substitute the Zenit camera applied on the triocular LOMO MFN-11 head of my optical microscope by a WebCam or a DigitalCam. IŽd like to have a high resolution. Could someone give me a suggestion about the best device (as brand) to apply? Thank you. Best Regards, Massimo
From MicroscopyL-request-at-microscopy.com Mon Sep 15 08:35:20 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FDZA4n007095 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 08:35:10 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FDZAbc007091 for L1; Mon, 15 Sep 2003 08:35:10 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FDYT4n007058 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 08:34:50 -0500 Received: from smtp2.libero.it (smtp2.libero.it [193.70.192.52]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FDYE4n007055 for {microscopy-at-MSA.microscopy.com} ; Mon, 15 Sep 2003 08:34:24 -0500 Received: from libero.it (193.70.192.91) by smtp2.libero.it (7.0.020-DD01) id 3F60342D0006378B for microscopy-at-MSA.microscopy.com; Mon, 15 Sep 2003 15:34:38 +0200
Hi all,
IŽd like to substitute the Zenit camera applied on the triocular LOMO MFN-11 head of my optical microscope by a WebCam or a DigitalCam. IŽd like to have a high resolution. Could someone give me a suggestion about the best device (as brand) to apply? Thank you. Best Regards, Massimo
From MicroscopyL-request-at-microscopy.com Mon Sep 15 11:27:20 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FGR94n008529 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 11:27:09 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FGR9IZ008524 for L1; Mon, 15 Sep 2003 11:27:09 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FGQT4n008492 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 11:26:49 -0500 Received: from earth-ox.its.caltech.edu (SteeleMR-loadb-NAT-49.caltech.edu [131.215.49.69]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FGQ84n008486 for {microscopy-at-msa.microscopy.com} ; Mon, 15 Sep 2003 11:26:19 -0500 Received: from fire-dog (fire-dog [192.168.1.4]) by earth-ox-postvirus (Postfix) with ESMTP id 10EE9109A1F for {microscopy-at-msa.microscopy.com} ; Mon, 15 Sep 2003 09:25:48 -0700 (PDT) Received: from water-ox ([192.168.1.10]) by fire-dog (MailMonitor for SMTP v1.2.2 ) ; Mon, 15 Sep 2003 09:25:47 -0700 (PDT) Received: from caltech.edu (pix-1.caltech.edu [131.215.2.21]) by water-ox.its.caltech.edu (Postfix) with ESMTP id 286CD26AC10 for {microscopy-at-msa.microscopy.com} ; Mon, 15 Sep 2003 09:25:47 -0700 (PDT)
On Friday, September 12, 2003, at 08:06 PM, by way of Ask-A-Microscopist wrote:
} Name: Gilbert Weiss } } Organization: Imaging Analysis } } Education: Graduate College } } Location: Stamford, CT } } Question: What is the practical use of circular polarization in } microscopy? What physical phenomena initiate it? I have not found a } good working source of info on the subject. } } Thanks for the help. } Dear Gilbert, I'll let the LM experts answer your first question. As for the second, molecules that do not have a center of symmetry will interact differently with right-hand and left-hand circularly polarized light. The usual description is to consider a molecule that has a carbon atom singly bonded to four distinct groups, such as H, F, Cl, and Br. There are two possible configurations, which cannot be superimposed, but which are mirror images of each other. (By rotating any configuration it can be superimposed onto one of the two possible configurations.) Any optics text should have a description of this phenomenon, so I am surprised you have not found one. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-microscopy.com Mon Sep 15 12:30:25 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FHUF4n009190 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 12:30:15 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FHUFru009185 for L1; Mon, 15 Sep 2003 12:30:15 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FHTZ4n009152 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 12:29:55 -0500 Received: from mail.dixie-net.com (r160h9.dixie-net.com [64.89.160.9]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FHTD4n009144 for {microscopy-at-msa.microscopy.com} ; Mon, 15 Sep 2003 12:29:24 -0500 Received: from bgrosso (r56h10.dixie-net.com [64.49.56.10]) by mail.dixie-net.com (8.12.9/8.12.9) with SMTP id h8FHPSTx056595 for {microscopy-at-msa.microscopy.com} ; Mon, 15 Sep 2003 12:25:30 -0500 (CDT) Message-ID: {027f01c37bbf$a857c440$0a383140-at-bgrosso}
I have a Zeiss Confocal LSM-510 for sale which has the Argon Helium lasers and I am looking to sell it.
Photos and a complete configuration are available by request. It is located in Bohemia NY and is able to be inspected with an appointment.
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
From MicroscopyL-request-at-microscopy.com Mon Sep 15 12:55:55 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FHtj4n009402 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 12:55:45 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FHtj9u009397 for L1; Mon, 15 Sep 2003 12:55:45 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FHt54n009370 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 12:55:25 -0500 Received: from usryim04.merck.com (wily.merck.com [155.91.6.40]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FHsm4n009363 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 15 Sep 2003 12:54:59 -0500 Received: from 155.91.2.6 by usryim04.merck.com with ESMTP (SMTP Relay ( MMS v5.5.3)); Mon, 15 Sep 2003 13:53:43 -0400 Received: from usrygw03.merck.com (usrygw03.merck.com [54.3.102.169]) by mmh1.merck.com (PMDF V6.0-24 #37353) with ESMTP id {01L0P08EUH14001FV9-at-mmh1.merck.com} for Microscopy-at-MSA.Microscopy.Com; Mon, 15 Sep 2003 12:54:22 -0500 (EST) Received: by usrygw03.merck.com with Internet Mail Service (5.5.2656.59) id {SVPDW52P} ; Mon, 15 Sep 2003 13:54:21 -0400 Content-return: allowed
Is anyone aware of the fluorescent properties of 5-(4-dimethylaminobenzylidene)rhodanine....not rhodamine. This is a histochemical stain fro copper usually used for brightfield microscopy, but I am also seeing a great fluorescent signal. Thanks, Brett
Brett M. Connolly, Ph.D. Merck & Co.,Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly-at-merck.com
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From MicroscopyL-request-at-microscopy.com Mon Sep 15 13:33:32 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FIXB4n009726 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 13:33:11 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FIXBr2009721 for L1; Mon, 15 Sep 2003 13:33:11 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FIWL4n009688 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 13:32:41 -0500 Received: from mail.helwigcp.com (mail.helwigcp.com [66.162.230.102]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FIW14n009679 for {microscopy-at-MSA.microscopy.com} ; Mon, 15 Sep 2003 13:32:11 -0500 Received: from wolf (office.helwigcp.com [66.162.230.100]) by mail.helwigcp.com (8.9.3/8.9.3) with ESMTP id NAA03144; Mon, 15 Sep 2003 13:29:58 -0500
Massimo;
"WebCam" and "High Resolution" are generally mutually exclusive. The required bandwidth for sending high res video over the Internet (and many "internal" networks) makes it unworkable. As a result, depending on what you're after, you may need two cameras. (Even many "high end" video cameras don't have a very high resolution, at least as compared to "still" digital cameras.)
WebCams, because of the low resolution are available from a number of companies, and there isn't much difference between them. They all offer at least a couple of models. The differences tend to be in zoom, microphone, and the level of "remote control". It may take some ingenuity to get one of these mounted.
As far as High Res, that subject comes up often in this group, and the Nikon CoolPix line seems to be most popular. There are microscope adapters available for the Nikon's and some of the other more popular choices.
Thank you;
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
-----Original Message----- } From: max_gra-at-libero.it [mailto:max_gra-at-libero.it] Sent: Monday, September 15, 2003 8:34 AM To: MSA Microscopy
Hi all,
IŽd like to substitute the Zenit camera applied on the triocular LOMO MFN-11 head of my optical microscope by a WebCam or a DigitalCam. IŽd like to have a high resolution. Could someone give me a suggestion about the best device (as brand) to apply? Thank you. Best Regards, Massimo
From MicroscopyL-request-at-microscopy.com Mon Sep 15 13:55:19 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FIt94n009955 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 13:55:09 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FIt9st009951 for L1; Mon, 15 Sep 2003 13:55:09 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FIsT4n009917 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 13:54:49 -0500 Received: from ucsc.edu (cats-mx2.ucsc.edu [128.114.129.35]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FIs84n009911 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 15 Sep 2003 13:54:18 -0500 Received: from [128.114.25.139] (dhcp-25-50.ucsc.edu [128.114.25.139]) by ucsc.edu (8.10.1/8.10.1) with SMTP id h8FIqDQ07228 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 15 Sep 2003 11:52:13 -0700 (PDT) X-Sender: jmkrupp-at-cats-po-1.ucsc.edu Message-Id: {v01550100bb8bb9ecc8ba-at-[128.114.25.139]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Hi:
Is there a quick and dirty method for looking at TMV?
I have never done it, a person from the local community wants to see if that's what killing his plants and insists on 'seeing' it with his own eyes.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-microscopy.com Mon Sep 15 16:48:39 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FLmO4n011511 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 16:48:24 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FLmOSX011507 for L1; Mon, 15 Sep 2003 16:48:24 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FLlc4n011478 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 16:48:04 -0500 Received: from outbound.lightshipmail.net (outbound.lightshipmail.net [216.204.0.39]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FLlC4n011475 for {microscopy-at-msa.microscopy.com} ; Mon, 15 Sep 2003 16:47:28 -0500 Received: from Barb500.mme1.com (ipn36372-b70930.net-resource.net [216.204.89.146]) by outbound.lightshipmail.net (Postfix) with ESMTP id 428CD57457 for {microscopy-at-msa.microscopy.com} ; Mon, 15 Sep 2003 17:51:08 -0400 (EDT) Message-Id: {5.1.0.14.0.20030915163820.04789ec0-at-postoffice.lightshipmail.net} X-Sender: pop3750-at-postoffice.lightshipmail.net X-Mailer: QUALCOMM Windows Eudora Version 5.1
Dear Gilbert,
Your question is not only a good one, it offers many people a new way do image analysis, strangely enough!
We have to start with regular polarized light. The difference between "ordinary light" and "polarized light" has to do with how the electric vectors are vibrating. Because light is electro-magnetic radiation, it has both an electrical component and a magnetic component. Most microscopists are only interested in the electrical function.
We often use a sine wave to describe light. The sine wave is actually the path traveled by the tip of light's electric vector as it builds up in a positive direction, then decreases, then builds up in a negative direction then decreases. This process gives light several key features: a. a direction of travel (you see an object because light is traveling from it to you) b. a direction of vibration (the direction in which the vector increases and decreases; direction of vibration is always perpendicular to the direction of travel) c. its intensity (intensity is proportional to the square of the maximum displacement in a positive or negative direction) c. its wavelength (which microscopists typically associate with color... all apologies to the physicists in the audience).
Ordinary light is comprised of waves of light traveling toward you, vibrating at all angles. The typical analogy: if you looked back along the direction of travel, you would see all the vectors like arrows vibrating N-S, E-W, and all angles in between.
To convert ordinary light to polarized light, you need to have it interact with something which preferentially absorbs all but one direction of vibration. That "something" could be the process of reflection at the specific angle known as "Brewster's angle" (dependent on the properties of a given material), absorption through something like a polarizing filter or polarizing sunglasses, or the process of beam splitting which happens when the beam enters a material which is birefringent. As this term implies, these materials have two ("bi") characteristic refractive ("refr") indices ("in"). (The "gent" means "having the property of..."). The refractive indices are the result of differences in electric field which result from the arrangement of atoms, ions, or elecrons. In the simplest term, birefringence is the electrical equivalent of wood grain: one direction is "with the grain" and the direction at right angles is "against the grain". Many minerals are birefringent, su ch as calcite and quartz. Also, many polymer fibers and films.
If you create polarized light by beam splitting, the beam will be split into two components: an "e" ray and an "o" ray. They will have directions of vibration at right angles to each other. Each will encounter a different electrical field. Both will slow down because of the interaction between the electrical field in light and the field in the object. The one which encounters the field represented by the higher refractive index ("against the grain") will slow down more and lag behind its partner. The "lag" is called "retardation" and is measured in wavelengths.
So, what does all of this have to do with circularly polarized light? This part is really tough without diagrams, but draw the following: A horizontal reference line On the left side of the line, draw rectangle to represent a polarizer which permits light to emerge vibrating at +/- 45 degrees = Electric Vector "E" At some distance along the line, draw another rectangle to represent a piece of birefringent material (ex: a polymer overhead transparency) In that rectangle, draw a short vector from the reference line pointing up (we'll call that "e") and another, again from the reference line, at 90 degrees ("o"). At a third point along the reference line, draw another short vector "e" and, a short distance further along, draw another "o".
Here is the explanation: a. When ordinary light passes through the polarizing filter, it absorbs all the electric vectors except "E". (We would have determined the direction of vibration by rotating the filter). b. When the polarized beam enters the birefringent material, it is split into "e" and "o", each polarized and each vibrating at right angles to each other. c. Because each interacts with a different electrical field, one will slow down and lag behind the other. In this case, we have drawn the example when "e" experiences the greater interaction and, therefore, lags behind "o". d. Analyzing the resultant electric vector will tell you the "State of Polarization": Again, a simple drawing helps. Draw two perpendicular lines which cross in their centers. Label the "Y" with + and - (top and bottom); Label the "X" with + and - (right and left). The next part takes a bit of imagination (there is a great diagram in Optimizing Light Microscopy, but I can't draw it here). First, keep in mind that "e" and "o" are vibrating at right angles. Let's say "e" is vibrating in the Y direction and "o", in the X direction. If "e" lags behind "o" by some multiple of whole ("e" goes to max in + direction when "o" goes to max in + ) or half wavelengths ("e" goes to max in + when "o" goes to max in -), the resultant vector would see-saw between +45 and -45 degrees, generating "linearly polarized light". (Direction changes, depending on whether a whole wave or half wave) If "e" lags behind "o" by odd multiples of quarter waves, the resultant vector travels in a corkscrew pattern. Quarter wave shifts produce clockwise rotation; three-quarter shift produces counter-clockwise rotation. This corkscrew path is what we call "circularly polarized light". Circular polarizing filters are actually sandwiches of (a) a regular polarizing filter and (b) a piece of plastic which causes a retardation of 1/4 wave (typically about 140nm).
Why is all this important? And what does it have to do with image analysis? The biggest challenge in image analysis is simplifying objects so that they can be recognized by an automated measuring system. In polarized light, birefringent objects viewed between crossed polars are bright against a dark background ... great for segmenting with an automated system. Unfortunately, the intensity changes with orientation (yet another discussion). The same crystal, for example, will appear dark if sitting N-S or E-W but bright at +/- 45 degrees! On the other hand, if you use crossed CIRCULAR polars (make sure the retarding plastic is toward the center of the "sandwich" and the sample is in between), all the orientation effects will be removed because of the corkscrew path of the light. The result: all birefringent objects which are behaving the same would appear the same intensity, regardless of orientation.
We used this trick very successfully, for example, with the Center for Urban Ecology, when they were trying to measure the % crystallinity of soil samples.
Note: if you are interested in trying circularly polarized light, you can ask your microscope rep to lend you a set of circular Pol filters or you can make your own, using 2" squares of polarizing filter and "quarter wave" material, typically available from companies like Edmund Scientific. (Caveat: no financial interest).
Whew! A long winded answer, but I hope that it is helpful.
If you are interested in copies of the diagrams described above, contact me off line.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
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From MicroscopyL-request-at-microscopy.com Mon Sep 15 18:06:25 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FN6F4n012091 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 18:06:15 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8FN6FTU012087 for L1; Mon, 15 Sep 2003 18:06:15 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8FN5Y4n012053 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 18:05:54 -0500 Received: from col-msxproto2.col.missouri.edu (col-msxproto2.col.missouri.edu [128.206.3.152]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8FN5A4n012042 for {Microscopy-at-msa.microscopy.com} ; Mon, 15 Sep 2003 18:05:21 -0500 Received: from phillips-dell.missouri.edu ([128.206.80.38]) by col-msxproto2.col.missouri.edu over TLS secured channel with Microsoft SMTPSVC(5.0.2195.6673); Mon, 15 Sep 2003 18:04:56 -0500 Message-Id: {5.1.1.5.2.20030915180330.02bab6c0-at-pop.missouri.edu} X-Sender: phillipst-at-pop.missouri.edu X-Mailer: QUALCOMM Windows Eudora Version 5.1.1
I am using Luft's technique for staining the glycocalyx with Ruthenium Red. I know the RR is supposed to be included in the aldehyde fix, the rinses and the osmium but how about the osmium rinses or ethanol dehydration series? Has anyone successfully done this procedure? Thanks, tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
From MicroscopyL-request-at-microscopy.com Mon Sep 15 19:41:20 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8G0et4n012751 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 19:40:55 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8G0etsH012747 for L1; Mon, 15 Sep 2003 19:40:55 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8G0dx4n012713 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 19:40:25 -0500 Received: from mailhost2.auckland.ac.nz (mailhost2.auckland.ac.nz [130.216.191.4]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8G0dN4n012709 for {microscopy-at-MSA.Microscopy.Com} ; Mon, 15 Sep 2003 19:39:38 -0500 Received: from glgnov2.auckland.ac.nz (glgnov2.auckland.ac.nz [130.216.59.90]) by mailhost2.auckland.ac.nz (8.12.9/8.12.9/8.12.9-ua) with ESMTP id h8G0d8s0002918; Tue, 16 Sep 2003 12:39:08 +1200 (NZST) Received: from GLGNOV2/SpoolDir by glgnov2.auckland.ac.nz (Mercury 1.48); 16 Sep 03 12:41:20 +1200 Received: from SpoolDir by GLGNOV2 (Mercury 1.48); 16 Sep 03 12:41:14 +1200
Don't bother even trying a webcam, they have insufficient resolution (unless all you want to do is to obtain images small enough to stream on the Net).
I've been there.
Buy a Nikon Coolpix 4500 or similar plus an adaptor from any of several vendors. I bought my adaptor from WPI (www.wpiinc.com).
It works well.
cheers
rtch
Date sent: Mon, 15 Sep 2003 15:34:03 +0200
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmolin-at-cooksonelectronics.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, September 15, 2003 at 13:43:45 ---------------------------------------------------------------------------
Email: dmolin-at-cooksonelectronics.com Name: Dick Molin
Organization: Cookson Electronics
Education: Graduate College
Location: Indianapolis, IN USA
Question: Our company specializes in applying a thin, transparent conformal coating called Parylene. I'm wondering if the use of DIC could be helpful in my analysis of the coating and the coating/substrate interface. I realize DIC is valuable for transparent biological specimens,and don't know if this usefulness translates for my application. Please let me know your opinion since I would want to justify the added cost for the prism and the optics.
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (echeung-at-eyetk.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 15, 2003 at 15:36:08 ---------------------------------------------------------------------------
Email: echeung-at-eyetk.com Name: Eunice Cheung
Organization: Eyetech Pharmaceuticals
Title-Subject: MListserver: Trouble with Formvar and glass coverslips
Question: I am doing a project in growing cells on Formvar covered grids. First, the film is made by floating the Formvar on water. Second, the 7 grids are put on the film. Third, a small circular glass coverslip is place covering on top of the grids. Fourth, the formvar, grids, and coverslip are picked up with a piece of parafilm. Afterwards, the coverslip is taken out with the grids in place and sterlized in UV. Later it is coated with gelatin. The problem: The Formvar with the grids falls off from the coverslip. Any suggestions in preventing this? Thanks.
From MicroscopyL-request-at-microscopy.com Mon Sep 15 20:28:27 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8G1SH4n013160 for {L1-at-ns.microscopy.com} ; Mon, 15 Sep 2003 20:28:17 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8G1SHmK013155 for L1; Mon, 15 Sep 2003 20:28:17 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8G1Ra4n013123 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 15 Sep 2003 20:27:56 -0500 Received: from hotmail.com (bay2-dav56.bay2.hotmail.com [65.54.246.191]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8G1RL4n013116 for {microscopy-at-MSA.microscopy.com} ; Mon, 15 Sep 2003 20:27:31 -0500 Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; Mon, 15 Sep 2003 18:27:10 -0700 Received: from 216.75.187.126 by bay2-dav56.bay2.hotmail.com with DAV; Tue, 16 Sep 2003 01:27:09 +0000 X-Originating-IP: [216.75.187.126] X-Originating-Email: [slakmon-at-hotmail.com]
Dear Collegues,
I am looking for people who have equipment to give away such as micrscopes that they would not mind parting with, that would serve as a teaching tool for high school students from grades 7 to 11. The ideal microscopes would be with trinocular tubes or without. I would also welcome video or digital cameras. Video microscopy would be a great help in teaching large groups of kids. Any other laboratory or imaging equipment or supplies would also be welcome.
Thank you in advance,
P. Slakmon
From MicroscopyL-request-at-microscopy.com Tue Sep 16 01:28:26 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8G6Rr4n015454 for {L1-at-ns.microscopy.com} ; Tue, 16 Sep 2003 01:27:53 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8G6RrIU015448 for L1; Tue, 16 Sep 2003 01:27:53 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8G6Qs4n015414 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 16 Sep 2003 01:27:22 -0500 Received: from mail.biomed.cas.cz (mail.biomed.cas.cz [147.231.40.5]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8G6QD4n015408 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 16 Sep 2003 01:26:30 -0500 Received: from 127.0.0.1 (mail [127.0.0.1]) by biomed.cas.cz (Postfix) with SMTP id A1A7A23D8A; Tue, 16 Sep 2003 08:54:46 +0200 (CEST) Received: from c117ob (mail.biomed.cas.cz [147.231.40.5]) by mail.biomed.cas.cz (Postfix) with ESMTP id D43A023E60; Tue, 16 Sep 2003 08:54:45 +0200 (CEST)
Hi, You can do a good job with negative staining. Best regards from Prague Oldrich On 15 Sep 2003 at 11:50, Jon Krupp wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } } Hi: } } Is there a quick and dirty method for looking at TMV? } } I have never done it, a person from the local community wants to see if } that's what killing his plants and insists on 'seeing' it with his own } eyes. } } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-241062399 Fax: +420-241062347 WEB: http://www.biomed.cas.cz/mbu/lem113/lem.htm
From MicroscopyL-request-at-microscopy.com Tue Sep 16 04:44:25 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8G9hq4n017049 for {L1-at-ns.microscopy.com} ; Tue, 16 Sep 2003 04:43:52 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8G9hqJq017045 for L1; Tue, 16 Sep 2003 04:43:52 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8G9gq4n017014 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 16 Sep 2003 04:43:17 -0500 Received: from hermes.sunderland.ac.uk (hermes.sunderland.ac.uk [157.228.37.117]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8G9gF4n017011 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 16 Sep 2003 04:42:31 -0500 Received: from sunderland.ac.uk (localhost [127.0.0.1]) by hermes.sunderland.ac.uk (iPlanet Messaging Server 5.2 Patch 1 (built Aug 19 2002)) with ESMTP id {0HLA00CBNWCG8R-at-hermes.sunderland.ac.uk} for Microscopy-at-sparc5.microscopy.com; Tue, 16 Sep 2003 10:43:28 +0100 (BST) Received: from [157.228.74.89] by hermes.sunderland.ac.uk (mshttpd); Tue, 16 Sep 2003 10:43:28 +0100
Heike
you haven't mentioned the size of your TEM negatives but I will assume that they are one of the larger formats (eg 83x84mm or 83x102mm).
I have used an Epson Perfection 1200 which produce fairly good results but I was always suspicious of its quoted top resolution of 1200 dpi. Negatives on this type of flatbed scanner have to be placed on the glass surface of the scanner to be scanned which creates several problems: Newton's rings; dust and other marks on the glass.
I now use a Microtek Scanmaker 8700 which you can buy with or without Silversoft image management software (I think its the PRO version in the US). It also has a glass panel for normal flatbed document scanning but importantly has a separate glassless drawer which can take several types of film holder. See website below: http://www.microtekusa.com/sm8700.html So far I have been very impressed because the scan quality is good and some of our staff have even used the 35mm film and slide adaptors to digitise their colour slides (1200dpi is adequate for 35mm if you're using a digital projector or displaying on screen). This model comes with USB 1.1 but more usefully Firewire with an adaptor card. In the UK you also get Adobe Photoshop Elements which is quite handy. There is sufficient room to place two large 83 x 102mm (3 1/4 x 4 inch) in the glassless film carriers. Although I am still just using the 5 x 4 inch adaptors I suspect that it would be possible to construct inserts.
There seem to be 3 main types of possible scanner: 1. Dedicated film scanners - these are often the best and highest resolution but large format film scanners are extremely expensive and rare (1,000 - 2,000 UK pounds and upwards). Make sure it really will do your film size. 2. Normal flatbed document scanners with a glass copying surface and a film/transparency adaptor light source either built into the lid of the scanner or replacing the lid. These can be extremely cheap with current resolutions in excess of 1200dpi but be sure that the transparency adaptor is not just for 35mm or medium format (costs start from below 100 UK pounds). A good way of trying out but the quality won't be as good as the other two types. 3. The hybrid flatbed scanner with separate glassless film carriers which slide into a drawer in the scanner body. These seem to strike a good balance in terms of quality, versatility and cost for large format film (600 UK pounds and upwards).
Basic things to look for are a genuine 1200 dpi or higher resolution (after all you may want to enlarge part of an image or examine in detail on the screen). Colour bit depth of 42 or 48 (ie 14 or 16 in greyshades). The ability to generate a variety of image formats (especially TIF, BMP and JPEG). If you have a lot of dense negatives check the D max or dynamic range - it should be at least 3.4 but 4.0 would be better for difficult film. Finally the connection to the computer should really be USB 2.0 or Firewire because if you scan a 4x3 inch negative at 1200dpi that's 17280000pixels (ie 17-18 Mb minimum for 8 bit B&W) which can be quite slow on USB 1.1.
Most e.m. film I scan is at 600 or 800dpi 8 bit grey-scale and stored as best quality JPEG. TIF and 14/16 bit greyscale would be nice but in the real world I still have the negatives and can store more digital images on CDs or hard disk. I can't remember the last time I produced an old style photographic print, although I do still offer the option for final publication quality
I hope this helps and if the 8700 isn't good enough I am sure either Microtek or a competitor will do higher resolution or better D max.
By the way if anyone has read this far I am seriously considering the Canon i950 bubble/inkjet printer as a successor to our Epson 750. Has anyone used the Canon for e.m. black and white printing because it is supposedly quiet, good resolution, faster and has a separate replaceable print head?
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK tel no: 0191 515 2872 / 3468 e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: hgabrisc-at-uno.edu (by way of Nestor J. Zaluzec)
Hello everyone,
If possible, I would like to find out the hourly rate for SEM/EDS work. Our management wants to increase the rate and we are trying to asses the "fare" rate for SEM work. I would like to find out the rate that you would charge and any other charges (extra copies, digital, WDS work, etc)
Thank you for the info.
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
From MicroscopyL-request-at-microscopy.com Tue Sep 16 07:51:39 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GCp94n018351 for {L1-at-ns.microscopy.com} ; Tue, 16 Sep 2003 07:51:09 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8GCp9Jv018346 for L1; Tue, 16 Sep 2003 07:51:09 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8GCoE4n018312 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 16 Sep 2003 07:50:34 -0500 Received: from Mail2.Millipore.com (mail2.millipore.com [157.93.11.15]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GCnf4n018299 for {microscopy-at-msa.microscopy.com} ; Tue, 16 Sep 2003 07:49:52 -0500 To: Barbara Foster {bfoster-at-mme1.com} Cc: microscopy-at-msa.microscopy.com
Barbara,
An enlightening discussion, as usual! Thanks for taking the time to put that all down in electrons!
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Barbara Foster {bfoster-at-mme1.com} 09/15/2003 08:50 PM
To: microscopy-at-msa.microscopy.com cc: Subject: Re: Ask-A-Microscopist: circular polarization in microscopy
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Dear Gilbert,
Your question is not only a good one, it offers many people a new way do image analysis, strangely enough!
We have to start with regular polarized light. The difference between "ordinary light" and "polarized light" has to do with how the electric vectors are vibrating. Because light is electro-magnetic radiation, it has both an electrical component and a magnetic component. Most microscopists are only interested in the electrical function.
We often use a sine wave to describe light. The sine wave is actually the path traveled by the tip of light's electric vector as it builds up in a positive direction, then decreases, then builds up in a negative direction then decreases. This process gives light several key features: a. a direction of travel (you see an object because light is traveling from it to you) b. a direction of vibration (the direction in which the vector increases and decreases; direction of vibration is always perpendicular to the direction of travel) c. its intensity (intensity is proportional to the square of the maximum displacement in a positive or negative direction) c. its wavelength (which microscopists typically associate with color... all apologies to the physicists in the audience).
Ordinary light is comprised of waves of light traveling toward you, vibrating at all angles. The typical analogy: if you looked back along the direction of travel, you would see all the vectors like arrows vibrating N-S, E-W, and all angles in between.
To convert ordinary light to polarized light, you need to have it interact with something which preferentially absorbs all but one direction of vibration. That "something" could be the process of reflection at the specific angle known as "Brewster's angle" (dependent on the properties of a given material), absorption through something like a polarizing filter or polarizing sunglasses, or the process of beam splitting which happens when the beam enters a material which is birefringent. As this term implies, these materials have two ("bi") characteristic refractive ("refr") indices ("in"). (The "gent" means "having the property of..."). The refractive indices are the result of differences in electric field which result from the arrangement of atoms, ions, or elecrons. In the simplest term, birefringence is the electrical equivalent of wood grain: one direction is "with the grain" and the direction at right angles is "against the grain". Many minerals are birefringent, su ch as calcite and quartz. Also, many polymer fibers and films.
If you create polarized light by beam splitting, the beam will be split into two components: an "e" ray and an "o" ray. They will have directions of vibration at right angles to each other. Each will encounter a different electrical field. Both will slow down because of the interaction between the electrical field in light and the field in the object. The one which encounters the field represented by the higher refractive index ("against the grain") will slow down more and lag behind its partner. The "lag" is called "retardation" and is measured in wavelengths.
So, what does all of this have to do with circularly polarized light? This part is really tough without diagrams, but draw the following: A horizontal reference line On the left side of the line, draw rectangle to represent a polarizer which permits light to emerge vibrating at +/- 45 degrees = Electric Vector "E" At some distance along the line, draw another rectangle to represent a piece of birefringent material (ex: a polymer overhead transparency) In that rectangle, draw a short vector from the reference line pointing up (we'll call that "e") and another, again from the reference line, at 90 degrees ("o"). At a third point along the reference line, draw another short vector "e" and, a short distance further along, draw another "o".
Here is the explanation: a. When ordinary light passes through the polarizing filter, it absorbs all the electric vectors except "E". (We would have determined the direction of vibration by rotating the filter). b. When the polarized beam enters the birefringent material, it is split into "e" and "o", each polarized and each vibrating at right angles to each other. c. Because each interacts with a different electrical field, one will slow down and lag behind the other. In this case, we have drawn the example when "e" experiences the greater interaction and, therefore, lags behind "o". d. Analyzing the resultant electric vector will tell you the "State of Polarization": Again, a simple drawing helps. Draw two perpendicular lines which cross in their centers. Label the "Y" with + and - (top and bottom); Label the "X" with + and - (right and left). The next part takes a bit of imagination (there is a great diagram in Optimizing Light Microscopy, but I can't draw it here). First, keep in mind that "e" and "o" are vibrating at right angles. Let's say "e" is vibrating in the Y direction and "o", in the X direction. If "e" lags behind "o" by some multiple of whole ("e" goes to max in + direction when "o" goes to max in + ) or half wavelengths ("e" goes to max in + when "o" goes to max in -), the resultant vector would see-saw between +45 and -45 degrees, generating "linearly polarized light". (Direction changes, depending on whether a whole wave or half wave) If "e" lags behind "o" by odd multiples of quarter waves, the resultant vector travels in a corkscrew pattern. Quarter wave shifts produce clockwise rotation; three-quarter shift produces counter-clockwise rotation. This corkscrew path is what we call "circularly polarized light". Circular polarizing filters are actually sandwiches of (a) a regular polarizing filter and (b) a piece of plastic which causes a retardation of 1/4 wave (typically about 140nm).
Why is all this important? And what does it have to do with image analysis? The biggest challenge in image analysis is simplifying objects so that they can be recognized by an automated measuring system. In polarized light, birefringent objects viewed between crossed polars are bright against a dark background ... great for segmenting with an automated system. Unfortunately, the intensity changes with orientation (yet another discussion). The same crystal, for example, will appear dark if sitting N-S or E-W but bright at +/- 45 degrees! On the other hand, if you use crossed CIRCULAR polars (make sure the retarding plastic is toward the center of the "sandwich" and the sample is in between), all the orientation effects will be removed because of the corkscrew path of the light. The result: all birefringent objects which are behaving the same would appear the same intensity, regardless of orientation.
We used this trick very successfully, for example, with the Center for Urban Ecology, when they were trying to measure the % crystallinity of soil samples.
Note: if you are interested in trying circularly polarized light, you can ask your microscope rep to lend you a set of circular Pol filters or you can make your own, using 2" squares of polarizing filter and "quarter wave" material, typically available from companies like Edmund Scientific. (Caveat: no financial interest).
Whew! A long winded answer, but I hope that it is helpful.
If you are interested in copies of the diagrams described above, contact me off line.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Optimizing Light Microscopy is still available through MME. For information on ordering single or small quantities, visit our website. For discounts on classroom-sized, call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&
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From MicroscopyL-request-at-microscopy.com Tue Sep 16 08:12:52 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GDCg4n018581 for {L1-at-ns.microscopy.com} ; Tue, 16 Sep 2003 08:12:42 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8GDCgZW018577 for L1; Tue, 16 Sep 2003 08:12:42 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8GDC14n018549 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 16 Sep 2003 08:12:22 -0500 Received: from f04n07.cac.psu.edu (f04s07.cac.psu.edu [128.118.141.35]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GDBe4n018542 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 16 Sep 2003 08:11:50 -0500 Received: from rwalsh.psu.edu (tnt1-161-110.cac.psu.edu [130.203.161.110]) by f04n07.cac.psu.edu (8.9.3p2/8.9.3) with ESMTP id JAA74632 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 16 Sep 2003 09:11:26 -0400 Message-Id: {5.1.0.14.2.20030916090548.009f8350-at-mail.psu.edu} X-Sender: rw9-at-mail.psu.edu X-Mailer: QUALCOMM Windows Eudora Version 5.1
Thanks to all who responded to the recent inquiry about forensic applications. Once again this is a wonderful site because of the people willing to share their knowledge.
There's a fingerprint exercise in Project MICRO's "Microscopic Explorations", which was written for grades 4-8. "Microscopic Explorations" is published in the Lawrence Hall of Science GEMS series, which also includes "Mystery Festival", (crime scene investigation) and "Fingerprints"; you'll find a link to GEMS on the MICRO website (URL below). The Minnesota Microscopy Society has an excellent outreach website, http://resolution.umn.edu/MMS/ProjectMicro/ : you'll find a link to fingerprints at NIST and images of several fabric types there. Don't miss Joe Neilly's delightful "Beanie Baby Mystery" in the Classroom Activities part of the MICRO site. 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 Ms. Walsh, This is not a reply to the general list but a remark that I hope will be taken as reflective. I know that interesting "hooks" are needed to get children interested in science with all the ramifications that a scientific grounding has for our future, but isn't the constant diet of mayhem, down to the most grotesque details of forensic science, being served up from enough sources already? I think that the constant stream of "forensic" programs and cop shows conditions children to an acceptance of a police state mentality. The same instructional subjects can be approached from the biological and earth sciences. (Microscopy to solve the mysteries of developing life; micro-fossils in common stones that explain the mystery of what happened on earth when there was no one yet to observe it, etc.) If you want to deal with fibers, how about the (relatively) benign world of counterfeit goods - the alpaca cape that turns out to be rayon, the shawl that turns out to be shanoosh from poaching of endangered antelope. In fact the whole area of wildlife forensics, the evidence gathered in the protection of endangered species, at least avoids more of the human-on-human violence that is made to seem the norm by its familiarity. (See the website of the federal wildlife forensics lab in Ashland, Oregon.) John Twilley
Rosemary, www.mccrone.com will lead you to some interesting things and Ted Clarke's pages have some information about asbestos http://www.couger.com/microscope/Ted-Clarke/ Good luck Gordon Gordon Couger gcouger-at-couger.com
Rosemary You should look up GSR (Gun Shot Residue Analysis), this is commonly used with a SEM and EDS. Try this URL: http://www.gunshotresidue.com/ JQ Jim Quinn {jquinn-at-/" EUDORA="AUTOURL"www.matscieng.sunysb.edu}
} From: "Ann St. Amand" {astamand-at-phycotech.com}
Jon, What you want to do is a leaf dip. Use a formvar + cabon grid held in a tweezer. Put a drop of uranyl acetate on the grid. Then take a piece of what appears to be an infected leaf, macerate an edge with a razor blade and drag it through the stain droplet a few times. This will deposit some of the sap containing some organelles and virus, if present, onto the grid. Wick off the remaining stain and view. TMV is very easily recognized as a rod shaped virus. You might find that the problem is another virus type so also look for round viruses or viruses that are rod shaped but of very unequal length. TMV is about 18nm wide. I forget exactly how long it is but probably not more than about 200nm. Particles are quite regular in length.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 9/15/03 1:50 PM, "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } } Hi: } } Is there a quick and dirty method for looking at TMV? } } I have never done it, a person from the local community wants to see if } that's what killing his plants and insists on 'seeing' it with his own } eyes. } } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
From MicroscopyL-request-at-microscopy.com Tue Sep 16 08:39:16 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GDcv4n018887 for {L1-at-ns.microscopy.com} ; Tue, 16 Sep 2003 08:38:57 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8GDcvNv018882 for L1; Tue, 16 Sep 2003 08:38:57 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8GDc74n018853 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 16 Sep 2003 08:38:27 -0500 Received: from mail.med.cornell.edu (mail.med.cornell.edu [140.251.3.3]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GDbf4n018848 for {Microscopy-at-msa.microscopy.com} ; Tue, 16 Sep 2003 08:37:51 -0500 Received: from [140.251.145.131] by mail.med.cornell.edu (Netscape Messaging Server 3.6) with ESMTP id AAA2BF40; Tue, 16 Sep 2003 09:37:29 -0400 Mime-Version: 1.0 X-Sender: lcgould-at-pop.med.cornell.edu Message-Id: {p05100304bb8cc213b5a8-at-[140.251.145.131]} In-Reply-To: {5.1.1.5.2.20030915180330.02bab6c0-at-pop.missouri.edu} References: {5.1.1.5.2.20030915180330.02bab6c0-at-pop.missouri.edu}
I've used RR to look at the extracellular matrix in cardiac muscle and to label the apical surface of cultured retinal pigment epithelial cells. You do not need to use it after the osmium step. don't be surprised if the labelling is patchy. 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
From MicroscopyL-request-at-microscopy.com Tue Sep 16 09:47:52 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GElp4n019962 for {L1-at-ns.microscopy.com} ; Tue, 16 Sep 2003 09:47:52 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8GElpZw019956 for L1; Tue, 16 Sep 2003 09:47:51 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8GEl64n019928 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 16 Sep 2003 09:47:28 -0500 Received: from rzaixsrv2.rrz.uni-hamburg.de (rzaixsrv2.rrz.uni-hamburg.de [134.100.32.71]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GEl34n019923 for {microscopy-at-MSA.Microscopy.Com} ; Tue, 16 Sep 2003 09:47:05 -0500 Received: from torsten (Laborwiese.zoologie.uni-hamburg.de [134.100.200.172]) by rzaixsrv2.rrz.uni-hamburg.de (8.12.9/8.12.9) with ESMTP id h8GEkx7m030720 for {microscopy-at-MSA.Microscopy.Com} ; Tue, 16 Sep 2003 16:46:59 +0200
Hi,
for more or less all more common digital cameras you can buy adapters for c-mount; pricing is about 100-400 Euros.
General information: http://www.shortcourses.com/how/microscope/microscope.htm http://www.microscopy-uk.org.uk/mag/artaug01/vrcoolpix.html
Another way is to build the adapter yourself. I did that myself with my Canon G3 and our old Leica microscope. It is equipped with an old analog camera, and the only thing I had to build was a small tubus which fits to the old eyepiece of the analog camera (and holds the camera in the right position above it). Leica sells a similar system for app. 2000 ... (Adapter and camera).
I have to admit that the Canon G3 is not the best camera for microscopy. It is a fine camera for normal pictures, and I really like it. "But" it's lense is quite large in diameter. That is nice, because it get's a lot of light, but the openings of eyepieces I know of are not large. Therefore, if you don't zoom in digitally (4x optical zoom is not enough), you always have circular black frames on the picture. The smaller the lens of the camera, the better for microscopy (I think). (But I bought the camera, b/c I can easily use a ring flash with it). Also, the part of the field in view is quite small. You have to take more then one picture to get all information you see through the eyepiece.
Another topic is the software you want to use to aquire pictures with. It is not nice to handle the camera itself when attached to the microscope (even if it has a display you can flip to the right direction).
All Canon cameras come with a software called "remote capture". With it, most functions of the camera can be controlled via USB from the computer. (The large hard disk is quite nice for time lapse recordings... Having space for } 10000 pictures (at 4 MP) is definitly different to a 36 picture film.) Another software from the original package (well, designed for preparing a panorama photo out of a number of single pictures) is quite nice to put single pictures together in a matrix - we just tried it out with 40 single C. elegans pictures - now we have a worm picture with app. 100 MP... Quite large and difficult to handle... And who want's to print that out??
:-) Torsten
Torsten Fregin
Universität Hamburg - Zoologisches Institut Abt. Neurophysiologie AG Wiese - Raum 413 Martin-Luther-King-Platz 3 20146 Hamburg, Germany Telefon *49-(0)40-42838-3931 Fax *49-(0)40-42838-3937 eMail Torsten.Fregin-at-zoologie.uni-hamburg.de Torsten-at-Fregin.de
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Hi, there,
The C2 apertures in our CM200 FEG has become very dirty and needs to be replaced as soon as possible. As you might be aware, the C2 apertures for FEG have to be very clear and need to take a special method for cleaning. Now I have got several options:
A.Buy from FEI directly but they obviously cost about more than three times from ordinary aperture of which if you purchase from Agar. B.Use special plasma cleaner to clear up those ordinary apertures so that apertures would be acceptable by FEG. C.Try to use the plasma from a carbon coater to clear up ordinary apertures.
My questions is either B or C would work? Alternatively, there are any good ideas for this purpose?
Many thanks!
Peiyi Manager of EM Krebs Institute for Biomolecular Research University of Sheffield Firth Court Western Bank Sheffield Yorkshire S10 2TN United Kingdom Tel: +44 (0)114 222 2000 Direct: +44 (0)114 222 2739 FAX: +44 (0)114 272 8697 E-mail: p.wang-at-sheffield.ac.uk
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Gary and Warren,
I have an additional remark. Sometimes it is useful to take the measured background (Bremsstrahlung) as reference to the beam current. That works only, if the unknown specimen and the standard have nearly the same mean atomic number. If not, the Bremsstrahlung depends from Z and this fact has to taken into account. If analyst takes the Bremsstrahlung reference, a very high precission is reachable, because charakteristic radiation and Bremsstrahlung intensities are both directly depend from beam current. It is useful, if a standard exists with nearly the same elements and concentrations as the expected in unknown.
Otherwise: If the beam current vary in percent units, the results must vary in same order of magnitude (in standard comparison analysis), too. Only normalization can help, like Warren remarked. But this works only if all elements are possible to measure and were measured in same time. That is why standard comparison analysis often do not have better results than standardless. Finally, to overcome the problem with normalization in standardless analysis, you need the Bremsstrahlung background, too.
Frank Eggert
By the way, for all who have got a Palm Handheld and need line energies (keV) or a periodic chart of elements for your Palm } } } try: http://www.microanalyst.net/software.phtml
______________________________________________________________________________ Zwei Mal Platz 1 mit dem jeweils besten Testergebnis! WEB.DE FreeMail und WEB.DE Club bei Stiftung Warentest! http://f.web.de/?mc=021183
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Massimo,
You may want to take a look at our webpage on connecting Coolpix cameras to microscopes: http://www.mvia.com/Coolpix/clpxadpt.htm
There is a lot of information there about different types of microscopes as well as various options for how to mount the camera onto your microscope.
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
"max_gra-at-libero.it" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } } Hi all, } } IŽd like to substitute the Zenit camera applied on the triocular LOMO MFN-11 head of my optical microscope by a WebCam or a DigitalCam. } IŽd like to have a high resolution. } Could someone give me a suggestion about the best device (as brand) to apply? } Thank you. } Best Regards, } Massimo
--
From MicroscopyL-request-at-microscopy.com Tue Sep 16 16:09:33 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GL955t004479 for {L1-at-ns.microscopy.com} ; Tue, 16 Sep 2003 16:09:05 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8GL95L1004474 for L1; Tue, 16 Sep 2003 16:09:05 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8GL885t004442 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 16 Sep 2003 16:08:34 -0500 Received: from mtiwmhc12.worldnet.att.net (mtiwmhc12.worldnet.att.net [204.127.131.116]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GL7V5t004430 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 16 Sep 2003 16:07:47 -0500 Received: from theduke (203.syracuse-05-10rs.ny.dial-access.att.net[12.89.4.203]) by mtiwmhc12.worldnet.att.net (mtiwmhc12) with SMTP id {200309162107171120061e2ie} ; Tue, 16 Sep 2003 21:07:17 +0000 Message-ID: {000701c37c96$8afa1650$7e81fea9-at-TheDuke} Reply-To: "Ladd Research" {sales-at-laddresearch.com}
Is there a third edition of "Scanning Electron Microscopy and X-Ray Microanalysis" by Goldstein et al? Does anyone know the publisher?
Thanks, Deb Sicard
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
From MicroscopyL-request-at-microscopy.com Tue Sep 16 17:05:59 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GM5n5t005030 for {L1-at-ns.microscopy.com} ; Tue, 16 Sep 2003 17:05:49 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8GM5nOW005025 for L1; Tue, 16 Sep 2003 17:05:49 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8GM585t004994 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 16 Sep 2003 17:05:28 -0500 Received: from ucsc.edu (cats-mx2.ucsc.edu [128.114.129.35]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8GM4l5t004986 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 16 Sep 2003 17:04:57 -0500 Received: from [128.114.25.139] (dhcp-25-50.ucsc.edu [128.114.25.139]) by ucsc.edu (8.10.1/8.10.1) with SMTP id h8GM2CQ16316 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 16 Sep 2003 15:02:12 -0700 (PDT) X-Sender: jmkrupp-at-cats-po-1.ucsc.edu Message-Id: {v01550100bb8d32c06ffa-at-[128.114.25.139]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Hi:
I got plenty of advice about how to do a quick survey for TMV. Most of it was what I had suspected, a simple leaf drag through some negative stain. Though I had never done it before, it was reassuring to know I was on the right track.
The guy came to the lab today, we did not see any TMV in his leaves. The leaves looked mottled, like TMV, but I did not see any in the EM. I tried 3 grids, got lots of other junk on the grids, but either no TMV or there was so much of it that it made a solid mat that made seeing individual particles impossible.
Now, here's the good part. He came in with a couple of species of tobacco leaves, they looked like they had TMV. I said "Looks like TMV, nothing you can do about it, doesn't matter if we can see virus or not, the plants are toast."
He said he really wanted to know if it was TMV because he had other plants growing and was afraid they might get infected too. As we talked over other ways of confirming what was wrong with his plants, he started to tell me the whole story.
Seems he is growing these plants for 'ceremonial' purposes. Got the seeds from a shamans garden in New Mexico and one of the species he had contains up to 15% nicotine, he says. He was concerned that he might have to 'sacrifice' the rest of the plants if this was TMV, he was hoping there might be some alternative to starting over.
Only in Santa Cruz ................
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-microscopy.com Wed Sep 17 02:31:56 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8H7Va5t008995 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 02:31:36 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8H7Va12008990 for L1; Wed, 17 Sep 2003 02:31:36 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8H7Ub5t008960 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 02:31:02 -0500 Received: from mail.neuro.mpg.de (mail.neuro.mpg.de [130.183.250.53]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8H7U55t008950 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 02:30:21 -0500 X-MimeOLE: Produced By Microsoft Exchange V6.0.6249.0 content-class: urn:content-classes:message MIME-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Dear Jon,
The leaf dip technique is valuable for locating plant viruses but not always 100% trustworthy, even when plenty of virus is present. Back in the 1960s a colleague of mine was studying plant viruses and used PTA as the negative stain. He saw plenty of rods (it was a rod-virus, but not TMV), however nothing else. For some now unknown reason he did another leaf dip but this time with a different negative stain, (uranyl acetate or ammonium molybdate?), and saw lots of small icosahedral virus particles but no rods. After much speculation about the reason for these "spherical rods" he concluded that this was a mixed (double) viral infection. This was later proved correct. The lesson here is not to rely solely on one negative stain when examining an unknown infection, especially when no virus can be seen. It could be that the stain is for some reason not capable of contrasting the virus; try another stain to be sure. Best wishes,
Jim
-----Original Message----- } From: Jon Krupp [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Wednesday, September 17, 2003 12:00 AM To: Microscopy-at-sparc5.microscopy.com
Hi:
I got plenty of advice about how to do a quick survey for TMV. Most of it was what I had suspected, a simple leaf drag through some negative stain. Though I had never done it before, it was reassuring to know I was on the right track.
The guy came to the lab today, we did not see any TMV in his leaves. The leaves looked mottled, like TMV, but I did not see any in the EM. I tried 3 grids, got lots of other junk on the grids, but either no TMV or there was so much of it that it made a solid mat that made seeing individual particles impossible.
Now, here's the good part. He came in with a couple of species of tobacco leaves, they looked like they had TMV. I said "Looks like TMV, nothing you can do about it, doesn't matter if we can see virus or not, the plants are toast."
He said he really wanted to know if it was TMV because he had other plants growing and was afraid they might get infected too. As we talked over other ways of confirming what was wrong with his plants, he started to tell me the whole story.
Seems he is growing these plants for 'ceremonial' purposes. Got the seeds from a shamans garden in New Mexico and one of the species he had contains up to 15% nicotine, he says. He was concerned that he might have to 'sacrifice' the rest of the plants if this was TMV, he was hoping there might be some alternative to starting over.
Only in Santa Cruz ................
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-microscopy.com Wed Sep 17 03:04:07 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8H83q5t009219 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 03:03:52 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8H83qu1009214 for L1; Wed, 17 Sep 2003 03:03:52 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8H8365t009183 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 03:03:31 -0500 Received: from tytan.inmat.pw.edu.pl (www.inmat.pw.edu.pl [194.29.176.2]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8H82d5t009176 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 03:02:55 -0500 Received: from inmat.pw.edu.pl (mailer.inmat.pw.edu.pl [148.81.84.68]) by tytan.inmat.pw.edu.pl (8.9.3/8.9.3) with ESMTP id IAA06065 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 08:41:36 +0200 Received: from MMM/SpoolDir by inmat.pw.edu.pl (Mercury 1.48); 17 Sep 03 10:01:33 +0100 Received: from SpoolDir by MMM (Mercury 1.48); 17 Sep 03 10:01:27 +0100
*Send reply to: "Ladd Research" {sales-at-laddresearch.com} *From: "Ladd Research" {ladres-at-worldnet.att.net} *To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} *Subject: 3rd edition - Goldstein et al *Date sent: Tue, 16 Sep 2003 17:07:30 -0400
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Best regards,
Witold Zielinski * *Is there a third edition of "Scanning Electron Microscopy and X-Ray *Microanalysis" by Goldstein et al? Does anyone know the publisher? * *Thanks, *Deb Sicard * *Ladd Research *83 Holly Court *Williston, VT 05495 * *On-line Catalog: http://www.laddresearch.com * *tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) *fax: 1-802-660-8859 *e-mail: sales-at-laddresearch.com * * * * :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) ;)
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From MicroscopyL-request-at-microscopy.com Wed Sep 17 07:22:37 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HCMR5t011142 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 07:22:27 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HCMRSE011137 for L1; Wed, 17 Sep 2003 07:22:27 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HCLk5t011104 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 07:22:07 -0500 Received: from hotmail.com (law12-f71.law12.hotmail.com [64.4.19.71]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HCLU5t011098 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 07:21:41 -0500 Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; Wed, 17 Sep 2003 05:21:14 -0700 Received: from 136.167.54.99 by lw12fd.law12.hotmail.msn.com with HTTP; Wed, 17 Sep 2003 12:21:13 GMT X-Originating-IP: [136.167.54.99] X-Originating-Email: [boronnitride-at-hotmail.com]
Dear colleage,
We are in the process of purchasing a 4K x 4k or 2k x 2k CCD camera for our JEOL 2010F TEM. The microscope is attached with Gatan's Digipeels, therefore the CCD camera has to be compatible with the existing Peels system. Our questions are:
1) Is Gatan the only provider for such kind of CCD camera? If there is other providers, we would like to consider.
2) Is the technology for the 4k x 4k camera still premature?
Any other input regarding CCD camera are welcome.
Thanks a lot!
James
_________________________________________________________________ Compare Cable, DSL or Satellite plans: As low as $29.95. https://broadband.msn.com
From MicroscopyL-request-at-microscopy.com Wed Sep 17 07:38:14 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HCc45t011388 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 07:38:04 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HCc4dt011383 for L1; Wed, 17 Sep 2003 07:38:04 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HCbO5t011353 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 07:37:44 -0500 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HCbD5u011350 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 17 Sep 2003 07:37:13 -0500 Mime-Version: 1.0 X-Sender: (Unverified) Message-Id: {p05111a00bb8e0577a8c3-at-[206.69.208.21]}
Dear colleage,
We are in the process of purchasing a 4K x 4k or 2k x 2k CCD camera for our JEOL 2010F TEM. The microscope is attached with Gatan's Digipeels, therefore the CCD camera has to be compatible with the existing Peels system. Our questions are:
1) Is Gatan the only provider for such kind of CCD camera? If there is other providers, we would like to consider.
2) Is the technology for the 4k x 4k camera still premature?
Any other input regarding CCD camera are welcome.
Thanks a lot!
James
From MicroscopyL-request-at-microscopy.com Wed Sep 17 07:52:54 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HCqi5t011515 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 07:52:44 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HCqitE011510 for L1; Wed, 17 Sep 2003 07:52:44 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HCq35t011483 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 07:52:24 -0500 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HCpp5u011478 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 17 Sep 2003 07:51:52 -0500 Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) Message-Id: {p05111a02bb8e09519073-at-[206.69.208.21]}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jjjani-at-gauanand.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Tuesday, September 16, 2003 at 22:43:21 ---------------------------------------------------------------------------
Email: jjjani-at-gauanand.com Name: Janardan Jani
Organization: Gujarat Agricultural University
Education: Graduate College
Location: Anand, Gujarat State, India
Question: Respected Sir/ Madam,
I wish to carry out SEM for my Bacterial isolates which are probably Bacillus thuringiensis. I wish to observe the spore and crystal the delta endotoxin which is bipyramidal in shape. I have access to Hitachi S3000 SEM but do not have protocol for the sample preparation. I would be obliged if i am given a protocol about the sample preparation and staining if any for the purpose.
From MicroscopyL-request-at-microscopy.com Wed Sep 17 11:19:26 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HGJF5t013255 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 11:19:15 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HGJF5F013251 for L1; Wed, 17 Sep 2003 11:19:15 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HGIZ5t013218 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 11:18:55 -0500 Received: from col-msxproto1.col.missouri.edu (col-msxproto1.col.missouri.edu [128.206.3.151]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HGID5t013214 for {Microscopy-at-msa.microscopy.com} ; Wed, 17 Sep 2003 11:18:24 -0500 Received: from phillips-dell.missouri.edu ([128.206.80.38]) by col-msxproto1.col.missouri.edu over TLS secured channel with Microsoft SMTPSVC(5.0.2195.6673); Wed, 17 Sep 2003 11:17:59 -0500 Message-Id: {5.1.1.5.2.20030917111540.02a2fca0-at-pop.missouri.edu} X-Sender: phillipst-at-pop.missouri.edu X-Mailer: QUALCOMM Windows Eudora Version 5.1.1
I keep my ethanol series (50, 70, 95, and 100%) in squirt bottles that have a long tube like shaft coming out the top of them. If you pull the shaft upwards, it seals off the bottle and prevents evaporation and minimizes humidity contamination of the ethanol. Unfortunately, these bottles are getting quite old (16 years) and need replacement. I can't remember where I got them. Does anyone use a similar type or have a better alternative? Thanks, tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
From MicroscopyL-request-at-microscopy.com Wed Sep 17 11:47:08 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HGkj5t013484 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 11:46:45 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HGkjWf013479 for L1; Wed, 17 Sep 2003 11:46:45 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HGjp5t013446 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 11:46:14 -0500 Received: from exchangesrvr.qdots.com ([12.152.176.130]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HGjN5t013437 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 11:45:35 -0500 Received: by EXCHANGESRVR with Internet Mail Service (5.5.2653.19) id {P5ZDYZ8D} ; Wed, 17 Sep 2003 09:36:40 -0700 Message-ID: {6570F6B26F36D511885100B0D0AA3DDB3C555A-at-EXCHANGESRVR}
Dear James,
Another CCD camera company worth considering is TVIPS. They are in Germany and they also have representatives in the USA. They make very good cameras and easy to install and use. The contact details are:
Hans Tietz Email: hans.tietz-at-tvips.com Fax: +(49)-89-8509488
Sincerely,
Thearith Ung
-----Original Message----- } From: Boron nitride [mailto:boronnitride-at-hotmail.com] Sent: Wednesday, September 17, 2003 5:21 AM To: Microscopy-at-sparc5.microscopy.com
Dear colleage,
We are in the process of purchasing a 4K x 4k or 2k x 2k CCD camera for our JEOL 2010F TEM. The microscope is attached with Gatan's Digipeels, therefore
the CCD camera has to be compatible with the existing Peels system. Our questions are:
1) Is Gatan the only provider for such kind of CCD camera? If there is other
providers, we would like to consider.
2) Is the technology for the 4k x 4k camera still premature?
Any other input regarding CCD camera are welcome.
Thanks a lot!
James
_________________________________________________________________ Compare Cable, DSL or Satellite plans: As low as $29.95. https://broadband.msn.com
From MicroscopyL-request-at-microscopy.com Wed Sep 17 12:40:59 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HHem5t013840 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 12:40:48 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HHemmr013835 for L1; Wed, 17 Sep 2003 12:40:48 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HHe85t013805 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 12:40:28 -0500 Received: from tisch.mail.mindspring.net (tisch.mail.mindspring.net [207.69.200.157]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HHdk5t013793 for {Microscopy-at-msa.microscopy.com} ; Wed, 17 Sep 2003 12:39:56 -0500 Received: from 1cust238.tnt1.mln6.ita.da.uu.net ([213.116.64.238]) by tisch.mail.mindspring.net with smtp (Exim 3.33 #1) id 19zgH2-0006bE-00 for Microscopy-at-MSA.Microscopy.com; Wed, 17 Sep 2003 13:39:29 -0400 To: MICROSCOPY BB {Microscopy-at-msa.microscopy.com}
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Peiyi Wang wrote the following: ================================================================= } The C2 apertures in our CM200 FEG has become very dirty and needs to be } replaced as soon as possible. As you might be aware, the C2 apertures for FEG } have to be very clear and need to take a special method for cleaning. Now I } have got several options: } } A.Buy from FEI directly but they obviously cost about more } than three times from ordinary aperture of which if you } purchase from Agar. } B.Use special plasma cleaner to clear up those ordinary } apertures so that apertures would be acceptable by FEG. } C.Try to use the plasma from a carbon coater to clear up } ordinary apertures. } } My questions is either B or C would work? Alternatively, there are any good } ideas for this purpose? =================================================================== You always have the option of purchasing new apertures from either the original equipment manufacturer or from other sources, such as Agar (as you indicated) or SPI Supplies or yet others.
A plasma cleaner operates at very low power (supposedly under 10 watts) whereas a plasma etcher operates typically at 100 watts or higher. The quality of the cleaning will depend on the power being used, and since there is no reason not to use 100 watts, you could clean the apertures in a few minutes or less in a plasma etcher using oxygen in a unit such as the SPI Plasma Prep II plasma etcher. See URL www.2spi.com/catalog/instruments/etchers1.shtml
If you were using a plasma cleaner, such as the SPI Plasma Cleaner, see URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml you could use a mixture of oxygen and argon, which would be more effective. You don't want to use argon at 100 watts since it could possibly etch the aperture itself, which obviously would be undesireable.
If you use this approach, your apertures will last nearly forever (well nothing is forever) since you are not heat cycling them as you would in a vacuum evaporator. Of course, users eventually (accidentally) damage the edges of the hole with the beam so apertures really don't last forever. But cleaning them this way will enable them to last far longer than would otherwise be possible.
We don't see how you could use the plasma in a carbon coater to clean apertures, we have never heard of anyone doing that. Are you sure you are not referring to the use of a vacuum evaporator (with diffusion or turbo pump) to clean apertures, but in that case, it would be the heat instead of a plasma responsible for the cleaning.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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From MicroscopyL-request-at-microscopy.com Wed Sep 17 12:48:43 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HHmR5t013948 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 12:48:27 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HHmRtO013944 for L1; Wed, 17 Sep 2003 12:48:27 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HHle5t013914 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 12:48:06 -0500 Received: from nightshade.noc.ucla.edu (nightshade.noc.ucla.edu [169.232.48.18]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HHl65t013904 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 12:47:23 -0500 Received: from azalea.noc.ucla.edu (azalea.noc.ucla.edu [169.232.47.12]) by nightshade.noc.ucla.edu (8.12.8p1/8.12.8) with ESMTP id h8HHkqW9021247 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 10:46:52 -0700 Received: from emi.ucla.edu ([149.142.241.180]) (authenticated bits=0) by azalea.noc.ucla.edu (8.12.9/8.12.9) with ESMTP id h8HHkp7T007203 (version=TLSv1/SSLv3 cipher=DES-CBC3-SHA bits=168 verify=NOT) for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 10:46:51 -0700 Message-Id: {5.2.0.9.2.20030917102256.01b30cf8-at-mail.ucla.edu} X-Sender: sryazant-at-pop.bol.ucla.edu X-Mailer: QUALCOMM Windows Eudora Version 5.2.0.9
Dear James Gatan is not only producer of high resolution digital cameras for TEM. You may shop around to find other manufacturers (or they will find you). Personally, I do have camera from Gatan and really happy with it. A few things to consider:
- CCD cameras with fiber-optics coupling (most modern cameras including Gatan's UltraScan 4x4 or 2x2) are slow. It means, you don't have real video-mode. It does not matter what manufacturer claims, in reality you do have a few frames per second and that's it. Again, as more pixels you have, camera is slower (in general). To overcome this problem, Gatan virtually split their 4x4 chip on 4 quadrants and read them simultaneously increasing speed 4 times. You have to pay a fortune for such trick.
- as far as I know, UltraScan 4x4K is pretty mature camera; they install them on new FEI TEMs as a part of standard package. If I will have money, I'll definetely get it (camera and FEI microscope as well).
- Another things to consider is customer service.
- Before you made final decision, ask company for the demo. When I did shopping for my camera, I had a few offers from different companies. The cameras were pretty similar on the paper and so different when I took pictures and compare them side-by-side. So, ask for the demo, bring your own samples, took your own pictures and compare.
Good luck, Sergey.
At 05:37 AM 9/17/2003, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
From MicroscopyL-request-at-microscopy.com Wed Sep 17 12:54:03 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HHrq5t014036 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 12:53:52 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HHrqHV014032 for L1; Wed, 17 Sep 2003 12:53:52 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HHrC5t014004 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 12:53:32 -0500 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HHr05u013999 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 17 Sep 2003 12:53:01 -0500 Mime-Version: 1.0 X-Sender: (Unverified) Message-Id: {p05111a01bb8e4fe62a85-at-[206.69.208.21]}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bart-at-lasalle.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 17, 2003 at 09:52:23 ---------------------------------------------------------------------------
Email: bart-at-lasalle.edu Name: Henry Bart
Organization: La Salle University
Title-Subject: MListserver:
Question: I have a TOPCOM SM-300 SEM and would like to know what company,if any, has picked up that line after RJ LEE stopped supporting it. I need some parts and advice. Thanks
From MicroscopyL-request-at-microscopy.com Wed Sep 17 15:32:10 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HKVe5t015211 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 15:31:40 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HKVetv015206 for L1; Wed, 17 Sep 2003 15:31:40 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HKUd5t015166 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 15:31:06 -0500 Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HKTx5t015156 for {Microscopy-at-msa.microscopy.com} ; Wed, 17 Sep 2003 15:30:15 -0500 Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 1.16 (built May 14 2003)) id {0HLD00E01KWWZQ-at-Polaris.umdnj.edu} for Microscopy-at-msa.microscopy.com; Wed, 17 Sep 2003 16:29:52 -0400 (EDT) Received: from rwja.umdnj.edu (rwja.umdnj.edu [130.219.4.100]) by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 1.16 (built May 14 2003)) with ESMTP id {0HLD00E4LKXRZR-at-Polaris.umdnj.edu} ; Wed, 17 Sep 2003 16:29:52 -0400 (EDT) Received: from umdnj.edu ([130.219.170.159]) by rwja.umdnj.edu (8.9.3 (PHNE_28809)/8.9.3) with ESMTP id QAA17672; Wed, 17 Sep 2003 16:29:40 -0400 (EDT)
Hi Tom:
I agree with Lee's answer, RR in the glut, rinses, osmium and after that no RR. I also used the Luft protocol. I seem to remember using para-phenylene diamine (0.5% in alcohol during dehydration) to intensify (reduce) the osmium-RR complex on at least some of my samples. I had fantastic contrast, almost too much. Since I was looking at cells in culture evenness of penetration/staining was not a problem. By some strange coincidence I also used RR on cultured retinal pigment epithelium just as Lee did.
Geoff
Tom Phillips wrote:
} I am using Luft's technique for staining the glycocalyx with Ruthenium } Red. I know the RR is supposed to be included in the aldehyde fix, } the rinses and the osmium but how about the osmium rinses or ethanol } dehydration series? Has anyone successfully done this procedure? } Thanks, tom } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-microscopy.com Wed Sep 17 15:37:22 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HKbC5t015316 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 15:37:12 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8HKbCQ8015311 for L1; Wed, 17 Sep 2003 15:37:12 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8HKaW5t015278 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 15:36:52 -0500 Received: from smtp3.doit.wisc.edu (blitzen.doit.wisc.edu [144.92.197.183]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8HKaA5t015272 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Sep 2003 15:36:20 -0500 Received: from conversion-daemon.smtp3.doit.wisc.edu by smtp3.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) id {0HLD00K01KKUVM-at-smtp3.doit.wisc.edu} for Microscopy-at-sparc5.microscopy.com; Wed, 17 Sep 2003 15:33:04 -0500 (CDT) Received: from [10.25.50.161] (ansci.wisc.edu [144.92.132.175]) by smtp3.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) with ESMTP id {0HLD004NQL34WK-at-smtp3.doit.wisc.edu} for Microscopy-at-sparc5.microscopy.com; Wed, 17 Sep 2003 15:33:04 -0500 (CDT)
Micromavens,
We need a hint or two about cryosectioning nylon. The cryocutter in our lab is used to sectioning complex hydrated heterocopolymers (i.e. biological specimens), and the nylon is being recalcitrant. Any tips (I checked the U. Florida Tips & Tricks site) for cutting temperature, cutting speed and so forth? This is for routine TEM study of a 2-phase nylon, and a nylon with silicon dioxide nanoparticles. Nothing extraordinary. Thanks for any hints.
Phil
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-microscopy.com Wed Sep 17 22:13:47 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8I3DM5t018246 for {L1-at-ns.microscopy.com} ; Wed, 17 Sep 2003 22:13:22 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8I3DMjL018242 for L1; Wed, 17 Sep 2003 22:13:22 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8I3CQ5t018208 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 17 Sep 2003 22:12:51 -0500 Received: from webmail.sas.upenn.edu (webmail2.sas.upenn.edu [128.91.55.36]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8I3Bo5t018202 for {Microscopy-at-msa.microscopy.com} ; Wed, 17 Sep 2003 22:12:06 -0500 Received: from webmail.sas.upenn.edu (localhost.localdomain [127.0.0.1]) by webmail.sas.upenn.edu (8.12.8/8.12.8) with ESMTP id h8I3BZwt020927; Wed, 17 Sep 2003 23:11:35 -0400 Received: (from nobody-at-localhost) by webmail.sas.upenn.edu (8.12.8/8.12.8/Submit) id h8I3BZYF020925; Wed, 17 Sep 2003 23:11:35 -0400 X-Authentication-Warning: webmail.sas.upenn.edu: nobody set sender to psconnel-at-sas.upenn.edu using -f Received: from pcp04013366pcs.walngs01.pa.comcast.net (pcp04013366pcs.walngs01.pa.comcast.net [68.80.33.179]) by webmail.sas.upenn.edu (IMP) with HTTP for {psconnel-at-192.0.3.3} ; Wed, 17 Sep 2003 23:11:35 -0400 Message-ID: {1063854695.3f692267330d7-at-webmail.sas.upenn.edu}
Tom, As a substitute for your ethanol bottles I would suggest that you consider the pull-up type caps that are available on some juice and water bottles. I have noticed that there is a wide variety of sizes and quality to the different suppliers. Perhaps there is one that fits glass bottles or better plastic ones, like the pint bottles that the ethanol comes in.
I hadn't thought of using these before but it seems like a good idea to try.
Pat Connelly U of P Biology Philadelphia, PA
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I have not been able to get the latest version (Beta 4.0.2) of Scion Image (on a machine running Win XP) to open or import digital images. I've tried to open and import both .tifs and .bmps to no avail. The program just crashes. Scion does not provide technical support unless you have purchased equipment from them, which I have not. Can anybody suggest a solution? Or, alternatively, do you know of any other image analysis software that's free/inexpensive? Thanks!
From MicroscopyL-request-at-microscopy.com Thu Sep 18 02:52:02 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8I7pliO020100 for {L1-at-ns.microscopy.com} ; Thu, 18 Sep 2003 02:51:47 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8I7plNZ020095 for L1; Thu, 18 Sep 2003 02:51:47 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8I7p1iO020068 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 18 Sep 2003 02:51:26 -0500 Received: from amdext2.amd.com (amdext2.amd.com [163.181.251.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8I7oZiO020062 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 Sep 2003 02:50:51 -0500 Received: from SAUSGW01.amd.com (sausgw01.amd.com [163.181.250.21]) by amdext2.amd.com (8.12.8/8.12.8/AMD) with ESMTP id h8I7msPw010403 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 Sep 2003 02:50:18 -0500 (CDT) Received: from 163.181.250.1 by SAUSGW02.amd.com with ESMTP (AMD SMTP Relay (MMS v5.5.3)); Thu, 18 Sep 2003 02:50:12 -0500 Received: from deexmta4.amd.com (deexmta4.amd.com [172.20.3.104]) by amdint2.amd.com (8.12.8/8.12.8/AMD) with ESMTP id h8I7o71i021434 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 Sep 2003 02:50:08 -0500 (CDT) Received: by deexmta4.amd.com with Internet Mail Service (5.5.2653.19) id {QSCATK7W} ; Thu, 18 Sep 2003 09:50:07 +0200 Message-ID: {E540DF203FFED21182EB0008C7287560146DFF53-at-deexmta4.amd.com}
ImageJ is a free Java port of NIH Image/Scion Image. Runs on any platform. http://rsb.info.nih.gov/ij/
---------------------------------------------------- Dr. Heiko Stegmann AMD Saxony LLC & Co. KG Materials analysis/TEM e-mail heiko.stegmann-at-amd.com ----------------------------------------------------
From MicroscopyL-request-at-microscopy.com Thu Sep 18 03:25:27 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8I8P2iO020297 for {L1-at-ns.microscopy.com} ; Thu, 18 Sep 2003 03:25:02 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8I8P22Y020292 for L1; Thu, 18 Sep 2003 03:25:02 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8I8O6iO020262 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 18 Sep 2003 03:24:27 -0500 Received: from lakemtao06.cox.net (lakemtao06.cox.net [68.1.17.115]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8I8NUiO020257 for {microscopy-at-msa.microscopy.com} ; Thu, 18 Sep 2003 03:23:41 -0500 Received: from home ([68.12.42.17]) by lakemtao06.cox.net (InterMail vM.5.01.06.05 201-253-122-130-105-20030824) with SMTP id {20030918082316.ENMC4662.lakemtao06.cox.net-at-home} ; Thu, 18 Sep 2003 04:23:16 -0400 Message-ID: {058001c37dbe$1c3576f0$112a0c44-at-home}
ImageJ put out by NIS is free and works on almost anything. http://rsb.info.nih.gov/ij/ It is the successor of Sicon image.
Gordon } From: "Diana Greenlee" {greenlee-at-u.washington.edu}
: : I have not been able to get the latest version (Beta 4.0.2) of Scion Image : (on a machine running Win XP) to open or import digital images. I've : tried to open and import both .tifs and .bmps to no avail. The program : just crashes. Scion does not provide technical support unless you have : purchased equipment from them, which I have not. Can anybody suggest a : solution? Or, alternatively, do you know of any other image analysis : software that's free/inexpensive? Thanks! :
From MicroscopyL-request-at-microscopy.com Thu Sep 18 08:34:03 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8IDXriO022888 for {L1-at-ns.microscopy.com} ; Thu, 18 Sep 2003 08:33:53 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8IDXrhM022884 for L1; Thu, 18 Sep 2003 08:33:53 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8IDXCiO022846 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 18 Sep 2003 08:33:33 -0500 Received: from web13408.mail.yahoo.com (web13408.mail.yahoo.com [216.136.175.66]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8IDWuiO022838 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 18 Sep 2003 08:33:07 -0500 Message-ID: {20030918133241.91747.qmail-at-web13408.mail.yahoo.com} Received: from [24.129.146.80] by web13408.mail.yahoo.com via HTTP; Thu, 18 Sep 2003 06:32:41 PDT
I need to purchase a frame grabber card for Image Analysis to use either NIH Images Analysis, Scion program or the Image Java Program but I need to know what boards will work with an Amray 1000 SEM. I have both a MAC G3 and PC Windows platforms.
john
From MicroscopyL-request-at-microscopy.com Thu Sep 18 12:30:44 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8IHURiO024939 for {L1-at-ns.microscopy.com} ; Thu, 18 Sep 2003 12:30:27 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8IHURkJ024934 for L1; Thu, 18 Sep 2003 12:30:27 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8IHTaiO024903 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 18 Sep 2003 12:29:56 -0500 Received: from mailhub-3.iastate.edu (mailhub-3.iastate.edu [129.186.140.13]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8IHSxiO024898 for {Microscopy-at-msa.microscopy.com} ; Thu, 18 Sep 2003 12:29:12 -0500 Received: from mailout-1.iastate.edu (mailout-1.iastate.edu [129.186.140.1]) by mailhub-3.iastate.edu (8.9.3p2/8.9.3) with SMTP id MAA04439; Thu, 18 Sep 2003 12:28:42 -0500 Received: from pdp2.marl.iastate.edu(129.186.227.2) by mailout-1.iastate.edu via csmap id 22927; Thu, 18 Sep 2003 17:26:41 +0000 (UTC) Message-Id: {5.2.0.9.2.20030918122246.047e5190-at-wesaia.mail.iastate.edu} X-Sender: wesaia-at-wesaia.mail.iastate.edu X-Mailer: QUALCOMM Windows Eudora Version 5.2.0.9
The short answer is I don't know. But I would ask you first, are you sure a frame grabber will provide you enough resolution?
I have nothing against Scion or any other manufacturer of frame grabbers, but I am not sure that you really want a frame grabber for SEM applications. The limitations of the video signal, RS-170, prevent you from digitizing more than about 640 pixels across the image, and that assumes that the Amray uses RS-170. A frame grabber may be adequate for basic imaging, but the image will be somewhat grainy. I would not be inclined to publish such images or to attempt image analysis on them.
I think most SEM imaging systems resort to either active or passive digitization to collect images at higher resolution.
Warren
At 06:32 AM 9/18/2003 -0700, you wrote:
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From MicroscopyL-request-at-microscopy.com Thu Sep 18 12:53:41 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8IHrUiO025134 for {L1-at-ns.microscopy.com} ; Thu, 18 Sep 2003 12:53:30 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8IHrUe5025130 for L1; Thu, 18 Sep 2003 12:53:30 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8IHqoiO025098 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 18 Sep 2003 12:53:10 -0500 Received: from muaix02.mcs.muohio.edu (muaix02.mcs.muohio.edu [134.53.6.35]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8IHq6iO025089 for {microscopy-at-MSA.Microscopy.com} ; Thu, 18 Sep 2003 12:52:17 -0500 Received: from muw2k05 (muw2k05.mcs.muohio.edu [134.53.6.19]) by muaix02.mcs.muohio.edu (Switch-3.0.4/Switch-3.0.0) with SMTP id h8IHpkq2193556 for {microscopy-at-MSA.Microscopy.com} ; Thu, 18 Sep 2003 13:51:46 -0400 Received: From mcsaix05.mcs.muohio.edu ([134.53.253.27]) by muw2k05 (WebShield SMTP v4.5 MR1a); id 1063907314703; Thu, 18 Sep 2003 13:48:34 -0400 Received: from emf03 (emf03.emf.muohio.edu [134.53.9.119]) by mcsaix05.mcs.muohio.edu (Switch-3.0.4/Switch-3.0.0) with ESMTP id h8IHpjPk120058 for {microscopy-at-MSA.Microscopy.com} ; Thu, 18 Sep 2003 13:51:45 -0400
Lookingfor a method of determining live vs dead via microscopy BUT not using a fluorescent stain. Organism is a level 2 pathogen and cannot be accomodated safely under our fluorescent scopes.
The samples do not need to be live at time of observation.
If there is a live dead fluorescent stain which retains live dead after fixation/killing that would work as well.
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-microscopy.com Thu Sep 18 16:42:35 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8ILgCiO026746 for {L1-at-ns.microscopy.com} ; Thu, 18 Sep 2003 16:42:12 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8ILgCiF026741 for L1; Thu, 18 Sep 2003 16:42:12 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8ILfIiO026713 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 18 Sep 2003 16:41:41 -0500 Received: from sidney.skcc.org (66-192-151-2.gen.twtelecom.net [66.192.151.2]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8ILeniO026704 for {microscopy-at-MSA.Microscopy.com} ; Thu, 18 Sep 2003 16:41:02 -0500 Received: by mail.skcc.org with Internet Mail Service (5.5.2653.19) id {QDA8SHSH} ; Thu, 18 Sep 2003 14:40:09 -0700 Message-ID: {8C585FEB9085D6119A4C0002A5EB792AD8BA3A-at-mail.skcc.org}
Years ago, when working on viability of ovine articular cartilage, I noticed that fresh tissue stained with acridine orange maintained its viability status (as judged by colors of the dye) after it had been kept in 70% ethanol at 4oC for several days.
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOHIO.EDU] Sent: Thursday, September 18, 2003 10:51 AM To: microscopy-at-MSA.Microscopy.com
Lookingfor a method of determining live vs dead via microscopy BUT not using a fluorescent stain. Organism is a level 2 pathogen and cannot be accomodated safely under our fluorescent scopes.
The samples do not need to be live at time of observation.
If there is a live dead fluorescent stain which retains live dead after fixation/killing that would work as well.
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-microscopy.com Thu Sep 18 17:57:08 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8IMuwiO027282 for {L1-at-ns.microscopy.com} ; Thu, 18 Sep 2003 17:56:58 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8IMuwYr027277 for L1; Thu, 18 Sep 2003 17:56:58 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8IMuHiO027214 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 18 Sep 2003 17:56:37 -0500 Received: from smtp2.doit.wisc.edu (donner.doit.wisc.edu [144.92.197.184]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8IMu2iQ027202 for {microscopy-at-MSA.Microscopy.com} ; Thu, 18 Sep 2003 17:56:12 -0500 Received: from conversion-daemon.smtp2.doit.wisc.edu by smtp2.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) id {0HLF00J01M61PK-at-smtp2.doit.wisc.edu} for microscopy-at-MSA.Microscopy.com; Thu, 18 Sep 2003 17:55:48 -0500 (CDT) Received: from m.Wisc.Edu (66-188-109-142.mad.wi.charter.com [66.188.109.142]) by smtp2.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) with ESMTPA id {0HLF006R4MD0S5-at-smtp2.doit.wisc.edu} for microscopy-at-MSA.Microscopy.com; Thu, 18 Sep 2003 17:55:48 -0500 (CDT)
We are very interested in buying a cryo-ultramicrotome below $30k. Vendors and refurbishing specialists very welcome!
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We are very interested in buying a cryo-attachment for Reichert Ultracut. Vendors and refurbishing specialists very welcome!
From MicroscopyL-request-at-microscopy.com Fri Sep 19 16:35:17 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8JLZ7iO003529 for {L1-at-ns.microscopy.com} ; Fri, 19 Sep 2003 16:35:07 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8JLZ7Ka003525 for L1; Fri, 19 Sep 2003 16:35:07 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8JLYRiO003494 for "MListServerFilteredEmail_5-at-microscopy.com"; Fri, 19 Sep 2003 16:34:47 -0500 Received: from ITEVS.uchc.net (itevs.uchc.edu [155.37.91.41] (may be forged)) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8JLYGiO003490 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 19 Sep 2003 16:34:26 -0500 X-MimeOLE: Produced By Microsoft Exchange V6.0.6487.1 content-class: urn:content-classes:message MIME-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1"
Zeiss TEM 10A has to be moved ASAP. It was in use and working well in July 2002. It was serviced in June 2002. Contact Dr. Cartun at the following address: Rcartun-at-harthosp.org It is located in Connecticut.
From MicroscopyL-request-at-microscopy.com Mon Sep 22 01:14:13 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8M6DwiO028246 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 01:13:58 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8M6DwKb028241 for L1; Mon, 22 Sep 2003 01:13:58 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8M6DCiO028215 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 01:13:37 -0500 Received: from cassandra.net.lu.se (cassandra.net.lu.se [130.235.128.120]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8M6CpiO028210 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 01:13:07 -0500 Received: from materialkemi.lth.se (cinderella.materialkemi.lth.se [130.235.24.32]) by cassandra.net.lu.se (iPlanet Messaging Server 5.2 HotFix 1.14 (built Mar 18 2003)) with ESMTP id {0HLL00DR5QKU9C-at-cassandra.net.lu.se} for Microscopy-at-MSA.Microscopy.Com; Mon, 22 Sep 2003 08:12:30 +0200 (MEST)
Dear Microscopy Listserver members,
I wonder if it is possible to use 6 nm-gold-tagged antibodies for pre-embedding immunolabeling, or if it is necessary to use smaller ones? I want to do immunolabeling of betacells from the islets of Langerhans (rat).
Sincerely, Pernilla Nevsten
........................................................................................................................................... Pernilla Nevsten, PhD-student Materials Chemistry Lund University, Sweden
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Question: I¥we used Ruthenium Red to stain the capsule of vibro colerae successfully. I use RR in the fixation, rinse and osmium but not in the osmium rinse or the ethanol dehydration steps. Good luck!
From MicroscopyL-request-at-microscopy.com Mon Sep 22 09:16:00 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MEFoiO031706 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 09:15:50 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8MEFowC031702 for L1; Mon, 22 Sep 2003 09:15:50 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8MEF9iO031675 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 09:15:29 -0500 Received: from lamont.ldeo.columbia.edu (ldgo.columbia.edu [129.236.10.30]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MEEriO031668 for {microscopy-at-msa.microscopy.com} ; Mon, 22 Sep 2003 09:15:04 -0500 Received: from [129.236.40.251] (dee-mac.ldgo.columbia.edu [129.236.40.251]) by lamont.ldeo.columbia.edu (8.12.9/8.12.3/LDEO-1.27) with ESMTP id h8MEESU7029739 for {microscopy-at-msa.microscopy.com} ; Mon, 22 Sep 2003 10:14:31 -0400 (EDT)
Dear colleagues,
We are seeking information from anyone who has knowledge of how long a critical-point dried specimen prepared for SEM is sufficiently stable for subsequent viewing. We anticipate receiving many samples of glutaraldehyde-fixed plankton with delicate protozoa and algae in them. Our colleagues sending the material will eventually sample only a small subset based on data they gather one year from now. So, we will be asked to examine the stubs approximately 12 months in the future.
Has anyone advice on the stability of specimens for SEM observation that have been placed on stubs one year prior to observation? Is it preferable to lightly sputter-coat the sample before storage and is there any secure hope of stabilizing delicate flagellar structures, etc. after one year? We understand the importance of storage in a dessicator and at fairly constant temperature, but we have never stored specimens for 1 year before observing them.
Many thanks, Dee
```````
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
From MicroscopyL-request-at-microscopy.com Mon Sep 22 09:23:55 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MENjiO031794 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 09:23:45 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8MENjD4031790 for L1; Mon, 22 Sep 2003 09:23:45 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8MEN4iO031762 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 09:23:25 -0500 Received: from night.its.uiowa.edu (night.its.uiowa.edu [128.255.56.106]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MEMliO031756 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 09:22:54 -0500 Received: from localhost (webmail1-maint.its.uiowa.edu [128.255.56.153]) by night.its.uiowa.edu (8.12.10/8.12.9/ns-mx-1.16) with ESMTP id h8MEMQas063214 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 09:22:26 -0500 Received: from dhcp80ff5982.dynamic.uiowa.edu (dhcp80ff5982.dynamic.uiowa.edu [128.255.89.130]) by webmail1.its.uiowa.edu (IMP) with HTTP for {rossjl-at-localhost} ; Mon, 22 Sep 2003 09:22:26 -0500 Message-ID: {1064240546.3f6f05a23e4c0-at-webmail1.its.uiowa.edu}
Hi everyone,
I would like some opinions about critical point dryers. What do you think about automated units vs. manual units. Or do you use HMDS and not use the CPD at all? If you could post to the list and not send them directly to me, I would appreciate it and everyone could read the responses.
Thanks, Jean Ross Central Microscopy Research Facility University of Iowa
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pfraundorf-at-umsl.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, September 20, 2003 at 08:45:53 ---------------------------------------------------------------------------
Email: pfraundorf-at-umsl.edu Name: Phil Fraundorf
Organization: UM-StL Physics and Astronomy/CME
Title-Subject: Education: a variable size-scale adventure
Question: We've been looking for web strategies to let non-microscopists share the experience of moving between macroscopic and nanoscopic size scales for some time. The link below* is a step in that direction. The puzzlers beneath the model are meant to be experimental in nature, rather than based on theory, so put on your Sherlock Holmes hat and enjoy.
Also, what are some: (i) learning challenges and storylines for use by students and teachers with an exploration tool like this; and (ii) ideas for scenarios to that end, e.g. different specimen configurations? The first model you encounter uses a fixed intersecting-axis goniometer. The beta model allows movement of that rotation point with respect to a "scope-based" cartesian system by hitting x,X,y,Y,z,Z. We also have a way to request lateral displacements accompanied by a dynamically redrawn heirarchy of grids. Although this can't yet be done at display rates (i.e. many times per second), current tools do allow us to generate "data" (e.g. simulated HREM images, diffraction patterns, X-ray or roughness spectra, etc.) from regions of interest. We're also interested in feedback from storyline field tests in university and high school classrooms, as well as in pooling resources for this between institutions.
Phil Fraundorf UM-StL Physics & Astronomy/CME Washington U. Physics/MCSS office: (314)516-5044 lab: (314)516-5024 pfraundorf-at-umsl.edu http://www.umsl.edu/~fraundor
From MicroscopyL-request-at-microscopy.com Mon Sep 22 09:48:38 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MEmSiO032083 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 09:48:28 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8MEmSTU032078 for L1; Mon, 22 Sep 2003 09:48:28 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8MElmiO032049 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 09:48:08 -0500 Received: from mail-na2.baxter.com (mail-na2.baxter.com [198.89.159.241]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MElWiO032043 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 09:47:42 -0500 Received: from ns-2.baxter.com ([10.8.1.25]) by mail-na2.baxter.com (Switch-2.2.8/Switch-2.2.0) with ESMTP id h8MElBi16801 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 09:47:12 -0500 (CDT) Received: from baxmms2.deerfield.baxter.com (baxmms2.deerfield.baxter.com [10.8.1.51]) by ns-2.baxter.com (Pro-8.9.3/Pro-8.9.3) with ESMTP id JAA11104 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 09:47:09 -0500 (CDT)
The Midwest Microscopy & Microanalysis Society would like to announce two upcoming meetings this fall. Please (!) circulate this notice to all your colleagues.
Friday, October 3rd University of Wisconsin, Room 235 USDA Dairy Forage Research Laboratory, 1925 Linden Drive West, Madison, Wisconsin 53706. 9:00AM to 4:30PM See the below link for the complete program: http://www.msa.microscopy.com/MSALAS/MMMS/MMMSOct2003.html
Changes to the posted program are as follows:
Ranier Guillery, Professor Emeritus UW Department of Anatomy, UW Madison will replace David Slautterback
In the afternoon Biological session: Sara Patterson's title is: "A look at cell separation in Arabidopsis using floral organ abscission as a model system"
In the afternoon Physical Sciences Session (moved to 3:05pm) : Mark Eriksson' s title is : " Novel scanning probe techniques: from off -angle tapping to dielectric imaging".
Tom Kelly from Imago Scientific will open this session (at 1:pm) with a talk entitled; "3D Atomic- Scale Compositional imaging with local electrode Atom Probes.
Friday, November 14th Harper College, Wojcik Conference Center, 1200 West Algonquin Road, W108, Palatine, Illinois 60067 SEM Workshop : Stuart McKernan - University of Minnesota: Environmental SEM John Mackenzie - North Carolina State University: Digital Imaging Robert Apkarian - Emory University: Cryo SEM in Biomedical Sciences Wayne Neimeyer- McCrone Associates- Forensic Microscopy
Details and www link to follow.
For further information please contact either:
Robert Mierzwa (MMMS President - Elect): Tel: 920-803-8945, email: mierzwa-at-jeol.com Arvid Casler (MMMS Program Coodinator): Tel: 847-674-7700, email: arvid_casler-at-fmo.com Jim DiOrio (MMMS President): Tel: 847-270-4676, email: jim_diorio-at-baxter.com
From MicroscopyL-request-at-microscopy.com Mon Sep 22 10:26:44 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MFQEiO032465 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 10:26:14 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8MFQE2u032460 for L1; Mon, 22 Sep 2003 10:26:14 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8MFPDiO032420 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 10:25:38 -0500 Received: from efsa3.efs-alsace.local (imap-efsa.u-strasbg.fr [130.79.248.7]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MFOaiO032413 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 10:24:52 -0500 content-class: urn:content-classes:message MIME-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1"
Dear Pernilla, In my opinion, you can use 6 nm gold tagged antibodies only if your cell is capable of endocytosis. Otherwise, you have to permeabilize your cell and use ultra small gold coupled to your antibody. Another alternative is to do a common immunolabelling, after permeabilization ( antibody of interest, then species specific ultra small gold.) In all case you have to amplify your small gold particles to visualize them in a electron microscope. Best regards danièle -------------------------------------------- Danièle Spehner, INSERM EPI 99-08 - EFS-Alsace 10 rue Spielmann - 67065 STRASBOURG tel : 03 88 21 25 25 - fax : 03 88 21 25 44 e-mail : daniele.spehner-at-efs-alsace.fr ------------------------------------
-----Message d'origine----- De : Pernilla Nevsten [mailto:pernilla.nevsten-at-materialkemi.lth.se] Envoyé : lundi 22 septembre 2003 08:16 À : Microscopy-at-MSA.Microscopy.Com Objet : pre-embedding immunolabeling
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Dear Microscopy Listserver members,
I wonder if it is possible to use 6 nm-gold-tagged antibodies for pre-embedding immunolabeling, or if it is necessary to use smaller ones? I want to do immunolabeling of betacells from the islets of Langerhans (rat).
Sincerely, Pernilla Nevsten
.......................................................................................................................................... Pernilla Nevsten, PhD-student Materials Chemistry Lund University, Sweden
From MicroscopyL-request-at-microscopy.com Mon Sep 22 12:28:05 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MHRjiO001004 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 12:27:45 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8MHRjUR000999 for L1; Mon, 22 Sep 2003 12:27:45 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8MHQsiO000966 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 12:27:14 -0500 Received: from smtp4.doit.wisc.edu (hermes.doit.wisc.edu [144.92.197.190]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MHQNiO000959 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 22 Sep 2003 12:26:33 -0500 Received: from conversion-daemon.smtp4.doit.wisc.edu by smtp4.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) id {0HLM00101LFUAR-at-smtp4.doit.wisc.edu} for Microscopy-at-sparc5.microscopy.com; Mon, 22 Sep 2003 12:26:02 -0500 (CDT) Received: from [10.25.50.161] (ansci.wisc.edu [144.92.132.175]) by smtp4.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) with ESMTP id {0HLM0090ULRDIE-at-smtp4.doit.wisc.edu} ; Mon, 22 Sep 2003 12:26:01 -0500 (CDT)
Dee,
As long as the samples are properly stored with good desiccant, they are archival. A year is no problem. I would suggest not sputter coating them, since you know you won't be looking at them for a year. Protection against mechanical shock -- jarring and the like -- is the best way to preserve delicate structures. But. Do you or your colleagues already know the proper and best methods for preparing the critters? Some might require OsO4, for example, and what's best for ciliates isn't necessarily best for dinoflagellates and so on. Meaning, it might be a good idea to process and examine some samples of each kind of wee beastie now, rather than waiting, to make sure they're actually processed right.
Phil
} Dear colleagues, } } We are seeking information from anyone who has knowledge of how long } a critical-point dried specimen prepared for SEM is sufficiently } stable for subsequent viewing. We anticipate receiving many samples } of glutaraldehyde-fixed plankton with delicate protozoa and algae in } them. Our colleagues sending the material will eventually sample } only a small subset based on data they gather one year from now. So, } we will be asked to examine the stubs approximately 12 months in the } future. } } Has anyone advice on the stability of specimens for SEM } observation that have been placed on stubs one year prior to } observation? Is it preferable to lightly sputter-coat the sample } before storage and is there any secure hope of stabilizing delicate } flagellar structures, etc. after one year? We understand the } importance of storage in a dessicator and at fairly constant } temperature, but we have never stored specimens for 1 year before } observing them. } } Many thanks, } Dee } *************************************************************** } Please do not publicly post any of my correspondence without permission } } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995)
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-microscopy.com Mon Sep 22 12:31:47 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MHVbiO001083 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 12:31:37 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8MHVbb4001079 for L1; Mon, 22 Sep 2003 12:31:37 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8MHUliO001037 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 12:31:16 -0500 Received: from smtp3.doit.wisc.edu (blitzen.doit.wisc.edu [144.92.197.183]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MHUQiO001029 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 22 Sep 2003 12:30:37 -0500 Received: from conversion-daemon.smtp3.doit.wisc.edu by smtp3.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) id {0HLM00I01LN6A5-at-smtp3.doit.wisc.edu} for Microscopy-at-sparc5.microscopy.com; Mon, 22 Sep 2003 12:30:03 -0500 (CDT) Received: from [10.25.50.161] (ansci.wisc.edu [144.92.132.175]) by smtp3.doit.wisc.edu (iPlanet Messaging Server 5.2 HotFix 1.18 (built Jul 28 2003)) with ESMTP id {0HLM00L9ILY3XM-at-smtp3.doit.wisc.edu} for Microscopy-at-sparc5.microscopy.com; Mon, 22 Sep 2003 12:30:03 -0500 (CDT)
Jean,
Manual, not automatic. The automatic CPDs I've seen are too inflexible, and I don't trust them anyway. Maybe I'm a Luddite, but I don't even like "semi-automatic" CPDs. And yes, I do use HMDS, but it's not appropriate for everything.
Phil
} Hi everyone, } } I would like some opinions about critical point dryers. What do you think } about automated units vs. manual units. Or do you use HMDS and not } use the CPD } at all? If you could post to the list and not send them directly to me, I } would appreciate it and everyone could read the responses. } } Thanks, } Jean Ross } Central Microscopy Research Facility } University of Iowa
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-microscopy.com Mon Sep 22 13:37:21 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MIauiO001644 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 13:36:56 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8MIauJp001639 for L1; Mon, 22 Sep 2003 13:36:56 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8MIa0iO001607 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 13:36:25 -0500 Received: from pony.its.uwo.ca (pony.its.uwo.ca [129.100.2.63]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MIZViO001601 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 13:35:49 -0500 Received: from spork.its.uwo.ca (hurdle.its.uwo.ca [129.100.254.76]) by pony.its.uwo.ca (8.12.10/8.12.9) with ESMTP id h8MIZ1cw013907 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 14:35:11 -0400 (EDT) Received: from uwo.ca (sparky.bg.biology.uwo.ca [129.100.68.235]) by spork.its.uwo.ca (8.12.10/8.12.9) with ESMTP id h8MIZ0Rd013426; Mon, 22 Sep 2003 14:35:00 -0400 Message-ID: {3F6F6DBB.3DC1D302-at-uwo.ca}
Hi all, I reply as an old foggie probably to set in my ways,but here goes. Over the past 30 or so years I have used several CPD's usually with acceptable results, however the machines I have had most unpredictable results with were those with automatic cycling and programmed temperture rise. The simple Polaron and others with manual valves and tap water temperature controlled by a shower mixing control have been most reliable and trouble free.
Good Luck,
Ron.
jean-ross-at-uiowa.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } } Hi everyone, } } I would like some opinions about critical point dryers. What do you think } about automated units vs. manual units. Or do you use HMDS and not use the CPD } at all? If you could post to the list and not send them directly to me, I } would appreciate it and everyone could read the responses. } } Thanks, } Jean Ross } Central Microscopy Research Facility } University of Iowa
-- Ronald J. Smith Department of Biology Room 235, Biological & Geological Sciences Bldg. U.W.O., London, Ontario N6A 5B7 (519) 661-2111 ext.86486
From MicroscopyL-request-at-microscopy.com Mon Sep 22 15:45:45 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MKjKiO002685 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 15:45:20 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8MKjKcC002680 for L1; Mon, 22 Sep 2003 15:45:20 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8MKiOiO002644 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 15:44:49 -0500 Received: from einsteinium.btinternet.com (einsteinium.btinternet.com [194.73.73.147]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MKhliO002631 for {microscopy-at-sparc5.microscopy.com} ; Mon, 22 Sep 2003 15:44:03 -0500 Received: from host81-152-214-198.range81-152.btcentralplus.com ([81.152.214.198] helo=MESH900) by einsteinium.btinternet.com with esmtp (Exim 3.22 #23) id 1A1XWc-0001zk-00; Mon, 22 Sep 2003 21:43:14 +0100 Message-ID: {000e01c3814a$27c39d00$c6d69851-at-ed.ac.uk} Reply-To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk}
Dee I have re-examined CPD virus specimens after 9 months to a year and they looked pretty much as they did at first. I think you have every chance of success, particularly if you store the specimens in a desiccator at a steady temperature as you suggest, and also preferably in the dark. I would recommend not coating them until you are going to examine them. Chris
University of Edinburgh Biological Sciences EM Facility
----- Original Message ----- } From: "Dee Breger" {micro-at-ldeo.columbia.edu} To: {microscopy-at-msa.microscopy.com} Sent: Monday, September 22, 2003 3:14 PM
For the last several years, we have been coating our glass microscope slides with aminopropyltriethoxysilane. We have found it superior to gelatin, chrome-gel, poly-D-lysine, etc for increasing adherence of both cryostat and plastic resin sections. We love this technique but after years of success, it has stopped working or at least fully working. One problem is decreased adhesion and lost sections during rigorous immunostaining protocols. Another easily seen difference is that water droplets no longer stay beaded up due to surface tension but spread over the surface more easily. We have tried two new bottles (different sources) and new acetone bottles. We tried doubling the APTS concentration to 4% but it made little difference. Does anyone have insights into this problem? I am not really looking for an alternative coating material but would rather solve the problem with this one. Our actual protocol is: Dip slides in 2% aminopropyltriethoxysilane in acetone for 1 min followed by 1 min in 100% acetone, then 1 min in dH20, allow to air dry. Thanks in advance for your comments. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
From MicroscopyL-request-at-microscopy.com Mon Sep 22 18:51:18 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MNp8iO004122 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 18:51:08 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8MNp8uT004117 for L1; Mon, 22 Sep 2003 18:51:08 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8MNoSiO004086 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 18:50:48 -0500 Received: from msg.ucsf.edu (msg.ucsf.edu [169.230.20.46]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8MNoCiO004078 for {Microscopy-at-msa.microscopy.com} ; Mon, 22 Sep 2003 18:50:22 -0500 Received: from [169.230.29.100] (account ackerman-at-msg.ucsf.edu) by msg.ucsf.edu (CommuniGate Pro WebUser 4.1.3) with HTTP id 1126784 for Microscopy-at-MSA.Microscopy.Com; Mon, 22 Sep 2003 16:50:00 -0700
I was discarding old files today when I noticed a paper from the old (undated) EMSA Bulletin by Christine Jeffery of Sothern Illinois University. She found that some osmium treated, non-coated SEM samples stored in one vendor's sample box with a soft plastic insert developed crystals after about six months. Metal coated samples did not exhibit the crystal formation. The implication was that an unknown mechanism involving osmium laden samples and a phenol compound in the box inserts resulted in the crystals. So perhaps it is wise to metal coat SEM samples if they will be retained for study after six or more months. Larry Ackerman Advanced Imaging Lab/Dr. John Sedat/Dr. David Agard Dept. of Biochemistry & Biophysics University of California San Francisco 600-16th Street, Box 2140, Room S101 San Francisco, CA 94143
415-476-4441 FAX 415-514-4142
From MicroscopyL-request-at-microscopy.com Mon Sep 22 22:05:33 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8N35NiO005356 for {L1-at-ns.microscopy.com} ; Mon, 22 Sep 2003 22:05:23 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8N35NmI005351 for L1; Mon, 22 Sep 2003 22:05:23 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8N34giO005319 for "MListServerFilteredEmail_5-at-microscopy.com"; Mon, 22 Sep 2003 22:05:02 -0500 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8N34WiP005316 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 22 Sep 2003 22:04:32 -0500 Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) Message-Id: {p05111a01bb956878d473-at-[206.69.208.21]}
We just acquired a used Leitz trinocular microscope (SM-LUX HL) and are searchhing for an instruction manual. This is purely for education.
Thanks if you can help us locate such an item.
Dad and son are beginners at this fantastic resource that is microscopy!
From MicroscopyL-request-at-microscopy.com Tue Sep 23 03:18:11 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8N8I1iO007730 for {L1-at-ns.microscopy.com} ; Tue, 23 Sep 2003 03:18:01 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8N8I0OY007725 for L1; Tue, 23 Sep 2003 03:18:00 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8N8HKiO007697 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 23 Sep 2003 03:17:40 -0500 Received: from pecka.isibrno.cz (pecka.isibrno.cz [195.178.70.10]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8N8GmiO007692 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 23 Sep 2003 03:16:59 -0500 Received: from aligator (aligator.isibrno.cz [195.178.70.170]) by pecka.isibrno.cz (8.11.6/8.11.6) with ESMTP id h8N8GPp24368 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 23 Sep 2003 10:16:26 +0200 (CEST)
Dear Microscopists,
An update of the list of meetings for microscopists in the year 2004 has been finished at the Petr's Microscopy Resources at the
Your submission of other meetings will be very appreciated (submission form is accessible from the page mentioned, the direct link is http://www.petr.isibrno.cz/microscopy/PMRform.php).
Petr Schauer +--------------------------------------------------------------------+ | Dr. Petr Schauer tel.: +420 541 514 313 | | Head of the Group of the Scintillation fax : +420 541 514 404 | | and Cathodoluminescent Systems +420 541 514 402 | | INSTITUTE OF SCIENTIFIC INSTRUMENTS | | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno petr-at-dp.fce.vutbr.cz | | Czech Republic www: http://www.petr.isibrno.cz/ | +--------------------------------------------------------------------+
From MicroscopyL-request-at-microscopy.com Tue Sep 23 07:20:44 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NCKYiO009529 for {L1-at-ns.microscopy.com} ; Tue, 23 Sep 2003 07:20:34 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8NCKYLH009524 for L1; Tue, 23 Sep 2003 07:20:34 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8NCJriO009486 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 23 Sep 2003 07:20:13 -0500 Received: from mailer.syr.edu (mailer.syr.edu [128.230.18.29]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NCJbiO009482 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 23 Sep 2003 07:19:48 -0500 Received: from [128.230.24.90] (www.geochemistry.syr.edu) by mailer.syr.edu (LSMTP for Windows NT v1.1b) with SMTP id {0.08CBCF77-at-mailer.syr.edu} ; Tue, 23 Sep 2003 8:17:38 -0400 Mime-Version: 1.0 Message-Id: {a05100315bb95ea138c85-at-[128.230.24.90]}
Hi, Is Gatan the only commercial vendor offering CL on SEM's? Have any of you heard of a company called KNE?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
From MicroscopyL-request-at-microscopy.com Tue Sep 23 07:43:41 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NChFiO009861 for {L1-at-ns.microscopy.com} ; Tue, 23 Sep 2003 07:43:15 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8NChFvV009857 for L1; Tue, 23 Sep 2003 07:43:15 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8NCgJiO009800 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 23 Sep 2003 07:42:39 -0500 Received: from sphinx.gsu.edu (sphinx.gsu.edu [131.96.2.23]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NCfViO009786 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 23 Sep 2003 07:41:41 -0500 Received: from DESKTOP (DESKTOP.gsu.edu [131.96.143.120]) by sphinx.gsu.edu (8.11.7+Sun/8.10.2) with SMTP id h8NCbaf23170; Tue, 23 Sep 2003 08:37:36 -0400 (EDT) Message-ID: {003901c381cf$b46f43f0$788f6083-at-DESKTOP} Reply-To: "Robert Simmons" {rsimmons-at-gsu.edu}
Dear Microscopy Listserver members,
I wonder if it is possible to use 6 nm-gold-tagged antibodies for pre-embedding immunolabeling, or if it is necessary to use smaller ones? I want to do immunolabeling of betacells from the islets of Langerhans (rat).
Sincerely, Pernilla Nevsten
........................................................................... .............................................................. Pernilla Nevsten, PhD-student Materials Chemistry Lund University, Sweden
I usually use something along the line of the Nanoprobes 1.4nm nanogold for pre-embedding labeling, the cells are permeabilized after primary fixation. You can enhance it after the labeling step (before embedding) to make it easier to locate in the microscope.
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-microscopy.com Tue Sep 23 09:51:35 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NEpEiO011264 for {L1-at-ns.microscopy.com} ; Tue, 23 Sep 2003 09:51:14 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8NEpE9E011259 for L1; Tue, 23 Sep 2003 09:51:14 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8NEoOiO011226 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 23 Sep 2003 09:50:44 -0500 Received: from dior.amnh.org (dior.amnh.org [209.2.164.10]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NEnriO011219 for {Microscopy-at-MSA.Microscopy.com} ; Tue, 23 Sep 2003 09:50:03 -0500 Received: from dell333.amnh.org (amnhhost251.amnh.org [209.2.165.251]) by dior.amnh.org (Postfix) with ESMTP id 3EF39B354B; Tue, 23 Sep 2003 10:47:27 -0400 (EDT) Message-Id: {5.1.0.14.0.20030923101705.03ec2720-at-mail.amnh.org} X-Sender: kfrisch-at-mail.amnh.org X-Mailer: QUALCOMM Windows Eudora Version 5.1
I believe Gatan may be the only commercial vendor of a well developed CL product with a presence in the U.S., however: There was an article/short paper in the May 2003, issue 60 (The Americas edition) Microscopy and Analysis that used a CL system from Japan (on a JEOL 6500F thermal FE-SEM) (the study was conducted at Kyoto Inst. of Tech., Japan) The CL device was a MP-32FE, Atago Bussan, Horiba Group, Tokyo The capabilities and features available as described in the paper seem similar to Gatan offerings, but I never investigated further than reading the paper. (We already own a Gatan MonoCL3 system)
Probing Nanoscopic Stresses in Glass using Luminescent Atoms Giuseppe Pezzotti, Ceramic Physics Laboratory & Research Inst. for Nanoscience, Kyoto Inst. of Tech., Japan
If you'd like the author's contact info, a copy of the paper, or have questions about our Gatan system, feel free to contact me.
------------------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 08:17 AM 9/23/03 -0400, Michael Cheatham {mmcheath-at-mailbox.syr.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-microscopy.com Tue Sep 23 10:35:43 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NFZMiO011606 for {L1-at-ns.microscopy.com} ; Tue, 23 Sep 2003 10:35:22 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8NFZMqd011602 for L1; Tue, 23 Sep 2003 10:35:22 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8NFYViO011571 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 23 Sep 2003 10:34:51 -0500 Received: from c000.snv.cp.net (h001.c000.snv.cp.net [209.228.32.65]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8NFY5iO011565 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 23 Sep 2003 10:34:16 -0500 Received: (cpmta 25959 invoked from network); 23 Sep 2003 08:33:42 -0700 Received: from 134.153.37.8 (HELO rarewolf) by smtp.shaffer.net (209.228.32.65) with SMTP; 23 Sep 2003 08:33:42 -0700 X-Sent: 23 Sep 2003 15:33:42 GMT
This reminds me I was looking for the next Microscopical Society of Canada meeting the other day. If anyone reading this is associated with that meeting, then please make note of the MSC wwwpage is way out of date. http://msc.rsvs.ulaval.ca/english/welcome.html ... and I didn't get a response from the executive secretary because the e-mail address is no good.
What I have been able to find, is that it is 'apparently' being held at Acadia University, Nova Scotia ... but consider it heresay.
} Dear Microscopists, } } An update of the list of meetings for microscopists in the year 2004 } has been finished at the Petr's Microscopy Resources at the } } http://www.petr.isibrno.cz/microscopy/meetings.php#2004 } } Your submission of other meetings will be very appreciated (submission } form is accessible from the page mentioned, the direct link is } http://www.petr.isibrno.cz/microscopy/PMRform.php). } } Petr Schauer } +--------------------------------------------------------------------+ } | Dr. Petr Schauer tel.: +420 541 514 313 | } | Head of the Group of the Scintillation fax : +420 541 514 404 | } | and Cathodoluminescent Systems +420 541 514 402 | } | INSTITUTE OF SCIENTIFIC INSTRUMENTS | } | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC e-mail: petr-at-isibrno.cz | } | Kralovopolska 147, CZ-612 64 Brno petr-at-dp.fce.vutbr.cz | } | Czech Republic www: http://www.petr.isibrno.cz/ | } +--------------------------------------------------------------------+ } } } }
From MicroscopyL-request-at-microscopy.com Tue Sep 23 11:33:35 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NGV4iO012021 for {L1-at-ns.microscopy.com} ; Tue, 23 Sep 2003 11:31:04 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8NGV4SN012017 for L1; Tue, 23 Sep 2003 11:31:04 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8NGS4iO011966 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 23 Sep 2003 11:28:25 -0500 Received: from smtp.mcn.org (mail2.mcn.org [204.189.12.25]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NGPOiO011950 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 23 Sep 2003 11:25:35 -0500 Received: from [63.193.12.97] (ha-1e-men-p2-m13.mcn.org [63.193.12.97]) (authenticated bits=0) by smtp.mcn.org (8.12.10/8.12.10) with ESMTP id h8NGOh0G020180; Tue, 23 Sep 2003 09:24:45 -0700 (PDT) Mime-Version: 1.0 Message-Id: {a05210600bb9622b91eb5-at-[63.193.12.97]}
I've just realized that the current frames format of the MICRO web page requires that you access all of its segments (the bibliography, for example) via the URL below. If you have a different bookmark stored, please change. -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-microscopy.com Tue Sep 23 13:21:26 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NIL4iO012847 for {L1-at-ns.microscopy.com} ; Tue, 23 Sep 2003 13:21:04 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8NIL4mu012843 for L1; Tue, 23 Sep 2003 13:21:04 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8NIKBiO012813 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 23 Sep 2003 13:20:32 -0500 Received: from vger.vgernet.net (vger.vgernet.net [63.246.192.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NIJaiO012799 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 23 Sep 2003 13:19:47 -0500 Received: from [63.246.194.15] (boston15.vgernet.net [63.246.194.15]) by vger.vgernet.net (8.12.4/8.12.6) with ESMTP id h8NIEn4P017563; Tue, 23 Sep 2003 14:14:50 -0400 User-Agent: Microsoft Outlook Express Macintosh Edition - 5.01 (1630)
Hi fellow microscopists
I think that Mike is referring to KE Developments in the UK. Their website with information regarding their detectors (including CL) is at http://www.kedev.co.uk/detectors.htm.
best regards, Steven Slap
on 9/23/03 8:17 AM, Michael Cheatham at mmcheath-at-mailbox.syr.edu wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } } Hi, } Is Gatan the only commercial vendor offering CL on SEM's? } Have any of you heard of a company called KNE? } } TIA } Mike
From MicroscopyL-request-at-microscopy.com Tue Sep 23 15:43:04 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NKgZiO013953 for {L1-at-ns.microscopy.com} ; Tue, 23 Sep 2003 15:42:35 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8NKgZ7A013948 for L1; Tue, 23 Sep 2003 15:42:35 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8NKfViO013920 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 23 Sep 2003 15:42:01 -0500 Received: from in.belttechnologies.com ([66.152.255.226]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8NKesiO013913 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 23 Sep 2003 15:41:14 -0500 Received: (qmail-ldap/ctrl 27436 invoked by uid 1103); 23 Sep 2003 20:40:37 -0000 Received: from MDufraine-at-ebsciences.com by postman by uid 64011 with qmail-scanner-1.20rc3 (clamuko: 0.60. sophie: 2.14/3.73. spamassassin: 2.55. Clear:RC:1:. Processed in 0.192951 secs); 23 Sep 2003 20:40:37 -0000 Received: from unknown (HELO mdufraine) ([10.0.0.139]) (envelope-sender {MDufraine-at-ebsciences.com} ) by postman.in.belttechnologies.com (qmail-ldap-1.03) with SMTP for {mmcheath-at-mailbox.syr.edu} ; 23 Sep 2003 20:40:35 -0000
Mike-
Your reference to KNE, may be KE Developments in the UK. Energy Beam Sciences has recently become the US distributor for KE Developments. You may check the KE Developments web-site www.kedev.com, contact me at 800-992-9037 X340, or by e-mail. for assistance on CL detectors
Regards,
Mike Dufraine Energy Beam Sciences, Inc.
-----Original Message----- } From: Michael Cheatham [mailto:mmcheath-at-mailbox.syr.edu] Sent: Tuesday, September 23, 2003 8:18 AM To: Microscopy-at-sparc5.microscopy.com
Hi, Is Gatan the only commercial vendor offering CL on SEM's? Have any of you heard of a company called KNE?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
From MicroscopyL-request-at-microscopy.com Tue Sep 23 19:45:03 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8O0j2iO015927 for {L1-at-ns.microscopy.com} ; Tue, 23 Sep 2003 19:45:02 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8O0j241015922 for L1; Tue, 23 Sep 2003 19:45:02 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8O0iMiO015883 for "MListServerFilteredEmail_5-at-microscopy.com"; Tue, 23 Sep 2003 19:44:42 -0500 Received: from [206.69.208.21] (anlvpn226.vpn.anl.gov [130.202.232.226]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8O0hwiP015876 for {microscopy-at-ns.microscopy.com} ; Tue, 23 Sep 2003 19:44:13 -0500 Mime-Version: 1.0 Message-Id: {p05111a06bb96964fda4b-at-[206.69.208.21]}
Colleagues
I believe AOL & Compuserve & Netscape.Net Email users are now restored to active participation and can now once again receive Microscopy Listserver Email. Sorry for the extended down time to those subscribers.
AOL in their infinite wisdom decided to block all Microscopy Listserver saying that it was a potential source of SPAM mail. A point that I would undoubtably argue to the contrary.
It has taken me several weeks to get them to reverse this decision and I had to jump through a lot of proverbial hoops. But as of this today mail appears to be flowing to blocked addresses at AOL.
I might point out that alot of ISP's, Companies and Universities have changed their SPAM software recently, and unilaterally block content or domains. For example I received rejected Email from several organizations from the (valid) Email posting asking about WebCams for microscopy. It was unilaterally decided that any Email message with the word Webcam was obviously SPAM mail. This was clearly not ture for this Email, but nevertheless the SPAM filters at various sites rejected it. This will be a continuing problem, for which I see little solution. So remember to choose your subject words carefully...
In any event, please don't be bashful of reporting outageous of MListserver Email which persist for several days. Your site may institute a domain or subject filter which blocks MListserver Email and I will not necessairly know about it.
For example,, AOL never actually notified me that they were blocking traffic. I only discovered it when testing a new filter and found that my AOL account was not receiving the test messages. That event (over 2 weeks ago) started me on search for the problem, which I erroneously thought I created, but happened to coincide with their system change.
BTW, I note with irony, that their blocking of the MListserver did not actually decrease the amount of SPAM mail I get on my (seldom used) AOL account, it only stopped me from using AOL, and also from my 80 year old Mother from receiving Email addressed from me via my normal microscopy.com account!
Sigh, now if I were running AOL things would be different.... ( Nestor grins )
;-)
Nestor Your Friendly Neighborhood SysOp.
From MicroscopyL-request-at-microscopy.com Wed Sep 24 04:01:08 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8O90wiO018932 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 04:00:58 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8O90wvT018927 for L1; Wed, 24 Sep 2003 04:00:58 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8O90HiO018899 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 04:00:37 -0500 Received: from pecka.isibrno.cz (pecka.isibrno.cz [195.178.70.10]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8O8xkiO018873 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 24 Sep 2003 03:59:56 -0500 Received: from aligator (aligator.isibrno.cz [195.178.70.170]) by pecka.isibrno.cz (8.11.6/8.11.6) with ESMTP id h8O8xLp03076 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 24 Sep 2003 10:59:23 +0200 (CEST)
The IFBLS'26 has been added to the list at the http://www.petr.isibrno.cz/microscopy/meetings.php#2004 . Other suggestions?
Petr Schauer
On 24 Sep 2003 at 7:51, Gareth Morgan wrote:
} That reminds me................ } International Federation of Biomedical Laboratory Science, IFBLS (formerly } IAMLT) will hold its 26th Congress in Stockholm, Sweden between the 13th } and 18th June, 2004. } } Take a look at: } http://www.vardforbundet.se/ifbls2004 } } } Med vänliga hälsningar/With best regards } } Gareth } } ***Come to Stockholm in 2004 to the 26th IFBLS congress*** Take a look at } http://www.vardforbundet.se/ifbls2004 } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-labmed.ki.se } } Tel +46 8 5858 1038 } Fax +46 8 5858 7730 } } Gareth Morgan MPhil MSc FIBMS, } Department of Laboratory Medicine (Labmed), } Karolinska Institutet, } Huddinge Universitetssjukhus, F46 } SE 141 86 Stockholm } Sweden } } OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet } för klinisk patologi och cytologi. } } NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. } Clinical Histo- and Cytopathology Laboratory. } }
From MicroscopyL-request-at-microscopy.com Wed Sep 24 04:47:29 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8O9lEiO019647 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 04:47:14 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8O9lEJt019643 for L1; Wed, 24 Sep 2003 04:47:14 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8O9kSiO019614 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 04:46:53 -0500 Received: from bonnie.med.univ-tours.fr (bonnie.med.univ-tours.fr [193.52.213.35]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8O9k6iO019608 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 24 Sep 2003 04:46:22 -0500 Received: from med.univ-tours.fr (bonnie [127.0.0.1]) by bonnie.med.univ-tours.fr (8.11.6/8.11.6) with ESMTP id h8O9jpa23721 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 24 Sep 2003 11:45:51 +0200 Message-ID: {3F716B12.98A6B0CF-at-med.univ-tours.fr}
Dear Members, I would like to try to observe cristalized KDP (KH2PO4) with a TEM and for this, I would like to obtain ultrathin sections of this cristalized KDP. However, I have no experience in cutting cristals with an ultramicrotome. Does anyone has ever try that? If so, what would be the best method? Thank you very much for any advice on this.
Pierre-Yves Sizaret
P-Y Sizaret Microscopy Facilities University of Tours, France
From MicroscopyL-request-at-microscopy.com Wed Sep 24 06:58:28 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OBw2iO020559 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 06:58:02 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OBw2M7020555 for L1; Wed, 24 Sep 2003 06:58:02 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OBv7iO020525 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 06:57:32 -0500 Received: from outbound.lightshipmail.net (outbound.lightshipmail.net [216.204.0.39]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OBuViO020519 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 24 Sep 2003 06:56:46 -0500 Received: from Barb500.mme1.com (ipn36372-b70930.net-resource.net [216.204.89.146]) by outbound.lightshipmail.net (Postfix) with ESMTP id 2CE805725F; Wed, 24 Sep 2003 07:59:08 -0400 (EDT) Message-Id: {5.1.0.14.0.20030924075753.009f4890-at-postoffice.lightshipmail.net} X-Sender: pop3750-at-postoffice.lightshipmail.net X-Mailer: QUALCOMM Windows Eudora Version 5.1
Petr,
I noticed that you have PITTCON listed now... It might also be worth noting that we run the American Chem Soc short course, "Applied Optical Microscopy" in conjunction with that meeting. It will be held March 5-7 in 2004.
Thanks, Barbara Foster
At 10:59 AM 9/24/03 +0200, Petr Schauer wrote:
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From MicroscopyL-request-at-microscopy.com Wed Sep 24 07:56:20 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OCu9iO020997 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 07:56:09 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OCu99h020992 for L1; Wed, 24 Sep 2003 07:56:09 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OCtSiO020960 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 07:55:49 -0500 Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OCtBiO020950 for {Microscopy-at-MSA.Microscopy.com} ; Wed, 24 Sep 2003 07:55:22 -0500 Received: from [209.120.201.218] (209-120-201-218.hosts.axs2000.net [209.120.201.218] (may be forged)) by diskless5.axs2000.net (8.11.2/8.11.2) with SMTP id h8OCsJ912768 for {Microscopy-at-MSA.Microscopy.com} ; Wed, 24 Sep 2003 08:54:19 -0400 Message-Id: {200309241254.h8OCsJ912768-at-diskless5.axs2000.net} To: MICROSCOPY BB {Microscopy-at-MSA.Microscopy.com}
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
P-Y Sizaret wrote: ====================================================== Dear Members, I would like to try to observe cristalized KDP (KH2PO4) with a TEM and for this, I would like to obtain ultrathin sections of this cristalized KDP. However, I have no experience in cutting cristals with an ultramicrotome. Does anyone has ever try that? If so, what would be the best method? Thank you very much for any advice on this. ====================================================== We have not tried cutting this particular crystal but we have thin sectioned , over the years, crystals that would seem to be not too different in properties.
You can not actually "section" with ultramicrotomy the entire crystal. Or at least we have never succeeded in doing that. But you can powder up some small amount of the crystal, we always would use a good mortar and pestle made from agate (not porcelain), see URL http://www.2spi.com/catalog/tools/mortar_pestles_agate.shtml and then embed the powder in one of the "Epon substitutes" such as our SPI- Pon 812 resin, see URL http://www.2spi.com/catalog/chem/spi-pon-812-epoxy-embedding-kit.shtml If you use a porcelain mortar and pestle, you will end up with porcelain powder in your ground sample, created a much more difficult time at interpreting your results.
If you experience "particle pull-out" then you have to either a) grind down to a smaller powder or b) use an adhesion promoter such as SPI-Chem 3GTMO, as shown on URL http://www.2spi.com/catalog/chem/trimethoxysilane.shtml
Since cutting crystals like this will be "hard"on your diamond knife, we would suggest as a money-saving measure, the use of a materials science diamond knife, but not one with a larger angle because compression effects would then make it much more difficult if not impossible to obtain usable sections of the particles (crystals). This is explained more fully on URL http://www.2spi.com/catalog/knives/materials.shtml This is a far more economical approach than to be using so-called "life science" diamond knives for this kind of work.
If you have other questions, let me know and I will try my best to answer them for you.
Disclaimer: SPI Supplies offers all three of the products mentioned in this posting so we would have a vested interest in having more people using these products.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-microscopy.com Wed Sep 24 08:50:52 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8ODoaiO021447 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 08:50:36 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8ODoaLG021443 for L1; Wed, 24 Sep 2003 08:50:36 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8ODnoiO021411 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 08:50:15 -0500 Received: from hickory.cmpcntr.tc.trincoll.edu (hickory.cc.trincoll.edu [157.252.15.139]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8ODnTiO021404 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 24 Sep 2003 08:49:45 -0500 X-MimeOLE: Produced By Microsoft Exchange V6.0.6375.0 content-class: urn:content-classes:message MIME-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Dear Listers,
I am working with a colleague to examine isolated Chodium chloroplasts by TEM. The chloroplasts were isolated according to a published protocol (I can get those details if pertinent), and their final solution was 0.9 M sorbitol/0.05 M Hepes buffer, pH 7.3. Some but not all were pelleted into 2% agarose in the same solution prior to fixation in Karnovsky's. Following washes in 0.1M cacodylate + 7% sucrose, they were post-fixed in buffered 1% OsO4, then were washed, enbloc stained in UA, and routinely dehydrated and embedded (Embed 812). Thin sections were stained with UA and Pb, then examined in the TEM.
The same protocols yield excellent results for mammalian tissues. (That may be the key here?)
Some questions:
1 - Is it usual for the chloroplasts to remain a bright green throughout processing? There was no obvious sign of Os reduction (blackening of the tissues).
2 - Missing membranes? The outer limiting membranes of the chloroplasts are missing. Thykaloid membranes are fairly well-preserved (a few ripples here and there). Either the outer membranes are utterly unstained and thus invisible? or, more likely, stripped away during isolation although the stroma maintains a high degree of integrity and does not look extracted. The stroma looks reasonably dense, starch granules are dark and prominent.
Any explanation for the persistent green coloration? Any thoughts on our missing outer membranes?
Thanks all. Ann
Ann Hein Lehman Assistant Director, Electron Microscopy Facility Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman
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Thanks to all who gave feedback on our SEM sample stability query. We can now initiate the project with confidence! -Dee
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
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Nestor, it never fails to astound me how ridiculous and anti-productive our modern life is getting. You're doing too fine and valuable a job to get mired in this kind of bureaucratic, er, um, stuff. My continued thanks for all your good work on our behalf. Dee
----------------------------------------------------------------------- } } Colleagues } } I believe AOL & Compuserve & Netscape.Net Email users are now restored } to active participation and can now once again receive Microscopy } Listserver Email. Sorry for the extended down time to those subscribers. } } AOL in their infinite wisdom decided to block all Microscopy Listserver } saying that it was a potential source of SPAM mail. A point that } I would undoubtably argue to the contrary. } } It has taken me several weeks to get them to reverse this decision and } I had to jump through a lot of proverbial hoops. But as of this today mail } appears to be flowing to blocked addresses at AOL. } } I might point out that alot of ISP's, Companies and Universities have changed } their SPAM software recently, and unilaterally block content or domains. } For example } I received rejected Email from several organizations from the (valid) Email } posting asking about WebCams for microscopy. It was unilaterally decided } that any Email message } with the word Webcam was obviously SPAM mail. This was clearly not ture for } this Email, but nevertheless the SPAM filters at various sites rejected } it. This will } be a continuing problem, for which I see little solution. So remember to } choose } your subject words carefully... } } In any event, please don't be bashful of reporting outageous of } MListserver Email which } persist for several days. Your site may institute a domain or subject filter } which blocks MListserver Email and I will not necessairly know about it. } } For example,, AOL never actually notified me that they were blocking traffic. } I only discovered it when testing a new filter and found that my AOL account } was not receiving the test messages. That event (over 2 weeks ago) } started me on search for the problem, which I erroneously thought I created, } but happened to coincide with their system change. } } BTW, I note with irony, that their blocking of the MListserver did not } actually } decrease the amount of SPAM mail I get on my (seldom used) AOL } account, it only stopped me from using AOL, and also from my 80 year old } Mother from receiving Email addressed from me via my normal microscopy.com } account! } } Sigh, now if I were running AOL things would be different.... ( Nestor } grins ) } } ;-) } } Nestor } Your Friendly Neighborhood SysOp.
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
From MicroscopyL-request-at-microscopy.com Wed Sep 24 10:02:11 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OF20iO022122 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 10:02:00 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OF201R022118 for L1; Wed, 24 Sep 2003 10:02:00 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OF1HiO022083 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 10:01:39 -0500 Received: from c000.snv.cp.net (h008.c000.snv.cp.net [209.228.32.72]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OF0viO022075 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 24 Sep 2003 10:01:11 -0500 Received: (cpmta 19970 invoked from network); 24 Sep 2003 08:00:30 -0700 Received: from 134.153.37.8 (HELO rarewolf) by smtp.shaffer.net (209.228.32.72) with SMTP; 24 Sep 2003 08:00:30 -0700 X-Sent: 24 Sep 2003 15:00:30 GMT
I was hoping someone may have some experience with a building's primary power transformer (1500kW), and installing an SEM.
Presently we're concerned with the anticipated location being directly above this transformer, but the facility design is also in the design phase, and can be moved. One question would be how far(?), and another would ask if there aren't any extenuating circumstances(?)
While the entire building is being rennovated, I cannot imagine any ability for anticipating field strengths or measuring them.
tia & cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com
From MicroscopyL-request-at-microscopy.com Wed Sep 24 11:46:34 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OGkMiO022932 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 11:46:22 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OGkMJu022927 for L1; Wed, 24 Sep 2003 11:46:22 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OGjfiO022890 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 11:46:02 -0500 Received: from usstp09.itcs.purdue.edu (usstp09.itcs.purdue.edu [128.210.5.248]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OGjTiO022886 for {microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 11:45:41 -0500 Received: from [128.210.121.247] (lsmf07.btny.purdue.edu [128.210.121.247]) by usstp09.itcs.purdue.edu (8.12.10/8.12.10/scan-smtp) with ESMTP id h8OGj4DU011536 for {microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 11:45:04 -0500 User-Agent: Microsoft-Entourage/9.0.2.4011
Hi there, We would like to locate intermediate filaments in mouse pancreas using immunocytochemistry. The samples have been fixed using 4% PAF and 0.5% glutaraldehyde and embedded in LR White. They have been successfully used for other ICC localization. Could anyone recommend an antibody to use against proteins specific to mammalian intermediate filaments. We would prefer to find a polyclonal antibody if possible.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
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Hello, microscopists,
I am looking for a couple of sturdy tables on which to place microscopy systems. I know that excellent vibration isolation is acieved by products like TMC tables, however, can anyone recommend good (one level lower than excellent) isolation with other products?
Any personal experience or manufacturer name would be highly appreciated.
Thank you. Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Queen 30 Bond St. Toronto, ON M5B 1W8 Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
From MicroscopyL-request-at-microscopy.com Wed Sep 24 12:51:31 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OHpJiO023520 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 12:51:19 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OHpJti023515 for L1; Wed, 24 Sep 2003 12:51:19 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OHociO023484 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 12:50:59 -0500 Received: from mnhgwia.si.edu (MNHGWIA.si.edu [160.111.81.23]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OHoMiO023481 for {microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 12:50:33 -0500 Received: from SIWP11-MTA by mnhgwia.si.edu with Novell_GroupWise; Wed, 24 Sep 2003 13:49:58 -0400 Message-Id: {sf71a106.026-at-mnhgwia.si.edu} X-Mailer: Novell GroupWise Internet Agent 6.0.2
Anyone care to comment on imaging samples in the SEM (LaB6) while they are offgassing naphthalene. The entomologists are still using the stuff and I have been kicking them out or transferring them to the ESEM, but is this really a problem for the instrument?? Since many of these samples are dried and completely unfixed they are offgassing all kinds of crap anyway, so should I even care? The policy was enforced as a result of the mess it makes of the sputter coater so I just banned it from the SEM's too. Maybe I don't need to though?? Thanks
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
From MicroscopyL-request-at-microscopy.com Wed Sep 24 14:07:29 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OJ79iO024106 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 14:07:09 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OJ79OU024101 for L1; Wed, 24 Sep 2003 14:07:09 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OJ6GiO024066 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 14:06:40 -0500 Received: from mail.interchange.ubc.ca (mail.interchange.ubc.ca [137.82.27.15]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OJ5eiO024053 for {microscopy-at-MSA.microscopy.com} ; Wed, 24 Sep 2003 14:05:52 -0500 Received: from desk.staff.mmat.ubc.ca ([137.82.16.178] helo=desk) by mail.interchange.ubc.ca with smtp (Exim 4.20) id 1A2Ewt-0001EY-VR; Wed, 24 Sep 2003 12:05:15 -0700 Message-ID: {039101c382ce$76451aa0$b2105289-at-staff.mmat.ubc.ca}
Dear Michael, Last year we had a proposal to renovate the basement of the research building for a high-resolution EM and SIMS lab, so I used my Gauss Mauss to check the fields. This is a hand-held field checker that I bought from Marivac on sale a few years ago (I think it was about $300CAD). It has proved its worth many times over. You can get total field and AC component, check in horizontal and vertical directions and it is still working on its first set of batteries. When we had the room surveyed by one of the EM vendors, the results were very close. We were concerned because the building power supply cable vault was just under the floor, but because it was a high voltage feed, the field was negligible three to four feet away. The room is planned around that. You can also pay an EM vendor service department to do a survey of both mag fields and vibration, so there is no obligation. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, September 24, 2003 8:00 AM
Jean,
I have a vintage 1991 Ted Pella CPD. It is as manual as they come and after 12 years it still performs reliably.
Regards, Evelyn
Scripps Institution of Oceanography University of California, San Diego Analytical Facility (858)534-2438
-----Original Message----- } From: jean-ross-at-uiowa.edu [mailto:jean-ross-at-uiowa.edu] Sent: Monday, September 22, 2003 7:22 AM To: Microscopy-at-MSA.Microscopy.Com
Hi everyone,
I would like some opinions about critical point dryers. What do you think about automated units vs. manual units. Or do you use HMDS and not use the CPD at all? If you could post to the list and not send them directly to me, I would appreciate it and everyone could read the responses.
Thanks, Jean Ross Central Microscopy Research Facility University of Iowa
From MicroscopyL-request-at-microscopy.com Wed Sep 24 16:15:10 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OLEliO025186 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 16:14:47 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OLElLF025181 for L1; Wed, 24 Sep 2003 16:14:47 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OLDqiO025148 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 16:14:14 -0500 Received: from mail.khri.med.umich.edu (mail2.khri.med.umich.edu [141.214.13.211]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OLDHiO025142 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 24 Sep 2003 16:13:30 -0500 Received: from Spooler by mail.khri.med.umich.edu (Mercury/32 v3.31) ID MO0008BC; 24 Sep 03 17:13:16 -0400 Received: from spooler by mail.khri.med.umich.edu (Mercury/32 v3.31); 24 Sep 03 17:12:40 -0400
I attempted pre-embed immunogold labeling of neurofilament in rodent cochlea using 10 nm gold and did not have much success. While 6 nm is smaller than what I used, I would probably suggest going smaller to help avoid the problems I encountered. I have since gone to post-embed immunolabeling with the 10 nm gold and have had success.
Good luck!
On 23 Sep 2003 at 8:39, Robert Simmons wrote:
} } ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } - } } Dear Microscopy Listserver members, } } I wonder if it is possible to use 6 nm-gold-tagged antibodies for } pre-embedding immunolabeling, or if it is necessary to use smaller } ones? I want to do immunolabeling of betacells from the islets of } Langerhans (rat). } } Sincerely, } Pernilla Nevsten } } } ...................................................................... } ..... .............................................................. } Pernilla Nevsten, PhD-student Materials Chemistry Lund University, } Sweden } } } } } I usually use something along the line of the Nanoprobes 1.4nm } nanogold for pre-embedding labeling, the cells are permeabilized after } primary fixation. You can enhance it after the labeling step (before } embedding) to make it easier to locate in the microscope. } } } Dr Robert Simmons } Program Director } Biological Imaging Core Laboratory } Georgia State University } Atlanta, GA 30302-4010 } } 404-651-3138 } 404-651-2509 FAX } } } }
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
From MicroscopyL-request-at-microscopy.com Wed Sep 24 16:35:55 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OLZWiO025364 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 16:35:32 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OLZWHU025359 for L1; Wed, 24 Sep 2003 16:35:32 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OLYciO025320 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 16:35:00 -0500 Received: from mail.khri.med.umich.edu (mail2.khri.med.umich.edu [141.214.13.211]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OLY6iO025311 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 24 Sep 2003 16:34:20 -0500 Received: from Spooler by mail.khri.med.umich.edu (Mercury/32 v3.31) ID MO0008CF; 24 Sep 03 17:34:06 -0400 Received: from spooler by mail.khri.med.umich.edu (Mercury/32 v3.31); 24 Sep 03 17:33:28 -0400
I've been taking on a post-embed immunogold staining project and I'm having a low section adhesion average. My staining periods and rinses are quite long so there is ample opportunity for the sections to float off. My latest attempt had 1 grid out of 6 that retained its sections, and that's a poor average even if I'm a hitter for the Detroit Tigers!
A colleague emailed me the grid adhesive described by Bozzola, 2nd Ed but wonder if there are any drawbacks I need to consider, both with the beam and any known reactivity with the gold reagent?
Any suggestions to increase my success are welcomed!
Thank you
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
From MicroscopyL-request-at-microscopy.com Wed Sep 24 17:09:49 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OM9ciO025634 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 17:09:38 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OM9cwe025629 for L1; Wed, 24 Sep 2003 17:09:38 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OM8wiO025600 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 17:09:18 -0500 Received: from earth-ox.its.caltech.edu (SteeleMR-loadb-NAT-49.caltech.edu [131.215.49.69]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OM8biO025595 for {microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 17:08:47 -0500 Received: from fire-dog (fire-dog [192.168.1.4]) by earth-ox-postvirus (Postfix) with ESMTP id 1B4A7109A63 for {microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 15:08:13 -0700 (PDT) Received: from water-ox ([192.168.1.10]) by fire-dog (MailMonitor for SMTP v1.2.2 ) ; Wed, 24 Sep 2003 15:08:12 -0700 (PDT) Received: from caltech.edu (pix-1.caltech.edu [131.215.2.21]) by water-ox.its.caltech.edu (Postfix) with ESMTP id 14C8926AC2E for {microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 15:08:12 -0700 (PDT)
On Wednesday, September 24, 2003, at 10:49 AM, Scott Whittaker wrote:
} Anyone care to comment on imaging samples in the SEM (LaB6) while they } are } offgassing naphthalene. The entomologists are still using the stuff } and I } have been kicking them out or transferring them to the ESEM, but is } this } really a problem for the instrument?? Since many of these samples are } dried } and completely unfixed they are offgassing all kinds of crap anyway, so } should I even care? The policy was enforced as a result of the mess it } makes } of the sputter coater so I just banned it from the SEM's too. Maybe I } don't } need to though?? Thanks } Dear Scott, When I was doing electron diffraction on naphthalene in a TEM, I put the grid in a cryo-holder and started putting in LN2 while the airlock was pumping down. That was sufficient to prevent the naphthalene from subliming. I don't know how cold you would need to get your specimen--i.e., I can't say whether a Peltier cooler would do it--but at ~ -185 C there is no observable effect on the vacuum in a TEM from the nanoliter quantities I had on the grid, and there was no noticeable loss of the specimen from the grid, although I'm sure that a higher electron dose would etch naphthalene away. YMMV. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-microscopy.com Wed Sep 24 17:16:47 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OMGaiO025718 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 17:16:36 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8OMGaNm025713 for L1; Wed, 24 Sep 2003 17:16:36 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8OMFuiO025684 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 17:16:16 -0500 Received: from water-ox.its.caltech.edu (SteeleMR-loadb-NAT-49.caltech.edu [131.215.49.69]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8OMFWiO025681 for {microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 17:15:45 -0500 Received: from fire-dog (fire-dog [192.168.1.4]) by water-ox-postvirus (Postfix) with ESMTP id 3EABD26AC8D for {microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 15:15:08 -0700 (PDT) Received: from water-ox ([192.168.1.10]) by fire-dog (MailMonitor for SMTP v1.2.2 ) ; Wed, 24 Sep 2003 15:15:07 -0700 (PDT) Received: from caltech.edu (pix-1.caltech.edu [131.215.2.21]) by water-ox.its.caltech.edu (Postfix) with ESMTP id 2D17526AC92 for {microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 15:15:06 -0700 (PDT)
On Wednesday, September 24, 2003, at 02:59 AM, Pierre-Yves Sizaret wrote:
} I would like to try to observe cristalized KDP (KH2PO4) with a TEM and } for this, I would like to obtain ultrathin sections of this cristalized } KDP. However, I have no experience in cutting cristals with an } ultramicrotome. Does anyone has ever try that? If so, what would be the } best method? } Thank you very much for any advice on this. } Dear Pierre-Yves, Do you need to observe the particular crystals you have, or would any KDP crystals do? If the latter, you could dry down a small volume of dilute solution--elevating the temperature will give smaller crystals. If the former, try to match the hardness of the resin to that of the KDP. Another concern is that if the substance is actually KH2PO4.xH2O, the structure can change in the TEM vacuum. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-microscopy.com Wed Sep 24 19:35:11 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8P0YuiO026713 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 19:34:56 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8P0YuUr026708 for L1; Wed, 24 Sep 2003 19:34:56 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8P0YAiO026681 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 19:34:36 -0500 Received: from dahlia.noc.ucla.edu (dahlia.noc.ucla.edu [169.232.46.11]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8P0XaiO026671 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 24 Sep 2003 19:33:51 -0500 Received: from tigerlily.noc.ucla.edu (tigerlily.noc.ucla.edu [169.232.46.12]) by dahlia.noc.ucla.edu (8.12.8p2/8.12.8) with ESMTP id h8P0XBbL013186 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 24 Sep 2003 17:33:11 -0700 Received: from emi.ucla.edu ([149.142.241.180]) (authenticated bits=0) by tigerlily.noc.ucla.edu (8.12.9p1/8.12.9) with ESMTP id h8P0XBNU032626 (version=TLSv1/SSLv3 cipher=DES-CBC3-SHA bits=168 verify=NOT) for {Microscopy-at-sparc5.microscopy.com} ; Wed, 24 Sep 2003 17:33:11 -0700 Message-Id: {5.2.0.9.2.20030924143106.01af6948-at-mail.ucla.edu} X-Sender: sryazant-at-pop.bol.ucla.edu X-Mailer: QUALCOMM Windows Eudora Version 5.2.0.9
I am not familiar what KDP is. I did oriented (perpendicular to the crystallographic axes) 10 nm thick sections of the 3D ribosomal crystals and some other biological macromolecules. I used to process them in the pretty standard way with embedding into Epon. If crystals was big enough (0.2-0.5 mm) I orient them before polymerization. If the crystals are small (less than 0.1 mm) it's practically impossible to orient them - so you will have a set of different planes. Some people used tannic acid to improve the preservation of the crystals during the dehydration and embedding. I had perfect sections from the whole crystal without huge problems. We used those sections for the crystal structure determination. The crystal cell unit parameters obtained from the sections were in great agreement with X-ray crystallography data obtained later. EM also helped to distinguish between two group of symmetry X-ray guys had problem with. As a joke, I determined 2D cell dimensions for G-factor crystals in two days with very good preciseness (+/- 1A) as X-ray guys. Nevertheless it took them 2 years to prove that my data was correct. Ironically, by mistake, my EM images of the cell unit were presented as a X-ray solution. We also used EM for crystal's quality determination. Sergey
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
From MicroscopyL-request-at-microscopy.com Wed Sep 24 19:44:58 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8P0iRiO026808 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 19:44:27 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8P0iR4h026804 for L1; Wed, 24 Sep 2003 19:44:27 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8P0hQiO026773 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 19:43:51 -0500 Received: from larkspur.noc.ucla.edu (larkspur.noc.ucla.edu [169.232.46.16]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8P0geiO026761 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 24 Sep 2003 19:42:55 -0500 Received: from tigerlily.noc.ucla.edu (tigerlily.noc.ucla.edu [169.232.46.12]) by larkspur.noc.ucla.edu (8.12.8p2/8.12.8) with ESMTP id h8P0gDlZ026093; Wed, 24 Sep 2003 17:42:13 -0700 Received: from emi.ucla.edu ([149.142.241.180]) (authenticated bits=0) by tigerlily.noc.ucla.edu (8.12.9p1/8.12.9) with ESMTP id h8P0gCNU006851 (version=TLSv1/SSLv3 cipher=DES-CBC3-SHA bits=168 verify=NOT); Wed, 24 Sep 2003 17:42:12 -0700 Message-Id: {5.2.0.9.2.20030924174041.01b1b208-at-mail.ucla.edu} X-Sender: sryazant-at-pop.bol.ucla.edu X-Mailer: QUALCOMM Windows Eudora Version 5.2.0.9
O-o-o-o-ps-s-s I just post reply about ribosomal crystals, which is nothing to do to inorganic materials. I apologize for not paying attention. Sergey
At 02:59 AM 9/24/2003, you wrote:
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
From MicroscopyL-request-at-microscopy.com Wed Sep 24 19:48:17 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8P0m2iO026863 for {L1-at-ns.microscopy.com} ; Wed, 24 Sep 2003 19:48:02 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8P0m25w026859 for L1; Wed, 24 Sep 2003 19:48:02 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8P0lGiO026828 for "MListServerFilteredEmail_5-at-microscopy.com"; Wed, 24 Sep 2003 19:47:42 -0500 Received: from mailsmtp.siu.edu (mailsmtp.siu.edu [131.230.254.197]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8P0kjiO026822 for {Microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 19:47:01 -0500 Received: from mail.siu.edu (mail.siu.edu [131.230.254.198]) by mailsmtp.siu.edu (8.12.10/8.12.10) with ESMTP id h8P0hVCt025286 for {Microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 19:43:31 -0500 Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) by mail.siu.edu (8.11.6p2/8.11.6) with ESMTP id h8P0hSD736908 for {Microscopy-at-msa.microscopy.com} ; Wed, 24 Sep 2003 19:43:28 -0500 Mime-Version: 1.0 X-Sender: bozzola-at-saluki-mail.fiber2.siu.edu Message-Id: {v04210103bb97ea914e4a-at-[131.230.177.142]}
We have a Leica (formerly Wild) M650 surgical microscope on the floor stand. Beautiful instrument.
We would like to have an alternate stand so that the optics could be used on a table top.
I believe such a stand is used in the Leica M651 MSD. I would appreciate hearing from anyone who has information on this topic (vendors, users, etc.)
Thank you.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (brian.kirkmeyer-at-iff.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 24, 2003 at 09:15:33 ---------------------------------------------------------------------------
Email: brian.kirkmeyer-at-iff.com Name: Brian Kirkmeyer, Ph.D.
Organization: IFF
Title-Subject: Replication from skin for OM
Question: I am conducting an experiment in which a substance is applied to skin and washed away (like with soap), and we need to examine the residue remaining on the skin. My arm, which is the skin in question, does not fit in either of our optical microscopes, so I thought replication might be the way to go.
What is the best way to replicate the residue on the surface of my skin for analysis with OM? Is something simple like scotch tape the way to go? For the record, I would prefer to use benign substances on my skin...I like my arm and would like to keep it. :)
From MicroscopyL-request-at-microscopy.com Thu Sep 25 04:00:34 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8P90JiO030056 for {L1-at-ns.microscopy.com} ; Thu, 25 Sep 2003 04:00:19 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8P90JuJ030052 for L1; Thu, 25 Sep 2003 04:00:19 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8P8xYiO030021 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 25 Sep 2003 03:59:59 -0500 Received: from tytan.inmat.pw.edu.pl (www.inmat.pw.edu.pl [194.29.176.2]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8P8x7iO030010 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 25 Sep 2003 03:59:23 -0500 Received: from inmat.pw.edu.pl (mailer.inmat.pw.edu.pl [148.81.84.68]) by tytan.inmat.pw.edu.pl (8.9.3/8.9.3) with ESMTP id JAA03528 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 25 Sep 2003 09:37:38 +0200 Received: from MMM/SpoolDir by inmat.pw.edu.pl (Mercury 1.48); 25 Sep 03 10:57:54 +0100 Received: from SpoolDir by MMM (Mercury 1.48); 25 Sep 03 10:57:27 +0100
I'm not sure about your problem but we couldn't install SEM in the entire building which was located about 50m from the street train's stop. A change in the magnetic field was measured when the train was starting.
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} Dear Ann, } } To your problems with the chloroplasts I can recall that here in } the lab we fix with glutaraldehyde 2 % in the same buffer (the same } molarity and pH) used for isolation. Post fixed in 2% Osmiun T.- The } Karnovsky is too much hypertonic for plants cells and tissues. The } chloroplast do not have to much lipid materials to reduce the Osmiun, } however 2 or 3 hours will rend enough to get a good conservation and } contrast.
In my hummble opinion! Have a good luck! Francisco
Francisco Freire, F.R.M.S. Head Service Electron Microscopy Service University Las Palmas de Gran Canaria Canary Islands Spain
} } } } Lehman, Ann wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi John,
You might try contacting Nuhsbaum Inc. They recently sold me a table top boom stand for my stereo microscope. web site is: http://www.nuhsbaum.com/
Best of luck,
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
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We have a Leica (formerly Wild) M650 surgical microscope on the floor stand. Beautiful instrument.
We would like to have an alternate stand so that the optics could be used on a table top.
I believe such a stand is used in the Leica M651 MSD. I would appreciate hearing from anyone who has information on this topic (vendors, users, etc.)
Thank you.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Amanda.Harman-at-newcastle.edu.au) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, September 25, 2003 at 00:30:17 ---------------------------------------------------------------------------
Organization: Reproductive Science Group, Newcastle University Australia
Title-Subject: Immunofluoresence with LR White Resin
Question: Hi,
I have been unsuccessfully trying to label 1 micron LR White resin sections of mouse epididymis,with a FITC tagged antibody. We are successfully gold labelling thin sections with the same antibody from the same blocks.
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Brian Kirkmeyer wrote: ============================================== Title-Subject: Replication from skin for OM
Question: I am conducting an experiment in which a substance is applied to skin and washed away (like with soap), and we need to examine the residue remaining on the skin. My arm, which is the skin in question, does not fit in either of our optical microscopes, so I thought replication might be the way to go.
What is the best way to replicate the residue on the surface of my skin for analysis with OM? Is something simple like scotch tape the way to go? For the record, I would prefer to use benign substances on my skin...I like my arm and would like to keep it. :) =============================================== I can refer you to one of my own (with R. Swayne and T. Nightingale) publications:
Characterizing Cosmetic Effects and Skin Morphology by Scanning Electron Microscopy", presented at SCC Annual Educational Symposium, May 1975, St. Louis; J. Soc. Cosmet. Chemists, 27, 509-531 (November 1976).
I don't have any reprints left but I trust you would have this paper in the IFF library.
The detection and measurement of materials that are substantive to skin (meaning that they tend to stick to the skin instead of being washed off such as active moisturizer materials that might be incorporated into a bar of soap) is not a straight forward analytical undertaking. We have found that if one can be happy with measuring the effect of the deposited material , rather than the material itself, if the work is done on cosmetically dry skin, and if you are doing replicas of the skin site (with the right replicating material), before vs. after product application, when there is a deposition of something, its effect can be measured by careful comparision with the before replicas. One can't do this kind of work on "normal" skin because there would not be present the uplifting (stratum corneum) layers from which one could measure such changes. The results can be analyzed semi-automatically with image analysis.
Although they are not images from the above referenced paper, examples of such skin replicas can be seen on URL http://www.2spi.com/catalog/spec_prep/a.html
These replicas were made with the SPI-Chem Wet Replica kit which uses a fast acting curing agent so that the true skin morphology is captured, and not that of a highly hydrated skin site (which would be the result if the replicating resin was curing over a longer time frame such as several minutes). See URL http://www.2spi.com/catalog/spec_prep/wet_rep_kits.shtml
Scotch tape strippings and even cyanoacrylate strippings (which do "hurt" when stripped off) don't seem to be very helpful in terms of measuring the amount present. We have tried measuring quantitatively, the amount of residue remaining by taking multiple Scotch tape strippings, each one removing a bit more of the residue, so that when all of the residue was removed, that last stripping really did "hurt". Counting the number of strippings needed to get down to that point, in theory at least, one could make some determination about the amount of residue present. But we could never get this approach to work very reproducibly either within our own laboratory or between laboratories using the same product application protocol.
Disclaimer: SPI Supplies is the manufacturer of the SPI-Chem Wet Replica Kit. The independent laboratory of Structure Probe, Inc. has been performing these kinds of studies for clients in the cosmetic and pharmaceutical industries since 1972.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
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Nestor, I agree keep up the good work. Now a little problem, for the last week or so all the communications I receive from the listserver comme in duplicate. Isi this a problem on the server or is my e-mail package doing this to me? Has anyone else had this problem.
Thanks,
Ron.
Dee Breger wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } } Nestor, it never fails to astound me how ridiculous and anti-productive our } modern life is getting. You're doing too fine and valuable a job to get } mired in this kind of bureaucratic, er, um, stuff. My continued thanks for } all your good work on our behalf. Dee } } ----------------------------------------------------------------------- } } } } Colleagues } } } } I believe AOL & Compuserve & Netscape.Net Email users are now restored } } to active participation and can now once again receive Microscopy } } Listserver Email. Sorry for the extended down time to those subscribers. } } } } AOL in their infinite wisdom decided to block all Microscopy Listserver } } saying that it was a potential source of SPAM mail. A point that } } I would undoubtably argue to the contrary. } } } } It has taken me several weeks to get them to reverse this decision and } } I had to jump through a lot of proverbial hoops. But as of this today mail } } appears to be flowing to blocked addresses at AOL. } } } } I might point out that alot of ISP's, Companies and Universities have changed } } their SPAM software recently, and unilaterally block content or domains. } } For example } } I received rejected Email from several organizations from the (valid) Email } } posting asking about WebCams for microscopy. It was unilaterally decided } } that any Email message } } with the word Webcam was obviously SPAM mail. This was clearly not ture for } } this Email, but nevertheless the SPAM filters at various sites rejected } } it. This will } } be a continuing problem, for which I see little solution. So remember to } } choose } } your subject words carefully... } } } } In any event, please don't be bashful of reporting outageous of } } MListserver Email which } } persist for several days. Your site may institute a domain or subject filter } } which blocks MListserver Email and I will not necessairly know about it. } } } } For example,, AOL never actually notified me that they were blocking traffic. } } I only discovered it when testing a new filter and found that my AOL account } } was not receiving the test messages. That event (over 2 weeks ago) } } started me on search for the problem, which I erroneously thought I created, } } but happened to coincide with their system change. } } } } BTW, I note with irony, that their blocking of the MListserver did not } } actually } } decrease the amount of SPAM mail I get on my (seldom used) AOL } } account, it only stopped me from using AOL, and also from my 80 year old } } Mother from receiving Email addressed from me via my normal microscopy.com } } account! } } } } Sigh, now if I were running AOL things would be different.... ( Nestor } } grins ) } } } } ;-) } } } } Nestor } } Your Friendly Neighborhood SysOp. } } *************************************************************** } Please do not publicly post any of my correspondence without permission } } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995)
-- Ronald J. Smith Department of Biology Room 235, Biological & Geological Sciences Bldg. U.W.O., London, Ontario N6A 5B7 (519) 661-2111 ext.86486
From MicroscopyL-request-at-microscopy.com Thu Sep 25 10:19:35 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8PFJ5iO001018 for {L1-at-ns.microscopy.com} ; Thu, 25 Sep 2003 10:19:05 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8PFJ55I001013 for L1; Thu, 25 Sep 2003 10:19:05 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8PFIAiO000962 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 25 Sep 2003 10:18:32 -0500 Received: from col-msxproto1.col.missouri.edu (col-msxproto1.col.missouri.edu [128.206.3.151]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8PFHWiO000940 for {microscopy-at-sparc5.microscopy.com} ; Thu, 25 Sep 2003 10:17:46 -0500 Received: from col-email4.col.missouri.edu ([128.206.52.22]) by col-msxproto1.col.missouri.edu with Microsoft SMTPSVC(5.0.2195.6673); Thu, 25 Sep 2003 10:17:00 -0500 X-MimeOLE: Produced By Microsoft Exchange V6.0.6375.0 Content-Class: urn:content-classes:message MIME-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
We routinely use grid pens from Electron Microscopy Sciences, due to terrible problems with losing sections and having sections tear apart during incubations for immuno work. This adhesive has almost completely solved our problems and even appears to have improved section stability under the beam.
As far as any effect on labeling, I have never done any side-by-side comparisons with and without adhesive, but I have not noticed any obvious effects on our labeling.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Christopher S. Zurenko [mailto:czurenko-at-mail.khri.med.umich.edu] Sent: Wednesday, September 24, 2003 4:33 PM To: Microscopy-at-MSA.Microscopy.Com
I've been taking on a post-embed immunogold staining project and I'm having a low section adhesion average. My staining periods and rinses are quite long so there is ample opportunity for the sections to float off. My latest attempt had 1 grid out of 6 that retained its sections, and that's a poor average even if I'm a hitter for the Detroit Tigers!
A colleague emailed me the grid adhesive described by Bozzola, 2nd Ed but wonder if there are any drawbacks I need to consider, both with the beam and any known reactivity with the gold reagent?
Any suggestions to increase my success are welcomed!
Thank you
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
From MicroscopyL-request-at-microscopy.com Thu Sep 25 10:59:59 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8PFxeiO001395 for {L1-at-ns.microscopy.com} ; Thu, 25 Sep 2003 10:59:40 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8PFxeY1001391 for L1; Thu, 25 Sep 2003 10:59:40 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8PFwqiO001354 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 25 Sep 2003 10:59:20 -0500 Received: from mailsmtp.siu.edu (mailsmtp.siu.edu [131.230.254.197]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8PFwFiO001350 for {Microscopy-at-msa.microscopy.com} ; Thu, 25 Sep 2003 10:58:33 -0500 Received: from mail.siu.edu (mail.siu.edu [131.230.254.198]) by mailsmtp.siu.edu (8.12.10/8.12.10) with ESMTP id h8PFstCt039018 for {Microscopy-at-msa.microscopy.com} ; Thu, 25 Sep 2003 10:54:55 -0500 Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) by mail.siu.edu (8.11.6p2/8.11.6) with ESMTP id h8PFsrD362102 for {Microscopy-at-msa.microscopy.com} ; Thu, 25 Sep 2003 10:54:53 -0500 Mime-Version: 1.0 X-Sender: bozzola-at-saluki-mail.fiber2.siu.edu Message-Id: {v04210100bb98bd909c7e-at-[131.230.177.142]} In-Reply-To: {3F71D55C.3100.B8FE46-at-localhost} References: {3F71D55C.3100.B8FE46-at-localhost}
} Question from: } } Christopher S. Zurenko } Kresge Hearing Research Institute, Otopathology } The University of Michigan Medical School
} I've been taking on a post-embed immunogold staining project and I'm } having a low } section adhesion average. My staining periods and rinses are quite } long so there is } ample opportunity for the sections to float off. My latest attempt } had 1 grid out of 6 } that retained its sections, and that's a poor average even if I'm a } hitter for the Detroit } Tigers! } } A colleague emailed me the grid adhesive described by Bozzola, 2nd } Ed but wonder } if there are any drawbacks I need to consider, both with the beam } and any known } reactivity with the gold reagent?
Comments:
Poor adhesion of sections is generally due to either dirty grids or grids that are not flat (bent or wavy). Before using grid glue, try cleaning the grids by sonication in a solvent such as acetone. Alternatively, you might use an acid such as 1M HCl for 5-10 minutes. If cleaning alone does not solve the adherence problem, then place the grids onto a filter paper (with the side that will hold the sections facing up) and place droplets of 0.25% Formvar or Collodion onto the grids. This puts a plastic coating over the meshwork of the grids without obstructing the open areas. The plastic coating enhances adherence of the plastic sections significantly. Also, after the sections are put onto the grids, place them overnight in a 60 C oven prior to immuno staining. This works wonders.
If all of these fail, maybe then I would consider grid glue as the last resort. But, really, you should not need it.
Let us know what solves your problem.
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 Email: bozzola-at-siu.edu ##############################################################
From MicroscopyL-request-at-microscopy.com Thu Sep 25 11:15:19 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8PGEmiO001564 for {L1-at-ns.microscopy.com} ; Thu, 25 Sep 2003 11:14:48 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8PGEmlW001559 for L1; Thu, 25 Sep 2003 11:14:48 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8PGDmiO001523 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 25 Sep 2003 11:14:10 -0500 Received: from imo-r04.mx.aol.com (imo-r04.mx.aol.com [152.163.225.100]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8PGDJiO001517 for {microscopy-at-msa.microscopy.com} ; Thu, 25 Sep 2003 11:13:30 -0500 Received: from DrJohnRuss-at-aol.com by imo-r04.mx.aol.com (mail_out_v36_r1.1.) id h.1e2.107f500d (4238); Thu, 25 Sep 2003 12:12:16 -0400 (EDT)
In a message dated 9/25/03 12:02:40 PM, rsmith-at-uwo.ca writes:
} I agree keep up the good work. Now a little problem, for the last week } or so all } the communications I receive from the listserver comme in duplicate. Isi } this a } problem on the server or is my e-mail package doing this to me? Has anyone } else had } this problem.
This happens to me as well, but (strangely) only on about half of the messages, and the second copy arrives quite some time after the first one. I was blaming my email client, since I have messages being forwarded from one address to another, but have no real explanation. John Russ
From MicroscopyL-request-at-microscopy.com Thu Sep 25 21:02:57 2003 Return-Path: {Microscopy-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8Q22viO005594 for {L1-at-ns.microscopy.com} ; Thu, 25 Sep 2003 21:02:57 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8Q22vAH005589 for L1; Thu, 25 Sep 2003 21:02:57 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to Microscopy-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8Q22HiO005558 for "MListServerFilteredEmail_5-at-microscopy.com"; Thu, 25 Sep 2003 21:02:37 -0500 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8Q22GiP005555 for {microscopy-at-ns.microscopy.com} ; Thu, 25 Sep 2003 21:02:16 -0500 Mime-Version: 1.0 Message-Id: {p05111a01bb9941cc67cf-at-[206.69.208.21]}
Colleagues
There are several possible explainations for duplicate messages.. If this problem persists by all means contact me, but don't post this problem to the MicroscopyListserver, send an off-line message to me explaining the problem and if at all possible include an example. and I'll try to help you sort it out.
Here are a few tips, if you would like to do some detective work ahead of time.
1.) If every message you receive is a duplicated and this persists for a long time (i.e. days) then it is likely you actually receiving multiple seperate mailings. There can be 2 possibilities here
a.) You are really subscribed multiple times.
The important information is in your Email header are your two subscriptions both the same email address or not. Some people have inadvertently subscribed with similiar (yet different) addresses..
These may all be you, but the server treats these as 3 different addresses and will dutifully sent out 3 copies of the message, arent' computers wonderful, they do what you tell them not what you mean't. Check your mail headers and if you are receving posts at different addresses unsubscribe the one you don't want. Use the WWW based form at
b.)Someone is (or has started) forwarding mail from their address to you, or you are forwarding mail to yourself from another system.
This happens sometimes when, for example, an employee leaves a company and email is forwarded from say an info-at-xyz.com account to another person in the company. Again you need to check your header lines to see how the mail is getting to you.
In both cases above I'll try to help, but I'll also need the header info to sort it out (at least to sort it out more easily).
2.)Occassional duplicate messages...
a). Check the time stamps in the headers
If the time stamps are different then it is likely that the sender just mailed it twice. This happens occassionally
b.)Time stamps identical, but duplicate messages are an occasional occurance.
Likely due to a mail delivery problem. All mail servers attempt to insure delivery. If an error is detected during the sending process the mail server may resend the whole message. The severity of the error is also sometimesirrelevant, it could just be a missing character even a space at the end of the message. It will be resent depending upon the error codes sent back an forth between the mail delivery agents (MDA's). This can happen because of network failures, congestion and or server errors. This problem can occur anywhere along the transmission path. It could be at this server, a relay point along the net, your ISP's server, or your client program having problems connecting and requesting a download to your local computer.. Unless this persists for a long time, there is not much that can be done, since the error is probably not reproducible.
3.) The whole list starts getting duplicates... I'll catch this, (it has happened a few times) but remember even I ocassionally have another life, and I might not catch the problem until later in the evening when I do system checks.
Also remember that I closely manage the subscription and unsubscription process so that spam/junk mailers have a difficult time getting in. New subscriptions and unsubscriptions must be approved, i.e. it is not an automatic process. This is done only once/day, so don't expect instanteous changes.
This minimizes junk mailers getting into the system, but it does delay things.
I'll also remind you that it takes time for all the mail queues to process. Postings don't happen immediately, there are delays in processing and delivery. On average the delay is on the order of a few hours. If you attempt to post a message and you don't see it within a day something is likely wrong.
All rejected messages which are spam suspects are automatically sent back to the sender with an explaination of the problem. However, I note that lately, some sites are flagging the MListserver warning messages as SPAM. It's a catch 22 situation and there have been a few subscribers caught in the middle with both servers rejecting each others Email, meaning the person who posted the message is out of the loop and doesn't know what is happening. In this case (i.e. you don't see your posting after about 24-48 +hours), contact me off-line or try using the on-line WWW submission form also at
as with the subsription WWW page this one is closely monitored to limit spam and there will be delays since these forms requires my personal "blessing" to be forwarded to the list.
Cheers.
Nestor Your Friendly Neighborhood SysOp.
From MicroscopyL-request-at-microscopy.com Fri Sep 26 09:35:44 2003 Return-Path: {MicroscopyL-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8QEZQiO011584 for {LL1-at-ns.microscopy.com} ; Fri, 26 Sep 2003 09:35:26 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8QEZQLa011579 for LL1; Fri, 26 Sep 2003 09:35:26 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to MicroscopyL-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8QEYbiO011545 for "MListServerFilteredEmail_6-at-microscopy.com"; Fri, 26 Sep 2003 09:35:06 -0500 Received: from mail1.hrz.uni-bielefeld.de (mail1.hrz.uni-bielefeld.de [129.70.4.88]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8QEXhiO011537 for {microscopy-at-msa.microscopy.com} ; Fri, 26 Sep 2003 09:34:22 -0500 Received: from conversion-daemon.mail1.hrz.uni-bielefeld.de by mail1.hrz.uni-bielefeld.de (iPlanet Messaging Server 5.2 HotFix 1.15 (built Apr 28 2003)) id {0HLT00H01SF3WO-at-mail1.hrz.uni-bielefeld.de} for microscopy-at-msa.microscopy.com; Fri, 26 Sep 2003 16:33:09 +0200 (MEST) Received: from UNI-BIELEFELD.DE ([129.70.82.150]) by mail1.hrz.uni-bielefeld.de (iPlanet Messaging Server 5.2 HotFix 1.15 (built Apr 28 2003)) with ESMTPPA id {0HLT00MUUSF8YM-at-mail1.hrz.uni-bielefeld.de} for microscopy-at-msa.microscopy.com; Fri, 26 Sep 2003 16:33:08 +0200 (MEST)
Dear Colleagues, we are searching for an antibody to analyze mitotic activity in mouse tissue (cryo sections, formaldehyde-prefixed, 4% pFA). Does anybody have recommendations for a nice ki67 or pcna (proliferating cell nuclear antigen) antibody to detect proliferation? Any experiences, tricks, tips and recommendations are welcome.
Thanks in advance
Daniel
-- (-)-(-) --------------------------- \"/ --- Dr. Daniel Eberhard =V=
From MicroscopyL-request-at-microscopy.com Fri Sep 26 10:34:28 2003 Return-Path: {MicroscopyL-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8QFYHiO012182 for {LL1-at-ns.microscopy.com} ; Fri, 26 Sep 2003 10:34:17 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8QFYHpU012177 for LL1; Fri, 26 Sep 2003 10:34:17 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to MicroscopyL-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8QFXWiO012140 for "MListServerFilteredEmail_6-at-microscopy.com"; Fri, 26 Sep 2003 10:33:57 -0500 Received: from VL-MO-MR005.ip.videotron.ca (relais.videotron.ca [24.201.245.36]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8QFWoiO012126 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 26 Sep 2003 10:33:02 -0500 Received: from laptop ([24.200.151.59]) by VL-MO-MR005.ip.videotron.ca (iPlanet Messaging Server 5.2 HotFix 1.16 (built May 14 2003)) with ESMTP id {0HLT007JAV5QR1-at-VL-MO-MR005.ip.videotron.ca} for Microscopy-at-MSA.Microscopy.Com; Fri, 26 Sep 2003 11:32:23 -0400 (EDT)
any suggestions of TEM spec prep for free flowing polymers ?
Ephram
From MicroscopyL-request-at-microscopy.com Sun Sep 28 10:57:45 2003 Return-Path: {MicroscopyL-request-at-microscopy.com} Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8SFvje2010247 for {LL1-at-ns.microscopy.com} ; Sun, 28 Sep 2003 10:57:45 -0500 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id h8SFvjHq010242 for LL1; Sun, 28 Sep 2003 10:57:45 -0500 X-Authentication-Warning: ns.microscopy.com: mail set sender to MicroscopyL-request-at-microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id h8SFv4e2010211 for "MListServerFilteredEmail_6-at-microscopy.com"; Sun, 28 Sep 2003 10:57:24 -0500 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id h8SFv4e3010208 for {microscopy-at-ns.microscopy.com} ; Sun, 28 Sep 2003 10:57:04 -0500 Mime-Version: 1.0 Message-Id: {p05111a00bb9cab68b25d-at-[206.69.208.21]}
Colleagues
Over the weekend I have modified the Microscopy Listserver Email filters which I hope should help you all in the constant SPAM/Virus battles.
As you know there is alot of Address spoofing (i.e. faking) going on lately, where a false sender's address is inserted into an Email to make you think it is coming from somewhere different than the actual source.
For many years the MListserver has been inserting a header line into the body of all Emails which indicates it is being sponsored by the Microscopy Society of America, and with a bit of reminder info on how to subscribe/unsubscribe. This is all handled by the SPAM filter, and was intended to help you weed out junk mail.
Many people filter email based upon the content of Email headers and/or the Email text in the message body. I have modified the SPAM filters so that all Email which now goes through the Filter now also should have it's SUBJECT modified and the phrase
[MicroscopyListserver]
prepended to the actual text of the original subject. This should help you all differentiate spoofed email from items that have actually been processed by the ML filter.
This makes the Subject line abit longer, but it adds abit of insurance that the message "might" be real ;-)
Of course, I have every confidence that this will be defeated at some time in the not to distant future, but it should help for awhile.
Remember, just because an Email appears to be coming from Microscopy.Com, you should not trust it implicitly. There was a virus sent out at the end of last week purportedly about Microscoft security updates. Had this gone through the ML Filter it would have been blocked because of the attachment which it contained.
The battles never end...
Cheers
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Sun Sep 28 16:31:04 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (docM_ph-at-hotmail.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, September 27, 2003 at 00:31:12 ---------------------------------------------------------------------------
Email: docM_ph-at-hotmail.com Name: Martin Moreno
Organization: St} Luke's Medical Center
Education: Graduate College
Location: Quezon City,Philippines
Question: Im trying to visualize viruses by Negative Stain on a transmission EM. BUT,I cant seem to get the right stain. Casn you please help me on this? Id appreciate it very much. Thanks you in advance.
I am looking for any used, but operational equipments, either as a donation or with a small charge. The equipments include: Microscopes (LOM, TEM, SEM, SPM) HPLC DSC/TGA FTIR GC-MS AA Thanks! Pat Sadhukhan
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 10:32:04 2003
The final notice for the Midwest Microscopy & Microanalysis Society (MMMS) Meeting in Madison, Wisconsin on October 3rd, can be found at the following URL: http://www.msa.microscopy.com/MSALAS/MMMS/MMMSOct2003.pdf
The MMMS Society in association with the ASM Chicago Chapter is holding an SEM workshop at Harper College in Palatine, Illinois on November 14th. The first announcement for this meeting can be found at the following URL: http://www.msa.microscopy.com/MSALAS/MMMS/MMMSNov2003.html
For further information please contact either:
Robert Mierzwa (MMMS President - Elect): Tel: 920-803-8945, email: mierzwa-at-jeol.com Arvid Casler (MMMS Program Coodinator): Tel: 847-674-7700, email: arvid_casler-at-fmo.com Jim DiOrio (MMMS President): Tel: 847-270-4676, email: jim_diorio-at-baxter.com
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 11:52:26 2003
The Connecticut Microscopy Society announces plans for its Fall Meeting October 23rd at Wesleyan University in Middletown, CT, featuring:
Dr. David Hall, from Albert Einstein School of Medicine. TITLE: "The Complete Nematode: Exploring C. elegans Tissues by EM"
Dr. Ron Anderson, former MSA President and now Editor of Microscopy Today. TITLE: "TEM Analysis of Thin Films: Specimen Preparation"
Colleagues
It was pointed out to me that the [MicroscopyListserver] label that I added over the weekend was far too long, and upon reflection I must agree. Chalk it up to too many hours in front of a computer screen.
I've shortened it to just [Microscopy]. Let's see how well this works.
Cheers...
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 14:39:50 2003
I am looking for recommendations for oxygen sensors. We have some small rooms where liquid nitrogen is used in amounts that require some safety measures including an oxygen monitor. I have found units ranging from about $600 to $1800 with varying costs and 1 year to 2 year lifetimes for the active sensor.
Larry Ackerman Advanced Imaging Lab/Dr. John Sedat/Dr. David Agard Dept. of Biochemistry & Biophysics University of California San Francisco 600-16th Street, Box 2140, Room S101 San Francisco, CA 94143
415-476-4441 FAX 415-514-4142
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 19:18:23 2003
I would suggest "ListServ" or "ListServer" is better, because "Microscopy is too much common word and may not be effectively used in any filters. For instance, "electron microscopy" would be filtered as a message from listserver even if it's not. Another suggestion: if it possible, it would be better to add it to the end of the subject line, otherwise, I have difficulties to read subject line, it's mostly occupied by "Microscopy"... Thanks, Sergey
At 10:20 AM 9/29/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I think that the current [Microscopy] preface is very good. "ListServer" is too generic...there are at least 15,000 of them. A filter should only look at the Subject line. If it wants to dig deeper, it can look at the sub-headers. But for Nestor's purpose, and ours, I think that [Microscopy] is a great advancement. Plus, this keeps me from wrongfully posting to the XL-30 ListServer.
Thanks Nestor.
gary g.
At 05:21 PM 9/29/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 29 23:08:31 2003
I would agree that [Microscopy] would be more appropriate than [Listserver]; I think Sergey's idea of putting the tag at the end of the subject line instead is a good one, though. Sergey: for purposes of filtering, why not include the [] brackets in the string to match for the filter? Any thoughts/comments on using [MSA], or is that getting carried away?
------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I think that the current [Microscopy] preface } is very good. "ListServer" is too generic...there } are at least 15,000 of them. A filter should only } look at the Subject line. If it wants to dig deeper, } it can look at the sub-headers. But for Nestor's } purpose, and ours, I think that [Microscopy] is a } great advancement. Plus, this keeps me from } wrongfully posting to the XL-30 ListServer. } } Thanks Nestor. } } gary g. } } } } At 05:21 PM 9/29/2003, you wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } I would suggest "ListServ" or "ListServer" is better, because "Microscopy } } is too much common word and may not be effectively used in any } } filters. For instance, "electron microscopy" would be filtered as a } } message from listserver even if it's not. Another suggestion: if it } } possible, it would be better to add it to the end of the subject line, } } otherwise, I have difficulties to read subject line, it's mostly occupied } } by "Microscopy"... Thanks, Sergey } } } } At 10:20 AM 9/29/2003, you wrote: } } } } } } } ------------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------- } } } } } } Colleagues } } } } } } It was pointed out to me that the [MicroscopyListserver] label that } } } I added over the weekend was far too long, and upon reflection I must agree. } } } Chalk it up to too many hours in front of a computer screen. } } } } } } I've shortened it to just [Microscopy]. Let's see how well this works. } } } } } } Cheers... } } } } } } Nestor } } } Your Friendly Neighborhood SysOp } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } 10833 Le Conte Ave, Room 33-089 } } Los Angeles, CA 90095 } } } } Phone: (310) 825-1144 (office) } } (310) 206-1029 (Lab) } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 10:59:29 2003
In tracking the logs (and my email) for the last few days I am starting to see that an increasing a number of postings are being rejected. Rejections in themselves are not a new event as it happens alot.
This is happening more and more often, because of the fact that people have changed their addresses or have a different return address than their sending address. The problem is that the warning message which is telling the sender that there is a possible problem with their post (and is automatically sent out by the Listserver ) are being rejected by many receipents hosts.
This means, of course, that the sender doesn't know there is a problem.
If you are having a problem posting a message, for the sort term please use the WWW based form at:
Sorry for the hassel, I'll investigate this latest problem and try to sort it out.
Nestor Your Friendly Neighborhood SysOp
-- ---===[|]===--- Nestor J. Zaluzec
Email: Zaluzec-at-Microscopy.Com {--------at- Home
Email: Zaluzec-at-aaem.amc.anl.gov {-------at- Argonne National Lab.
---===[|]===---
--------------------------------------------------------- The box said "This program requires Win 95/98/NT or better..." so I bought a G3 Mac ---------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 12:05:33 2003
We have replaced the Link eXL (now Oxford) automation that controlled our Cameca SX50, and the computer and its peripherals can be made available to anyone interested and who can pay the packing and shipping charges. I imagine there may be someone who may need it for parts(?)
I, personally, do not know a whole lot about this automation for the SX50, but it has controllers for 3 WDX, a second terminal (no monitors), PC-Link software/hardware, and EDX control (we are keeping the detector). It would be too much work to cannibalize, so please do not ask.
Those interested should contact me direct ... {michael -at- shaffer.net}
Hi all, I am looking for a non-autoflourescent resin. I am labeling specific cells that are not synchronized. The cells grow on ACLAR, and they are fixed with glut, and OsO4, embedded on a piece of ACLAR with the cells facing down into the resin. This creates a thin layer of resin with cells close or directly on the surface. I'm using DAPI to stain the nucleus / chromosomes to identify cells of interest. I can see cells (although it is had to focus) using a light microscope, but the resin seems to be auto-fluorescent. Currently i have only tried Epon-812. Has anybody tried this before? Thanks in advance.
Joel Mancuso Department of Molecular and Cell Biology 345 LSA University of California Berkeley, CA 94720-3200
p - 510.643.8277 email - jmancuso-at-uclink.berkeley.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 12:50:17 2003
We have the DuoScan T2500 and the computer it was on died (Macintosh OS 8.?), so we tried reinstalling it on a different computer (Macintosh (9.1).
We cannot get it to scan higher than 600 PPI. Before it died we could scan at 2500 PPI.
Any help in installing/reinstalling to drive it at full resolution would be great.
Thanks.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 15:06:07 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yan.328-at-osu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 30, 2003 at 10:55:27 ---------------------------------------------------------------------------
Email: yan.328-at-osu.edu Name: Yanping Yan
Title-Subject: [Microscopy] EM, need help with serial reconstructions
Question: Hi, all:
We are attempting serial reconstruction the axon of a cultured neuron. The axons are just about 200 nm thick.
The problem that we have run into is that when we try to line up the axons in the two sections using unambiguous landmarks, such as large organelles or varicosities (which are large enough to be present in both sections), they simply don't line up. In the one case that we have examined so far, the distance between the two landmarks is about 18% greater in one section than in the other. This is definitely not due to errors in our montaging, because we have confirmed the discrepancy within single images of the two sections taken at low mag.
Some possibly relevant technical info is that we are embedding in a pretty standard PolyBed 812 formulation and cutting silver sections.
We guess the sections must have been compressed or expanded differentially during processing. What surprises us is that the discrepancy is quite large. Moreover, we would actually expect the amount of compression to be greatest perpendicular to the knife edge, yet the axon is actually running close to parallel to the knife edge (maybe about 30 degrees off parallel) in this case.
Do you ever encounter this problem when you were doing serial reconstructions? Do you do anything to avoid this problem? If we stretch the sections with chloroform or heat, we don't how much it will help since sections can react differently.
I am wondering if you can give us some suggestions.
Thanks,
Yanping Yan
Neuroscience Graduate Studies Program Rightmire Hall, Room 056 The Ohio State University Center for Molecular Neurobiology 1060 Carmack Road Columbus, OH 43210
Greetings Joel, Do you need to achieve good image quality for fluorescence or are you just screening for cells to perform EM analysis on? I'm not sure what the refractive index of Epon-812 is, and refractive index will have an impact on image quality. Glutaraldehyde is known to cause extensive background fluorescence, but you can probably still discriminate DAPI from the greenish glutaraldehyde signal. You might try epon/araldite as outlined in:
Sun, Tolbert and Hildebrand. (1995). Using laser scanning confocal microscopy as a guide for electron microscopic study: a simple method for correlation of light and electron microscopy. J. Histochem. Cytochem. 43(3),329-355.
Other methods to correlate images from photonic microscopy and electron microscopy include the use of cryosections, light microscope imaging in agarose followed by embedding for EM. Pertinent references are below:
Robinson, Takizawa, Pombo and Cook. (2001). Correlative Fluorescence and Electron Microscopy on Ultrathin Cryosections: Bridging the Resolution Gap. J. Histochem. Cytochem. 48, 471-480.
Sun, Tolbert, Hildebrand and Meinertzhagen. (1998). A Rapid Method for Combined Laser Scanning Confocal Microscopic and Electron Microscopic Visualization of Biocytin or Neurobiotin-labeled Neurons. J. Histochem. Cytochem. 46, 263-274.
Deitch, Smith, Swann and Turner. (1991). Ultrastructural investigation of neurons identified and localized using the confocal scanning laser microscope. J. Electron Microsc. Tech. 18, 82-90.
Sun, Tolbert and Hildebrand (1997). Synaptic organization of the uniglomerular neurons of the antennal lobe of the moth Manduca sexta: a laser scanning confocal and electron microscopic study. J. Comp. Neurol. 379,2-20.
Regards, Karl G.
jmancuso wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 30 17:44:30 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (beth-at-plantbio.uga.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 30, 2003 at 16:16:36 ---------------------------------------------------------------------------
Question: Hi all, We are trying to cryostat section root-knot nematodes and soybean cyst nematodes that have been fixed in 2% paraformaldehyde/PBS, etc. but even with a new blade in place the 'todes and roots are often tearing up rather than sectioning. Any help/advice would be greatly appreciated. We'd like to see other people's protocol for fixation and hear any thoughts on 10% DMSO vs 30% sucrose? We would esp. like to know how long you leave the samples in each step of your protocol. thanks!!! Beth
Question: I need info on microscopist as a career. I want a job description and the training needed. The different types or specialties would be great also. Please include other info that would be great for a research paper.
Sasaki, S., J.K. Steven & N. Bodick (1983). Serial reconstruction of microtubula array within dendrite of the cat retinal ganglion cell: the cytoskeleton of a vertebrate dendrite. Brain Res. 259, 193-206.
Stevens, J.K. & J.T. Trogadi (1984). Computer-assisted reconstruction from serial electron micrographs: a tool fo the systemati stud of neuronal form and function. Adv. Cell. Neurobiol. 5, 341-369.
Yours, -Dick Gordon
At 3:09 P -0500 9/30/03, b wa of MicroscopyListserve wrote: } ------------------------------------------------------------------------------ } The Microscop ListServe -- Sponsor: The Microscop Societ of America
-- .................................................................................................. Dr. Richard Gordon, Depts. of Radiology/Electrical & Compute Engineering, U.Manitoba, Room GA216, Health Science Centre, 820 Sherbrook Street, Winnipeg, MB R3A 1R9 Canada Adjunct Scientist: TRLabs, Mentor: IEEE/EMBS Student Chapter Scientist, Manitoba Institute of Child Health. Editorial Board: Journal of Mechanic in Medicine and Biology; Home page: http://www.umanitoba.ca/faculties/medicine/radiology/stafflist/rgordon.html Lab: (204) 789-3828, Fax: (204) 787-2080, E-mail: GordonR-at-ms.UManitoba.ca Reprint Hunter/Professo Finder: http://www.umanitoba.ca/faculties/medicine/radiology/search/universities.html
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 08:19:54 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rsimmons-at-gsu.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 1, 2003 at 07:59:29 ---------------------------------------------------------------------------
Question: You might have a look at the LR resins (LR White / LR Gold) from London Resin Co. I believe that they will work well with your fluorescence application and you can thin section them as well if you need to.
Joel, It may not be your resin that is fluorescing, but the glutaraldehyde you fixed with. For light microscopy, we quench that fluorescence with one of the following used after the post-fix rinse: 0.1 M glycine 50 mM ammonium chloride or sodium borohydride (I hate using that one, so I don't have a recipe at my finger tips) 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
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 09:13:38 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jwilkinson-at-seton.org) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, October 1, 2003 at 08:59:11 ---------------------------------------------------------------------------
Question: I am currently working in a EM lab in a hospital. I need a procedure on how to process ciliary biopsies. After we receive the brushing from surgery. Can you help me?
Hi there, I am looking for spare screens for my JEOL 2010F, they are JEOL part numbers 607071 (small screen - focussing screen) and 607072 (large screen - viewing screen) respectively. If anyone has any spare I would be happy to buy them. JEOL have none in stock and have a 45 day lead time on them and so I was wondering if anyone is sat on a spare pair. Reply off line please.
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 09:31:01 2003
Hi there, I'm trying again to see if anyone is interested in starting a JEOL 2010F user group. We have some questions on how other users configure things on their microscope and also have some tips that we would like to share with other users. I tried doing this before but we were tied up in the installation of the scope and I never pursued it. Let me know if you are interested offline and I will see if I can get a mailing list together at least. Thanks in advance.
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 09:34:36 2003
} Subject: [Microscopy] IASTED Modelling and Simulation Newsletter - Sept. 2003 } } IASTED International Newsletter on Modelling and Simulation } September 30, 2003 } } RECENTLY ANNOUNCED MODELLING AND SIMULATION CONFERENCES } 1. The 15th IASTED International Conference on Modelling and Simulation - MS 2004 } March 1-3, 2004, Marina Del Rey, CA, USA } } Important Deadlines: } Submissions Due: Oct. 15, 2003 } Notification of Acceptance: Nov. 15, 2003 } Registration Deadline: Dec. 15, 2003 } } To submit a paper, tutorial or special session or for registration information visit our website at http://www.iasted.com/conferences/2004/marina/ms.htm } } 2. The 4th IASTED International Conference on Modelling, Simulation, and Optimization - MSO 2004 } August 16-18, 2004, Kauai, Hawaii, USA } Important Deadlines: } Submissions Due: Feb. 15, 2004 } Notification of Acceptance: Apr. 15, 2004 } Registration Deadline: June 1, 2004 } To submit a paper, tutorial or special session or for registration information visit our website at http://www.iasted.com/conferences/2004/hawaii/mso.htm } } UPCOMING CONFERENCE DEADLINES } The submission deadline for the 23rd IASTED International Conference on Modelling, Identification and Control - MIC 2004 is Oct. 15, 2003. MIC 2004 is being held Feb. 23-25, 2004 in Grindelwald, Switzerland. Delegates without papers are also welcome to register until Dec. 15, 2003. http://www.iasted.com/conferences/2004/switzerland/mic.htm } } } FUTURE CONFERENCES } Mark your calendars! The IASTED International Conference on Environmental Modelling and Simulation will be held Nov. 22-24, 2004 in St. Thomas, Virgin Islands, USA. } } } MODELLING AND SIMULATION JOURNALS FROM ACTA PRESS } The International Journal of Modelling and Simulation - First published in 1981, this journal covers all aspects of modelling, simulation, languages, software, hardware, methodology, numerical and graphical methods, virtual reality, statistical techniques, tutorials, surveys, and applications. It al so includes book reviews, conference notices, call for papers, and new publications. } Editor-in-Chief: Prof. A. Houshyar } Frequency: 4 issues per year } 2003 Rate: US$256.00 } Postage & Handling: US$25.00 } ISSN: 0228-6203 (205) } http://www.actapress.com/journals/journals.htm#Modelling } } } CONFERENCE PROCEEDINGS AVAILABLE } Past conference proceedings in the area of modelling and simulation are available for purchase from ACTA Press - http://www.actapress.com/proceedings/proceedings.htm } } For more information, to be removed from our mailing list, or to join one of the following mail groups: Power, Telecommunication, Bio-Medicine, Signal and Image Processing, Artificial Intelligence, Business, Software Engineering, Education, Databases and Knowledge Engineering, Internet and Applications, Parallel and Distributed Computing, please contact: } IASTED } #80, 4500 - 16th Avenue N.W. } Calgary, Alberta } Canada T3B 0M6 } Tel: 403-288-1195 } Fax: 403-247-6851 } E-mail: calgary-at-iasted.com } Web site: http://www.iasted.org }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 09:36:34 2003
The Fall Meeting of the Michigan Microscopy and Microanalysis Society will be held October 17, 2003 at the Kellogg Hotel and Conference Center in Lansing, Michigan.
Oral and poster sessions will be held in this one day conference. Abstract submission deadline was September 19 but late submissions are welcome. You are encouraged to attend or submit a paper. For further information or to submit an abstract, contact:
Ginam Kim President Michigan Microscopy and Microanalysis Society (MMM) Tel. 989-496-5077 or e-mail to g.kim-at-dowcorning.com
Kevin Battjes MMM Secretary Impact Analytical Voice 989-832-5555, ext 556 Michigan Molecular Institute Fax 989-832-5560 1910 W. St Andrews Road e-mail: battjes-at-mmi.org Midland MI 48640 battjes-at-impactanalytical.com
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 09:37:42 2003
Sasaki, S., J.K. Stevens & N. Bodick (1983). Serial reconstruction of microtubular arrays within dendrites of the cat retinal ganglion cell: the cytoskeleton of a vertebrate dendrite. Brain Res. 259, 193-206.
Stevens, J.K. & J.T. Trogadis (1984). Computer-assisted reconstruction from serial electron micrographs: a tool for the systematic study of neuronal form and function. Adv. Cell. Neurobiol. 5, 341-369.
Yours, -Dick Gordon
At 3:09 PM -0500 9/30/03, by way of MicroscopyListserver wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- .................................................................................................. Dr. Richard Gordon, Depts. of Radiology/Electrical & Computer Engineering, U.Manitoba, Room GA216, Health Sciences Centre, 820 Sherbrook Street, Winnipeg, MB R3A 1R9 Canada Adjunct Scientist: TRLabs, Mentor: IEEE/EMBS Student Chapter Scientist, Manitoba Institute of Child Health. Editorial Board: Journal of Mechanics in Medicine and Biology; Home page: http://www.umanitoba.ca/faculties/medicine/radiology/stafflist/rgordon.html Lab: (204) 789-3828, Fax: (204) 787-2080, E-mail: GordonR-at-ms.UManitoba.ca Reprint Hunter/Professor Finder: http://www.umanitoba.ca/faculties/medicine/radiology/search/universities.html
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 10:07:33 2003
I didn't see, or prematurely deleted, the original post but if glut is being used, that will cause problems with fluorescence. We use paraformaldehyde for all of our tissue fixation/fluorescent applications with no trouble.
Cheers
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 10:34:54 2003
The way that I process nasal brushing is to pellet the cells in microcentrifuge tubes, (many if necessary), overlay with 3% glutaraldehyde for a hour or so, gently unstick the pellet and process as a tissue block. You will continually lose some cells, but the majority will stay together. The trouble with this method is that ciliated cells will be far and few between, thus many grids will be necessary to obtain an adequate number of cells.
Best of Luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } by way of Ask-A-Microscopist {jwilkinson-at-seton.org} 10/01/03 10:12AM } } }
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jwilkinson-at-seton.org) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, October 1, 2003 at 08:59:11 ---------------------------------------------------------------------------
Question: I am currently working in a EM lab in a hospital. I need a procedure on how to process ciliary biopsies. After we receive the brushing from surgery. Can you help me?
Thanks for all the replies about the autofluorescence. I did quench glutaraldehyde with sodium borohydride. Any other suggestions? Thanks
jmancuso wrote: Hi all, I am looking for a non-autoflourescent resin. I am labeling specific cells that are not synchronized. The cells grow on ACLAR, and they are fixed with glut, and OsO4, embedded on a piece of ACLAR with the cells facing down into the resin. This creates a thin layer of resin with cells close or directly on the surface. I'm using DAPI to stain the nucleus / chromosomes to identify cells of interest. I can see cells (although it is had to focus) using a light microscope, but the resin seems to be auto-fluorescent. Currently i have only tried Epon-812. Has anybody tried this before? Thanks in advance.
Joel Mancuso Department of Molecular and Cell Biology 345 LSA University of California Berkeley, CA 94720-3200
p - 510.643.8277 email - jmancuso-at-uclink.berkeley.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 11:18:54 2003
Dear Joel, We've been doing something similar with Spurr's epoxy for quite a while with paraformaldehyde fixed tissue, avoiding the use of glut. Both plastics seem to have as little autofluorescence as any others I've tried. I've gotten around the autofluorescence problem in two ways. One, collect a z-series through your sample by widefield fluorescence and use deconvolution. A nearest neighbors can be sufficient for this purpose, as well as the most time efficient algorithm. the other approach is confocal, provided you can get access to a system with UV or a 405 nm laser diode. Even excitation at 351 nm didn't present much autofluorescence by confocal.
The glut. will add problems with autofluorescence that are worst when exciting in the UV or blue wavelengths. perhaps you could use a nuclear stain excitable in the visible range.
We have a paper accepted by Brain Research Protocols related to this, but don't yet have a publication date.
Regards, Glen
On Tuesday, September 30, 2003, at 10:43 AM, jmancuso wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Hi all, } I am looking for a non-autoflourescent resin. I am labeling specific } cells } that are not synchronized. The cells grow on ACLAR, and they are fixed } with } glut, and OsO4, embedded on a piece of ACLAR with the cells facing } down into } the resin. This creates a thin layer of resin with cells close or } directly on } the surface. I'm using DAPI to stain the nucleus / chromosomes to } identify } cells of interest. } I can see cells (although it is had to focus) using a light } microscope, but } the resin seems to be auto-fluorescent. Currently i have only tried } Epon-812. } Has anybody tried this before? } Thanks in advance. } } Joel Mancuso } Department of Molecular and Cell Biology } 345 LSA } University of California } Berkeley, CA 94720-3200 } } p - 510.643.8277 } email - jmancuso-at-uclink.berkeley.edu } } } Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 11:42:25 2003
Joyce, we also use the centrifugation method, then mix the pellet with a drop of warm 2% agar, let harden and cut into pieces for osmication and embedding. You may find of value my "cilia principles" when scoping:
IMMOTILE CILIA SYNDROME (ICS)
Key Info for diagnosis (Dec 2001)
Based on my reading of Mierau et al 1992 (MIE92) and other pertinent articles which point out the controversies and problems, I have reached the following conclusions re making a diagnosis one way or the other on a cilia specimen where ICS may be suspected:
1. For a diagnosis of ICS, the issues of "Universality and Permanence" must be met i.e. several sites and a repeat bx are required since there are cases where dynein arms have been absent one time but present another.
2. For a diagnosis of "complete dynein arm absence" (ICS according to some authors), negative findings i.e. absence of all arms on 50-100 cilia examined in perfect cross section allow only for a "consistent with" type of statement since some pts may have normal axonemes on repeat bx (MIE92)
3. For a "rule out" diagnosis i.e. reporting the occurrence of normal cilia, several well preserved cilia with some inner and outer arms must be seen. Inner arms are usually blurred and less distinct than outer arms. (DIC00). Rarely are all arms seen in a given cross section since successive arms within (i.e. through the depth of) the thin section must be superimposed to be seen (MIE92). Under the best circumstances, only about 20% of inner arms are seen. Dickersin (Dic00) p771 contends that the normal average number of outer/inner arms per axoneme is two or more. But, keep in mind that Ghadially (GHA88) reports that in ICS pts, occasional cilia can show some arms, usually short. Therefore, I conclude that it is best to find some arms in all or most of a dozen or more cilia examined in perfect cross section.
REFERENCES
MIE92: Mierau GW, et al 1992. The role of electron microscopy in evaluating ciliary dysfunction: report of a workshop. Ultrastructural Pathology 16:245-254.
DIC00: Dickersin GR, 2000. Diagnostic Electron Microscopy 2nd ed, Springer
GHA88: Ghadially F, 1988 Ultrastructural Pathology of the Cell and Matrix. erd ed, p1200, Butterworths
Tom Christensen Director EM Lab Pathology Boston U Med Center
} --------------------------------------------------------------------------- } } Email: jwilkinson-at-seton.org } Name: Joyce Wilkinson } } Organization: Brackenridge Hospital } } Education: Undergraduate College } } Location: Austin, Texas USA } } Question: I am currently working in a EM lab in a hospital. } I need a procedure on how to process ciliary biopsies. After we receive the brushing from surgery. Can you help me? } } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 12:02:48 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (william.treasurer-at-exeloncorp.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 1, 2003 at 10:50:37 ---------------------------------------------------------------------------
Email: william.treasurer-at-exeloncorp.com Name: William Treasurer
Organization: Exelon PowerLabs
Title-Subject: [Microscopy] MListserver: HNU EDS System
Question: We have a HNU Model 5000, Ser # AX2001 EDS System. Is there any companies that still provide service on these systems?
We are located in Willington, IL. My phone number is 815-458-7654.
I gather from your note that you are embedding with the ACLAR. Is it possible that this is the source of your autofluorescence?
Many years ago, when growing cells on polystyrene culture dishes, I experimented with fixing the cells, and then dissolving the plastic with ethylene dichloride. The wisps of cells were repelleted and embedded. It might work for you.
} } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Hello again, } } Thanks for all the replies about the autofluorescence. I did quench } glutaraldehyde with sodium borohydride. Any other suggestions? Thanks } } } jmancuso wrote: } Hi all, } I am looking for a non-autoflourescent resin. I am labeling specific } cells that are not synchronized. The cells grow on ACLAR, and they are } fixed with glut, and OsO4, embedded on a piece of ACLAR with the cells } facing down into the resin. This creates a thin layer of resin with } cells close or directly on the surface. I'm using DAPI to stain the } nucleus / chromosomes to identify cells of interest. I can see cells } (although it is had to focus) using a light microscope, but the resin } seems to be auto-fluorescent. Currently i have only tried Epon-812. } Has anybody tried this before? } Thanks in advance. } } Joel Mancuso } Department of Molecular and Cell Biology } 345 LSA } University of California } Berkeley, CA 94720-3200 } } p - 510.643.8277 } email - jmancuso-at-uclink.berkeley.edu } }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 15:20:02 2003
.. dusting off a very old Email message out of the archives....
} } --------------------------------------------- } } Date: Fri, 1 Oct 1993 15:18:05 -0500 (CDT) } } From: ZALUZEC-at-ANLEMC.MSD.ANL.GOV (Nestor J. Zaluzec (708)-252-5075, -4964) } } Message-Id: {931001151805.20201c44-at-ANLEMC.MSD.ANL.GOV} } } Subject: [Microscopy] Open for Business } } To: Microscopy-at-anlemc.msd.anl.gov } } X-Vmsmail-To: MICROSCOPY } } } } } } The Microscopy Listserver/Mailreflector is } } now running you may begin posting messages } } } } Cheers- Nestor } } } } ---------------------------------------------
Well after a decade of (very nearly) continuous operation, we've survived/outlived a number of servers (the original VAX-785, several Sun's, and now a PC running Linux), a few domain names and address changes, and had several different sponsors (MSA has obviously been the longest running), and I'll have to admit alot of headaches.
Many of you will recall that we delivered our 100 Millionth Email message about two years ago, and the searchable archives occupy ~ 120 Mbytes of on-line storage. Lately have been averaging ~ 18 Gbits/month of Microscopy Email (and that is not counting the junk mail!).
There was no junk/spam mail back in '93, today the ratio is around over 10:1, fortunately junk is for the most part blocked, albeit at the expense of occassionally rejecting some real messages in error.
A number of the original subscribers from back in Oct of 93 are still members of this list. I glad this means I'm not the only die hard in group. I unfortunately did not keep a complete copy of that initial list of subscribers, at least not on any media that I can still read and process here at home. I'll have to go back into my old files at ANL and see if I can resurrect a very old tape drive for that job. It would be interesting to see how many of the original members are still subscribed. I certainly have grown to recognize alot of the Email addresses on the list, and I'll have to admit having met alot of you over the years.
I do, however, remember that Arthur Day from ANSTO (ard-at-ansto.gov.au) was the first subscriber, while john chandler (chandler-at-lamar.ColoState.EDU) replied to the first question on the list. Want to guess who posted that first question ? ;-)
I'll have to admit that back in '93 I would have never predicted that I would still be running this server 10 years later.
In any event, as I hoist a virtual pint in the air ... onto the next decade..
Cheers....
Nestor Your (suddenly feeling tired) Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 15:25:35 2003
I have done astrocytes stained with GFAP+fluorescine and embedded in EMBed 812, no problems. Fixation was with 4% paraformaldehyde though, not glut.
Geoff
Joel Mancuso wrote:
} Hello again, } } Thanks for all the replies about the autofluorescence. I did quench } glutaraldehyde with sodium borohydride. Any other suggestions? Thanks } } } jmancuso wrote: } Hi all, } I am looking for a non-autoflourescent resin. I am labeling specific cells } that are not synchronized. The cells grow on ACLAR, and they are fixed with } glut, and OsO4, embedded on a piece of ACLAR with the cells facing down into } the resin. This creates a thin layer of resin with cells close or directly on } the surface. I'm using DAPI to stain the nucleus / chromosomes to identify } cells of interest. } I can see cells (although it is had to focus) using a light microscope, but } the resin seems to be auto-fluorescent. Currently i have only tried Epon-812. } Has anybody tried this before? } Thanks in advance. } } Joel Mancuso } Department of Molecular and Cell Biology } 345 LSA } University of California } Berkeley, CA 94720-3200 } } p - 510.643.8277 } email - jmancuso-at-uclink.berkeley.edu } } } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 15:39:18 2003
On Friday, September 26, 2003, at 08:32 AM, Ephram Shizgal wrote:
} any suggestions of TEM spec prep for free flowing polymers ? } Dear Ephram, I don't know what the viscosity of your polymers might be, but cryo-TEM may be possible if you can blot enough of the polymer to leave a sufficiently thin layer. Alternatively, you could cool the polymer until it is solid, then thin it or microtome it as one would with other solid polymers. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 15:50:05 2003
Nestor, Looking back over the last 10 years with all the long hours, headaches and God knows what else I'd like to suggest that you hoist not a virtual pint but a real one. If you haven't already done so I'm sure you're planning it!
Cheers, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-microscopy.com] Sent: Wednesday, October 01, 2003 4:19 PM To: microscopy-at-ns.microscopy.com
Nestor: I think I can speak for most all of us subscribers in saying that's it's been a decade of extremely GREAT service by our Friendly Neighborhood SysOp.
"Nestor J. Zaluzec" wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } .. dusting off a very old Email message out of the archives.... } } } } } --------------------------------------------- } } } Date: Fri, 1 Oct 1993 15:18:05 -0500 (CDT) } } } From: ZALUZEC-at-ANLEMC.MSD.ANL.GOV (Nestor J. Zaluzec (708)-252-5075, -4964) } } } Message-Id: {931001151805.20201c44-at-ANLEMC.MSD.ANL.GOV} } } } Subject: [Microscopy] Open for Business } } } To: Microscopy-at-anlemc.msd.anl.gov } } } X-Vmsmail-To: MICROSCOPY } } } } } } } } } The Microscopy Listserver/Mailreflector is } } } now running you may begin posting messages } } } } } } Cheers- Nestor } } } } } } --------------------------------------------- } } Well after a decade of (very nearly) continuous operation, we've } survived/outlived a number of servers (the original VAX-785, } several Sun's, and now a PC running Linux), a few domain } names and address changes, and had several different sponsors } (MSA has obviously been the longest running), } and I'll have to admit alot of headaches. } } Many of you will recall that we delivered our 100 Millionth Email } message about two years ago, and the searchable archives occupy } ~ 120 Mbytes of on-line storage. Lately have been averaging } ~ 18 Gbits/month of Microscopy Email (and that is not counting the } junk mail!). } } There was no junk/spam mail back in '93, today the ratio is around } over 10:1, fortunately junk is for the most part blocked, albeit at } the expense of occassionally rejecting some real messages in error. } } A number of the original subscribers from back in Oct of 93 } are still members of this list. I glad this means I'm not the only } die hard in group. I unfortunately did not keep a complete copy of that } initial list of subscribers, at least not on any media that I can still read } and process here at home. I'll have to go back into my old files at ANL } and see if I can resurrect a very old tape drive for that job. } It would be interesting to see how many of the original members } are still subscribed. I certainly have grown to recognize alot of the Email } addresses on the list, and I'll have to admit having met alot of you over } the years. } } I do, however, remember that Arthur Day from ANSTO (ard-at-ansto.gov.au) } was the first subscriber, while john chandler (chandler-at-lamar.ColoState.EDU) } replied to the first question on the list. Want to guess who posted that } first question ? ;-) } } I'll have to admit that back in '93 I would have never predicted that I } would still be running this server 10 years later. } } In any event, as I hoist a virtual pint in the air ... onto the next decade.. } } Cheers.... } } Nestor } Your (suddenly feeling tired) Friendly Neighborhood SysOp } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 16:28:23 2003
So come Savannah, let's each of us buy Nestor a pint. Phil
} .. dusting off a very old Email message out of the archives.... } } } } } --------------------------------------------- } } } Date: Fri, 1 Oct 1993 15:18:05 -0500 (CDT) } } } From: ZALUZEC-at-ANLEMC.MSD.ANL.GOV (Nestor J. Zaluzec } } } (708)-252-5075, -4964) } } } Message-Id: {931001151805.20201c44-at-ANLEMC.MSD.ANL.GOV} } } } Subject: [Microscopy] Open for Business } } } To: Microscopy-at-anlemc.msd.anl.gov } } } X-Vmsmail-To: MICROSCOPY } } } } } } } } } The Microscopy Listserver/Mailreflector is } } } now running you may begin posting messages } } } } } } Cheers- Nestor } } } } } } --------------------------------------------- } } Well after a decade of (very nearly) continuous operation, we've } survived/outlived a number of servers (the original VAX-785, } several Sun's, and now a PC running Linux), a few domain } names and address changes, and had several different sponsors } (MSA has obviously been the longest running), } and I'll have to admit alot of headaches. } } Many of you will recall that we delivered our 100 Millionth Email } message about two years ago, and the searchable archives occupy } ~ 120 Mbytes of on-line storage. Lately have been averaging } ~ 18 Gbits/month of Microscopy Email (and that is not counting the } junk mail!). } } There was no junk/spam mail back in '93, today the ratio is around } over 10:1, fortunately junk is for the most part blocked, albeit at } the expense of occassionally rejecting some real messages in error. } } A number of the original subscribers from back in Oct of 93 } are still members of this list. I glad this means I'm not the only } die hard in group. I unfortunately did not keep a complete copy of that } initial list of subscribers, at least not on any media that I can still read } and process here at home. I'll have to go back into my old files at ANL } and see if I can resurrect a very old tape drive for that job. } It would be interesting to see how many of the original members } are still subscribed. I certainly have grown to recognize alot of the Email } addresses on the list, and I'll have to admit having met alot of you over } the years. } } I do, however, remember that Arthur Day from ANSTO (ard-at-ansto.gov.au) } was the first subscriber, while john chandler (chandler-at-lamar.ColoState.EDU) } replied to the first question on the list. Want to guess who posted that } first question ? ;-) } } I'll have to admit that back in '93 I would have never predicted that I } would still be running this server 10 years later. } } In any event, as I hoist a virtual pint in the air ... onto the } next decade.. } } Cheers.... } } Nestor } Your (suddenly feeling tired) Friendly Neighborhood SysOp } } }
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 16:36:51 2003
Are you a classroom volunteer? Or might you try being one, if you knew a bit about how to really stimulate youngsters' interest? Project MICRO's "Microscopic Explorations" is one approach, and another very effective one is "Private Eye", which introduces observation of the microworld with cheap jeweler's loupes. If you live in the Los Angeles area, you can attend a "Private Eye" workshop at the California State Teachers Association meeting at the Long Beach Convention Center - Friday October 10, 12-1, room 301, or Saturday October 11, 10:30-11, room Seaside A; you don't need to register for the meeting.
The "Private Eye" workshops are excellent, and there will be future opportunities in other cities. If you're interested, contact me, or "Private Eye" directly at dmelody-at-theprivateeye.com. -- 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/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 16:36:19 2003
An editor of one of the National Science Teacher's Association magazines has just contacted me with a request for a short article; I know that several of you listers enjoy writing short essays, so how about it? She has an unreasonably short deadline - Oct. 15. Please remember that the readers will be elementary school teachers! Contact the editor directly {valynda_m-at-nsta.org} . ----------------------------------- At 11:55 AM -0400 10/1/03, Valynda Mayes wrote: } I am writing from the National Science Teachers Association in search of a } scientist to write a short piece for our journal for elementary teachers, } Science and Children. "Science 101" explains a topic in a question and } answer format and is written by a specialist working in the chosen field. } } In our next issue, we are publishing an article about an upper elementary } classroom using a digital microscope to observe the reproductive parts of } plants. The question we'd like to address in "Science 101" is "How do } microscopes work?" We are looking for a brief explanation appropriate for } the elementary science classroom, with a few interesting tidbits thrown in, } such as examples of ways microscopes are used in science. In research on } this topic, I came across the Project MICRO website and thought you might be } able to assist us in finding someone interested in writing this piece. } } I usually try to locate scientists with an interest in children and } education who have an affinity for this sort of thing. The piece is short } (about 400 words) and we offer an honorarium. It is our hope that "Science } 101" will introduce professions to our readers and their students in } addition to providing interesting background content. We include a short bio } and photograph of the scientist at the end of each article. If you can be of } help, please be in touch. } } Thank you for your time, } } Valynda Mayes } Assistant Editor } Science and Children
-- 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/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 17:02:03 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lesleykelley11-at-hotmail.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 1, 2003 at 15:34:52 ---------------------------------------------------------------------------
Email: lesleykelley11-at-hotmail.com Name: Lesley
Organization: ExxonMobil Chemical
Title-Subject: [Microscopy] MListserver:
Question: We are looking at purchasing a new FESEM next year. Any input on current models would be greatly appeciated.
We are currently looking at SEMs from LEO, JEOL and Hitachi.
Please respond to lesley.a.kelley-at-exxonmobil.com
Good job Nestor. Many thanks from all of us. If you will be at UCLA I'll provide beer or vodka (your choice). Sergey.
At 02:27 PM 10/1/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Joel Sheffield wrote: ====================================================== I gather from your note that you are embedding with the ACLAR. Is it possible that this is the source of your autofluorescence?
Many years ago, when growing cells on polystyrene culture dishes, I experimented with fixing the cells, and then dissolving the plastic with ethylene dichloride. The wisps of cells were repelleted and embedded. It might work for you. ================================================================ We have been selling ACLAR® cut sheets (as have been others as well) for some number of years and a substantial number of those using it find one of its main advantage to be its virtual freedom from any autofluorescence. This is described on our URL http://www.2spi.com/catalog/photo/aclar-film.shtml
Obviously there could in theory be some low level autofluorescence from the ACLAR film, but it is apparently too low to be measured (at least by most users of the film).
Chuck
===========================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 03:51:02 2003
I haven't used the Brunel version but it looks like a generic Cambridge rocking microtome only grey instead of black. If it is built to the Cambridge standard then it will probably last forever. The only thing that I've ever had to replace on our 15 or so Cambridge units is the occasional spring but that seems to be after about 15 or more years of student use. Our models also have a cord to attach the end of the arm to the specimen feed ratchet mechanism and this can wear out after many years but we found that we could get a replacement locally.
These microtomes are very simple and will happily cut between 4 and 10+ um wax (and I assume plastic) sections. Specimen orientation is rudimentary you rotate or move the specimen backwards or forwards to line up by releasing the front clamp. I found that a disposable blade system was more convenient than the solid steel blade but that might depend on your facilities for sharpening microtome knives.
If this microtome is for student use, research or irregular use then it will be ideal but if you need to cut hundreds of sections in long 'ribbons' then a rotary microtome would be better but much more expensive.
One further issue is that because the knife is exposed at the front it would be advisable to get into a strict routine of removing it from the microtome when not in use or fashioning a cover to prevent accidents.
Please again note that my comments are about Cambridge rocking microtomes (many of ours must be 30+ years old by now). But these are easily repaired and maintained because of their simple mechanisms.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: "max_gra-at-libero.it" {max_gra-at-libero.it}
We are in the process of sorting out the safety of the lab. This was a question I could not answer. A good website I used was: http//www.healthcouncil.nl
Thanks
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 05:03:51 2003
I am a new user in EM and I am having a difficult time getting my Epoxy sections to stick to the copper grids. I have tried Acetic acid washes, sonicators, HCl wash etc. I have not tried BSA, I am waiting for the order to come in. Does anyone have any more suggestions?
Thank you in Advance. Sue
-- Sue Tyler Biologist Cooperative Oxford Laboratory Center for Coastal Environmental Health& Biomolecular Research at Charleston ( CCHEBR) USDOC/NOAA/NOS/NCCOS 904 S. Morris St. Oxford, Maryland 21654-9724 410-226-5193 Fax: 410- 226-5925 Sue.Tyler-at-noaa.gov
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 08:31:41 2003
I realize that bone is bone, but I'm trying to differentiate between new bone and old bone in paraffin sections of bone with implants. I'm looking for a method of determining which areas are "old bone" and which are "new" (the new being those areas where bone has started to grow from the insertion of the implant). I have tried basic H&E, 0.5% aq. toluidine blue, and a Van Gieson's method. I thought somebody might have a suggestion, but if not, it was worth a try!
Thanks so much,
Elke Pravda Biostructure Core Facility Forsyth Institute Boston, MA 02115
epravda-at-forsyth.org
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 11:23:57 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (slevinso-at-ic.sunysb.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 2, 2003 at 10:14:53 ---------------------------------------------------------------------------
Email: slevinso-at-ic.sunysb.edu Name: Sarah Zimov
Organization: Stony Brook University
Title-Subject: [Microscopy] MListserver: Ultrastructural localization of Ca
Question: Hello, I was just wondering if anyone was familiar with the oxalate precipitation method or the osmium/potassium-bichromate method for visualizing calcium- at the EM level. Particularly is it important to fix in Glutaraldehyde? I am trying to visualize calcium in an intracellular store in retinal/nervous tissue. If anyone has any suggestions on the best method to use and/or a good working protocol it would be greatly appreciated!!!!!
Sarah Zimov Neurobiology and Behavior Stony Brook University Stony Brook, NY 11794
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (opto-at-klughammer.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 2, 2003 at 10:07:15 ---------------------------------------------------------------------------
Central States Microscopy and Microanalysis Society Fall Meeting: October 24, 2003
The fall meeting of the Central States Microscopy and Microanalysis Society will be held at the Donald Danforth Plant Science Center in St. Louis, on Friday, October 24, 2003. Our keynote speaker will be Dr. James Pawley of the University of Wisconsin Department of Zoology, speaking on "The 3-methods of 3D microscopy: which one to choose". Dr. Pawley's presentation will be sponsored by the Microscopy Society of America Tour Speaker Program and by CSMMS.
We are also soliciting presentations on all aspects of microscopy and microanalysis, including electron microscopy, confocal and fluorescence microscopy, x-ray microanalysis, materials, geological, and biological microscopy applications, immunocytochemistry, and related topics. Students, faculty, and staff are all welcome to make short presentations of about 10-30 minutes in length, and we are planning a competition with cash prizes for best student presentation.
Details on the meeting schedule, as well as directions to the Danforth Plant Science Center and links to information about the Center, can be found at: http://www.danforthcenter.org/imf/csmms/index.htm. As the schedule firms up, more information will be added.
Many thanks to Howard Berg of the Danforth Center and to Heather Ford, previously of the Center, for their efforts at organizing the meeting at this great venue. A tour of the Center will be offered at the meeting. Dr. Berg may be contacted at: RHBerg-at-danforthcenter.org.
Randy Tindall, President Central States Microscopy and Microanalysis Society
Electron Microscopy Core 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/
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 13:51:39 2003
On Thursday, October 2, 2003, at 02:55 AM, Coetzee, Mr S. H Physics Science wrote:
} We are in the process of sorting out the safety of the lab. This was } a question I could not answer. A good website I used was: } http//www.healthcouncil.nl } Dear Mr. Coetzee, OsO4 is very toxic as a vapor, so heating it would, presumably, increase the concentration of vapor, hence increase toxicity. If, however, the fire is in a reducing condition, i.e., there is so much flammable material that oxygen content is very low, then the OsO4 could be reduced to a less toxic compound. I would not base safety considerations on speculation about the state of the fire, but I'd treat heated OsO4 as a very serious safety hazard. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 14:17:28 2003
I posed a similar question recently. I too have had section adhesion issues.
I didn't have a EMS grid coating pen (which I have since purchased) so I tried dissolving different types of tape and adhesive in chloroform (I didn't have any benzene or ethylene dichloride).
My adhesives are untested as for actual section adhesion (maybe next week), but I did view naked grids with a dried drop of adhesive solution under a brightfield/light scope to see how well the grid was coated. I felt that 3-4 Avery Spot-O-Glue tabs transferred to something else (I used a SEM stub) then dissolved in 10 mL chloroform produced the most clean, uniform coating of adhesive.
The pen is a $33 item and a few people suggest it but I think I'm still going to try my home brew to see if it works.
Regards,
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 15:12:15 2003
Sue, I have found that after picking up sections if I put the grids in the 50-60 degree oven for several hours to overnight, the sections always stick. It isn't necessary to clean them first either. (I use the thin bar copper grids from EMS) Good luck, Mary Gail Engle
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
phone 859-323-6108 fax 859-257-9700
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 15:12:59 2003
I don't use this type of equipment, so I can't comment on it, but Gatan sent me some literature a while back with SEM hot stages that I still happen to have around. 3 models: H1001, H1002, H1005 Max temp.: 500, 750, 1500 (deg. C), respectively Hope this helps ------------------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 11:21 AM 10/2/03 -0500, opto-at-klughammer.de wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 15:20:54 2003
Krzysztof Herman wrote: ========================================================== Does somebody know direct contact to manufacturer of good dry boxes for instruments like cameras, lenses, prec, mechanics....
But this is probably from fifth hand.... ====================================================== The Secador line of desiccating cabinets represents an economical yet very highly effective means of storage of delicate optical parts be they parts for LM or even EM instrumentation or cameras. They can be found on URL http://www.2spi.com/catalog/supp/desiccator-cabinets-secador.html
The larger models come with an innovative electrically driven desiccant regeneration system so that one never needs to worry about desiccant replacement.
One of these cabinets should work quite nicely in your intended application depending on how much capacity you require. And how much automation you need. The 220v units are CE certified.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 15:50:48 2003
The Fall meeting of the New England Society for Microsocpy (NESM) will be held on Tuesday, October 14th at the headquarters of Boston Scientific Corporation in Natick, MA.
The meeting will feature dinner and two technical presentations in materials science. Dr. Francesco Stellacci will speak on "The Use of the Scanned Probe Microscope as a Lithographic Tool" and Steve Stokowski will speak on "Black Granite from Hell". Preregistration is strongly encouraged.
For complete program, registration deadline, etc. re: this meeting, please go to NESM's website: http://prism.mit.edu:8083/Newsletters/September2003Newsletter.pdf
Peggy Sherwood Corresponding Secretary, NESM
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 16:13:56 2003
You are invited to participate to a Workshop on Advanced Techniques in Scanning Electron Microscopy & Microanalysis for materials Characterization. That workshop will be held from May 17 to 21, 2004 at McGill University, Montreal, Canada.
The lecturers are:
Raynald Gauvin, McGill University, Canada
Pierre Hovington, Hydro-Québec Research Center, Canada David C. Joy, University of Tennessee and Oak Ridge National Laboratory, USA Marin Lagacé, Hydro-Québec Research Center, Canada Eric Lifshin, State University of New York at Albany, USA
For more information:
http://www.minmet.mcgill.ca/ebeamworkshop
Since there is a limit in the number of participants, please register early if you are interested.
Very best regards
Raynald GAUVIN ************************************************************ Professor Raynald Gauvin Department Mining, Metals and Materials Engineering McGill University M.H. Wong Building 3610 University Street Montreal, H3A 2B2 Canada Tel: (514) 398-8951 FAX: (514) 398-4492 http://www.minmet.mcgill.ca EMAIL:Raynald.Gauvin-at-McGill.ca ************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 22:27:16 2003
Hello all I am interested in following the fate of our biodegradable microparticles over time, when incubated in aqueous solutions such as plasma. We have a laser diffraction particle sizer, but I would like to be able to look at structural changes in wet mounts by microscopy, ideally monitoring particle diameter with good enough statistics to correlate the LM result with the laser diffraction result. I am interested not just in average size, but the distribution of sizes. Thus ideally I'd like to automate the measurements of particle diameter. Our particles are ~ 15 microns diameter on average, encapsulating aqueous solutions, and the particles (being denser than the suspending medium, which must be aqueous and isotonic) sink to the bottom of a wet mount. Just like latex beads they act as lenses in brightfield, so becke lines (bright & dark rings) are visible and vary with the plane of focus, which makes it hard to define the actual particle's edge. Using a low NA objective & closing the iris to maximize depth of field doesn't seem to eliminate the becke lines. There can be a range of sizes of particles present simultaneously in a field, so (at least when using higher NA optics) when I am focused on the equator of a 30 micron particle I am far away from the equator of a 10 micron particle, and vice versa. I may be able to increase the refractive index of the suspending medium to minimize the becke lines, so the bigger problem in my mind is the variation in height (above the surface of the slide) of the particles present.
Because of these considerations, I wonder if anyone has devised a way to accurately size a _mixture_ of, say, 10 um & 30 um calibrated latex beads in water, settled on the slide, by microscopy. It seems tricky to do; I can't imagine doing it with a single image (containing many particles) from a single focal plane. I can imagine 3 ways: 1) autofocus on each bead, find the plane of the equator & image that; 2) take a z-series of a field, for each particle find the slice with greatest diameter and take measurements for that particle only from that slice; or 3) full-on 3D reconstruction & characterization. I would need to accumulate data from many hundreds of particles, but with many particles per field it may not require that many fields. It doesn't seem that MetaMorph, Image-Pro Plus, or ImageJ is up to this level of automation or analysis but I could certainly be mistaken; I'm not familiar with any of these. I would appreciate suggestions for how to approach this, using either conventional or confocal microscopy.
Thanks! Richard
Richard Thrift SkyePharma, Inc 10450 Science Center Drive San Diego, CA, 92121 USA (858) 625-2424
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 3 05:35:06 2003
A simple way to do this is to give tetracycline to the animals in the period which you think the new bone is being formed. The tetracycline will be deposed in the new bone and will be fluorescente in green (FITC filter) under a fluorescent microscopy and you will be able to differentiate the new bone from the old bone.
I wish you good luck. Sincerely Prof. Dr. Francisco Javier Hernandez Blazquez Universidade de São Paulo Fac. de Medicina Veterinária e Zootecnia Departamento de Cirurgia - Setor de Anatomia Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) - Brasil Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
----- Original Message ----- } From: "Pravda, Elke" {epravda-at-forsyth.org} To: {Microscopy-at-msa.microscopy.com} Sent: Thursday, October 02, 2003 10:30 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (karla.schuster-at-delta.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, October 3, 2003 at 13:14:27 ---------------------------------------------------------------------------
Email: karla.schuster-at-delta.com Name: karla schuster
Education: Graduate College
Location: Marietta, GA, USA
Question: I'm thinking of a microscope as a birthday present for my daughter (8 yrs). Can you recommend 2-3 good microscopes?
I've read several sites on that tell you what to look for, but I'm not familiar with good manufacturers.
Since it is pretty clear that that you will not be measuring the same particle every time, is there some reason why you cannot take a "representative" sampling of the particles remove them from the liquid and then image them dry?
Once they are dry then you have a variety of approaches (possibly even low voltage SEM) that might improve things.
Nestor Your Friendly Neighborhood SysOp
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From MicroscopyL-request-at-ns.microscopy.com Sat Oct 4 00:27:21 2003
It isn't a must do now, fast track, immediate deal, but I need to begin assembling a list (particularly in the NE, USA) of qualified confocal service companies, individuals, technicians, and/or contract people capable of entering into a service agreement with a major university on a Zeiss confocal META LSM-510 for a long term contract.
If anyone has any suggestions, referrals, or underpaid or out of work Zeiss confocal tekkies please let me know
Please forward them or refer them to me.
Gratefully,
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 4 06:30:41 2003
I received an answer at my questions about the Brunel microtome. Unlikely I deleted that mail before I read it completely. I remember that he was telling me that he had bought the microtome but he had not jet used it because he have had to finish the wax inclusion. I apologize but could who answer me to re-mail his answer. Thank you, Best Regards,
Massimo T.
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 4 18:05:35 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (florea-lupu-at-omrf.ouhsc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, October 4, 2003 at 09:07:29 ---------------------------------------------------------------------------
Organization: Oklahoma Medical Research Foundation
Title-Subject: [Microscopy] Techni G2 12TWEEN vs Hitachi H7600
Question: We are interested to buy a new electron microscope and we have short listed two instruments: Techni G2 12TWEEN and Hitachi H7600. I would appreciate opinions about the reliability and performances of these microscopes from colleagues who used both instruments (not sales representatives!) A swift response would be very helpful,
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kolb-at-uni-mainz.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, October 5, 2003 at 13:18:52 ---------------------------------------------------------------------------
Dear Sarah, Oxalate precipitation method is quite good for visualizing calcium. You have to fix the tissue no matter what you use- glut or paraformaldehyde. You can only avoid osmium. The protocol is-- Infuse or perfuse your tissue with oxalate sol and fix it with GLu or paraformaldehyde. Dehydrate and embedd in resin. cut section and scan grids without staining with UA and L Citrate. with regards Shashi CCMB, Hyderabad INDIA
--- by way of MicroscopyListserver {slevinso-at-ic.sunysb.edu} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form } (NJZFM-ultra-55). It was submitted by } (slevinso-at-ic.sunysb.edu) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html } on Thursday, October 2, 2003 at 10:14:53 } --------------------------------------------------------------------------- } } Email: slevinso-at-ic.sunysb.edu } Name: Sarah Zimov } } Organization: Stony Brook University } } Title-Subject: [Microscopy] MListserver: } Ultrastructural localization of Ca } } Question: Hello, } I was just wondering if anyone was familiar with the } oxalate precipitation method or the } osmium/potassium-bichromate method for visualizing } calcium- at the EM level. Particularly is it } important to fix in Glutaraldehyde? I am trying to } visualize calcium in an intracellular store in } retinal/nervous tissue. If anyone has any } suggestions on the best method to use and/or a good } working protocol it would be greatly } appreciated!!!!! } } Sarah Zimov } Neurobiology and Behavior } Stony Brook University } Stony Brook, NY 11794 } } --------------------------------------------------------------------------- }
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 08:32:23 2003
Hello everyone, I've recently inherited a LKB glass knife maker. I've used these beast before, but not the Type 7801B. If anyone has a set of instructions for this unit, would you please contact me and fax me a copy?
Thanks!!!!
Frank Karl Degussa Corp. frank.karl-at-degussa.com Fax 330-668-3846
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 09:42:17 2003
At 01:53 PM 10/5/2003 -0500, you wrote: } Email: kolb-at-uni-mainz.de } Name: Ute Kolb } } Organization: University Mainz Germany } } Title-Subject: [Microscopy] MListserver: HAADF image quantification } } Question: Dear Electronmicroscopists, } } I wonder if anyone has tried to quantify HAADF images and if there is } anything published on this subject.
There certainly is. The earliest papers on ADF-STEM by Albert Crewe (Science 168, 1338 (1970)) contain attempts at quantification. More recently, M. M. J. Treacy and Rice have quantified the contrast of atoms and small particles on a support (J. Microscopy 156, 211 (1989)). And in a series of several papers, the Silcox group at Cornell has studied quantification of high-resolution HAADF images of zone axis crystals. Here's a more or less random collection of papers:
Hillyard, S. E. & Silcox, J. (1995). Detector geometry, thermal diffuse scattering and strain effects in ADF STEM imaging. Ultramicroscopy 58, 6-17.
Kirkland, E. J., Loane, R. F. & Silcox, J. (1987). Simulation of annular dark field STEM images using a modified multislice method. Ultramicroscopy 23, 77-96.
Loane, R. F., Kirkland, E. J. & Silcox, J. (1988). Visibility of single heavy atoms on thin crystalline silicon in simulated annular dark field. Acta. Cryst. A44, 912-927.
Loane, R. F., Xu, P. & Silcox, J. (1991). Thermal vibrations in convergent-beam electron diffraction. Acta. Cryst. A47, 267-278.
Muller, D. A., Edwards, B., Kirkland, E. J. & Silcox, J. (2001). Simulation of thermal diffuse scattering including a detailed phonon dispersion curve. Ultramicroscopy 86, 371-380.
My own recent efforts in that direction (Voyles, Muller, & Kirkland, Ultramicroscopy 96, 251 (2003) and to appear in the December issue of Microscopy & Microanalysis) are a direct extension of the Silcox results.
Best wishes, Paul Voyles
Paul Voyles Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 09:44:16 2003
The oxalate precipitation method does show calcium deposits but one has no way of assessing their physiological relevance. Oxalate doesn't precipitate the calcium until it is inside the cell and it doesn't get inside the cell until it is fixed by the aldehydes (which takes many seconds to many minutes depending on the tissue, fixative and cellular location). This allows massive re-distribution of calcium. The only valid way to look at calcium distribution in fixed tissues is in quick-frozen (high pressure freezing or metal mirror slam freezing) followed by freeze-drying. Freeze-substitution might be an acceptable alternative depending on the solvent and processing conditions. Even with this rigorous approach, redistribution can occur if you cut your thin sections on a water medium; sectioning on glycerol can help (see Dudek & Boyne 1986 Am J Anat 175:217). It is not a trivial problem to localize physiological ions in fixed cells.
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I came across a problem concerning some specific sample preparation for TEM. The samples include (1) Precipitated Silica, (2) Silica sol and (3) Silica dispersed in polystyrene. As I am more familiar with only biological sample preparation, I would really appreciated your help in this regards.
Thanking you in advance.
Jos Nongkynrih
Sophisticated Analytical Instrumentation Facility
North Eastern Hills University
Bijni complex
Shillong 793003
Meghalaya, India
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 13:47:08 2003
Hi, I've just been informed by JOEL that the JOEL field service engineer in our area may be tied up with a major new installation for the next 2 months. I have a JOEL6300 SEM that I'm hoping to have installed in a newly renovated lab beginning in a about two or three weeks. Are there any independent SEM service engineers out there that are qualified to install JOEL SEM's, particularly the 6300/6400 series?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 15:45:15 2003
On Thu, 2 Oct 2003, Mary Gail Engle wrote: } I have found that after picking up sections if I put the grids in the 50-60 } degree oven for several hours to overnight, the sections always stick. It } isn't necessary to clean them first either. (I use the thin bar copper } grids from EMS)
I use the same grids and find that 10 minutes in my 60-65 degree oven is long enough to do the trick to help in adhesion.
Karen Bovard Creighton University Medical Center Department of Pathology Omaha, Nebraska
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 18:50:38 2003
Another factor that affects section adhesion is the method in which the sections are picked up. If the sections are picked up by immersing the grid and coming from underneath, or picked up in a loop and placed on top of the grid, they adhere much better than if the grid is pushed down on them from above.
Ralph Common Electron Microscopist Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824 517-355-7558; fax 517-432-1053 ralph.common-at-ht.msu.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 20:06:21 2003
PLEASE CONTACT ME IMMEDIATELY IF YOU HAVE EXPERIENCE SERVICING THE
ZESS LSM-510 META CONFOCAL MICROSCOPE.
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 21:26:30 2003
Our clients, a college of medicine and a Zeiss LSM service and repair facility have asked us to screen potential candidates to work in the Columbia, Maryland area as a service and repair technician for the Zeiss LSM-510 META Microscope being placed in a college of medicine in the greater New York City area.
We are looking for, on behalf of our client, a fully trained and qualified service and repair technician familiar with all aspects of the maintenance, repair, software, installation and upkeep of the LSM-510.
The screening is being done by AssetRecovery.Net, Inc. due to the fact that we are providing a; what is a, now fully functional, scope to our buyer and as a part of that agreement we need to locate with the intention of recruiting an LSM-510 level service and repair technician to the service company being asked to deliver a service contract to the School for this scope.
All qualified applicants will be immediately referred to the service contractor for final evaluation.
Duties include operation, maintenance, installation and upgrading of light microscope instrumentation; utilization of associated digital imaging system; specimen preparation; and methods development.
This individual will manage a centralized LSM (currently, a new Zeiss LSM 510) located in the New York area, and will provide confocal services, consultation, training, and access for user operation to researchers and other Zeiss qualified LSM service technicians.
Please submit resume to:
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 21:27:34 2003
Our clients, a college of medicine and a Zeiss LSM service and repair facility have asked us to screen potential candidates to work in the Columbia, Maryland area as a service and repair technician for the Zeiss LSM-510 META Microscope being placed in a college of medicine in the greater New York City area.
We are looking for, on behalf of our client, a fully trained and qualified service and repair technician familiar with all aspects of the maintenance, repair, software, installation and upkeep of the LSM-510.
The screening is being done by AssetRecovery.Net, Inc. due to the fact that we are providing a; what is a, now fully functional, scope to our buyer and as a part of that agreement we need to locate with the intention of recruiting an LSM-510 level service and repair technician to the service company being asked to deliver a service contract to the School for this scope.
All qualified applicants will be immediately referred to the service contractor for final evaluation.
Duties include operation, maintenance, installation and upgrading of light microscope instrumentation; utilization of associated digital imaging system; specimen preparation; and methods development.
This individual will manage a centralized LSM (currently, a new Zeiss LSM 510) located in the New York area, and will provide confocal services, consultation, training, and access for user operation to researchers and other Zeiss qualified LSM service technicians.
Please submit resume to:
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 08:05:45 2003
Dear TEM friends, Thank you so much for all of the grid adhesion suggestions I can now write a book! I will give them all a try and let you know what works for me. Thank you, Sue -- Sue Tyler Biologist Cooperative Oxford Laboratory Center for Coastal Environmental Health& Biomolecular Research at Charleston ( CCHEBR) USDOC/NOAA/NOS/NCCOS 904 S. Morris St. Oxford, Maryland 21654-9724 410-226-5193 Fax: 410- 226-5925 Sue.Tyler-at-noaa.gov
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 08:59:55 2003
I have the following position available in St. Louis MO. It is a one year contract position. Position is with a major Bio-Tech firm.
The contract researcher will be responsible for the in-house transmission electron microscopy experiments on nanophase materials/heterogeneous catalysts, including acquiring proper TEM images, diffraction data, and spectra as well as the interpretation of these experimental results in relation to catalyst structure, performance, and synthesis parameters/methodologies. The selected candidate will interact with multifunctional groups of scientists working on various materials/catalysts projects. This is a post-doctoral position.
Thank you,
Dennis Madden 314-432-5833 800-351-5369 Q & C International, Inc. 11330 Olive Boulevard Suite 218 St. Louis, MO 63141 dmadden-at-qcintl.com http://www.qcintl.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 10:05:54 2003
The University of Wisconsin Materials Science Center is looking for a staff member to see to the care and feeding of our TEMs and TEM users. UW is a great place to work, and Madison is a great place to live (if you don't mind cold), so I encourage qualified people to apply. The official ad with all the details is given below.
If you would like to apply, DO NOT reply to this email. Instead, contact Rick Noll (noll-at-engr.wisc.edu, or the information given below).
Sincerely, Paul Voyles --------------
The Materials Science Center at the University of Wisconsin invites applications for an electron microscopist to oversee its transmission electron microscopy facility. The facility includes high resolution and energy filtered imaging, STEM, EDS, EELS, and sample preparation. It supports a broad range of research by students, staff, and faculty with interests in materials characterization. The candidate's responsibilities will include user training and assistance, instrument maintenance, and instrument development.
Candidates must have a strong background in the theory and practice of TEM microcharacterization of materials with a minimum of 2 years post-graduate experience. The candidate should also have a strong background in vacuum technology, electronics, and computers. Strong interpersonal and communications skills are essential. Candidates should have an MS (PhD preferred) in a physical science or engineering discipline. Candidates with a BS in physical science or engineering with professional experience and knowledge in the required field equivalent to an MS will also be considered. Candidates are encouraged to view additional information at http://www.ohr.wisc.edu/pvl/pv_045842.html.
Interested candidates should send a resume and cover letter referring to Position Vacancy Listing #45842 to Richard Noll, 1509 University Ave., Madison, WI 53706, or email to noll-at-engr.wisc.edu.
To ensure full consideration, all information should be received by November 30, 2003. Note that unless confidentiality is requested in writing, information regarding the names of applicants must be released upon request. Finalists cannot be guaranteed confidentiality.
UW-Madison is an equal opportunity/ affirmative action employer. We promote excellence through diversity and encourage all qualified individuals to apply.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 10:10:30 2003
Hello all, A friend recently acquired a dinosaur of a cryostat and is interested in trying to find a manual or someone with knowledge of the instrument. Any help would be appreciated. All relevant info is below and responses may be addressed directly to him, or I will forward if posted on the list. Thanks much, Jay
} International Equipment Company (IEC) } Needham Heights, Mass. } Model CTD - International - Harris - Cryostat } } Model CTD serial No. 69835M-5 } H.P. 1/5 Volts 115 AC 60 cycles } 4 amps } Refrigerant R-12 10 oz. } } International minot custom microtome } No. 69835M - 5 } } Please see what information you can find on this equipment and if you } know someone that we could hire to come in here and help us set it up. } } Thanks, } } Steven Hahn } Materials Engineer } Kelch Corporation } A Bemis Manufacturing Company } N85 W12545 Westbrook Crossing } P.O. Box 9025 } Menomonee Falls, WI 53052-9025 } Phone: 262.257.7029 } Fax: 262.257.7128 } email: stevehahn-at-kelch.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 10:35:17 2003
Does anyone have an Olympus BX51 with a phototube? We don't around here, and the Olympus information is unclear as to: mount, C-mount or bayonet? at the top, and if there is a projection/transfer lens within the tube, and if so (there must be), where exactly and what power? 1? 0.32? etc. Thanks.
Phil
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 11:03:17 2003
We need a double tilt holder for our JEM-100CXII. If you want to sell your one or give it away, or if you know the website on which I can look for the used TEM stuffs, please let me know. I remembered that someone mentioned the website for second-hands TEM Stuffs, but I forgot to keep it. Many thanks.
Regards, Qi
*************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu qizhang-at-email.unc.edu
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From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 12:46:08 2003
Howdy. I recently began a new job in which I am the scientist responsible for our TA Instruments micro-TA 2990 AFM. I have never used an AFM before, let alone this particular one (my background is in STEM/TEM), so I don't know how or where to begin. I have been instructed by the person previously in this position (at a new company now) not to "play" with the AFM until I have been trained, which won't be until first quarter 2004. We will need to use this equipment in the next three months, so it is imperitive that I get up to speed on this technique.
Does someone have experience with this particular AFM that might be able to help me get started? Alternatively, I could arrange to visit a relatively local lab (I am in Union Beach, NJ, generally in the NYC metro area) that has a micro-TA for at least rudimentary instruction on aligning the system and such. As a third option, I could buy lunch for a user able to come here to get me started.
Anyone who can assist, I look forward to hearing from you. Thanks.
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Materials Characterization Lab International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 16:49:23 2003
The BX51 with a trinoc head has a female dovetail port for photos. Depending on what type of camera you want to use, that will determine the camera port adapter that you would need.
If you use a SLR bayonet (Nikon, etc.) camera, then there is no photo eyepiece (phototube). If you use the old or new PM-series film camera systems, they mount directly and require a photo eyepiece. These are from 2.5, 3.3, 4 and 5X. 3.3 is the normal mag photo eyepiece. If your camera is C-mount and is a typical 1/2" CCD, then get the Olympus U-TVO.5XC adapter. It allows setting parfocality. Other types of adapters for different cameras may also require a photo eyepiece.
What type of camera are you going to use?
gary g.
At 08:33 AM 10/7/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 18:14:12 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (evdargelis-at-esi-il.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 7, 2003 at 16:48:50 ---------------------------------------------------------------------------
Email: evdargelis-at-esi-il.com Name: Ed Dargelis
Organization: Engingeering Systems, Inc., 3851 Exchange Ave., Aurora, IL 60504
Title-Subject: [Microscopy] MListserver: SEM Impression Materials
Question: Hello Listers, I'm interested to know if anyone has experience using dental impression materials (vinyl polysiloxanes) or mold/impression brands such as Struers or GE for replicating surfaces for SEM examination. Thanks kindly for your time and info.
Ed Dargelis evdargelis-at-esi-il.com
Engineering Systems, Inc. 3851 Exchange Ave. Aurora, IL 60504
At last I have finished to build my hand microtome. It works quite well, although the slices are not so thin as I expected. I think I have to recover the clearance between the cylinder, moved by a micrometer screw, and its seat. For cutting IŽm using an USA razor blade. But all that concerns the improvement of the device. For testing it, I tried to cut a little wax block without speciemen included. The trouble is that the thin wax film rolls up when the blade goes forward in the cutting direction. The microtome worked at room temperature, about 20°C. The lukewarm water doesn't stretch the wax roll, and if I try to open it with needles the wax crumbles. So I wonder if someone could give me some suggestion to overcame this problem. Thank you. Best Regards,
Massimo T.
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 10:06:01 2003
I have used the polymer molding materials extensively for making replicas of surfaces for microscopic examination. The Struers material is very easy to use and makes an excellent reproduction down to submicron features. You can even make a good positive image by making a replica of the replica with this material.
I have also used a product called ReproRubber, which is marketed to tool makers. This material is much cheaper than the Struers product, so may be preferable for replicating larger areas/volumes. I have done comparisons of the surface features on the replicas from this material to the original surface using SEM and quantitative surface profiling with interferometry. Comparisons were good at least to the micron level, but I did not evaluate smaller features. Hope this helps.
One disadvantage of these silicone materials is vacuum compatibility. Even moderately large replicas may outgas excessively for observation in high vacuum instruments (FESEM at 10-7 Torr), but I have not had a problem at standard SEM vacuum (10-5 Torr).
--
Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
} Question: Hello Listers, } I'm interested to know if anyone has experience using dental impression materials (vinyl polysiloxanes) or mold/impression brands such as Struers or GE for replicating surfaces for SEM examination. } Thanks kindly for your time and info. } } Ed Dargelis } evdargelis-at-esi-il.com } } Engineering Systems, Inc. } 3851 Exchange Ave. } Aurora, IL 60504
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 10:57:31 2003
Yes, I have a great deal of experience in this. What did you want to know?
Lesley Weston.
n 07/10/2003 4:12 PM, by way of MicroscopyListserver at evdargelis-at-esi-il.com wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted } by (evdargelis-at-esi-il.com) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October } 7, 2003 at 16:48:50 } --------------------------------------------------------------------------- } } Email: evdargelis-at-esi-il.com } Name: Ed Dargelis } } Organization: Engingeering Systems, Inc., 3851 Exchange Ave., Aurora, IL } 60504 } } Title-Subject: [Microscopy] MListserver: SEM Impression Materials } } Question: Hello Listers, } I'm interested to know if anyone has experience using dental impression } materials (vinyl polysiloxanes) or mold/impression brands such as Struers or } GE for replicating surfaces for SEM examination. } Thanks kindly for your time and info. } } Ed Dargelis } evdargelis-at-esi-il.com } } Engineering Systems, Inc. } 3851 Exchange Ave. } Aurora, IL 60504 } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 13:35:20 2003
} I am a new user in EM and I am having a difficult time getting my Epoxy } sections to stick to the copper grids. I have tried Acetic acid washes, } sonicators, HCl wash etc. I have not tried BSA, I am waiting for the } order to come in. Does anyone have any more suggestions? } } Thank you in Advance. } Sue
Here's my methodology for collecting epoxy sections onto mesh grids and getting them to stay there. Works very well 99% of the time:
I always clean, by sonication for 30-60 seconds in dedicated clean 25ml glass beaker, my uncoated copper mesh grids (no support films, like Formvar) in the following solution:
10% concentrated HCl 20% acetone 70% distilled water
Note: Be careful mixing this up. Add HCl slowly to water, then add acetone. Store in clean capped glass bottle. pH paper tests gives pH =1.0.
After sonication in cleaning solution, rinse 2x in 99% acetone, air dry, invert beaker over clean filter paper and they will fall down as acetone rinse drys.
Always clean grids the day you section. Even after overnight, I find that some sections might tend to loosen up a bit on copper grids during staining procedures, I guess due to new oxidation layer forming within a day or two (rust never sleeps!).
I think the HCl removes oxidation layers from the grids, acetone breaks the surface tension of the aqueous solution so grids don't float, and also cleans, and facilitates quick air drying.
Also, during staining, minimize the turbulance of stains and rinses over section surfaces on grids. For that reason, I like to use the SynapTek(tm) GridStick(tm) pipet method from Ted Pella (I have no financial interest in Pella, just a satisfied user of GridStick system).
I prefer to pick up sections from the knife's water filled trough from above, rather than to slide grid underneath sections (underwater) and lift up. For me, collecting from above allows me to place a group of sections right where I want them on the grid, without them sliding off to one side or wrapping around the grid edge. Also, just as I place grid over sections, I press very slightly down, but not nearly enough to break surface of water, which resultant slight water pressure seems to push the sections onto grid bars so they stay put. Then after lifting grid off water surface, I blot the edge of the grid with a filter paper point to remove that drop of water that is hanging from the grid, air dry, or dry over slide warmer surface, but not too hot. Be sure to give adequate drying before you move on to the staining.
But I think the crucial step is the cleaning of the grids using above solution same day as they are used to collect sections. I've been using this method for many years with consistent success. Hope this helps you, and good luck!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra } -- } Sue Tyler } Biologist } Cooperative Oxford Laboratory } Center for Coastal Environmental Health& } Biomolecular Research at Charleston ( CCHEBR) } USDOC/NOAA/NOS/NCCOS } 904 S. Morris St. } Oxford, Maryland 21654-9724 } 410-226-5193 Fax: 410- 226-5925 } Sue.Tyler-at-noaa.gov } I am a new user in EM and I am having a difficult time getting my Epoxy } sections to stick to the copper grids. I have tried Acetic acid washes, } sonicators, HCl wash etc. I have not tried BSA, I am waiting for the } order to come in. Does anyone have any more suggestions? } } Thank you in Advance. } Sue
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 15:41:40 2003
We are setting up to develop Kodak SO163 film for low dose cryoEM. We are push processing it, so we are developing it for 12 minutes in straight D-19, we're using a nitrogen gas burst, for 1 second every 8 seconds.
I was wondering what the life of the chemistry might be. Kodak said that we could develop 60 8x10 negatives per gallon. This works out to about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to us, so we are wondering what other people are using for an exhaustion time, both in negative numbers and days in the tank.
Thanks for the help. Leslie Cummins
Leslie Gunther Cummins Analytical Imaging Facility Albert Einstein College of Medicine 1300 Morris Park Ave. Bronx, NY 10461 718-430-3547
http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 19:07:02 2003
Massimo- We put a water droplet on the slide. There is a device that heats the slide up to about 45 degrees C. When the wax roll is put onto the rounded-up surface of the droplet, it gradually unfolds and stretches out to occupy the greatest possible area on the droplet. Carol Heckman (Bowling Green State University)
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 21:17:15 2003
On Wednesday, October 8, 2003, at 10:55 AM, Leslie Cummins wrote:
} We are setting up to develop Kodak SO163 film for low dose cryoEM. We } are push processing it, so we are developing it for 12 minutes in } straight D-19, we're using a nitrogen gas burst, for 1 second every 8 } seconds. } } I was wondering what the life of the chemistry might be. Kodak said } that we could develop 60 8x10 negatives per gallon. This works out to } about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to } us, so we are wondering what other people are using for an exhaustion } time, both in negative numbers and days in the tank. } } Thanks for the help. } Dear Leslie, We've done 250 in the past, and that is our plan, if we have as many as 250 films in a short period--a week or two. If the solution is older that two weeks, we'll pitch it. You can also check by color; light brown is fresh, reddish orange is OK, and when the color intensifies to dark reddish brown, it's bad. We have covered tanks, and the developer is pretty well nitrogenated after use, but, if your tanks allow oxygen to dissolve in the developer over time, YMMV. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 23:43:41 2003
You can check out our website for the various Olympus trinocular ports: http://www.mvia.com/Coolpix/clpxadpt.htm#Olympus
As far as the mount that goes on top, it would be dictated by the camera you are using, but we have C-mount F-mount, bayonet mount, digital camera mounts, etc. that will all fit onto your scope.
Feel free to give me a call or email me if you need any further info.
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Philip Oshel wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Micromavens, } } Does anyone have an Olympus BX51 with a phototube? We don't around } here, and the Olympus information is unclear as to: mount, C-mount or } bayonet? at the top, and if there is a projection/transfer lens } within the tube, and if so (there must be), where exactly and what } power? 1? 0.32? etc. } Thanks. } } Phil } } -- } Philip Oshel } Supervisor, BBPIC microscopy facility } Department of Animal Sciences } University of Wisconsin } 1675 Observatory Drive } Madison, WI 53706 - 1284 } voice: (608) 263-4162 } fax: (608) 262-5157 (dept. fax)
--
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 04:21:26 2003
Ed A couple of artefacts are worth looking out for. Delicate surface structures can sometimes be stripped from the specimen surface when the silicone replica is removed. For example, silicone replicas will strip the epicuticular wax layer from plant surfaces. When this happens, direct examination of the uncleaned negative replica in the SEM may falsely suggest that the replica has failed to record the finest structures. Similarly, an epoxy resin casting will record the fracture surface at the base of the stripped structures, and not the original outer surface structure, again suggesting poor replica performance. If this happens to you, consider the chemistry of the stripped material and consider whether it can be removed using a solvent. in the case of epicuticular waxes, I have found that fidelity of the positive replica is improved if a hot-melt method is used, because the wax embedded in the silicone is melted and displaced by hot plastic. Polycarbonate plastic (melting point ~200oC) is good for this.
A second problem can arise if the specimen surface is pitted or very rough, especially if subsurface pits open to the surface via restricted apertures smaller than the pits themselves. An example would be the open stomatal apertures of plants, but a percentage of the cavities in foamed materials might also fit this model. In this case, the silicone compound may freely enter the cavities while fluid, but once polymerized will form a solid plug that may break off at the narrowest point of the neck when the replica is stripped. This may destroy evidence of the aperture, leading to underestimates of porosity or surface roughness.
Chris
} } Email: evdargelis-at-esi-il.com } Name: Ed Dargelis } } Organization: Engingeering Systems, Inc., 3851 Exchange Ave., Aurora, IL } 60504 } } Title-Subject: [Microscopy] MListserver: SEM Impression Materials } } Question: Hello Listers, } I'm interested to know if anyone has experience using dental impression } materials (vinyl polysiloxanes) or mold/impression brands such as Struers or } GE for replicating surfaces for SEM examination. } Thanks kindly for your time and info. } Ed Dargelis } evdargelis-at-esi-il.com } } Engineering Systems, Inc. } 3851 Exchange Ave. } Aurora, IL 60504 --------------------------------------------------------------------------
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 07:17:36 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nicholas.pearson-at-alcoa.com.au) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 9, 2003 at 00:24:31 ---------------------------------------------------------------------------
Email: nicholas.pearson-at-alcoa.com.au Name: Nick Pearson
Organization: Alcoa World Alumina
Title-Subject: [Microscopy] Video output to data projector
Question: I am trying to take the video output from a JEOL JSM 6400 and project it using a data projector (for a community display). The service engineer indicates that the signal is NTSC format, but having tried 4 projectors that are capable of detecting NTSC have not had any luck. 3 of the projectors indicate video signal not detected, and the other displays an unstable image. Has anybody else tried this on a JEOL instrument? Any suggestions?
(Another observation - Feeding the video signal to a monitor capable of displaying NTSC, the image displays momentarily (very small fraction of a second), then goes blank).
12 minutes in straight D-19 sounds like a huge push to me! On the other hand, I have no experience with cryo EM so ... Have you talked to Kodak about this? Long times in developer can lead to fogging. As to life of the chemistry in a tank. If you have a floating lid (that excludes most air) the chemistry will keep better than if the lid allows free access to air. Again, a call to Kodak is in order. As for the number of films I always stay on the conservative side, maybe 2/3 of the recommendation. D-19 is the cheapest part of the equation (animals, prep time, etc) so trying to squeez a few extra films out of a pot of developer is not the way to go. Also, D-19 does not last more than 2 years in the package on the shelf. I don't do much TEM anymore so I mix my D-19 from scratch using the individual chemicals, not the prepackaged Kodak mix. The recipe is widely published, Kodak will give it to you if you ask.
Geoff
Leslie Cummins wrote:
} Hello Listers } } We are setting up to develop Kodak SO163 film for low dose cryoEM. We } are push processing it, so we are developing it for 12 minutes in } straight D-19, we're using a nitrogen gas burst, for 1 second every 8 } seconds. } } I was wondering what the life of the chemistry might be. Kodak said } that we could develop 60 8x10 negatives per gallon. This works out to } about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to } us, so we are wondering what other people are using for an exhaustion } time, both in negative numbers and days in the tank. } } Thanks for the help. } Leslie Cummins } } } Leslie Gunther Cummins } Analytical Imaging Facility } Albert Einstein College of Medicine } 1300 Morris Park Ave. } Bronx, NY 10461 } 718-430-3547 } } http://www.aecom.yu.edu/aif/ } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 08:33:18 2003
I've never used SO163, but with the Kodak 4489 film (yes, I still have some left over!), our rule of thumb: 100 negatives and/or 2 weeks whichever comes first. Then we discard the D-19 in tank and refresh with new. (When we make it up, it usually makes more than a tank holds, so we store the rest in a brown (light-free) bottle. For use: mix 1 part D-19:2parts H20. Hope this helps. Note: the stock solution should be good until it gets brown--it should be relatively "clear".
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Leslie Cummins [mailto:gunther-at-aecom.yu.edu] Sent: Wednesday, October 08, 2003 1:55 PM To: Microscopy-at-MSA.Microscopy.Com
Hello Listers
We are setting up to develop Kodak SO163 film for low dose cryoEM. We are push processing it, so we are developing it for 12 minutes in straight D-19, we're using a nitrogen gas burst, for 1 second every 8 seconds.
I was wondering what the life of the chemistry might be. Kodak said that we could develop 60 8x10 negatives per gallon. This works out to about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to us, so we are wondering what other people are using for an exhaustion time, both in negative numbers and days in the tank.
Thanks for the help. Leslie Cummins
Leslie Gunther Cummins Analytical Imaging Facility Albert Einstein College of Medicine 1300 Morris Park Ave. Bronx, NY 10461 718-430-3547
http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 08:52:24 2003
We are currently changing the D19 in our low-dose darkroom after 280 films are processed and this usually happens within 3 weeks of preparation.
On a side note...has anyone ever come across a batch of D19 with brown crystals? It appeared very dark once prepared and was thrown out without attempting to use it.
cheers, paul
} } } } On Wednesday, October 8, 2003, at 10:55 AM, Leslie Cummins wrote: } } } We are setting up to develop Kodak SO163 film for low dose cryoEM. } } We are push processing it, so we are developing it for 12 minutes } } in straight D-19, we're using a nitrogen gas burst, for 1 second } } every 8 seconds. } } } } I was wondering what the life of the chemistry might be. Kodak } } said that we could develop 60 8x10 negatives per gallon. This } } works out to about 360 3 1/4 x 4 negatives. This seems like a lot } } of negatives to us, so we are wondering what other people are using } } for an exhaustion time, both in negative numbers and days in the } } tank. } } } } Thanks for the help. } } } Dear Leslie, } We've done 250 in the past, and that is our plan, if we have } as many as 250 films in a short period--a week or two. If the } solution is older that two weeks, we'll pitch it. You can also } check by color; light brown is fresh, reddish orange is OK, and when } the color intensifies to dark reddish brown, it's bad. We have } covered tanks, and the developer is pretty well nitrogenated after } use, but, if your tanks allow oxygen to dissolve in the developer } over time, YMMV. } Yours, } Bill Tivol } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 } tivol-at-caltech.edu
-- Paul Chipman Electron Microscopy Facility Manager Dept. of Biology, Purdue University Lilly Hall, Rm. B216 Phone: 765-494-1487 Fax:765-496-1189
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 08:56:20 2003
} I am a new user in EM and I am having a difficult time getting my Epoxy } sections to stick to the copper grids. I have tried Acetic acid washes, } sonicators, HCl wash etc. I have not tried BSA, I am waiting for the } order to come in. Does anyone have any more suggestions? } } Thank you in Advance. } Sue
Hi Sue, With the technique that we use, no known force in the universe will be able to remove the sections from your grids.
First of all, use clean grids. There are several ways to clean grids, but the way that I like, because it is fast, is to hold the grid in concentrated NaOH while I count to 6, and then briefly dip it repeatedly in water to rinse the grid. I do this just before I pick up the sections.
Then, most importantly, we dry down our grids in a drying oven. The oven is just hot enough to dry glassware, but not really superhot. In other words, I can put my hand on the bottom of the oven without burning it, but I really wouldn't want to keep my hand there for more than a second or so. And the temperature isn't hot enough to warp our plastic petri dishes. If it starts to warp the petri dishes, it's too hot, though it still probably won't hurt your sections. [we know that from experience] You can dry your sections there for about 5-10 minutes. But if you even forget about them overnight, it won't hurt them at all.
Then, I can assure you that your sections will NEVER NEVER NEVER wash off your grids no matter how vigorously you wash them during staining.
Garry
Garry Burgess Charge Technologist Electron Microscopy Department of Pathology Health Sciences Centre Winnipeg, Canada
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 08:59:43 2003
Forgive me if you've covered this possibility already but we once had some difficulty getting video out of our 6400 under similar circumstances. We eventually realised that it was because we had it synced to the line frequency (to null out the wobbles from room fields etc) and not the "standard" rate. Try toggling this feature on and off and see if it makes any difference. That is, at the bottom of the FIS-1 Functions menu (normally brought up with the F3 key), you will see the VIDO on/off item. With the scan going at TV rate scroll down and toggle the VIDO setting on and off and see if one of the settings (presumably the one that gives video operating at the conventional sync rate) results in a signal that your data projector can understand.
Good luck.
} Email: nicholas.pearson-at-alcoa.com.au } Name: Nick Pearson } } Organization: Alcoa World Alumina } } Title-Subject: [Microscopy] Video output to data projector } } Question: I am trying to take the video output from a JEOL JSM 6400 } and project it using a data projector (for a community display). } The service engineer indicates that the signal is NTSC format, but } having tried 4 projectors that are capable of detecting NTSC have } not had any luck. 3 of the projectors indicate video signal not } detected, and the other displays an unstable image. } Has anybody else tried this on a JEOL instrument? } Any suggestions? } } (Another observation - Feeding the video signal to a monitor capable } of displaying NTSC, the image displays momentarily (very small } fraction of a second), then goes blank). } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 09:05:44 2003
Forgive me if you've covered this possibility already but we once had some difficulty getting video out of our 6400 under similar circumstances. We eventually realised that it was because we had it synced to the line frequency (to null out the wobbles from room fields etc) and not the "standard" rate. Try toggling this feature on and off and see if it makes any difference. That is, at the bottom of the FIS-1 Functions menu (normally brought up with the F3 key), you will see the VIDO on/off item. With the scan going at TV rate scroll down and toggle the VIDO setting on and off and see if one of the settings (presumably the one that gives video operating at the conventional sync rate) results in a signal that your data projector can understand.
Good luck.
} Email: nicholas.pearson-at-alcoa.com.au } Name: Nick Pearson } } Organization: Alcoa World Alumina } } Title-Subject: [Microscopy] Video output to data projector } } Question: I am trying to take the video output from a JEOL JSM 6400 } and project it using a data projector (for a community display). } The service engineer indicates that the signal is NTSC format, but } having tried 4 projectors that are capable of detecting NTSC have } not had any luck. 3 of the projectors indicate video signal not } detected, and the other displays an unstable image. } Has anybody else tried this on a JEOL instrument? } Any suggestions? } } (Another observation - Feeding the video signal to a monitor capable } of displaying NTSC, the image displays momentarily (very small } fraction of a second), then goes blank). } } ---------------------------------------------------------------------------
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/
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 11:12:54 2003
} I've recently inherited a LKB glass knife maker. I've used these beast } before, but not the Type 7801B. If anyone has a set of instructions for } this unit, would you please contact me and fax me a copy? } } Frank Karl } Degussa Corp. } frank.karl-at-degussa.com } Fax 330-668-3846 ----- Frank, I have a copy of the instructions and will fax them shortly.
Patricia Stranen Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 11:40:42 2003
We use 2 parts water, 1 part full strength D-19 and develop using 1 second nitrogen burst every 8 seconds for 10 minutes. With this recipe we get about 400 micrographs per gal. Incidentally, we have more problems with cross contamination of solutions than chemical exhaustion. Therefore we test the developer solution before each run using a test sheet of kodabromide print paper by immersing for 2 minutes to check the paper then lights on and back into the developer. The paper should turn black relatively quickly if the solution is good.
Fran Laabs
Ames Lab, ISU
On Wednesday, October 8, 2003, at 10:55 AM, Leslie Cummins wrote:
We are setting up to develop Kodak SO163 film for low dose cryoEM. We are push processing it, so we are developing it for 12 minutes in straight D-19, we're using a nitrogen gas burst, for 1 second every 8 seconds.
I was wondering what the life of the chemistry might be. Kodak said that we could develop 60 8x10 negatives per gallon. This works out to about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to us, so we are wondering what other people are using for an exhaustion time, both in negative numbers and days in the tank.
Thanks for the help.
Dear Leslie, We've done 250 in the past, and that is our plan, if we have as many as 250 films in a short period--a week or two. If the solution is older that two weeks, we'll pitch it. You can also check by color; light brown is fresh, reddish orange is OK, and when the color intensifies to dark reddish brown, it's bad. We have covered tanks, and the developer is pretty well nitrogenated after use, but, if your tanks allow oxygen to dissolve in the developer over time, YMMV. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 11:39:43 2003
I have used brown developer made with brown crystals before in my home darkroom (not in the lab!) and have had satisfactory results. As I recall, the developers were Microdol-X for film and Dektol for paper. This doesn't mean I would ever risk anyone else's films in it, but for the replaceable stuff I was doing at the time, it was worth the experiment.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Paul Chipman [mailto:paulrc-at-bilbo.bio.purdue.edu] Sent: Thursday, October 09, 2003 8:51 AM To: microscopy-at-msa.microscopy.com
Hi Leslie,
We are currently changing the D19 in our low-dose darkroom after 280 films are processed and this usually happens within 3 weeks of preparation.
On a side note...has anyone ever come across a batch of D19 with brown crystals? It appeared very dark once prepared and was thrown out without attempting to use it.
cheers, paul
} } } } On Wednesday, October 8, 2003, at 10:55 AM, Leslie Cummins wrote: } } } We are setting up to develop Kodak SO163 film for low dose cryoEM. } } We are push processing it, so we are developing it for 12 minutes } } in straight D-19, we're using a nitrogen gas burst, for 1 second } } every 8 seconds. } } } } I was wondering what the life of the chemistry might be. Kodak } } said that we could develop 60 8x10 negatives per gallon. This } } works out to about 360 3 1/4 x 4 negatives. This seems like a lot } } of negatives to us, so we are wondering what other people are using } } for an exhaustion time, both in negative numbers and days in the } } tank. } } } } Thanks for the help. } } } Dear Leslie, } We've done 250 in the past, and that is our plan, if we have } as many as 250 films in a short period--a week or two. If the } solution is older that two weeks, we'll pitch it. You can also } check by color; light brown is fresh, reddish orange is OK, and when } the color intensifies to dark reddish brown, it's bad. We have } covered tanks, and the developer is pretty well nitrogenated after } use, but, if your tanks allow oxygen to dissolve in the developer } over time, YMMV. } Yours, } Bill Tivol } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 } tivol-at-caltech.edu
-- Paul Chipman Electron Microscopy Facility Manager Dept. of Biology, Purdue University Lilly Hall, Rm. B216 Phone: 765-494-1487 Fax:765-496-1189
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 12:56:05 2003
I just wash my grids in 100% Acetone and then water to rinse and let them dry off over a hot plate.... has worked for me for the past 4 years.. no problems...
Eric
================================================= } Hi Sue, } } You wrote: } } } I am a new user in EM and I am having a difficult time getting my Epoxy } } sections to stick to the copper grids. I have tried Acetic acid washes, } } sonicators, HCl wash etc. I have not tried BSA, I am waiting for the } } order to come in. Does anyone have any more suggestions?
} Hi Sue, } With the technique that we use, no known force in the universe will be able } to remove the sections from your grids. } } First of all, use clean grids. There are several ways to clean grids, but } the way that I like, because it is fast, is to hold the grid in concentrated } NaOH while I count to 6, and then briefly dip it repeatedly in water to } rinse the grid. I do this just before I pick up the sections. } } Then, most importantly, we dry down our grids in a drying oven. The oven is } just hot enough to dry glassware, but not really superhot. In other words, } I can put my hand on the bottom of the oven without burning it, but I really } wouldn't want to keep my hand there for more than a second or so. And the } temperature isn't hot enough to warp our plastic petri dishes. If it starts } to warp the petri dishes, it's too hot, though it still probably won't hurt } your sections. [we know that from experience] You can dry your sections } there for about 5-10 minutes. But if you even forget about them overnight, } it won't hurt them at all. } } Then, I can assure you that your sections will NEVER NEVER NEVER wash off } your grids no matter how vigorously you wash them during staining. } } Garry } } Garry Burgess } Charge Technologist } Electron Microscopy } Department of Pathology } Health Sciences Centre } Winnipeg, Canada }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 18:49:52 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dsd-at-ogpnet.com; d_scott_davis-at-yahoo.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, October 9, 2003 at 13:57:46 ---------------------------------------------------------------------------
Email: dsd-at-ogpnet.com; d_scott_davis-at-yahoo.com Name: Scott Davis
Organization: Optical Gaging Products, Inc.
Education: Graduate College
Location: Rochester, NY 14621 USA
Question: Where would you suggest that I look for information on the current state of the art in X-ray Photoelectron Spectroscopy for use in semiconductor dielectric thickness and nitrogen doping concentration measurement? Is NIST current or lagging? How about SEMATECH? Or is this technology really in the hands of metrology companies and therefore proprietary information?
Basically, the life of unused (even diluted) developer is quite long (without air). The clock starts ticking as soon as you develop even one piece of the film (similar but less dramatic for paper developers). The amount of film company recommends to develop usually mean that you supposed to develop all those films in relatively short period of time. Classical example is quite popular in amateur photography D-76 developer. In 1 liter of the fresh developer you could develop about 5 rolls of 35 mm film, but developer will die in about 4-5 hours does not matter how many films you manage to develop, so hurry up! Similarly, it's true for D-19 especially non-diluted. It will not die in few hours, but perhaps two weeks. The exposure to air is very critical here also. So, the bottom line here is: if you need to develop a lot of films in quite short period of time - you may do it to extend company's recommendation. As soon as you start use developer, it will gradually lost capacity in the matter of few weeks and will die even if you just developed 10 films. Note: this does not applied to the fixer - as long as you do not contaminate it with developer, it will works to the end of capacity, does not matter how long it will take (6 mo for sure and even longer). Fixer will die soon if you contaminate it with developer. By the way - if you are using those test drops to check fixer - they are too sensitive, you may use fixer much longer and for more films without any harm to the film. Experimentally I determined that when those "drops" indicate, the fixer is bad, it actually used about 50% of capacity. I hope it helps. Sergey
At 06:11 AM 10/9/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
While I don't use SO163 film or compatible developers, I disagree in general about the life of film developer mix. The film developer is the most important step in processing silver halide media. At least, this is how I see it.
I use (or would have used) fresh developer for each batch of film that I developed. And Kodak explicitly details (as do others) how many rolls or sheets of film can be processed with a fresh batch before refreshing or dumping a pot of developer. I never refreshed developer. The results are just not the same as with a fresh batch.
It may be that this TEM film is pathetically ignorant of basic photo chemistry. However, for such a small cost of chemicals, why take the chance? If the shots are worth shooting, they are worth proper processing. That means new chemicals. And, these chemicals must be at the correct temperature to be consistent and effective. I've seen too many instances of no temperature control. Consequently, the results vary wildly. Duh.
Furthermore, I dumped stop bath and fixer routinely. Why cut corners after so much work on the instrument?
gary g.
At 05:12 PM 10/9/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 22:51:18 2003
Garry You are right and aren't. Kodak spent a lot of time (and perhaps money) to develop chemical formulations, which are stable over some period of time being in use. It, actually, made a revolution in the photographic process: all C-41 process machines in our drug-stores used the solutions, which stable for quite a while. It dramatically reduces the cost of developing and maintenance. Similarly they invented "long-play" solutions to develop X-rays films in the hospitals (we do have such machine - technician changed solutions ones a month). Because in TEM it's just impossible to use fresh developer every time you need to develop a few films and Kodak suggest to use "used" developer over some period of time, I suspect D-19 has "extended life" formulation as well. Based on my personal experience, it's clear to me that diluted solution of D-19 (used or non-used) deliver about the same results for at least 3 weeks if properly stored without exposure to air. Usually we develop about 100 films in 1.5 liter. I think, the decent quality of the TEM film developed in used developer is also because of special film formulation (bye-bye old 4489!). It's called "electron microscopy film" and I assume it separates this film from other type of the films. As for price for chemicals - please count your technician time to make fresh solutions 3 times a day and utilize used ones, it's not cheap as you think. On the positive side, I do agree with you that it's good idea in general to use fresh components everywhere: in photography, biochemistry, cooking etc. Sergey
P.S. You don't need to change Stop-bath and fixer so often. I do believe, that Kodak suggests that stop-bath solution is good for 1000+ films... The things about fixer - it could not spoil your film as long as you careful do not mix it with developer. You may left film in the fixer for hours and it does not harm film. You need to wash it after all by the way. The bottom line here is that you may extend fixation time if your fixer start working slowly saving money and doing good for environment (less chemical waste).
At 07:46 PM 10/9/2003, you wrote: } While I don't use SO163 film or compatible developers, } I disagree in general about the life of film developer mix. } The film developer is the most important step in } processing silver halide media. At least, this is } how I see it. } } I use (or would have used) fresh developer for each } batch of film that I developed. And Kodak explicitly } details (as do others) how many rolls or sheets of film } can be processed with a fresh batch before refreshing } or dumping a pot of developer. I never refreshed } developer. The results are just not the same as with } a fresh batch. } } It may be that this TEM film is pathetically ignorant } of basic photo chemistry. However, for such a small } cost of chemicals, why take the chance? If the shots } are worth shooting, they are worth proper processing. } That means new chemicals. And, these chemicals must } be at the correct temperature to be consistent and } effective. I've seen too many instances of no temperature } control. Consequently, the results vary wildly. Duh. } } Furthermore, I dumped stop bath and fixer routinely. } Why cut corners after so much work on the instrument? } } gary g. } } } } At 05:12 PM 10/9/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
The FocusOnMicroscopy FOM2004 international conference, announced to take place in Singapore, will now be held in Philadelphia, PA, USA, April 4 to April 7. Various organizational issues and travel concerns made this move necessary.
The 2004 meeting will be hosted by Drexel University, School of Biomedical Engineering, Science and Health Systems. The conference and exhibition will be located at the Sheraton University City Hotel, central on the Philadelphia campus. As the next in a series of unique interdisciplinary meetings on advanced multidimensional light microscopy and image processing, the 2004 meeting will pay special attention to the conjunction of multidimensional microscopies with the areas of bioinformatics, bio-nanotechnology and bioengineering.
Deadline for the submission of abstracts will be January 30, 2004. Further information on location, registration, and abstract submission will soon be made available at the Focus on Microscopy webpage: www.focusonmicroscopy.org. We invite you to participate in this conference on behalf the FOM 2004 organizing committee: A. Kriete, Pittsburgh; F. Brakenhoff, Amsterdam; P.C. Cheng, Buffalo; Banu Onaral, Philadelphia. -- +-----------------------------------------+ Prof. Dr. G.J. Brakenhoff University of Amsterdam Institute for Molecular Cell Biology Section Molecular Cytology Kruislaan 316 1098 SM Amsterdam, The Netherlands
Sergey Sodium thiosulphate has a mild solvent effect on silver, and this is accelerated at low pH in acid and hardening fixer formulations. The consequences are negligible during standard fixing times for film or paper, but prolonged over-fixing will produce noticeable bleaching, reducing overall negative or print density and bleaching out low density areas so that shadow detail in negatives and highlight detail in prints are lost.
An old rule of thumb to determine the required fix time for EM film in old or partially-used fixer is twice the time taken to clear the film. Another rule of thumb is to discard the fixer when the clearing time is twice that for a fresh solution. Development is stopped very quickly in a fix bath unless it is completely exhausted, and it is perfectly safe to inspect the negatives as they clear in safelight or even in subdued white light. The most serious downside of stretching fixer life to the limit is that it increases the risk of silver staining of the negatives, and may reduce their archival permanence.
I gave a formula for D19 on this list some months back. Here it is again, derived from British Journal of Photography almanac.
However, other sources quote markedly different proportions of the ingredients, e.g. this one from Leica Users group http://mejac.palo-alto.ca.us/leica-users/v03/msg13172.html D19: water 500 cc Metol 2.0 grams sodium sulfite, desc. 90.0 grams Hydroquinone 8.0 grams sodium carbonate, monohyd. 52.5 grams potassium bromide 5 grams cold water to make 1.0 liter and this one from http://astro.umsystem.edu/apml/ARCHIVES/MAR01/msg01118.html You can make D19 yourself by using its formula: Metol 2 g } } Sodium Sulfite 90 g } } Hydroquinone 8 g } } Sodium Carbonate (anhydrous) 45 g } } Potassium Bromide 5 g } } Water 1 liter
What is the official formula of Kodak D19 as currently marketed. Anyone have authoritative information?
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Faciility
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, October 10, 2003 4:49 AM
The Analytical Instrumentation Facility at NC State University has an opening for a staff member. Please respond directly to Prof. Phil Russell. Do not respond to this email.
NC State University Analytical Instrumentation Facility SEM Laboratory Supervisor
The North Carolina State University Analytical Instrumentation Facility (AIF) is seeking a qualified person to join AIF's professional team of microscopists and microanalysts as the Laboratory Supervisor of the Variable Pressure SEM laboratory. Variable Pressure SEM Laboratory Supervisor is a full time permanent research staff position. Duties and responsibilities include: Management of AIF's Variable Pressure SEM/EDS laboratory; operation and maintenance of Variable Pressure SEM instrumentation and related sample preparation and analysis equipment; scheduling of access to and oversight of the above instrumentation; user training and assistance; assistance with the teaching of electron microscopy laboratory classes; and analysis of a wide variety of samples including development of new analytical techniques when applicable. Duties also include responsibility for providing back up support for the FESEM and SPM laboratories. Qualifications must include a BS or higher in a Materials Science related discipline. Required skills must include: one or more years HANDS ON experience with SEM and related techniques and accessories (e.g. EDS, specimen prerparation and other associated analytical tools) and a record of relavent technical publications and presentations. Preferred qualifications include: MS or higher degree or equivalent, teaching of SEM and SEM user training; familiarity with modern electronics; experience with vacuum systems and some experience with SPM and/or surface analytical techniques. Opportunities for professional growth include opportunities to learn new techniques including STEM, FIB, surface analysis techniques and advanced sample preparation techniques.
Please send resume and three letters of reference to: Phillip E. Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 318A EGRC; 2410 Campus Shore Drive; Raleigh, NC 27695-7531 or email prussell-at-ncsu.edu.
North Carolina State University is an Equal Opportunity and Affirmative Action Employer. ADA Accommodations: Dieter Griffis, 919-515-2128. In its commitment to diversity and equity, North Carolina State University seeks applications from women, minorities, and persons with disabilities. In addition, NC State welcomes all persons without regard to sexual orientation.
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 10 15:04:16 2003
From Kodak publication J-1, "Processing Chemicals and Formulas for Black and White Photography"
D-19
Water at 50 degrees C/ 125 degrees F 500 ml Elon (Kodak's name for Metol*) 2.0 grams Sodium sulfite, anhydrous 90.0 grams Hydroquinone 8.0 grams Sodium carbonate, monohydrated 52.5 grams Potassium bromide, anhydrous 5.0 grams water to make one liter.
* metol is p-methylamino-phenol sulfate, C.A.S 55-55-0
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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 10 18:38:38 2003
On Thursday, October 9, 2003, at 06:50 AM, Paul Chipman wrote:
} On a side note...has anyone ever come across a batch of D19 with brown } crystals? It appeared very dark once prepared and was thrown out } without attempting to use it.
Dear Paul, Yes, the xtals will turn yellow, then brown. They have been oxidized, and discarding the package is best. As Geoff McAuliffe said, D-19 is the cheapest item in the procedure. If, however, you have no other D-19 available, and the solution is not dark brown when mixed, you can safely develop some film with it in a pinch. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 10 22:40:05 2003
Chris Thanks for your message. It forced me to refresh my memory in chemistry. The process of dissolving of metal silver is reduction. Metal Ag in the film is already reduced in compare with silver halide. The conditions in the fixer are again reducing, so there is no chemical direct way to reduce metal silver in the fixer. It meant that directly sodium thiosulphate could not dissolve the metal silver in the film but silver halide. There are couple of things you need to keep in mind: if for some reason silver is oxidized, then you could reduce it with sodium thiosulphate and therefore "dissolve it". Because general reducing conditions in the fixer, oxygen from air may not be effective. You need to add something like bleach into the fixer to oxidize silver and then you may reduce it with sodium thiosulphate. Another thing comes to my mind is chlor gas. So, you may keep film in the fixer for quite while and silver will not dissolved (if only you will not add the bleach). Professional photographers usually do not recommend to keep film in fixer for so long for another reason: sodium thiosulphate is slowly decomposing with creation of elementary sulphur, which colored film in yellowish color. If you will keep your film in the fixer for couple of days, elementary sulfur will be deposited in the gelatin layer, which made film yellowish irreversibly. Similar thing is happening when you do not wash film well after fixer - the residue of sodium thiosulphate will decompose with sulphur creation. So, it's good practice to ensure your film is washed well after fixer. Have a great wekend, Sergey
At 03:08 AM 10/10/2003, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Lithpingthnake-at-aol.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, October 11, 2003 at 08:43:43 ---------------------------------------------------------------------------
Email: Lithpingthnake-at-aol.com Name: Henry Behrens
Organization: N.C.C. Long Island N.Y.
Education: Undergraduate College
Location: Garden City,N.Y. Nassau
Question: I am looking for a resource that will help me understand and get supplies related to.Can you help? Thankyou for your attention.
We had though quite good results with CD31 and vWF in thick sections and whole mounts, as sec. antibody one of the Alexa's from Probes. But what thickness are you talking about? How about inserting eGFP?
Sven _______________________________________________________________ Sven Terclavers LM/CLSM Research Assistent Center for Transgene Technology & Gene Therapy Campus Gasthuisberg O&N Herestraat 49 3000 Leuven Belgium Tel.: +32 (0)16 34 63 71 Fax: +32 (0)16 34 59 90 Email: Sven.Terclavers-at-med.kuleuven.ac.be Web: http://www.kuleuven.ac.be/mcm & http://browse.to/microscopy (only intern) _______________________________________________________________
-------Original Message-------
} From: Michael Herron
I would like suggestions for a reasonably bright marker for mouse endothelium viewed in thick section confocal.
We have tried CD31, Ulex and von Willebrand's factor, and they were all fairly worthless.
Suggestions?!
Thanks, Mike
---
Michael J. Herron, U of MN, Dept. of Entomology herro001-at-umn.edu 612-624-3688 (office) 612-625-5299 (FAX)
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 12 16:59:26 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gilbert.scheffer-at-wanadoo.fr) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, October 12, 2003 at 10:47:53 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sergey-at-seas.ucla.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, October 10, 2003 at 20:16:22 ---------------------------------------------------------------------------
Dear all, What books are you using in the undergrad teaching of electron microscopy? I am in a materials department so we are most concerned with EM of inorganic materials, although polymers might be worth covering too. Also, I do mostly TEM, but I do also talk more generally about SEM and other electron beam instruments too. Does anyone have particular recommendations for textbooks for the undergraduates in this area, they will be in the fifth semester (3rd year, 1st term)?
Much as I like Williams and Carter, this is still a bit big and detailed for the target audience. Mike Loretto's book used to be quite good for this sort of thing, but times have moved on. Maybe, there are newer books on the market that cover the target audience better, with some covereage of recent developments.
Thanks in advance for any useful advice you can give.
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 04:16:53 2003
Sergey I don't pretend to be able to account for this effect, but it happens anyway. Try this simple test: stand an unwanted negative on edge with lower half immersed in a bath of your favourite acid or rapid fixer overnight. Wash and dry. Compare density of immersed and non-immersed areas.
See quotes below from Grant Haist (1979) Modern Photographic Processing; John Wiley & Sons, Inc.; New York; Vol. 1; pps. 564-565 Another danger of long fixing bath immersions is the direct attack on the silver image by the combination of oxygen and the acid fixing bath. It is believed that oxygen from the air dissolves in the fixing bath and attacks the very finely divided silver particles of the image. Oxygen converts these metallic silver particles to silver ions by removing electrons. The silver ions are then complexed by the thiosulfate and removed, resulting in the loss of image silver. Fine-grained films and paper images are especially susceptible to image reduction by acid thiosulfate solutions. The extent of the reducing effect of fixing baths on the silver image during the process of fixation is greater than has been generally supposed. This attack does not occur in alkaline thiosulfate solutions" Best wishes Chris
On 10 Oct 03, at 20:38, Sergey Ryazantsev wrote:
} } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Chris } Thanks for your message. It forced me to refresh my memory in } chemistry. The process of dissolving of metal silver is reduction. } Metal Ag in the film is already reduced in compare with silver halide. } The conditions in the fixer are again reducing, so there is no } chemical direct way to reduce metal silver in the fixer. It meant that } directly sodium thiosulphate could not dissolve the metal silver in } the film but silver halide. There are couple of things you need to } keep in mind: if for some reason silver is oxidized, then you could } reduce it with sodium thiosulphate and therefore "dissolve it". } Because general reducing conditions in the fixer, oxygen from air may } not be effective. You need to add something like bleach into the } fixer to oxidize silver and then you may reduce it with sodium } thiosulphate. Another thing comes to my mind is chlor gas. So, you } may keep film in the fixer for quite while and silver will not } dissolved (if only you will not add the bleach). Professional } photographers usually do not recommend to keep film in fixer for so } long for another reason: sodium thiosulphate is slowly decomposing } with creation of elementary sulphur, which colored film in yellowish } color. If you will keep your film in the fixer for couple of days, } elementary sulfur will be deposited in the gelatin layer, which made } film yellowish irreversibly. Similar thing is happening when you do } not wash film well after fixer - the residue of sodium thiosulphate } will decompose with sulphur creation. So, it's good practice to ensure } your film is washed well after fixer. Have a great wekend, Sergey } } At 03:08 AM 10/10/2003, you wrote: } } } } --------------------------------------------------------------------- } } --------- The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } --------------------------------------------------------------------- } } ---------- } } } } Sergey } } Sodium thiosulphate has a mild solvent effect on silver, and this is } } accelerated at low pH in acid and hardening fixer formulations. The } } consequences are negligible during standard fixing times for film or } } paper, but prolonged over-fixing will produce noticeable bleaching, } } reducing overall negative or print density and bleaching out low } } density areas so that shadow detail in negatives and highlight detail } } in prints are lost. } } } } An old rule of thumb to determine the required fix time for EM film } } in old or partially-used fixer is twice the time taken to clear the } } film. Another rule of thumb is to discard the fixer when the clearing } } time is twice that for a fresh solution. Development is stopped very } } quickly in a fix bath unless it is completely exhausted, and it is } } perfectly safe to inspect the negatives as they clear in safelight or } } even in subdued white light. The most serious downside of stretching } } fixer life to the limit is that it increases the risk of silver } } staining of the negatives, and may reduce their archival permanence. } } } } I gave a formula for D19 on this list some months back. Here it is } } again, derived from British Journal of Photography almanac. } } } } Metol 2.2g, Hydroquinone 8.8g, Sodium sulphite 144g, Sodium carbonate } } 130g Potassium bromide 4g, H2O to 1litre } } } } However, other sources quote markedly different proportions of the } } ingredients, e.g. this one from Leica Users group } } http://mejac.palo-alto.ca.us/leica-users/v03/msg13172.html D19: water } } 500 cc Metol 2.0 grams sodium sulfite, desc. 90.0 grams Hydroquinone } } 8.0 grams sodium carbonate, monohyd. 52.5 grams potassium bromide 5 } } grams cold water to make 1.0 liter and this one from } } http://astro.umsystem.edu/apml/ARCHIVES/MAR01/msg01118.html You can } } make D19 yourself by using its formula: Metol 2 g } } Sodium Sulfite } } 90 g } } Hydroquinone 8 g } } Sodium Carbonate (anhydrous) 45 g } } } } Potassium Bromide 5 g } } Water 1 liter } } } } What is the official formula of Kodak D19 as currently marketed. } } Anyone have authoritative information? } } } } } } Dr. Chris Jeffree } } University of Edinburgh } } Biological Sciences EM Faciility } } } } ----- Original Message ----- } } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Friday, October 10, 2003 4:49 AM } } Subject: [Microscopy] Re: SO163 Development } } } } } } } } } } } } } ------------------------------------------------------------------ } } } -- } } ---------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------ } } } -- } } ----------- } } } } } } Garry } } } You are right and aren't. Kodak spent a lot of time (and perhaps } } money) to } } } develop chemical formulations, which are stable over some period } } } of } } time } } } being in use. It, actually, made a revolution in the photographic } } process: } } } all C-41 process machines in our drug-stores used the solutions, } } which } } } stable for quite a while. It dramatically reduces the cost of } } developing } } } and maintenance. Similarly they invented "long-play" solutions to } } develop } } } X-rays films in the hospitals (we do have such machine - } } } technician } } changed } } } solutions ones a month). Because in TEM it's just impossible to } } } use } } fresh } } } developer every time you need to develop a few films and Kodak } } suggest to } } } use "used" developer over some period of time, I suspect D-19 has } } "extended } } } life" formulation as well. Based on my personal experience, it's } } clear to } } } me that diluted solution of D-19 (used or non-used) deliver about } } the same } } } results for at least 3 weeks if properly stored without exposure } } } to air. Usually we develop about 100 films in 1.5 liter. I } } } think, the } } decent } } } quality of the TEM film developed in used developer is also } } } because } } of } } } special film formulation (bye-bye old 4489!). It's called } } } "electron microscopy film" and I assume it separates this film } } } from other type } } of the } } } films. As for price for chemicals - please count your technician } } time to } } } make fresh solutions 3 times a day and utilize used ones, it's not } } cheap as } } } you think. On the positive side, I do agree with you that it's } } good idea } } } in general to use fresh components everywhere: in photography, } } } biochemistry, cooking etc. Sergey } } } } } } P.S. You don't need to change Stop-bath and fixer so often. I do } } believe, } } } that Kodak suggests that stop-bath solution is good for 1000+ } } films... The } } } things about fixer - it could not spoil your film as long as you } } careful do } } } not mix it with developer. You may left film in the fixer for } } } hours } } and it } } } does not harm film. You need to wash it after all by the way. } } } The } } bottom } } } line here is that you may extend fixation time if your fixer } } } start } } working } } } slowly saving money and doing good for environment (less chemical } } waste). } } } } } } } } } At 07:46 PM 10/9/2003, you wrote: } } } } While I don't use SO163 film or compatible developers, } } } } I disagree in general about the life of film developer mix. } } } } The film developer is the most important step in } } } } processing silver halide media. At least, this is } } } } how I see it. } } } } } } } } I use (or would have used) fresh developer for each } } } } batch of film that I developed. And Kodak explicitly } } } } details (as do others) how many rolls or sheets of film } } } } can be processed with a fresh batch before refreshing } } } } or dumping a pot of developer. I never refreshed } } } } developer. The results are just not the same as with } } } } a fresh batch. } } } } } } } } It may be that this TEM film is pathetically ignorant } } } } of basic photo chemistry. However, for such a small } } } } cost of chemicals, why take the chance? If the shots } } } } are worth shooting, they are worth proper processing. } } } } That means new chemicals. And, these chemicals must } } } } be at the correct temperature to be consistent and } } } } effective. I've seen too many instances of no temperature } } } } control. Consequently, the results vary wildly. Duh. } } } } } } } } Furthermore, I dumped stop bath and fixer routinely. } } } } Why cut corners after so much work on the instrument? } } } } } } } } gary g. } } } } } } } } } } } } } } } } At 05:12 PM 10/9/2003, you wrote: } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } -- } } ---------- } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } } } To Subscribe/Unsubscribe -- } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } ------------------------------------------------------------------ } } } } -- } } ----------- } } } } } } } } } } Basically, the life of unused (even diluted) developer is quite } } long } } } } } (without air). The clock starts ticking as soon as you develop } } even one } } } } } piece of the film (similar but less dramatic for paper } } developers). The } } } } } amount of film company recommends to develop usually mean that } } } } } you supposed to develop all those films in relatively short } } } } } period of time. Classical example is quite popular in amateur } } } } } photography } } D-76 } } } } } developer. In 1 liter of the fresh developer you could develop } } about 5 } } } } } rolls of 35 mm film, but developer will die in about 4-5 hours } } does not } } } } } matter how many films you manage to develop, so hurry up! } } Similarly, } } } } } it's true for D-19 especially non-diluted. It will not die in } } } } } few } } hours, } } } } } but perhaps two weeks. The exposure to air is very critical } } } } } here also. So, the bottom line here is: if you need to develop } } } } } a lot } } of films } } } } } in quite short period of time - you may do it to extend } } } } } company's recommendation. As soon as you start use developer, it } } } } } will } } gradually } } } } } lost capacity in the matter of few weeks and will die even if } } } } } you } } just } } } } } developed 10 films. Note: this does not applied to the fixer - } } } } } as } } long } } } } } as you do not contaminate it with developer, it will works to } } } } } the } } end of } } } } } capacity, does not matter how long it will take (6 mo for sure } } } } } and } } even } } } } } longer). Fixer will die soon if you contaminate it with } } developer. By } } } } } the way - if you are using those test drops to check fixer - } } } } } they } } are too } } } } } sensitive, you may use fixer much longer and for more films } } without any } } } } } harm to the film. Experimentally I determined that when those } } "drops" } } } } } indicate, the fixer is bad, it actually used about 50% of } } capacity. I } } } } } hope it helps. Sergey } } } } } } } } } } } } } } } At 06:11 AM 10/9/2003, you wrote: } } } } } } } } } } } } } } } } } } ----------------------------------------------------------------- } } } } } -- } } ----------- } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } } } } } of } } America } } } } } } To Subscribe/Unsubscribe -- } } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } ----------------------------------------------------------------- } } } } } -- } } ------------ } } } } } } } } } } } } Hi Leslie: } } } } } } } } } } } } 12 minutes in straight D-19 sounds like a huge push to me! } } } } } } On } } the } } } } } } other hand, I have no experience with cryo EM so ... Have you } } talked to } } } } } } Kodak about this? Long times in developer can lead to fogging. } } } } } } As to life of the chemistry in a tank. If you have a } } } } } } floating } } lid } } } } } } (that excludes most air) the chemistry will keep better than } } } } } } if } } the } } } } } } lid allows free access to air. Again, a call to Kodak is in } } order. As } } } } } } for the number of films I always stay on the conservative } } } } } } side, } } maybe } } } } } } 2/3 of the recommendation. } } } } } } D-19 is the cheapest part of the equation (animals, prep } } time, etc) } } } } } } so trying to squeez a few extra films out of a pot of } } } } } } developer } } is not } } } } } } the way to go. Also, D-19 does not last more than 2 years in } } } } } } the package on the shelf. I don't do much TEM anymore so I mix } } } } } } my } } D-19 from } } } } } } scratch using the individual chemicals, not the prepackaged } } Kodak mix. } } } } } } The recipe is widely published, Kodak will give it to you if } } } } } } you } } ask. } } } } } } } } } } } } Geoff } } } } } } } } } } } } Leslie Cummins wrote: } } } } } } } } } } } } } Hello Listers } } } } } } } } } } } } } } We are setting up to develop Kodak SO163 film for low dose } } cryoEM. We } } } } } } } are push processing it, so we are developing it for 12 minutes } } in } } } } } } } straight D-19, we're using a nitrogen gas burst, for 1 second } } every 8 seconds. } } } } } } } } } } } } } } I was wondering what the life of the chemistry might be. } } } } } } } Kodak } } said } } } } } } } that we could develop 60 8x10 negatives per gallon. This } } } } } } } works } } out to } } } } } } } about 360 3 1/4 x 4 negatives. This seems like a lot of } } negatives to } } } } } } } us, so we are wondering what other people are using for an } } exhaustion } } } } } } } time, both in negative numbers and days in the tank. } } } } } } } } } } } } } } Thanks for the help. } } } } } } } Leslie Cummins } } } } } } } } } } } } } } } } } } } } } Leslie Gunther Cummins } } } } } } } Analytical Imaging Facility } } } } } } } Albert Einstein College of Medicine } } } } } } } 1300 Morris Park Ave. } } } } } } } Bronx, NY 10461 } } } } } } } 718-430-3547 } } } } } } } } } } } } } } http://www.aecom.yu.edu/aif/ } } } } } } } } } } } } -- } } } } } } -- } } } } } } ********************************************** } } } } } } 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 } } } } } } ********************************************** } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } Electron Microscopy } } } } } UCLA School of Medicine } } } } } Department of Biological Chemistry } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } Los Angeles, CA 90095 } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } (310) 206-1029 (Lab) } } } } } FAX (departmental): (310) 206-5272 } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } Sergey Ryazantsev Ph. D. } } } Electron Microscopy } } } UCLA School of Medicine } } } Department of Biological Chemistry } } } 10833 Le Conte Ave, Room 33-089 } } } Los Angeles, CA 90095 } } } } } } Phone: (310) 825-1144 (office) } } } (310) 206-1029 (Lab) } } } FAX (departmental): (310) 206-5272 } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } ------- End of forwarded message ------- } } ========================================== } } Dr. Chris Jeffree } } University of Edinburgh } } BIOSEM - Biological Sciences Electron Microscope Facility } } Institute of Cell and Molecular Biology } } Daniel Rutherford Building } } King's Buildings, Mayfield Road } } EDINBURGH, EH9 3JH, Scotland, UK } } Tel. #44 (0) 131 650 5554 } } FAX. #44 (0) 131 650 5392 } } Mobile 07710 585 401 } } email c.jeffree-at-ed.ac.uk } } ========================================= } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-089 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 09:51:00 2003
I'm seeking a "lamp housing mirror" for a Zeiss Axiomat microscope or information on someone who may have one. Please contact me off line if you can help.
Thanks,
Owen Mills
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 12:45:18 2003
There is an older book by Ian Watt. I don't know whether it is still in print. Carol Heckman
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 15:37:57 2003
I am attempting to ultracryosection a cell monolayer grown on a filter. I find the filter resists infiltration with sucrose/PVP, and shatters when I try sectioning. I have tried varying the cutting temperature and speed to no avail. I wonder if anyone in the field has successfully sectioned cells grown on a filter for cryo-immunoEM, and if so whether you would be willing to share your techniques (type of filter, cutting speed/temp, etc.) with me? Thanks so much. Angela Welford, EMT Dept of Pathology UNM-HSC Albuquerque, NM, 87131
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 17:18:33 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (scott.watson-at-granite.k12.ut.us) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, October 13, 2003 at 17:11:52 ---------------------------------------------------------------------------
Email: scott.watson-at-granite.k12.ut.us Name: Scott Watson
Organization: Hunter High School
Education: 9-12th Grade High School
Location: West Valley City, Utah
Question: Hello. I am the electronics, engineering, and scanning electron microscopy instructor at Hunter High School in West Valley City, Utah. Here at Hunter we have an old Amray 1200C2 SEM which has been extensively modified since Amray went out of business. The microscope has had 99.9 percent of electronic circuitry replaced with new/redesigned circuitry which transformed the SEM into an ESEM. Recently several of the universal swivel joints used for adjusting the X,Y,Z, and T axises of the specimen chamber have broken. I was wondering if you knew of a source for these joints? I only need to find a source for joints - they do not have to be original manufacturer? Do you know if another SEM manufacturers specimen chambers were similar in design to the Amray 1200 and could have their specimen axis adjustment joints placed into the Amray?
Ian M. Watt, The principles and practice of Electron Microscopy, 2.ed. Cambridge University Press 1997, ISBN 0521 43591 9
very low on theory!
Philip
----- Original Message ----- } From: "Carol Heckman" {heckman-at-bgnet.bgsu.edu} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Monday, October 13, 2003 10:46 PM
Scott, a long while back I had a conversation with a microscope designer. He mentioned that the trend is to go away from mechanical stage manipulation and use manipulation by electric motors. One conspicuous benefit was fewer mechanical intrusions into the vacuum chamber that needed sealing and maintenance. The other was overall lower cost. Should you reach a dead end with your search for a swivel joint replacement, you may want to consider an upgrade to electrical stage manipulation.
Stu Smalinskas, P.E. SKF USA Plymouth, Michigan (734) 414-6862
Scott Watson wrote:
Question: Hello. I am the electronics, engineering, and scanning electron microscopy instructor at Hunter High School in West Valley City, Utah. Here at Hunter we have an old Amray 1200C2 SEM which has been extensively modified since Amray went out of business. The microscope has had 99.9 percent of electronic circuitry replaced with new/redesigned circuitry which transformed the SEM into an ESEM. Recently several of the universal swivel joints used for adjusting the X,Y,Z, and T axises of the specimen chamber have broken. I was wondering if you knew of a source for these joints? I only need to find a source for joints - they do not have to be original manufacturer? Do you know if another SEM manufacturers specimen chambers were similar in design to the Amray 1200 and could have their specimen axis adjustment joints placed into the Amray?
Sincerely
Scott S. Watson
Email: scott.watson-at-granite.k12.ut.us Name: Scott Watson Organization: Hunter High School Education: 9-12th Grade High School Location: West Valley City, Utah
__________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 08:46:10 2003
We did a survey on available textbooks about five years ago for an undergraduate course on SEM, TEM and analytical methods and we decided to use "Electron Microscopy and Analysis" by Goodhew and Humphreys. It is now published in a 3rd edition with Beanland as coauthor:
P. J. Goodhew, J. Humphreys and R. Beanland: "Electron Microscopy and Analysis" Taylor & Francis.
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: 13. oktober 2003 10:09 To: Microscopy Listserver
Dear all, What books are you using in the undergrad teaching of electron microscopy? I am in a materials department so we are most concerned with EM of inorganic materials, although polymers might be worth covering too. Also, I do mostly TEM, but I do also talk more generally about SEM and other electron beam instruments too. Does anyone have particular recommendations for textbooks for the undergraduates in this area, they will be in the fifth semester (3rd year, 1st term)?
Much as I like Williams and Carter, this is still a bit big and detailed for the target audience. Mike Loretto's book used to be quite good for this sort of thing, but times have moved on. Maybe, there are newer books on the market that cover the target audience better, with some covereage of recent developments.
Thanks in advance for any useful advice you can give.
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 09:23:38 2003
Postdoctoral position: Gold labels will be developed to target genetic tags on expressed proteins for identification of subunits and alignment for image processing of cryoelectron micrographs. Three areas will be coordinated: 1) Chemical design and synthesis of specialized gold nanoparticles, 2) genetic expression of tagged proteins, 3) cryoEM using JEOL 2010F 200keV field emission with 2k CCD and STEM attachment. Position will involve all areas, but the major emphasis will be in using the EM and performing reconstructions. Contact Dr. James Hainfeld, Brookhaven National Lab, Biology Dept., Long Island, NY, hainfeld-at-bnl.gov.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 15:06:27 2003
Dear All: When I use Lowicryl HM20, it sometimes turns to light pink color after polymerization. Has anyone else experienced this? Does anyone know why it changes color? Thank you much?
Hong
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 16:07:59 2003
I have recently collected some large (13.8M) 16 bit grayscale tiff images from my SEM. From the SEM software they look very good and print well to my inkjet printer. The problem I am having is reading the files into any 3rd party imaging software (ie Photoshop, Photo Impact) for resizing and saving in additional formats such as jpeg. Is there software out there that can handle such files or am I missing something about the tiff file format?
Thanks
Roy Beavers Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, Tx. 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 17:43:01 2003
Have you tried the good old IrfanView (free from www.irfanview.com)?
cheers
rtch
Is there anybody out there successfully sectioning large Immunobed or JB-4 blocks with Ralph knives or Tungsten Carbide knives? What is the practical limit to the specimen width?
Ralph Common Electron Microscopist Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824 517-355-7558; fax 517-432-1053 ralph.common-at-ht.msu.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 19:33:34 2003
I've occasionally encountered problems opening tif files in Photoshop, perhaps because of header info that did not conform in some way to what was expected. In those cases I was able to open them in UTHSCSA Image Tool (U. of Texas) or Irfanview and immediately re-save them from those applications. Once saved from one of those applications I could open them in Photoshop without further problems. I never pursued the issue further.
John Twilley
Beavers, Roy wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Group, } } I have recently collected some large (13.8M) 16 bit grayscale tiff images from my SEM. From the SEM software they look very good and print well to my inkjet printer. The problem I am having is reading the files into any 3rd party imaging software (ie Photoshop, Photo Impact) for resizing and saving in additional formats such as jpeg. Is there software out there that can handle such files or am I missing something about the tiff file format? } } Thanks } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 19:42:40 2003
ImageJ, the free program from NIH, should have no trouble reading the file, but it depends on the available memory in your system. You can get ImageJ here: http://rsb.info.nih.gov/ij/
When you install it, read the section on memory management. You will have to tweak the command line in the shortcut to make all of your machine's memory available.
Good luck
Joel
Hi Chris I think you are right. When I immersed half film in the fixer - after a few hours I did see noticeable bleached line on the border between fixer and air. This line is about 1 mm thick by the way. It clearly indicates for me that air is involved (most noticeable on the water-air interface). Except that interface line, the effect of bleaching was about 3% from untouched film for 1.5 h exposure to the fixer. My guess is that at fixer-air interface the metal Ag is more available for the oxidation and then bleaching by the sodium thiosulphate as it correctly cited in your last posting. 3% bleaching effect inside the solution is not big deal: variation in temperature/time perhaps will benefit more pronounced effect on the film. But still, I am impressed: this is a pure example how real life interfere with our basic knowledge without any respect to the "theory"... Anyway, thanks for pointing out this problem. Sergey.
At 02:16 AM 10/13/2003, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
14MB is not all that large of a TIFF file. If it is 16-bits, then it is actually 7MB as 8-bit.
Load the file into PS and do a Image/Mode change from 16-bit to 8-bit and you can do most anything you want to. Make sure it is not indexed RGB but pure and simple grey scale.
gary g.
At 02:09 PM 10/14/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 23:01:55 2003
The price is right but the performance is not. I get too many "out of memory" errors.
I suppose that 2GB RAM is not enough. IJ is OK for small files but not for big or larger ones, IMOH.
gary g.
At 05:47 PM 10/14/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 03:05:56 2003
I am trying to prepare some samples for TEM analysis, lattice-fringes images. I have different suspensions which include Wyoming montmorillonite and iron oxides. I tried to obtain oriented aggregates by filtration, but I did not managed. I also tried the sedimentation way, but it needs time and at the end, the mineral phases are not the same (the iron oxides may transform in time and may also interact with the clay mineral). I am trying now to use the supercentrifuge, but I do not know if I will manage to take the oriented aggregate without disturbing them. I should need an advice. I should need at the end to use the L.R.White resin to keep the montmorillonite interlayer hydrated. Thank you
----------------------------------------------------------------------------------- Prof. Elena BELLUSO - Ph.D. Mineralogy and Crystallography Dipartimento di Scienze Mineralogiche e Petrologiche Università degli Studi di Torino Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel: (39) 011 670 71 35 - fax: (39) 011 670 71 28 e-mail: elena.belluso-at-unito.it {http://www.dsmp.unito.it/} http://www.dsmp.unito.it ----------------------------------------------------------------------------------- "I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain." Blade Runner
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 05:43:33 2003
Hello Hong, For HM20 it is just the normal color whe block is polymerized. The K4M is jelowish and the HM20 reddish when exposed to air Best regards Danièle
-----Message d'origine----- De : Hong Yi [mailto:hyi-at-emory.edu] Envoyé : mardi 14 octobre 2003 22:07 À : Microscopy-at-msa.microscopy.com Objet : Lowicryl HM20
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Dear All: When I use Lowicryl HM20, it sometimes turns to light pink color after polymerization. Has anyone else experienced this? Does anyone know why it changes color? Thank you much?
Hong
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 06:37:02 2003
Is it possible to explain how you would like to keep the hydraded stucture in TEM? From my experience the resin embedding will modify the structure. Unless you need to do cryo-sectioning and cryo-TEM.
My suggestion is that if your suspension is not very dilute, you could try a direct collection on a Cu-grid with a carbon film. For TEM application this would be perfectly sufficient and you will not induce any interaction of the two constituents. After collection of the sample, if you obtain a very dense distribution you can embed directly and immediatedly with the grid.
The other alternative is to do cryo-TEM using the suspension directly on the grid followed by rapid cooling and cryo-TEM. For that you need specialised laboratories and if you were interested I could inform you.
Hope this helps, Sousan
Dr. Sousan Abolhassani Laboratory for Materials Behaviour Paul Scherrer Institut 5232 Villigen PSI Switzerland
Elena Belluso wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear All, } } I am trying to prepare some samples for TEM analysis, lattice-fringes } images. I have different suspensions which include Wyoming montmorillonite } and iron oxides. } I tried to obtain oriented aggregates by filtration, but I did not managed. } I also tried the sedimentation way, but it needs time and at the end, the } mineral phases are not the same (the iron oxides may transform in time and } may also interact with the clay mineral). } I am trying now to use the supercentrifuge, but I do not know if I will } manage to take the oriented aggregate without disturbing them. } I should need an advice. I should need at the end to use the L.R.White } resin to keep the montmorillonite interlayer hydrated. } Thank you } } ----------------------------------------------------------------------------------- } Prof. Elena BELLUSO - Ph.D. Mineralogy and Crystallography } Dipartimento di Scienze Mineralogiche e Petrologiche } Università degli Studi di Torino } Via Valperga Caluso, 35 } I-10125 TORINO - ITALIA } tel: (39) 011 670 71 35 - fax: (39) 011 670 71 28 } e-mail: elena.belluso-at-unito.it } {http://www.dsmp.unito.it/} http://www.dsmp.unito.it } ----------------------------------------------------------------------------------- } "I've... seen things you people wouldn't believe. } Attack ships on fire off the shoulder of Orion. } I watched C-beams... glitter in the dark near the Tanhauser Gate. } All those... moments will be lost... in time..., } like... tears... in... rain." } Blade Runner } } _____________________________________________________________________ } For your security, this mail has been scanned and protected by Inflex
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 06:47:05 2003
Laboratory Manager position opening in NYC in multi-user microscopy lab.
Responsibilities include assisting and training staff in scanning electron microscopy (SEM), x-ray microanalysis (EDS), confocal laser scanning microscopy (CLSM), and cathodoluminescence spectroscopy (CLS). Additional responsiblities include assisting and training staff with image processing, and printing for publication and presentation. General lab duties include PC/MAC hardware and software maintenance/upgrades, ordering consumables, and scheduling equipment maintenance.
The ideal candidate will have a B.S. in a natural science, 2-3 yrs experience with SEM theory and operation, and experience with computer hardware/software. Good interpersonal skills are essential.
Contact Dr. Angela V. Klaus via email at avklaus-at-amnh.org. EOE.
----------------------------------------- Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-769-5977 Fax: 212-496-3480 -----------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 07:04:03 2003
} I have recently collected some large (13.8M) 16 bit } grayscale tiff images from my SEM. From the SEM software } they look very good and print well to my inkjet printer. } The problem I am having is reading the files into any 3rd } party imaging software (ie Photoshop, Photo Impact) for resizing } and saving in additional formats such as jpeg. } Is there software out there that can handle such files or am } I missing something about the tiff file format?
I am surprised Photoshop couldn't open them! ... although there is always that possibility, but I also imagine other softwares would have a similar problem with an inaccurate TIFF format. If you know the size of the image, then try opening them "as raw", and then filling in the appropriate info ... e.g., 1024 pixels horizontal, 807 pixels vertical, 16bits deep, and have PS guess at the header size.
If the header size is wrong, then you'll see an image with a border down the middle, and the left & right parts of the image opposite. You can then reopen and guess at the header again, and the correct guess should be accurate for all other images. There is a more systematic approach to guessing the header size, and you can e-mail me as I need to find it ... but the clue is in the row of pixels at the top of the image.
I'll admit that I'm frustrated with my TEM vendor (out of MA). I have an "older" 200KV TEM, but have faithfully kept it under service contract for over 10 years. Given its age it is experiencing some failures, as one would expect. My frustration is that the vendor seems to be trying to fix the same problem over and over without coming to a conclusion. This is causing a huge amount of downtime (since the summer i've not had more than a week of contiguous uptime). When it fails usually it takes 3-4 days or a week to get someone on-site, although they often give me some things to check or "try". The net result is that my time is chewed up and still I have no operating TEM. To be honest, my financial situation requires that I purchase a time-and-parts limited contract at discount, but it seems that the current situation would be unacceptable even with a full contract. With the current uptime percentage I can't even break even on reimbursement for the limited contract from my userbase. Using automobile service as a comparison, I'd be very upset if it was in the shop or awaiting service (for the same or related failures) as much as this TEM is. My question to the group is: what can reasonably be expected for a generation-or-two old TEM for uptime, and how can i work with the vendor to maximize the impact of service calls? Thanks! Brian
_________________________________________________ Brian McIntyre Univ. of Rochester The Institute of Optics River Campus EMLab 585-275-3058/4875 585-244-4936 (fax)
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 10:31:13 2003
Thank you for your answer. I do not know if I can obtain my sample using your first proposed method. I am trying to explain you better what I need. I must prepare some systems composed by iron oxides and Wy-montmorillonite, using different ratios iron oxide/montmorillonite in systems with 10 g solid/1 l suspension. I intend to study the mineral phases modifications in time. By TEM, I hope to observe the behavior of d(001) in different moments of the reaction. My intention was to separate the solid, obtaining first an oriented aggregate. After I would want to embed this aggregate with L.R. White resin, following the procedure described by Kim J.-W., Peacor D.R., Tessier D., Elsass F. (1995) A technique for maintaining texture and permanent expansion of smectite interlayer for TEM observations, Clays Clay Miner., 43, 51-57. This procedure allows to preserve the d spacing of montmorillonite unmodified. Thus, after the embedding, I would use a microtome to obtain the TEM samples from slides perpendicular on the 001 planes. Do you think that using your first method I could arrive at this? I already tried to use the sedimentation on glass. I managed obtain a small disc, but very, very thin in time. I put some drops, after I waited to dry, I put another drops etc. I tried also to put directly a greater quantity, limiting my disc with a vertical tube, but an important quantity of solid (the finest) remained on the walls. Anyway, to obtain a disc of 3 mm thickness I would need much time and the initial suspension will not be the same at the end. I need to separate the solid quickly to preserve this composition. I am trying now the supercentrifuge method. I hope to recover the solid, cutting the plastic tube. I read about the rapid cooling, but using these technique I will have crystals randomly oriented. I hope that now I managed to explain better my problem.
Thank you again
----------------------------------------------------------------------------------- Prof. Elena BELLUSO - Ph.D. Mineralogy and Crystallography Dipartimento di Scienze Mineralogiche e Petrologiche Università degli Studi di Torino Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel: (39) 011 670 71 35 - fax: (39) 011 670 71 28 e-mail: elena.belluso-at-unito.it {http://www.dsmp.unito.it/} http://www.dsmp.unito.it ----------------------------------------------------------------------------------- "I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain." Blade Runner
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 10:33:24 2003
I got pink color with HM 20 after polymeration too. The color fades in a few days. I asked around about the reason and didn't get the answer yet.
Shanling
-----Message d'origine----- De : Hong Yi [mailto:hyi-at-emory.edu] Envoyé : mardi 14 octobre 2003 22:07 À : Microscopy-at-msa.microscopy.com Objet : Lowicryl HM20
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Dear All: When I use Lowicryl HM20, it sometimes turns to light pink color after polymerization. Has anyone else experienced this? Does anyone know why it changes color? Thank you much?
Hong
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 10:53:20 2003
If you're using a Mac, there's an excellent shareware programme called Graphic Converter, which can read anything and translate it into anything else.
http://www.lemkesoft.com/
Hope this helps.
Lesley Weston.
on 14/10/2003 3:44 PM, Ritchie Sims at r.sims-at-auckland.ac.nz wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } } Have you tried the good old IrfanView (free from www.irfanview.com)? } } cheers } } rtch } } } Subject: [Microscopy] Image reader } Date sent: Tue, 14 Oct 2003 16:09:02 -0500 } } From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} } To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} } } } } } } } ---------------------------------------------------------------------- } } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } --------- } } } } Group, } } } } } } I have recently collected some large (13.8M) 16 bit grayscale tiff } } images from my SEM. From the SEM software they look very good and } } print well to my inkjet printer. The problem I am having is reading } } the files into any 3rd party imaging software (ie Photoshop, Photo } } Impact) for resizing and saving in additional formats such as jpeg. } } Is there software out there that can handle such files or am I } } missing something about the tiff file format? } } } } Thanks } } } } Roy Beavers } } Southern Methodist University } } Department of Geological Sciences } } P.O. Box 750395 } } Dallas, Tx. 75275 } } Voice: 214-768-2756 } } Fax: 214-768-2701 } } Email: rbeavers-at-mail.smu.edu } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 11:19:49 2003
I'm looking for a vacuum gauge power supply for an Hitachi HU125E similar to the HU11E series built in the late 60s early 70s (you remember, back when music was good). The supply was typically housed in a rack along with the beam deflector/stigmator module.
I'm also interested in other parts from the HU125E series, all going to support undergraduate EM training.
Contact me off list if you can lend a hand.
Thanks!
Robert -- Robert Fitton Teaching Associate/Director of Laboratories Luther College Department of Biology 700 College Drive Decorah, IA 52101 fittonro-at-luther.edu Voice 563-387-1559 FAX 563-387-1080
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 13:07:25 2003
For grayscale image, most standard image format can support only 256 greyness scale(that means the image depth can only reach 8 bits). Tiff is a flexible format and it can be extended very easily. Many people just extend it to 16 bits or 32 bits. However, this extension can not be recognized by most image processing software.
I think your SEM manufactuer should provide you some ways to change it to other file formats or standard tiff format. If they didn't do it, they were very irresponsible.
If there were no other ways to do it. I suggest you write some programs yourself. You don't need to start from scratch. There are many tiff libraries which can be downloaded from web. Besides, in the book "Advanced computation in Electron Microscopy", Earl Kirkland gave some source code to cope with 32 bits extended tiff file format. I guess you can just revise it to cope with 16 bits.
By the way, opening tiff file is not related to the memory of your computer, even though some software told you that your system lacks memory.
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------ - } } Group, } } } I have recently collected some large (13.8M) 16 bit grayscale tiff images } from my SEM. From the SEM software they look very good and print well to my } inkjet printer. The problem I am having is reading the files into any 3rd } party imaging software (ie Photoshop, Photo Impact) for resizing and saving } in additional formats such as jpeg. Is there software out there that can } handle such files or am I missing something about the tiff file format? } } Thanks } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu } } }
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 13:36:43 2003
Most imaging applications do not support 16-bit TIFF files, including Adobe Photoshop (which is strange, because Adobe maintains the TIFF standard). You could use our software, Image-Pro Plus version 5.0. It supports 16- and 48-bit TIFF. You can convert them to 12-, 8-, 24-, 36-bit images as well as JPEG, TGA (Targa), BMP, PICT, CUT (the Media Cybernetics standard from 20 years ago- still popular), PCX, EPS, flat file, and AVI (for multi-TIFF files).
Best Regards,
Dean Sequera V.P. Marketing & Product Development Media Cybernetics, Inc. 8484 Georgia Ave. Suite 200 Silver Spring, MD 20910 (301) 495-3305 x 262 (301) 495-5964 FAX http://www.mediacy.com
} Subject: [Microscopy] Image reader } Date sent: Tue, 14 Oct 2003 16:09:02 -0500 } From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} } To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} } } } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Group, } } } I have recently collected some large (13.8M) 16 bit grayscale tiff } images from my SEM. From the SEM software they look very good and } print well to my inkjet printer. The problem I am having is reading } the files into any 3rd party imaging software (ie Photoshop, Photo } Impact) for resizing and saving in additional formats such as jpeg. } Is there software out there that can handle such files or am I } missing something about the tiff file format? } } Thanks } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 14:47:39 2003
Paul--I am using a Falcon cell culture insert with an 0.4micron pore size. It incorporates polyethylene terephthalate (PET) into the membrane. Can you recommend a more preferable one from your experience?
Thanks, Angela Welford
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 15:13:05 2003
I would love to locate an additional wehnelt cylinder for our JSM-840. Please keep me in mind if you have an additional one you could part with or are planning to mothball or replace this model SEM.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 18:06:57 2003
Hi Chris I think, you guys have some special oxygen, more aggressive, I guess. So, your butter should be softer and brain healthier. You right, it's 7 levels of grey. I don't understand what you meant talking about accuracy? Of coarse I did average for the area about 2cm^2 in both control and treated with fixer. But still, I did not expect to have even such effect. Thanks for the good point. Sergey.
At 12:10 AM 10/15/2003, you wrote: } Sergey } I estimated the effect as a bit stronger than that, but maybe the } oxygen concentration is greater in this part of the world. That could } account for all sorts of stuff - the sheer quality of Scottish } science, the frequency of Glasgow house fires... } To be serious for a moment, 3-5% reduction may not always be important } pictorially if it takes place uniformly, but that is unlikely to } occur in a neglected fixer bath. Also converting the effect of 3-5% } reduction into grey levels you might expect a density change of } about -7 to -13 which won't contribute any accuracy to your } densitometry. } Chris } } Dr. Chris Jeffree } Edinburgh University } Biological Sciences EM Facility } } ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, October 15, 2003 3:29 AM } Subject: [Microscopy] Re: Re: Fixer life SO163 Development } } } } } } } } -------------------------------------------------------------------- } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ----------- } } } } Hi Chris } } I think you are right. When I immersed half film in the fixer - } after a } } few hours I did see noticeable bleached line on the border between } fixer } } and air. This line is about 1 mm thick by the way. It clearly } indicates } } for me that air is involved (most noticeable on the water-air } } interface). Except that interface line, the effect of bleaching was } about } } 3% from untouched film for 1.5 h exposure to the fixer. My guess is } that } } at fixer-air interface the metal Ag is more available for the } oxidation } } and then bleaching by the sodium thiosulphate as it correctly cited } in your } } last posting. 3% bleaching effect inside the solution is not big } deal: } } variation in temperature/time perhaps will benefit more pronounced } effect } } on the film. But still, I am impressed: this is a pure example how } real } } life interfere with our basic knowledge without any respect to the } } "theory"... Anyway, thanks for pointing out this problem. Sergey. } } } } At 02:16 AM 10/13/2003, you wrote: } } } } } } } } --------------------------------------------------------------------- } --------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } --------------------------------------------------------------------- } ---------- } } } } } } Sergey } } } I don't pretend to be able to account for this effect, but it } happens } } } anyway. Try } } } this simple test: stand an unwanted negative on edge with lower } half immersed } } } in a bath of your favourite acid or rapid fixer overnight. Wash and } dry. } } } Compare } } } density of immersed and non-immersed areas. } } } } } } See quotes below from } } } Grant Haist (1979) Modern Photographic Processing; John Wiley & } Sons, } } } Inc.; New York; Vol. 1; pps. 564-565 } } } "Another danger of long fixing bath immersions is the direct attack } on the } } } silver image by the combination of oxygen and the acid fixing bath. } It is } } } believed that oxygen from the air dissolves in the fixing bath and } attacks } } } the } } } very finely divided silver particles of the image. Oxygen converts } these } } } metallic silver particles to silver ions by removing electrons. The } silver } } } ions } } } are then complexed by the thiosulfate and removed, resulting in the } loss of } } } image silver. Fine-grained films and paper images are especially } susceptible } } } to image reduction by acid thiosulfate solutions." } } } "The extent of the reducing effect of fixing baths on the silver } image during } } } the process of fixation is greater than has been generally } supposed. This } } } attack does not occur in alkaline thiosulfate solutions" } } } Best wishes } } } Chris } } } } } } } } } On 10 Oct 03, at 20:38, Sergey Ryazantsev wrote: } } } } } } } } } } } } } } } } -------------------------------------------------------------------- } -- } } } } -------- The Microscopy ListServer -- Sponsor: The Microscopy } Society } } } } of America To Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------------- } -- } } } } --------- } } } } } } } } Chris } } } } Thanks for your message. It forced me to refresh my memory in } } } } chemistry. The process of dissolving of metal silver is } reduction. } } } } Metal Ag in the film is already reduced in compare with silver } halide. } } } } The conditions in the fixer are again reducing, so there is no } } } } chemical direct way to reduce metal silver in the fixer. It } meant that } } } } directly sodium thiosulphate could not dissolve the metal silver } in } } } } the film but silver halide. There are couple of things you need } to } } } } keep in mind: if for some reason silver is oxidized, then you } could } } } } reduce it with sodium thiosulphate and therefore "dissolve it". } } } } Because general reducing conditions in the fixer, oxygen from } air may } } } } not be effective. You need to add something like bleach into } the } } } } fixer to oxidize silver and then you may reduce it with sodium } } } } thiosulphate. Another thing comes to my mind is chlor gas. So, } you } } } } may keep film in the fixer for quite while and silver will not } } } } dissolved (if only you will not add the bleach). Professional } } } } photographers usually do not recommend to keep film in fixer for } so } } } } long for another reason: sodium thiosulphate is slowly } decomposing } } } } with creation of elementary sulphur, which colored film in } yellowish } } } } color. If you will keep your film in the fixer for couple of } days, } } } } elementary sulfur will be deposited in the gelatin layer, which } made } } } } film yellowish irreversibly. Similar thing is happening when you } do } } } } not wash film well after fixer - the residue of sodium } thiosulphate } } } } will decompose with sulphur creation. So, it's good practice to } ensure } } } } your film is washed well after fixer. Have a great wekend, } Sergey } } } } } } } } At 03:08 AM 10/10/2003, you wrote: } } } } } } } } } } } } } } --------------------------------------------------------------------- } } } } } --------- The Microscopy ListServer -- Sponsor: The Microscopy } } } } } Society of America To Subscribe/Unsubscribe -- } } } } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } --------------------------------------------------------------------- } } } } } ---------- } } } } } } } } } } Sergey } } } } } Sodium thiosulphate has a mild solvent effect on silver, and } this is } } } } } accelerated at low pH in acid and hardening fixer formulations. } The } } } } } consequences are negligible during standard fixing times for } film or } } } } } paper, but prolonged over-fixing will produce noticeable } bleaching, } } } } } reducing overall negative or print density and bleaching out } low } } } } } density areas so that shadow detail in negatives and highlight } detail } } } } } in prints are lost. } } } } } } } } } } An old rule of thumb to determine the required fix time for EM } film } } } } } in old or partially-used fixer is twice the time taken to clear } the } } } } } film. Another rule of thumb is to discard the fixer when the } clearing } } } } } time is twice that for a fresh solution. Development is stopped } very } } } } } quickly in a fix bath unless it is completely exhausted, and it } is } } } } } perfectly safe to inspect the negatives as they clear in } safelight or } } } } } even in subdued white light. The most serious downside of } stretching } } } } } fixer life to the limit is that it increases the risk of silver } } } } } staining of the negatives, and may reduce their archival } permanence. } } } } } } } } } } I gave a formula for D19 on this list some months back. Here it } is } } } } } again, derived from British Journal of Photography almanac. } } } } } } } } } } Metol 2.2g, Hydroquinone 8.8g, Sodium sulphite 144g, Sodium } carbonate } } } } } 130g Potassium bromide 4g, H2O to 1litre } } } } } } } } } } However, other sources quote markedly different proportions of } the } } } } } ingredients, e.g. this one from Leica Users group } } } } } http://mejac.palo-alto.ca.us/leica-users/v03/msg13172.html D19: } water } } } } } 500 cc Metol 2.0 grams sodium sulfite, desc. 90.0 grams } Hydroquinone } } } } } 8.0 grams sodium carbonate, monohyd. 52.5 grams potassium } bromide 5 } } } } } grams cold water to make 1.0 liter and this one from } } } } } http://astro.umsystem.edu/apml/ARCHIVES/MAR01/msg01118.html You } can } } } } } make D19 yourself by using its formula: Metol 2 g } } Sodium } Sulfite } } } } } 90 g } } Hydroquinone 8 g } } Sodium Carbonate (anhydrous) 45 g } } } } } } } Potassium Bromide 5 g } } Water 1 liter } } } } } } } } } } What is the official formula of Kodak D19 as currently } marketed. } } } } } Anyone have authoritative information? } } } } } } } } } } } } } } } Dr. Chris Jeffree } } } } } University of Edinburgh } } } } } Biological Sciences EM Faciility } } } } } } } } } } ----- Original Message ----- } } } } } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } } } } } To: {Microscopy-at-sparc5.microscopy.com} } } } } } Sent: Friday, October 10, 2003 4:49 AM } } } } } Subject: [Microscopy] Re: SO163 Development } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } -- } } } } } ---------- } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of } } } } } America } } } } } } To Subscribe/Unsubscribe -- } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } } On-Line Help } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } ------------------------------------------------------------------ } } } } } } -- } } } } } ----------- } } } } } } } } } } } } Garry } } } } } } You are right and aren't. Kodak spent a lot of time (and } perhaps } } } } } money) to } } } } } } develop chemical formulations, which are stable over some } period } } } } } } of } } } } } time } } } } } } being in use. It, actually, made a revolution in the } photographic } } } } } process: } } } } } } all C-41 process machines in our drug-stores used the } solutions, } } } } } which } } } } } } stable for quite a while. It dramatically reduces the cost } of } } } } } developing } } } } } } and maintenance. Similarly they invented "long-play" } solutions to } } } } } develop } } } } } } X-rays films in the hospitals (we do have such machine - } } } } } } technician } } } } } changed } } } } } } solutions ones a month). Because in TEM it's just } impossible to } } } } } } use } } } } } fresh } } } } } } developer every time you need to develop a few films and } Kodak } } } } } suggest to } } } } } } use "used" developer over some period of time, I suspect } D-19 has } } } } } "extended } } } } } } life" formulation as well. Based on my personal experience, } it's } } } } } clear to } } } } } } me that diluted solution of D-19 (used or non-used) deliver } about } } } } } the same } } } } } } results for at least 3 weeks if properly stored without } exposure } } } } } } to air. Usually we develop about 100 films in 1.5 liter. I } } } } } } think, the } } } } } decent } } } } } } quality of the TEM film developed in used developer is also } } } } } } because } } } } } of } } } } } } special film formulation (bye-bye old 4489!). It's called } } } } } } "electron microscopy film" and I assume it separates this } film } } } } } } from other type } } } } } of the } } } } } } films. As for price for chemicals - please count your } technician } } } } } time to } } } } } } make fresh solutions 3 times a day and utilize used ones, } it's not } } } } } cheap as } } } } } } you think. On the positive side, I do agree with you that } it's } } } } } good idea } } } } } } in general to use fresh components everywhere: in } photography, } } } } } } biochemistry, cooking etc. Sergey } } } } } } } } } } } } P.S. You don't need to change Stop-bath and fixer so often. } I do } } } } } believe, } } } } } } that Kodak suggests that stop-bath solution is good for } 1000+ } } } } } films... The } } } } } } things about fixer - it could not spoil your film as long as } you } } } } } careful do } } } } } } not mix it with developer. You may left film in the fixer } for } } } } } } hours } } } } } and it } } } } } } does not harm film. You need to wash it after all by the } way. } } } } } } The } } } } } bottom } } } } } } line here is that you may extend fixation time if your } fixer } } } } } } start } } } } } working } } } } } } slowly saving money and doing good for environment (less } chemical } } } } } waste). } } } } } } } } } } } } } } } } } } At 07:46 PM 10/9/2003, you wrote: } } } } } } } While I don't use SO163 film or compatible developers, } } } } } } } I disagree in general about the life of film developer mix. } } } } } } } The film developer is the most important step in } } } } } } } processing silver halide media. At least, this is } } } } } } } how I see it. } } } } } } } } } } } } } } I use (or would have used) fresh developer for each } } } } } } } batch of film that I developed. And Kodak explicitly } } } } } } } details (as do others) how many rolls or sheets of film } } } } } } } can be processed with a fresh batch before refreshing } } } } } } } or dumping a pot of developer. I never refreshed } } } } } } } developer. The results are just not the same as with } } } } } } } a fresh batch. } } } } } } } } } } } } } } It may be that this TEM film is pathetically ignorant } } } } } } } of basic photo chemistry. However, for such a small } } } } } } } cost of chemicals, why take the chance? If the shots } } } } } } } are worth shooting, they are worth proper processing. } } } } } } } That means new chemicals. And, these chemicals must } } } } } } } be at the correct temperature to be consistent and } } } } } } } effective. I've seen too many instances of no temperature } } } } } } } control. Consequently, the results vary wildly. Duh. } } } } } } } } } } } } } } Furthermore, I dumped stop bath and fixer routinely. } } } } } } } Why cut corners after so much work on the instrument? } } } } } } } } } } } } } } gary g. } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 05:12 PM 10/9/2003, you wrote: } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } } -- } } } } } ---------- } } } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of } } } } } America } } } } } } } } To Subscribe/Unsubscribe -- } } } } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } } } } On-Line Help } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } } -- } } } } } ----------- } } } } } } } } } } } } } } } } Basically, the life of unused (even diluted) developer is } quite } } } } } long } } } } } } } } (without air). The clock starts ticking as soon as you } develop } } } } } even one } } } } } } } } piece of the film (similar but less dramatic for paper } } } } } developers). The } } } } } } } } amount of film company recommends to develop usually mean } that } } } } } } } } you supposed to develop all those films in relatively } short } } } } } } } } period of time. Classical example is quite popular in } amateur } } } } } } } } photography } } } } } D-76 } } } } } } } } developer. In 1 liter of the fresh developer you could } develop } } } } } about 5 } } } } } } } } rolls of 35 mm film, but developer will die in about 4-5 } hours } } } } } does not } } } } } } } } matter how many films you manage to develop, so hurry up! } } } } } Similarly, } } } } } } } } it's true for D-19 especially non-diluted. It will not } die in } } } } } } } } few } } } } } hours, } } } } } } } } but perhaps two weeks. The exposure to air is very } critical } } } } } } } } here also. So, the bottom line here is: if you need to } develop } } } } } } } } a lot } } } } } of films } } } } } } } } in quite short period of time - you may do it to extend } } } } } } } } company's recommendation. As soon as you start use } developer, it } } } } } } } } will } } } } } gradually } } } } } } } } lost capacity in the matter of few weeks and will die even } if } } } } } } } } you } } } } } just } } } } } } } } developed 10 films. Note: this does not applied to the } fixer - } } } } } } } } as } } } } } long } } } } } } } } as you do not contaminate it with developer, it will works } to } } } } } } } } the } } } } } end of } } } } } } } } capacity, does not matter how long it will take (6 mo for } sure } } } } } } } } and } } } } } even } } } } } } } } longer). Fixer will die soon if you contaminate it with } } } } } developer. By } } } } } } } } the way - if you are using those test drops to check } fixer - } } } } } } } } they } } } } } are too } } } } } } } } sensitive, you may use fixer much longer and for more } films } } } } } without any } } } } } } } } harm to the film. Experimentally I determined that when } those } } } } } "drops" } } } } } } } } indicate, the fixer is bad, it actually used about 50% of } } } } } capacity. I } } } } } } } } hope it helps. Sergey } } } } } } } } } } } } } } } } } } } } } } } } At 06:11 AM 10/9/2003, you wrote: } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ----------------------------------------------------------------- } } } } } } } } -- } } } } } ----------- } } } } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy } Society } } } } } } } } } of } } } } } America } } } } } } } } } To Subscribe/Unsubscribe -- } } } } } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } } } } } On-Line Help } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } } } } } } } ----------------------------------------------------------------- } } } } } } } } -- } } } } } ------------ } } } } } } } } } } } } } } } } } } Hi Leslie: } } } } } } } } } } } } } } } } } } 12 minutes in straight D-19 sounds like a huge push } to me! } } } } } } } } } On } } } } } the } } } } } } } } } other hand, I have no experience with cryo EM so ... } Have you } } } } } talked to } } } } } } } } } Kodak about this? Long times in developer can lead to } fogging. } } } } } } } } } As to life of the chemistry in a tank. If you have a } } } } } } } } } floating } } } } } lid } } } } } } } } } (that excludes most air) the chemistry will keep better } than } } } } } } } } } if } } } } } the } } } } } } } } } lid allows free access to air. Again, a call to Kodak } is in } } } } } order. As } } } } } } } } } for the number of films I always stay on the } conservative } } } } } } } } } side, } } } } } maybe } } } } } } } } } 2/3 of the recommendation. } } } } } } } } } D-19 is the cheapest part of the equation (animals, } prep } } } } } time, etc) } } } } } } } } } so trying to squeez a few extra films out of a pot of } } } } } } } } } developer } } } } } is not } } } } } } } } } the way to go. Also, D-19 does not last more than 2 } years in } } } } } } } } } the package on the shelf. I don't do much TEM anymore so } I mix } } } } } } } } } my } } } } } D-19 from } } } } } } } } } scratch using the individual chemicals, not the } prepackaged } } } } } Kodak mix. } } } } } } } } } The recipe is widely published, Kodak will give it to } you if } } } } } } } } } you } } } } } ask. } } } } } } } } } } } } } } } } } } Geoff } } } } } } } } } } } } } } } } } } Leslie Cummins wrote: } } } } } } } } } } } } } } } } } } } Hello Listers } } } } } } } } } } } } } } } } } } } } We are setting up to develop Kodak SO163 film for low } dose } } } } } cryoEM. We } } } } } } } } } } are push processing it, so we are developing it for 12 } minutes } } } } } in } } } } } } } } } } straight D-19, we're using a nitrogen gas burst, for 1 } second } } } } } every 8 seconds. } } } } } } } } } } } } } } } } } } } } I was wondering what the life of the chemistry might be. } } } } } } } } } } Kodak } } } } } said } } } } } } } } } } that we could develop 60 8x10 negatives per gallon. } This } } } } } } } } } } works } } } } } out to } } } } } } } } } } about 360 3 1/4 x 4 negatives. This seems like a lot of } } } } } negatives to } } } } } } } } } } us, so we are wondering what other people are using for } an } } } } } exhaustion } } } } } } } } } } time, both in negative numbers and days in the tank. } } } } } } } } } } } } } } } } } } } } Thanks for the help. } } } } } } } } } } Leslie Cummins } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Leslie Gunther Cummins } } } } } } } } } } Analytical Imaging Facility } } } } } } } } } } Albert Einstein College of Medicine } } } } } } } } } } 1300 Morris Park Ave. } } } } } } } } } } Bronx, NY 10461 } } } } } } } } } } 718-430-3547 } } } } } } } } } } } } } } } } } } } } http://www.aecom.yu.edu/aif/ } } } } } } } } } } } } } } } } } } -- } } } } } } } } } -- } } } } } } } } } ********************************************** } } } } } } } } } 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 } } } } } } } } } ********************************************** } } } } } } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } } } } Electron Microscopy } } } } } } } } UCLA School of Medicine } } } } } } } } Department of Biological Chemistry } } } } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } } } } Los Angeles, CA 90095 } } } } } } } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } } } } (310) 206-1029 (Lab) } } } } } } } } FAX (departmental): (310) 206-5272 } } } } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } } Electron Microscopy } } } } } } UCLA School of Medicine } } } } } } Department of Biological Chemistry } } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } } Los Angeles, CA 90095 } } } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } } (310) 206-1029 (Lab) } } } } } } FAX (departmental): (310) 206-5272 } } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------- End of forwarded message ------- } } } } } ========================================== } } } } } Dr. Chris Jeffree } } } } } University of Edinburgh } } } } } BIOSEM - Biological Sciences Electron Microscope Facility } } } } } Institute of Cell and Molecular Biology } } } } } Daniel Rutherford Building } } } } } King's Buildings, Mayfield Road } } } } } EDINBURGH, EH9 3JH, Scotland, UK } } } } } Tel. #44 (0) 131 650 5554 } } } } } FAX. #44 (0) 131 650 5392 } } } } } Mobile 07710 585 401 } } } } } email c.jeffree-at-ed.ac.uk } } } } } ========================================= } } } } } } } } _____________________________________ } } } } } } } } Sergey Ryazantsev Ph. D. } } } } Electron Microscopy } } } } UCLA School of Medicine } } } } Department of Biological Chemistry } } } } 10833 Le Conte Ave, Room 33-089 } } } } Los Angeles, CA 90095 } } } } } } } } Phone: (310) 825-1144 (office) } } } } (310) 206-1029 (Lab) } } } } FAX (departmental): (310) 206-5272 } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } ========================================== } } } Dr. Chris Jeffree } } } University of Edinburgh } } } BIOSEM - Biological Sciences Electron Microscope Facility } } } Institute of Cell and Molecular Biology } } } Daniel Rutherford Building } } } King's Buildings, Mayfield Road } } } EDINBURGH, EH9 3JH, Scotland, UK } } } Tel. #44 (0) 131 650 5554 } } } FAX. #44 (0) 131 650 5392 } } } Mobile 07710 585 401 } } } email c.jeffree-at-ed.ac.uk } } } ========================================= } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } 10833 Le Conte Ave, Room 33-089 } } Los Angeles, CA 90095 } } } } Phone: (310) 825-1144 (office) } } (310) 206-1029 (Lab) } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } } } }
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
We are replacing our working Philips 410. It has been under continuous service contract since 1985. If interested in acquiring it, please contact me ASAP.
Thanks in advance
Steve ************************************************** * Join us for our annual * * * * INNERSPACE/OUTERSPACE OPEN HOUSE * * * * SATURDAY NOVEMBER 15, 2003 * * * * 4-8 PM , FREE ADMISSION * * * * www.sci.sdsu.edu/EM_Facility/ioflyer.html * * * ************************************************** 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/
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 01:18:11 2003
We also use Graphic converter as well as Corel Draw. Alby On Mercoledì, ott 15, 2003, at 17:55 Europe/Rome, Lesley Weston wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } If you're using a Mac, there's an excellent shareware programme called } Graphic Converter, which can read anything and translate it into } anything } else. } } http://www.lemkesoft.com/ } } Hope this helps. } } Lesley Weston. } } } on 14/10/2003 3:44 PM, Ritchie Sims at r.sims-at-auckland.ac.nz wrote: } } } } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ----------------------------------------------------------------------- } ----- } --} - } } } } } } Have you tried the good old IrfanView (free from www.irfanview.com)? } } } } cheers } } } } rtch } } } } } } Subject: [Microscopy] Image reader } } Date sent: Tue, 14 Oct 2003 16:09:02 -0500 } } } From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} } } To: "Microscopy Listserver" } } {Microscopy-at-sparc5.microscopy.com} } } } } } } } } } } } --------------------------------------------------------------------- } } } - } } } -------- The Microscopy ListServer -- Sponsor: The Microscopy } } } Society } } } of America To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } --------------------------------------------------------------------- } } } - } } } --------- } } } } } } Group, } } } } } } } } } I have recently collected some large (13.8M) 16 bit grayscale tiff } } } images from my SEM. From the SEM software they look very good and } } } print well to my inkjet printer. The problem I am having is reading } } } the files into any 3rd party imaging software (ie Photoshop, Photo } } } Impact) for resizing and saving in additional formats such as jpeg. } } } Is there software out there that can handle such files or am I } } } missing something about the tiff file format? } } } } } } Thanks } } } } } } Roy Beavers } } } Southern Methodist University } } } Department of Geological Sciences } } } P.O. Box 750395 } } } Dallas, Tx. 75275 } } } Voice: 214-768-2756 } } } Fax: 214-768-2701 } } } Email: rbeavers-at-mail.smu.edu } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------------ -------------------------- Alberto Diaspro Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa Italy voice +390103536426/480, facsimile +39010314218 visit http://www.lambs.it; http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ -------------------------- RESISTER! RESISTERE! RESISTERE! ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 03:12:31 2003
I was wondering if anybody of you know, where I could get a gold coated insect! It would be very nice to have such a toy when presenting our SEMs to freshers or pupils.
Regards Dirk Kirch
-- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 03:40:28 2003
Sergey I was thinking about accuracy in densitometry applied to quantitative estimation of something, e.g. in cytochemistry, where density of the negative is calibrated to be equivalent to some amount of material. In quantitative autoradiography for example one would expect to have to control the exposure time and the development conditions - chemistry, concentration, temperature, time, agitation. But you should also standardise the conditions under which the negatives are fixed, and not leave them in the fix while you go off to the pub. Chris
ps round here most of the aggressive oxygen seems to be bound up in C2H5OH
} Hi Chris } I think, you guys have some special oxygen, more aggressive, I guess. } So, your butter should be softer and brain healthier. You right, it's } 7 levels of grey. I don't understand what you meant talking about } accuracy? Of coarse I did average for the area about 2cm^2 in both } control and treated with fixer. But still, I did not expect to have } even such effect. Thanks for the good point. Sergey. } } At 12:10 AM 10/15/2003, you wrote: } } Sergey } } I estimated the effect as a bit stronger than that, but maybe the } } oxygen concentration is greater in this part of the world. That could } } account for all sorts of stuff - the sheer quality of Scottish } } science, the frequency of Glasgow house fires... To be serious for a } } moment, 3-5% reduction may not always be important pictorially if it } } takes place uniformly, but that is unlikely to occur in a neglected } } fixer bath. Also converting the effect of 3-5% reduction into grey } } levels you might expect a density change of about -7 to -13 which } } won't contribute any accuracy to your densitometry. Chris } } } } Dr. Chris Jeffree } } Edinburgh University } } Biological Sciences EM Facility } } } } ----- Original Message ----- } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Wednesday, October 15, 2003 3:29 AM } } Subject: [Microscopy] Re: Re: Fixer life SO163 Development } } } } } } } } } } } } } ------------------------------------------------------------------ } } } -- } } ---------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------ } } } -- } } ----------- } } } } } } Hi Chris } } } I think you are right. When I immersed half film in the fixer - } } after a } } } few hours I did see noticeable bleached line on the border between } } fixer } } } and air. This line is about 1 mm thick by the way. It clearly } } indicates } } } for me that air is involved (most noticeable on the water-air } } } interface). Except that interface line, the effect of bleaching } } } was } } about } } } 3% from untouched film for 1.5 h exposure to the fixer. My guess } } } is } } that } } } at fixer-air interface the metal Ag is more available for the } } oxidation } } } and then bleaching by the sodium thiosulphate as it correctly } } } cited } } in your } } } last posting. 3% bleaching effect inside the solution is not big } } deal: } } } variation in temperature/time perhaps will benefit more pronounced } } effect } } } on the film. But still, I am impressed: this is a pure example } } } how } } real } } } life interfere with our basic knowledge without any respect to } } } the "theory"... Anyway, thanks for pointing out this problem. } } } Sergey. } } } } } } At 02:16 AM 10/13/2003, you wrote: } } } } } } } } } } } } ------------------------------------------------------------------- } } } -- } } --------- } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } } To Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } ------------------------------------------------------------------- } } } -- } } ---------- } } } } } } } } Sergey } } } } I don't pretend to be able to account for this effect, but it } } happens } } } } anyway. Try } } } } this simple test: stand an unwanted negative on edge with lower } } half immersed } } } } in a bath of your favourite acid or rapid fixer overnight. Wash } } } } and } } dry. } } } } Compare } } } } density of immersed and non-immersed areas. } } } } } } } } See quotes below from } } } } Grant Haist (1979) Modern Photographic Processing; John Wiley & } } Sons, } } } } Inc.; New York; Vol. 1; pps. 564-565 } } } } "Another danger of long fixing bath immersions is the direct } } } } attack } } on the } } } } silver image by the combination of oxygen and the acid fixing } } } } bath. } } It is } } } } believed that oxygen from the air dissolves in the fixing bath } } } } and } } attacks } } } } the } } } } very finely divided silver particles of the image. Oxygen } } } } converts } } these } } } } metallic silver particles to silver ions by removing electrons. } } } } The } } silver } } } } ions } } } } are then complexed by the thiosulfate and removed, resulting in } } } } the } } loss of } } } } image silver. Fine-grained films and paper images are especially } } susceptible } } } } to image reduction by acid thiosulfate solutions." } } } } "The extent of the reducing effect of fixing baths on the silver } } image during } } } } the process of fixation is greater than has been generally } } supposed. This } } } } attack does not occur in alkaline thiosulfate solutions" } } } } Best wishes } } } } Chris } } } } } } } } } } } } On 10 Oct 03, at 20:38, Sergey Ryazantsev wrote: } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } -- } } -- } } } } } -------- The Microscopy ListServer -- Sponsor: The Microscopy } } Society } } } } } of America To Subscribe/Unsubscribe -- } } } } } http://www.msa.microscopy.com/MicroscopyListserver On-Line } } } } } Help } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } ------------------------------------------------------------------ } } } -- } } -- } } } } } --------- } } } } } } } } } } Chris } } } } } Thanks for your message. It forced me to refresh my memory in } } } } } chemistry. The process of dissolving of metal silver is } } reduction. } } } } } Metal Ag in the film is already reduced in compare with silver } } halide. } } } } } The conditions in the fixer are again reducing, so there is no } } } } } chemical direct way to reduce metal silver in the fixer. It } } meant that } } } } } directly sodium thiosulphate could not dissolve the metal } } } } } silver } } in } } } } } the film but silver halide. There are couple of things you } } } } } need } } to } } } } } keep in mind: if for some reason silver is oxidized, then you } } could } } } } } reduce it with sodium thiosulphate and therefore "dissolve } } } } } it". Because general reducing conditions in the fixer, oxygen } } } } } from } } air may } } } } } not be effective. You need to add something like bleach into } } the } } } } } fixer to oxidize silver and then you may reduce it with sodium } } } } } thiosulphate. Another thing comes to my mind is chlor gas. } } } } } So, } } you } } } } } may keep film in the fixer for quite while and silver will not } } } } } dissolved (if only you will not add the bleach). Professional } } } } } photographers usually do not recommend to keep film in fixer } } } } } for } } so } } } } } long for another reason: sodium thiosulphate is slowly } } decomposing } } } } } with creation of elementary sulphur, which colored film in } } yellowish } } } } } color. If you will keep your film in the fixer for couple of } } days, } } } } } elementary sulfur will be deposited in the gelatin layer, } } } } } which } } made } } } } } film yellowish irreversibly. Similar thing is happening when } } } } } you } } do } } } } } not wash film well after fixer - the residue of sodium } } thiosulphate } } } } } will decompose with sulphur creation. So, it's good practice } } } } } to } } ensure } } } } } your film is washed well after fixer. Have a great wekend, } } Sergey } } } } } } } } } } At 03:08 AM 10/10/2003, you wrote: } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------- } } } -- } } } } } } --------- The Microscopy ListServer -- Sponsor: The } } } } } } Microscopy Society of America To Subscribe/Unsubscribe -- } } } } } } http://www.msa.microscopy.com/MicroscopyListserver On-Line } } } } } } Help } } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } ------------------------------------------------------------------- } } } -- } } } } } } ---------- } } } } } } } } } } } } Sergey } } } } } } Sodium thiosulphate has a mild solvent effect on silver, and } } this is } } } } } } accelerated at low pH in acid and hardening fixer } } } } } } formulations. } } The } } } } } } consequences are negligible during standard fixing times for } } film or } } } } } } paper, but prolonged over-fixing will produce noticeable } } bleaching, } } } } } } reducing overall negative or print density and bleaching out } } low } } } } } } density areas so that shadow detail in negatives and } } } } } } highlight } } detail } } } } } } in prints are lost. } } } } } } } } } } } } An old rule of thumb to determine the required fix time for } } } } } } EM } } film } } } } } } in old or partially-used fixer is twice the time taken to } } } } } } clear } } the } } } } } } film. Another rule of thumb is to discard the fixer when the } } clearing } } } } } } time is twice that for a fresh solution. Development is } } } } } } stopped } } very } } } } } } quickly in a fix bath unless it is completely exhausted, and } } } } } } it } } is } } } } } } perfectly safe to inspect the negatives as they clear in } } safelight or } } } } } } even in subdued white light. The most serious downside of } } stretching } } } } } } fixer life to the limit is that it increases the risk of } } } } } } silver staining of the negatives, and may reduce their } } } } } } archival } } permanence. } } } } } } } } } } } } I gave a formula for D19 on this list some months back. Here } } } } } } it } } is } } } } } } again, derived from British Journal of Photography almanac. } } } } } } } } } } } } Metol 2.2g, Hydroquinone 8.8g, Sodium sulphite 144g, Sodium } } carbonate } } } } } } 130g Potassium bromide 4g, H2O to 1litre } } } } } } } } } } } } However, other sources quote markedly different proportions } } } } } } of } } the } } } } } } ingredients, e.g. this one from Leica Users group } } } } } } http://mejac.palo-alto.ca.us/leica-users/v03/msg13172.html } } } } } } D19: } } water } } } } } } 500 cc Metol 2.0 grams sodium sulfite, desc. 90.0 grams } } Hydroquinone } } } } } } 8.0 grams sodium carbonate, monohyd. 52.5 grams potassium } } bromide 5 } } } } } } grams cold water to make 1.0 liter and this one from } } } } } } http://astro.umsystem.edu/apml/ARCHIVES/MAR01/msg01118.html } } } } } } You } } can } } } } } } make D19 yourself by using its formula: Metol 2 g } } Sodium } } Sulfite } } } } } } 90 g } } Hydroquinone 8 g } } Sodium Carbonate (anhydrous) 45 } } } } } } g } } } } } } } } Potassium Bromide 5 g } } Water 1 liter } } } } } } } } } } } } What is the official formula of Kodak D19 as currently } } marketed. } } } } } } Anyone have authoritative information? } } } } } } } } } } } } } } } } } } Dr. Chris Jeffree } } } } } } University of Edinburgh } } } } } } Biological Sciences EM Faciility } } } } } } } } } } } } ----- Original Message ----- } } } } } } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } } } } } } To: {Microscopy-at-sparc5.microscopy.com} } } } } } } Sent: Friday, October 10, 2003 4:49 AM } } } } } } Subject: [Microscopy] Re: SO163 Development } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } } -- } } } } } } ---------- } } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of } } } } } } America } } } } } } } To Subscribe/Unsubscribe -- } } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } } } On-Line Help } } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } ------------------------------------------------------------------ } } } } } } } -- } } } } } } ----------- } } } } } } } } } } } } } } Garry } } } } } } } You are right and aren't. Kodak spent a lot of time (and } } perhaps } } } } } } money) to } } } } } } } develop chemical formulations, which are stable over some } } period } } } } } } } of } } } } } } time } } } } } } } being in use. It, actually, made a revolution in the } } photographic } } } } } } process: } } } } } } } all C-41 process machines in our drug-stores used the } } solutions, } } } } } } which } } } } } } } stable for quite a while. It dramatically reduces the } } } } } } } cost } } of } } } } } } developing } } } } } } } and maintenance. Similarly they invented "long-play" } } solutions to } } } } } } develop } } } } } } } X-rays films in the hospitals (we do have such machine - } } } } } } } technician } } } } } } changed } } } } } } } solutions ones a month). Because in TEM it's just } } impossible to } } } } } } } use } } } } } } fresh } } } } } } } developer every time you need to develop a few films and } } Kodak } } } } } } suggest to } } } } } } } use "used" developer over some period of time, I suspect } } D-19 has } } } } } } "extended } } } } } } } life" formulation as well. Based on my personal } } } } } } } experience, } } it's } } } } } } clear to } } } } } } } me that diluted solution of D-19 (used or non-used) } } } } } } } deliver } } about } } } } } } the same } } } } } } } results for at least 3 weeks if properly stored without } } exposure } } } } } } } to air. Usually we develop about 100 films in 1.5 liter. } } } } } } } I think, the } } } } } } decent } } } } } } } quality of the TEM film developed in used developer is } } } } } } } also because } } } } } } of } } } } } } } special film formulation (bye-bye old 4489!). It's called } } } } } } } "electron microscopy film" and I assume it separates this } } film } } } } } } } from other type } } } } } } of the } } } } } } } films. As for price for chemicals - please count your } } technician } } } } } } time to } } } } } } } make fresh solutions 3 times a day and utilize used ones, } } it's not } } } } } } cheap as } } } } } } } you think. On the positive side, I do agree with you } } } } } } } that } } it's } } } } } } good idea } } } } } } } in general to use fresh components everywhere: in } } photography, } } } } } } } biochemistry, cooking etc. Sergey } } } } } } } } } } } } } } P.S. You don't need to change Stop-bath and fixer so } } } } } } } often. } } I do } } } } } } believe, } } } } } } } that Kodak suggests that stop-bath solution is good for } } 1000+ } } } } } } films... The } } } } } } } things about fixer - it could not spoil your film as long } } } } } } } as } } you } } } } } } careful do } } } } } } } not mix it with developer. You may left film in the fixer } } for } } } } } } } hours } } } } } } and it } } } } } } } does not harm film. You need to wash it after all by the } } way. } } } } } } } The } } } } } } bottom } } } } } } } line here is that you may extend fixation time if your } } fixer } } } } } } } start } } } } } } working } } } } } } } slowly saving money and doing good for environment (less } } chemical } } } } } } waste). } } } } } } } } } } } } } } } } } } } } } At 07:46 PM 10/9/2003, you wrote: } } } } } } } } While I don't use SO163 film or compatible developers, I } } } } } } } } disagree in general about the life of film developer mix. } } } } } } } } The film developer is the most important step in } } } } } } } } processing silver halide media. At least, this is how I } } } } } } } } see it. } } } } } } } } } } } } } } } } I use (or would have used) fresh developer for each } } } } } } } } batch of film that I developed. And Kodak explicitly } } } } } } } } details (as do others) how many rolls or sheets of film } } } } } } } } can be processed with a fresh batch before refreshing or } } } } } } } } dumping a pot of developer. I never refreshed developer. } } } } } } } } The results are just not the same as with a fresh batch. } } } } } } } } } } } } } } } } It may be that this TEM film is pathetically ignorant of } } } } } } } } basic photo chemistry. However, for such a small cost of } } } } } } } } chemicals, why take the chance? If the shots are worth } } } } } } } } shooting, they are worth proper processing. That means } } } } } } } } new chemicals. And, these chemicals must be at the } } } } } } } } correct temperature to be consistent and effective. I've } } } } } } } } seen too many instances of no temperature control. } } } } } } } } Consequently, the results vary wildly. Duh. } } } } } } } } } } } } } } } } Furthermore, I dumped stop bath and fixer routinely. Why } } } } } } } } cut corners after so much work on the instrument? } } } } } } } } } } } } } } } } gary g. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 05:12 PM 10/9/2003, you wrote: } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } } } -- } } } } } } ---------- } } } } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of } } } } } } America } } } } } } } } } To Subscribe/Unsubscribe -- } } } } } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } } } } } On-Line Help } } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } } } -- } } } } } } ----------- } } } } } } } } } } } } } } } } } } Basically, the life of unused (even diluted) developer } } } } } } } } } is } } quite } } } } } } long } } } } } } } } } (without air). The clock starts ticking as soon as you } } develop } } } } } } even one } } } } } } } } } piece of the film (similar but less dramatic for paper } } } } } } developers). The } } } } } } } } } amount of film company recommends to develop usually } } } } } } } } } mean } } that } } } } } } } } } you supposed to develop all those films in relatively } } short } } } } } } } } } period of time. Classical example is quite popular in } } amateur } } } } } } } } } photography } } } } } } D-76 } } } } } } } } } developer. In 1 liter of the fresh developer you could } } develop } } } } } } about 5 } } } } } } } } } rolls of 35 mm film, but developer will die in about 4-5 } } hours } } } } } } does not } } } } } } } } } matter how many films you manage to develop, so hurry } } } } } } } } } up! } } } } } } Similarly, } } } } } } } } } it's true for D-19 especially non-diluted. It will not } } die in } } } } } } } } } few } } } } } } hours, } } } } } } } } } but perhaps two weeks. The exposure to air is very } } critical } } } } } } } } } here also. So, the bottom line here is: if you need to } } develop } } } } } } } } } a lot } } } } } } of films } } } } } } } } } in quite short period of time - you may do it to extend } } } } } } } } } company's recommendation. As soon as you start use } } developer, it } } } } } } } } } will } } } } } } gradually } } } } } } } } } lost capacity in the matter of few weeks and will die } } } } } } } } } even } } if } } } } } } } } } you } } } } } } just } } } } } } } } } developed 10 films. Note: this does not applied to the } } fixer - } } } } } } } } } as } } } } } } long } } } } } } } } } as you do not contaminate it with developer, it will } } } } } } } } } works } } to } } } } } } } } } the } } } } } } end of } } } } } } } } } capacity, does not matter how long it will take (6 mo } } } } } } } } } for } } sure } } } } } } } } } and } } } } } } even } } } } } } } } } longer). Fixer will die soon if you contaminate it with } } } } } } developer. By } } } } } } } } } the way - if you are using those test drops to check } } fixer - } } } } } } } } } they } } } } } } are too } } } } } } } } } sensitive, you may use fixer much longer and for more } } films } } } } } } without any } } } } } } } } } harm to the film. Experimentally I determined that when } } those } } } } } } "drops" } } } } } } } } } indicate, the fixer is bad, it actually used about 50% } } } } } } } } } of } } } } } } capacity. I } } } } } } } } } hope it helps. Sergey } } } } } } } } } } } } } } } } } } } } } } } } } } } At 06:11 AM 10/9/2003, you wrote: } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ----------------------------------------------------------------- } } } } } } } } } -- } } } } } } ----------- } } } } } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society } } } } } } } } } } of } } } } } } America } } } } } } } } } } To Subscribe/Unsubscribe -- } } } } } } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } } } } } } On-Line Help } } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } } } } } } } } } } ----------------------------------------------------------------- } } } } } } } } } -- } } } } } } ------------ } } } } } } } } } } } } } } } } } } } } Hi Leslie: } } } } } } } } } } } } } } } } } } } } 12 minutes in straight D-19 sounds like a huge push } } to me! } } } } } } } } } } On } } } } } } the } } } } } } } } } } other hand, I have no experience with cryo EM so ... } } Have you } } } } } } talked to } } } } } } } } } } Kodak about this? Long times in developer can lead to } } fogging. } } } } } } } } } } As to life of the chemistry in a tank. If you have } } } } } } } } } } a floating } } } } } } lid } } } } } } } } } } (that excludes most air) the chemistry will keep } } } } } } } } } } better } } than } } } } } } } } } } if } } } } } } the } } } } } } } } } } lid allows free access to air. Again, a call to Kodak } } is in } } } } } } order. As } } } } } } } } } } for the number of films I always stay on the } } conservative } } } } } } } } } } side, } } } } } } maybe } } } } } } } } } } 2/3 of the recommendation. } } } } } } } } } } D-19 is the cheapest part of the equation (animals, } } prep } } } } } } time, etc) } } } } } } } } } } so trying to squeez a few extra films out of a pot of } } } } } } } } } } developer } } } } } } is not } } } } } } } } } } the way to go. Also, D-19 does not last more than 2 } } years in } } } } } } } } } } the package on the shelf. I don't do much TEM anymore } } } } } } } } } } so } } I mix } } } } } } } } } } my } } } } } } D-19 from } } } } } } } } } } scratch using the individual chemicals, not the } } prepackaged } } } } } } Kodak mix. } } } } } } } } } } The recipe is widely published, Kodak will give it to } } you if } } } } } } } } } } you } } } } } } ask. } } } } } } } } } } } } } } } } } } } } Geoff } } } } } } } } } } } } } } } } } } } } Leslie Cummins wrote: } } } } } } } } } } } } } } } } } } } } } Hello Listers } } } } } } } } } } } } } } } } } } } } } } We are setting up to develop Kodak SO163 film for low } } dose } } } } } } cryoEM. We } } } } } } } } } } } are push processing it, so we are developing it for 12 } } minutes } } } } } } in } } } } } } } } } } } straight D-19, we're using a nitrogen gas burst, for 1 } } second } } } } } } every 8 seconds. } } } } } } } } } } } } } } } } } } } } } } I was wondering what the life of the chemistry might } } } } } } } } } } } be. Kodak } } } } } } said } } } } } } } } } } } that we could develop 60 8x10 negatives per gallon. } } This } } } } } } } } } } } works } } } } } } out to } } } } } } } } } } } about 360 3 1/4 x 4 negatives. This seems like a lot } } } } } } } } } } } of } } } } } } negatives to } } } } } } } } } } } us, so we are wondering what other people are using } } } } } } } } } } } for } } an } } } } } } exhaustion } } } } } } } } } } } time, both in negative numbers and days in the tank. } } } } } } } } } } } } } } } } } } } } } } Thanks for the help. } } } } } } } } } } } Leslie Cummins } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Leslie Gunther Cummins } } } } } } } } } } } Analytical Imaging Facility } } } } } } } } } } } Albert Einstein College of Medicine } } } } } } } } } } } 1300 Morris Park Ave. } } } } } } } } } } } Bronx, NY 10461 } } } } } } } } } } } 718-430-3547 } } } } } } } } } } } } } } } } } } } } } } http://www.aecom.yu.edu/aif/ } } } } } } } } } } } } } } } } } } } } -- } } } } } } } } } } -- } } } } } } } } } } ********************************************** } } } } } } } } } } 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 } } } } } } } } } } ********************************************** } } } } } } } } } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } } } } } Electron Microscopy } } } } } } } } } UCLA School of Medicine } } } } } } } } } Department of Biological Chemistry } } } } } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } } } } } Los Angeles, CA 90095 } } } } } } } } } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } } } } } (310) 206-1029 (Lab) } } } } } } } } } FAX (departmental): (310) 206-5272 } } } } } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } } } Electron Microscopy } } } } } } } UCLA School of Medicine } } } } } } } Department of Biological Chemistry } } } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } } } Los Angeles, CA 90095 } } } } } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } } } (310) 206-1029 (Lab) } } } } } } } FAX (departmental): (310) 206-5272 } } } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------- End of forwarded message ------- } } } } } } ========================================== } } } } } } Dr. Chris Jeffree } } } } } } University of Edinburgh } } } } } } BIOSEM - Biological Sciences Electron Microscope Facility } } } } } } Institute of Cell and Molecular Biology Daniel Rutherford } } } } } } Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, } } } } } } Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 } } } } } } 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk } } } } } } ========================================= } } } } } } } } } } _____________________________________ } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } Electron Microscopy } } } } } UCLA School of Medicine } } } } } Department of Biological Chemistry } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } Los Angeles, CA 90095 } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } (310) 206-1029 (Lab) } } } } } FAX (departmental): (310) 206-5272 } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ========================================== } } } } Dr. Chris Jeffree } } } } University of Edinburgh } } } } BIOSEM - Biological Sciences Electron Microscope Facility } } } } Institute of Cell and Molecular Biology } } } } Daniel Rutherford Building } } } } King's Buildings, Mayfield Road } } } } EDINBURGH, EH9 3JH, Scotland, UK } } } } Tel. #44 (0) 131 650 5554 } } } } FAX. #44 (0) 131 650 5392 } } } } Mobile 07710 585 401 } } } } email c.jeffree-at-ed.ac.uk } } } } ========================================= } } } } } } _____________________________________ } } } } } } Sergey Ryazantsev Ph. D. } } } Electron Microscopy } } } UCLA School of Medicine } } } Department of Biological Chemistry } } } 10833 Le Conte Ave, Room 33-089 } } } Los Angeles, CA 90095 } } } } } } Phone: (310) 825-1144 (office) } } } (310) 206-1029 (Lab) } } } FAX (departmental): (310) 206-5272 } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-089 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 04:45:03 2003
Thank you for your answer. I try to explain what I want. Firstly, I have not hematite in my systems. I intend to prepare ferrihydrite in the presence of smectite. Some researches showed the Fe (and Al also) may be enter in different forms in the smectite interlayer, leading to as called hydroxy-Fe interlayered smectites. Thus, the reactions between disordered Fe oxides and smectites are complex, because these latter may be adsorbed also on the external surface of smectite. The XRD measurements may indicate the d spacing variation, but it would be possible that these Fe complexes enter into the smectite interlayer as islands, not as continuous layer. XRD can indicate in this case some peak modifications, but not details. I need TEM to see how it is happening. I would need to have an image of the interaction extension. I also try to have a chemical information from these smectite crystals. I intend to use L.R.White resin to maintain the original interlayer spacing to compare the crystals between them. I do not know if now I was clear enough. Maybe I expect too much from TEM; but I would wish to try. I already know that the centrifugation gives a differential settling of the phases. For this reason I wanted to try another method, but for moment I have not choice. The filtration would be the best, but it was impossible to obtain something. Maybe the pomp is not strong enough or the clay expansion and impermeability stops the process. Now I am trying to use the centrifugation, but I put each rotation time a small quantity to obtain 2-3 solid beds. Thus I am trying to keep in the sample all the phases. I know that this process will disturb also the relations between the mineral phases. For this reason I asked help. If I will discover at the end that my intention is utopical, I will have to change my idea. Now I am still waiting, hopping to find something.
Thank you again
----------------------------------------------------------------------------------- Prof. Elena BELLUSO - Ph.D. Mineralogy and Crystallography Dipartimento di Scienze Mineralogiche e Petrologiche Università degli Studi di Torino Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel: (39) 011 670 71 35 - fax: (39) 011 670 71 28 e-mail: elena.belluso-at-unito.it {http://www.dsmp.unito.it/} http://www.dsmp.unito.it
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 05:33:34 2003
} Most imaging applications do not support 16-bit TIFF files, including } Adobe Photoshop ...
Not true ... PS has been supporting 16bit (and 48bit) TIFFs since version 4. Granted, not every tool is available, but all tonal adjustments and noise removal is supported.
} You could use our software,
Of course! ... {TIC} ...
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 06:46:04 2003
My mistake. The current version of Photoshop supports reading 16-bit and 48-bit TIFF files. Maybe the application you have uses some special tags in the header that Photoshop doesn't recognize. Often times special lookup tables are used and embedded. Great liberties are taken with TIFF (the TIFF specification allows for great flexibility) and there is no organization that certifies that you write a genuine TIFF file.
Dean Sequera
-----Original Message----- } From: Dean Sequera [mailto:DSequera-at-mediacy.com] Sent: Wednesday, October 15, 2003 2:37 PM To: 'Microscopy Listserver'
I agree with Michael, Photoshop does support 16-bit TIF images. However, we ran into this problem a while ago: A user of our software could open a TIF image acquired with analySIS in PS, but it was all black. I don't know if this is the same problem, but the solution to the problem was fairly easy. A bit of background:
There are very few b/w cameras that actually provide 16 bit of data. Most cameras are 12 bit or 14 bit. As there is no 12 or 14 bit data format for storing (they come in multiples of 1 byte=8bit), a 12 bit image has to be stored as a 16 bit file. 16 bit span the range of 0 to 65,536, but the data only span 0 to 4,096. If PS does not do an automatic adjustment, the 12-bit data is displayed as very low intensity and the image appears black.
The solution to the problem was to simply have PS do a "histogram stretch" (I think that was the term PS uses, but I am not sure. I don't really use PS) and the image appeared in all its glory.
There is of course also a possibility that there is something wrong with the TIF file. If the original poster is still listening: If you want to send me the file I could try to open it in our software and convert it for you. Please contact me offline for instructions (mb-at-soft-imaging.com).
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: michael shaffer [mailto:michael-at-shaffer.net] Sent: Thursday, October 16, 2003 4:33 AM To: Dean Sequera Cc: Microscopy Listserver
Dean Sequera (of MediaCybernetics) writes ...
} Most imaging applications do not support 16-bit TIFF files, including } Adobe Photoshop ...
Not true ... PS has been supporting 16bit (and 48bit) TIFFs since version 4. Granted, not every tool is available, but all tonal adjustments and noise removal is supported.
} You could use our software,
Of course! ... {TIC} ...
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 12:02:35 2003
Hi, 1) Does anyone know the name of the company that makes the OV60 crystal and might you have a phone number, email address, or url for them? 2) What is the typical delivery pressure for P-10 gas to WDS detectors?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 12:17:38 2003
Dear Dirk, Most people who run SEM labs have a gold sputtering machine to coat non-conductive samples with a thin layer of gold or gold alloy. Samples that go into a conventional SEM must be electrically conductive. If I want to view an insect, I first dry it for two or three months, so that all the moisture inside is gone (otherwise the insect may explode when you try to pump it in a vacuum), then I cut it and mount it on an SEM stub so the eyes are up, then gold coat it several times. I have several that I use for school tours and have had them for years. I like to show the students the difference between insects and spiders. Regards,
----- Original Message ----- } From: "Dirk Kirch" {kirch-at-imm.rwth-aachen.de} To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 16, 2003 1:14 AM
Yes, Photoshop can open 16-bit images, but they will appear black. To get initial control of them, go to Image - Adjust - Auto Levels, then you can work with the histogram from there. You may have to change them to 8-bit for many Photoshop functions. We have had no problems with Photoshop handling the 16-bit (14-bit) images generated with our analySIS software on our EFTEM.
However, the images generated from our analySIS ADDA II digital image acquisition system on our FESEM (changed to 8-bit) do not not open with some other programs, such as LViewPro, because of header issues. It was easy to have our user who used LViewPro switch to another progam (Irfanview) that could handle the header. The advantage of these programs is that they open the image much faster than Photoshop if you just want to take a look.
The header issue and the 16-bit issue are two different things. I think.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 14:50:11 2003
You might try SemTech Solutions of Billerca, MA. TN is (978) 663-9822 and on the web at www.semtechsolutions.com. They are an SEM service company that specializes in Amray scopes. If they cannot help you themselves they will probably know what direction to point you in.
Good luck,
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Tel: 413 786-9322 Fax: 413 789-2786 mnesta-at-ebsciences.com www.ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:scott.watson-at-granite.k12.ut.us] Sent: Monday, October 13, 2003 6:20 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (scott.watson-at-granite.k12.ut.us) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, October 13, 2003 at 17:11:52 ---------------------------------------------------------------------------
Email: scott.watson-at-granite.k12.ut.us Name: Scott Watson
Organization: Hunter High School
Education: 9-12th Grade High School
Location: West Valley City, Utah
Question: Hello. I am the electronics, engineering, and scanning electron microscopy instructor at Hunter High School in West Valley City, Utah. Here at Hunter we have an old Amray 1200C2 SEM which has been extensively modified since Amray went out of business. The microscope has had 99.9 percent of electronic circuitry replaced with new/redesigned circuitry which transformed the SEM into an ESEM. Recently several of the universal swivel joints used for adjusting the X,Y,Z, and T axises of the specimen chamber have broken. I was wondering if you knew of a source for these joints? I only need to find a source for joints - they do not have to be original manufacturer? Do you know if another SEM manufacturers specimen chambers were similar in design to the Amray 1200 and could have their specimen axis adjustment joints placed into the Amray?
I have not had problems with Adobe Photoshop (R) handling 16 bit images in tif format - it works quite well even with images over 100MB. The problems I've seen arise when people attempt to save 16 bit images in jpg format.
When you first open your image in Photoshop (R), go into "Image" "Mode" and change your image from 16 bit to 8 bit. You should now be able to save your grayscale image in jpg format with no problem. It is also worth checking to be sure you really have a grayscale image - my SEM digitizing software defaults to an indexed color image which must be converted to grayscale, RGB, or CMYK before it can be saved as a jpg.
If Photoshop (R) complains about space on your computer, one of the first things to try is defragmenting the hard drive. Also check how you allocated space for Photoshop (R) when you set up the program.
You may also want to check that your files really are in tif format. I came across one program that "converted" tif to jpg by changing the extension to ".jpg" but did nothing else - needless to say the file size didn't change and other programs would not open the renamed file!
Good luck!
- Louise
(no ties to Adobe - just a happy user)
Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-337-9529 phone 508-339-4996 fax Louise_Harner-at-albint.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 16:25:31 2003
All, I am interested in your suggestions for vendors of phase separators for application in LN2 manual fill stations.
The last two products we used failed prematurely. In case of Cu sponge pressed on a SS fitting, use for about a month lead to Cu sponge increasing in diameter to fall of the fitting. In case of SS sponge welded to SS fitting, the weld heat affected zone in the sponge fails due to brittleness of the material during the first few fills (we might be contributing to it by dropping the hose from time to time). I am at the end of my rope. EHS requires us to have such devices installed.
Warm regards from Texas.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 512 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-amd.com ******************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 18:26:20 2003
Does anyone know where I can lay my hands on an anode assembly for an S360? I have a friends who needs one. It must be cheap.
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42! 16' 48" Long. 83! 43' 48"
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 18:48:41 2003
We've done this routinely for our visiting students and when we do the school tours. The drying time can actually be much quicker by placing the insect, etc. into a vial (although often we can find them already deceased)..
I've observed many of these creatures without the coating as well..simply using a low voltage can often work very well. For the higher magnifications, however, coating does result in better resolution.
Dirk, if you'd like some examples of the images, I can forward them over to you (let me know your preferred format).
Carol Jean Hirt Vice President Materials Research Laboratories, Inc. mrllab.com
} From: "Mary Mager" {mager-at-interchange.ubc.ca} } Date: Thu, 16 Oct 2003 10:15:59 -0700 } To: "Dirk Kirch" {kirch-at-imm.rwth-aachen.de} } Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} } Subject: [Microscopy] Re: coated insects } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Dear Dirk, } Most people who run SEM labs have a gold sputtering machine to coat } non-conductive samples with a thin layer of gold or gold alloy. Samples that } go } into a conventional SEM must be electrically conductive. If I want to view an } insect, I first dry it for two or three months, so that all the moisture } inside } is gone (otherwise the insect may explode when you try to pump it in a } vacuum), } then I cut it and mount it on an SEM stub so the eyes are up, then gold coat } it } several times. I have several that I use for school tours and have had them } for } years. I like to show the students the difference between insects and spiders. } Regards, } } ----- Original Message ----- } } From: "Dirk Kirch" {kirch-at-imm.rwth-aachen.de} } To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} } Sent: Thursday, October 16, 2003 1:14 AM } Subject: [Microscopy] coated insects } } } } } } } } ---------------------------------------------------------------------------- -} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- -} } - } - } } } } Hello Everybody! } } } } I was wondering if anybody of you know, where I could get } } a gold coated insect! It would be very nice to have such } } a toy when presenting our SEMs to freshers or pupils. } } } } Regards Dirk Kirch } } } } -- } } begin:vcard } } n:Kirch;Dirk } } tel;fax:+49(0)241-8022301 } } tel;work:+49(0)241-8026861 } } x-mozilla-html:FALSE } } url:www.imm.rwth-aachen.de } } org:Institute of Physical Metallurgy and Metal Physics;University of Aachen } } version:2.1 } } email;internet:kirch-at-imm.rwth-aachen.de } } title:Dipl.- Phys. Dirk Kirch } } adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; } } end:vcard } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 19:11:16 2003
I have an Amray 1700 Turbo that probably has the same stage mechanism. I have replaced several of the universal joints with ones purchased from Small Parts, Inc. www.smallparts.com. They offer delrin u-joints that fit the splines in my Amray at about $18. ea. They also sell the telescoping joints that appear identical to those on my stage.
Disclaimer: Edison Labs is a private analytical laboratory and has no interest in Small Parts other than as a satisfied customer.
James Carnahan
Edison Analytical Laboratories, Inc. 301 Nott Street Schenectady, NY 12305 (518) 393-2112
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 20:54:21 2003
I'm just getting into using my WDS system on my JXA-840A, setting up to look for low-level Zr in garnets, and I'm a bit disappointed with the apparent detection limits.
I'm using the Zr La, PET, 15kv, 15 nA beam current, 30 seconds on background, 100 seconds on peak, and 10 replicate analyses of a reputedly zero-Zr almandine garnet gave me mean 0.10% ZrO2, Excel STDEV 0.07.
If we take detection limit (whatever that means) as 3SDs, this is 0.21% ZrO2. I had hoped for better.
Other zero-concentartion elements gave (30 sec on peak, 10 sec on backgrounds) STDEVs of
These values are not a heck of an improvement on what I can get with EDS for 100- second counts at 1.5nA.
Is this about par for the WDS course?
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 21:58:32 2003
Can anyone supply me with useful information regarding the processing and sectioning of aortic stents embedded in plastic. I have tried several different resins (LX112 and LR White Grade Hard) as well as glass and diamond knifes with no success.
Thank you for your assistance. Sincerely,
Ken _______________________________________ Kenneth L. Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N Willamette Blvd. Portland, OR 97203 USA
Director, MicroImaging Dx Center Legacy Portland Hospitals Legacy Holladay Park Medical Center 1225 NE 2nd Avenue Portland, OR 97232 USA
Tel.: 503.413.5391
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 06:39:43 2003
} ... A user of our software could open a TIF } image acquired with analySIS in PS, but it was all black. } I don't know if this is the same problem, but the solution } to the problem was fairly easy. ...
Yes ... maybe its because because I work with x-ray counts, but this should be expected. I'll always acquire x-ray counts with, and write to, 16bits, but many times I'll get less than 500 counts max. This will be a very dark 16bit TIFF, and could be 'equalized' for presentation, but it would no longer be 'real' data.
In defense of Cybernetics & Dean Sequera, Image Pro Plus definitely provides some advantages. It can equalize for presentation without changing the data, and as Dean made me aware (personal conversation), Cybernetics has investigated many of the irregular TIFF tags and headers, and can open them properly. Clearly, some TIFFs cannot be opened properly with Photoshop at all ... you may get to see the data, but you may not be able to interpret it.
SEM images are normal grayscale images however, and irregular TIFF tags and headers are a real problem. Whenever we detect a problem we should insist the manufacturer fix it. An example would be to include the definition of magnification without also including the print size. Manufacturers vary, but I have worked with their post-acquisition software and found it totally inadequate for ending up with presentation quality prints ... they really should acknowledge many of us would prefer to use something different ... and write TIFF files we can use.
I would like to prepare some Giemsa stained blood smears and preserve them by mounting with a coverslip. I am told that when using mounting media (such as Canada balsam, per mount, poly-mount, poly-mount-Aqua etc.) there is a chance that the stain may leach into the medium, or the medium bleach out the stain. Does anyone have a suggestion or experience with this stain? Is there a way other than empirically testing, to determine the best mounting medium to use? Thank you
W. Roy Prescott HydasR Inc. PO Box 420 Hershey, PA 17033 FAX - +1-717-533-5548 Voice - +1-717-533-5583 Mobile - +1-717-554-2572 email(s) Roy-at-Hydas.com Info-at-Hydas.com
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 09:59:07 2003
I found this out and fixed it easily by opening the TIFF file in Photoshop and then performing an Image/Auto Levels or Image/Auto Contrast. Auto Levels fixes the dynamic range of the smaller number of bits to match full 16-bits. Image then looks just like the original captured image.
gary g.
At 10:00 AM 10/16/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 10:12:56 2003
Scott, There are small u-joints available through Berg www.wmberg.com and Reid Tool http://reidtool.com
You can also make your own with brass stock found in hobby shops. Square tubing stock is easier, although I believe AMRAY may have used round. The end can be ground to leave two ears sticking up about 3/16 on opposite sides. Drill a hole in each one. Cut a short (1/16 to 3/32) piece of the next smaller size solid brass stock. Drill 2 holes all the way through, large enough to either accept small brass brads or tapped for 0-80 screws. Drill a hole (up to 1/8) through the center where the previous holes intersect. If you use brass brads, put them each through one of the ears and into one of the holes in the spider. Trim the length of each so that they all meet in the center, where you drilled the larger hole, and solder them together. If you opted for screws, then trim four 0-80 brass or stainless screws so they will meet in the center and jam each other.
One of the chief advantages of this is that they can be repaired in the future, particularly easily if you opted for the screws.
Ken Converse owner QUALITY IMAGES 16 CREEK RD DELTA PA 17314 717-456-5491 FAX 717-456-7996 qualityimages-at-netrax.net
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 10:47:48 2003
you are now getting to the heart of the matter: TIF, as flexible as it may be, was not developed with microscopists in mind. Rather, the standard tags refer to parameters that a printer requires for a good rendering of the image. One tag you mention in your response:
"An example would be to include the definition of magnification without also including the print size."
First of all, magnification is not a very useful number. Let's say you print something at 100x and write the mag on the image. Next you take it to a copier and reduce it's size to 50%. The copy will still say 100x, but the image is only half the size as before, so it can't be 100x. Calibration is a much better way to do this. If you have a calibration bar, it scales with the image. So, magnification without specifying the image size is dangerous. Also, as far as I know, there is no public tag in the TIF format for magnification. You would have to convince whoever is responsible for the TIF format to put that in. That takes us to Calibration. There is actually a tag for that in TIF, but again, it is used differently than you'd expect. We had used that tag in the beginning to store image calibration (for example 1nm/pixel). That worked fine, until we tried to import the images into something like Word or other software. There the calibration value was interpreted as print size, so a 1kx1k image with a calibration value of 1nm/pixel was then interpreted to be printed with a size of 1 micron x 1 micron! Not very easy to see!!
What I want to say is, that even with the TIF format, as flexible as it is, we have to use non-standard headers in order to be able to use it.
Having said that, the TIF format does anticipate this and says, that "unknown" tags will be ignored by the standard. So, everybody can modify the header without interfering with the ability of other software to read the file, if done correctly. But it depends on both the writing software AND the reading software to adhere to the TIF formats. If they don't, all bets are off.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
An example would be to include the definition of magnification without also including the print size.
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 10:50:28 2003
On Thursday, October 16, 2003, at 02:25 PM, jerzy.gazda-at-amd.com wrote:
} I am interested in your suggestions for vendors of phase separators } for application in LN2 manual fill stations. } Dear Jerzy, I am not a vendor, but perhaps a simple solution would be to suspend a funnel from the fill line so the LN2 passes through the funnel while the gas does not. This worked well in many instances. Be sure to use a material that does not get brittle at 77 K, such as teflon. Good luck satisfying EHS. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 14:46:39 2003
I have used the same brass phase separator for over 14 years. I bought it at Liquid Carbonics, 1600 Perry Dr. SW, Canton, Ohio. The phone number then was 766-6900. The area code is probably 330 now. A lady named Glenda gave me part number LX0369; phase separator brass; 1-1/4" OD; 3/8" female NPT Nut. It looks like a pressed and sintered brass frit device. At the bottom of the order it says, "453-9904 Placed w/ plant but Canton will invoice".
The cost on April 3, 1989 was $38.50 but I am sure they cost more now. I measured the total length and it is 3.25 inches including the welded nut. The frit is a right circular cylinder 3 inches long.
This thing was real hard to get after someone broke my first one in 1989. For some reason Carbonics didn't like to bother with ordering them. I am sure Liquid Carbonics is another company by now. That's your problem.
A previous separator only failed when people dropped the thing onto concrete and cracked the bottom of the frit. I now take it with me--every time.
As for a funnel, I am using the same two funnels I bought from Fisher Scientific. They are polypropylene, Fisher number 10-371-F. None has ever exploded as was stated on the list awhile back. PE may be a problem but PP has worked for me for over 15 years. No warranty given.
Hope this helps you out and you find a source.
Sincerely,
Paul Beauregard Senior Research Associate PPG Industries Monroeville Technical Center 440 College Park Drive Monroeville, PA 15146
-----Original Message----- } From: jerzy.gazda-at-amd.com [mailto:jerzy.gazda-at-amd.com] Sent: Thursday, October 16, 2003 5:26 PM To: Microscopy-at-MSA.Microscopy.Com
All, I am interested in your suggestions for vendors of phase separators for application in LN2 manual fill stations.
The last two products we used failed prematurely. In case of Cu sponge pressed on a SS fitting, use for about a month lead to Cu sponge increasing in diameter to fall of the fitting. In case of SS sponge welded to SS fitting, the weld heat affected zone in the sponge fails due to brittleness of the material during the first few fills (we might be contributing to it by dropping the hose from time to time). I am at the end of my rope. EHS requires us to have such devices installed.
Warm regards from Texas.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 512 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-amd.com ******************************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 15:57:17 2003
Dear Ritchie, The formula I use for detection limit on my WDX system is: C1 = Cst x(3x(square root B)/P-B) where C1 is the detection limit in mass % Cst is the mass concentration of the analysed element in the standard (100% in Zr metal standard = 1, less if you use a compound standard) B is the background counts P is the peak counts. Make sure you normalize the peak and background counts to the same number of seconds or convert them to counts per second. As soon as you take your standard, you can calculate this. I don't know if this formula will give you a better or worse detection limit, but the WDX detection limits vary a lot more than EDX, because of the position of the different elements around the Roland Circle and how close to the sample the detector is for that particular element. You can then experiment with kV and nA settings to try to optimize the detection limit. Also, the other elements you tested were K lines and the Zr is an L line; this will also reduce the height of the peak and therefore the detection limit. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, October 16, 2003 6:56 PM
Jerry, I can't tell you where to get a good phase separator, but there are also good economic reasons for having one: you will get far more of the liquid into your container with one than without. It pretty much eliminates the atomization that occurs without one.
In my experience, a sintered bronze separator should be available and work well for years.
Ken Converse owner Quality Images third party SEM service Delta, PA
Bill Tivol wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } } On Thursday, October 16, 2003, at 02:25 PM, jerzy.gazda-at-amd.com wrote: } } } I am interested in your suggestions for vendors of phase separators } } for application in LN2 manual fill stations. } } } Dear Jerzy, } I am not a vendor, but perhaps a simple solution would be to } suspend a funnel from the fill line so the LN2 passes through the } funnel while the gas does not. This worked well in many instances. } Be sure to use a material that does not get brittle at 77 K, such as } teflon. Good luck satisfying EHS. } Yours, } Bill Tivol } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 } tivol-at-caltech.edu } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 17:01:28 2003
We are in great need of a negative film scanner used to scan HRTEM films with the size of 3.25x4 inches. Although we have a Nikon super coolscan 8000 film scanner, the film holder is kind of small. It is annoying to cut the films every time when we scan them. If you use scanner to scan the electron microscopy films, please give me your suggestion for the vendor and model. Actually, this question has been discussed before, so any new comment? Thanks.
Regards, Qi Zhang
*************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu qizhang-at-email.unc.edu
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From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 17:09:41 2003
I know about the theoretical, but my question was really about 'in-practice' detection limits ie what standard deviation do you really get from replicate measurements on zero-concentration standards. The latter includes influences like beam current instability and measurement imprecision, spectrometer irregularities, etc, as well as the counting stats (as well, of course, as sample inhomogeneity).
cheers
rtch
} From: "Mary Mager" {mager-at-interchange.ubc.ca} To: "Ritchie Sims" {r.sims-at-auckland.ac.nz} Copies to: "Microscopy" {microscopy-at-MSA.microscopy.com}
If you are really that upset about cutting TEM negs for the LS 8000, well, I doubt that something else will suffice. You have the most awesome scanner available today. Cutting negs is a small chore for super high quality scans. At least I think so. Just slice them and put them in the glass carrier with 6x7 mask.
you are not going to find a 4,000dpi scanner with this kind of D range for film without going to PMT drum scanners. This can be done. but bring along about $25,000 at least. And...some require coating the negs with oil. So, would you rather soak or cut--and/or have lower resolution and lower D range?
gary g.
At 03:02 PM 10/17/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 21:36:31 2003
Ted Clarke is requesting assistance with bloom in a Colorado lake.
"I would appreciate help from the group in identifying the species of organism causing a red film on our lake shore right after the ice melted in 1999. Our lake is considered non-polluted, but is a eutrophic lake. It has a marl bottom with a PH of 8 and a ferrous iron content of 1 ppm in the ground water. I have attached two photos showing the red film and the causing organism. The organism is unusual because it is birefringent, as shown in the images taken with crossed polars and a first order red compensator. The shots taken in the summer were from a bulk sample and no surface film was present. I think I found the same organism," Ted Clarke.
I made a full resoluion photo album at http://www.couger.com/microscope/Ted-Clarke/album/ I will make a better presentation when I am less pressed for time but the images are full resolution and all the information that I have is present. I will add more in as the discussion proceeds [gcc]
Ted can be reached buy email {tclarke-at-ligtel.com http://www.couger.com/microscope/Ted-Clarke/camera%20adapter/adapter.htm http://www.couger.com/microscope/Ted-Clarke/
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 02:14:23 2003
Hello all; The subject of scanning TEM negatives in "standard format" film scanners has once again raised its head. We bemoan the fact that TEM negatives do not conform with what scanner manufactures want to provide for the masses and slice and dice our negatives to conform. The solution lies in "older technology". For a number of years I have used a Leafscan 45 scanner to scan uncut TEM negatives with great success. This scanner is capable of scanning negatives from 35mm to 4x5 and all in.between. The Leaf uses a set of film holders that are the same as those used in Bezler sp? enlargers. To accommodate TEM negatives, I just had a machine shop modify a spare 35mm negative carrier I had to accommodate the TEM negative. Then I set the Leaf 45 to 4x5 setting and scan the whole negative. The scanner can be adjusted in software to scan the TEM negative area only. I get images of ~2500 real DPI. The file sizes are approximately 160mb. The scanner runs on both PC or Mac platforms. In Mac mode data is transferred via SCSI and in PC mode via a GPIB card. In Mac mode you can run the scanner from a Photoshop plug-in. Yay Mac. The downside to all this is that the LeafScan is no longer made but many are still available on EBAY or on the Leafscan listserver on YAHOO (egad). They generally go for about 600-1200 dollars depending on what options are offered at the time of sale. There is a chap in the Eastern US who sells refurbished scanners. I THINK his web address is www.leafstuff.com. He also carries a stock of all parts including the rather hard to find scanner lamp. The Leafscan 45 would be a much cheaper solution to scanning TEM negatives and the need for scissors would be eliminated. If anyone is interested in this solution and want more information, contact me off line.
Cheers; Jon McGovern
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 10:48:17 2003
I would like to purchase a negative scanner and have read the comments about the size problems with the negative carriers. I have not seen the Nikon 8000 but the specs certainly look great. Would it be feasible to have a machinist modify the existing carrier or make one that would fit the 3.25x4' film? Also has anyone approached NIKON about making a carrier for this size film?
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 10/17/03 6:39 PM, "Gary Gaugler" {gary-at-gaugler.com} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } If you are really that upset about cutting TEM } negs for the LS 8000, well, I doubt that } something else will suffice. You have the } most awesome scanner available today. Cutting } negs is a small chore for super high quality } scans. At least I think so. Just slice them } and put them in the glass carrier with 6x7 mask. } } you are not going to find a 4,000dpi scanner with } this kind of D range for film without going } to PMT drum scanners. This can be done. but } bring along about $25,000 at least. And...some } require coating the negs with oil. So, would } you rather soak or cut--and/or have lower resolution } and lower D range? } } gary g. } } } At 03:02 PM 10/17/2003, you wrote: } } } } ---------------------------------------------------------------------------- -} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------------- } } -- } } } } Dear colleagues, } } } } We are in great need of a negative film scanner used } } to scan HRTEM films with the size of 3.25x4 inches. } } Although we have a Nikon super coolscan 8000 film } } scanner, the film holder is kind of small. It is } } annoying to cut the films every time when we scan } } them. If you use scanner to scan the electron } } microscopy films, please give me your suggestion for } } the vendor and model. Actually, this question has been } } discussed before, so any new comment? Thanks. } } } } Regards, } } Qi Zhang } } } } *************************************************** } } Qi Zhang, Ph.D } } 164 Phillips Hall, CB#3255 } } Department of Physics and Astronomy } } the University of North Carolina at Chapel Hill } } Chapel Hill, NC 27599 } } Tel: 919-843-2407 } } Fax: 919-962-0480 } } E-mail: qizhang-at-physics.unc.edu } } qizhang-at-email.unc.edu } } } } } } __________________________________ } } Do you Yahoo!? } } The New Yahoo! Shopping - with improved product search } } http://shopping.yahoo.com } } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 11:04:54 2003
Out of the box, I don't think that any modification could be made to allow the scanner to do a full TEM neg. The reason is that the film adapter units for large frames is masked off using keyed plastic masks. Each mask has a set of holes that mate with stubs sticking out of the film holder. Each mask has a unique hole patter to tell the scanner what frame size is in the holder--6x4.5cm, 6x6cm, 6x7cm. I wind up cutting the neg to fit the width of 6x7cm film and use the 6x7cm mask. This costs about 1/4" on each side of the neg, or 1/2" overall. The length does not have to be reduced.
I suspect that Nikon would have little interest in making a TEM neg holder due to low demand. If one does not have this scanner already, then probably a high end flat bed is a better solution if you don't want to cut the negs. Take a look at the Microtek 6800 (I think that is the model) which has the transparency adapter option. It is lower resolution and lower D than the Nikon 8000 but may do a satisfactory job. My UMAX Powerlook III is needing replacement and am considering the Microtek scanner--not for TEM negs though.
gary g.
At 08:48 AM 10/18/2003, you wrote: } I would like to purchase a negative scanner and have read the comments about } the size problems with the negative carriers. I have not seen the Nikon } 8000 but the specs certainly look great. Would it be feasible to have a } machinist modify the existing carrier or make one that would fit the 3.25x4' } film? Also has anyone approached NIKON about making a carrier for this size } film? } } Debby } } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } } } } } On 10/17/03 6:39 PM, "Gary Gaugler" {gary-at-gaugler.com} wrote: } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } --} - } } } } If you are really that upset about cutting TEM } } negs for the LS 8000, well, I doubt that } } something else will suffice. You have the } } most awesome scanner available today. Cutting } } negs is a small chore for super high quality } } scans. At least I think so. Just slice them } } and put them in the glass carrier with 6x7 mask. } } } } you are not going to find a 4,000dpi scanner with } } this kind of D range for film without going } } to PMT drum scanners. This can be done. but } } bring along about $25,000 at least. And...some } } require coating the negs with oil. So, would } } you rather soak or cut--and/or have lower resolution } } and lower D range? } } } } gary g. } } } } } } At 03:02 PM 10/17/2003, you wrote: } } } } } } } } ---------------------------------------------------------------------------- } -} } - } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } ----------------------------------------------------------------------------- } } } -- } } } } } } Dear colleagues, } } } } } } We are in great need of a negative film scanner used } } } to scan HRTEM films with the size of 3.25x4 inches. } } } Although we have a Nikon super coolscan 8000 film } } } scanner, the film holder is kind of small. It is } } } annoying to cut the films every time when we scan } } } them. If you use scanner to scan the electron } } } microscopy films, please give me your suggestion for } } } the vendor and model. Actually, this question has been } } } discussed before, so any new comment? Thanks. } } } } } } Regards, } } } Qi Zhang } } } } } } *************************************************** } } } Qi Zhang, Ph.D } } } 164 Phillips Hall, CB#3255 } } } Department of Physics and Astronomy } } } the University of North Carolina at Chapel Hill } } } Chapel Hill, NC 27599 } } } Tel: 919-843-2407 } } } Fax: 919-962-0480 } } } E-mail: qizhang-at-physics.unc.edu } } } qizhang-at-email.unc.edu } } } } } } } } } __________________________________ } } } Do you Yahoo!? } } } The New Yahoo! Shopping - with improved product search } } } http://shopping.yahoo.com } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 11:09:36 2003
If this is for filling an EDS Dewar, check into a product called Safe Fill. It connects via a vacuum hose to a big LN2 Dewar or LN2 source and automatically fills the EDS Dewar when the level gets low. It is totally hands-off. I can get more info for you if you can't find the product.
gary g.
At 02:25 PM 10/16/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 17:53:51 2003
There is a simple way to modify the large negative holder to get most of a TEM negative in one scan. Machine away the raised portions of the holder on the right side to accommodate the negative. You have to get pretty close to the hinge so be careful but you can get enough space to that the negative has a bit of play so there is no fear of warping. Given the time and space requirements I doubt if we will want to scan full negatives. The machining is a very simple job. Just two or three straight passes with an end mill. Regarding another thread: Image J opens the 60 meg TIF I produced without a problem. Just don't open too many at once. You can pan and scroll and play with brightness and contrast to your hearts content.
I will update you on my next project: I plan to modify the 35mm mounted slide holder to hold an EM negative in the "portrait" orientation. Then you will almost never have to do any stitching.
Michael
Gary Gaugler wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Out of the box, I don't think that any modification } could be made to allow the scanner to do a full TEM } neg. The reason is that the film adapter units for } large frames is masked off using keyed plastic masks. } Each mask has a set of holes that mate with stubs } sticking out of the film holder. Each mask has a } unique hole patter to tell the scanner what frame } size is in the holder--6x4.5cm, 6x6cm, 6x7cm. } I wind up cutting the neg to fit the width of 6x7cm } film and use the 6x7cm mask. This costs about 1/4" } on each side of the neg, or 1/2" overall. The length } does not have to be reduced. } } I suspect that Nikon would have little interest in } making a TEM neg holder due to low demand. If one } does not have this scanner already, then probably } a high end flat bed is a better solution if you } don't want to cut the negs. Take a look at the } Microtek 6800 (I think that is the model) which } has the transparency adapter option. It is lower } resolution and lower D than the Nikon 8000 but } may do a satisfactory job. My UMAX Powerlook III } is needing replacement and am considering the } Microtek scanner--not for TEM negs though. } } gary g. } } } } At 08:48 AM 10/18/2003, you wrote: } } } I would like to purchase a negative scanner and have read the } } comments about } } the size problems with the negative carriers. I have not seen the Nikon } } 8000 but the specs certainly look great. Would it be feasible to have a } } machinist modify the existing carrier or make one that would fit the } } 3.25x4' } } film? Also has anyone approached NIKON about making a carrier for } } this size } } film? } } } } Debby } } } } } } Debby Sherman, Manager Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-052 Whistler Building } } 170 S. University Street } } West Lafayette, IN 47907 } } } } } } } } } } On 10/17/03 6:39 PM, "Gary Gaugler" {gary-at-gaugler.com} wrote: } } } } } } } } } } } } } ------------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } ---------------------------------------------------------------------------- } } } } --} - } } } } } } If you are really that upset about cutting TEM } } } negs for the LS 8000, well, I doubt that } } } something else will suffice. You have the } } } most awesome scanner available today. Cutting } } } negs is a small chore for super high quality } } } scans. At least I think so. Just slice them } } } and put them in the glass carrier with 6x7 mask. } } } } } } you are not going to find a 4,000dpi scanner with } } } this kind of D range for film without going } } } to PMT drum scanners. This can be done. but } } } bring along about $25,000 at least. And...some } } } require coating the negs with oil. So, would } } } you rather soak or cut--and/or have lower resolution } } } and lower D range? } } } } } } gary g. } } } } } } } } } At 03:02 PM 10/17/2003, you wrote: } } } } } } } } } } } } ---------------------------------------------------------------------------- } } } } -} } - } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } } To Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } ----------------------------------------------------------------------------- } } } } } } -- } } } } } } } } Dear colleagues, } } } } } } } } We are in great need of a negative film scanner used } } } } to scan HRTEM films with the size of 3.25x4 inches. } } } } Although we have a Nikon super coolscan 8000 film } } } } scanner, the film holder is kind of small. It is } } } } annoying to cut the films every time when we scan } } } } them. If you use scanner to scan the electron } } } } microscopy films, please give me your suggestion for } } } } the vendor and model. Actually, this question has been } } } } discussed before, so any new comment? Thanks. } } } } } } } } Regards, } } } } Qi Zhang } } } } } } } } *************************************************** } } } } Qi Zhang, Ph.D } } } } 164 Phillips Hall, CB#3255 } } } } Department of Physics and Astronomy } } } } the University of North Carolina at Chapel Hill } } } } Chapel Hill, NC 27599 } } } } Tel: 919-843-2407 } } } } Fax: 919-962-0480 } } } } E-mail: qizhang-at-physics.unc.edu } } } } qizhang-at-email.unc.edu } } } } } } } } } } } } __________________________________ } } } } Do you Yahoo!? } } } } The New Yahoo! Shopping - with improved product search } } } } http://shopping.yahoo.com } } } } } } } } } } } } } } } }
-- _____________________________ W. Michael Schoel PhD Research Associate Department of Physiology & Biophysics Faculty of Medicine, University of Calgary 3330 Hospital Drive, N.W. Calgary, Alberta, Canada T2N 4N1 Phone: Office (403) 220-3662 Lab (403) 220-6674 E-mail: schoel-at-ucalgary.ca
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 20 05:33:04 2003
I have replied to a similar query back in September but will repeat an extract of the message below in case you haven't seen it
{SNIP} I have used an Epson Perfection 1200 which produce fairly good results but I was always suspicious of its quoted top resolution of 1200 dpi. Negatives on this type of flatbed scanner have to be placed on the glass surface of the scanner to be scanned which creates several problems: Newton's rings; dust and other marks on the glass.
I now use a Microtek Scanmaker 8700 which you can buy with or without Silversoft image management software (I think its the PRO version in the US). It also has a glass panel for normal flatbed document scanning but importantly has a separate glassless drawer which can take several types of film holder. See website below: http://www.microtekusa.com/sm8700.html So far I have been very impressed because the scan quality is good and some of our staff have even used the 35mm film and slide adaptors to digitise their colour slides (1200dpi is adequate for 35mm if you're using a digital projector or displaying on screen). This model comes with USB 1.1 but more usefully Firewire with an adaptor card. In the UK you also get Adobe Photoshop Elements which is quite handy. There is sufficient room to place two large 83 x 102mm (3 1/4 x 4 inch) in the glassless film carriers. Although I am still just using the 5 x 4 inch adaptors I suspect that it would be possible to construct inserts. {SNIP}
I believe Microtek do some more powerful dedicated film scanners that will take the format you desire and manage a greater optical density.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Zhang Qi {qzhangtj-at-yahoo.com}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcervantes-at-bendres.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 16, 2003 at 11:10:02 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] MListserver: TEM Stain for LR-White
Question: Hello: I am working with a soft polymer dispersion, and embedding the sample in LR-White using the cold-cure method. Our sample prep is a little unconventional (I believe), as we want to avoid any alteration of the dispersion's "native" structure in order to observe evidence of phase separation, crystallization in nano-domains, etc. Thus, I do not use water, stains, etc, and microtome the blocks dry with a glass knife (what a pain), a technique used by Dr JoAn Hudson at the University of Oregon whom we have been collaborating with.
The difficulty arises in the TEM. I can not convince myself - or my colleagues - of the difference between the polymer in the dispersion and the embedding media itself (the polymer has extremely low contrast). I wondered if there was a way to label the resin (leaving the polymer alone), and came across a tip on the EMS website that suggested dying the LR-White with a fat-soluble dye (Sudan Black was listed as a good option). I wonder if anyone has experience with this sort of problem or any information on how to stain the media? I'd like to avoid the conventional techniques of staining the polymer for reasons already mentioned. Also, the second component in the dispersion is relatively electron dense and I am afraid I won't be able to tell the difference between the components in the dispersion.
Thanks in advance for any bones you can throw me!
Jessica Cervantes Bend Research, Inc Bend, OR 97701 (541) 382-0212 x240
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sergey-at-seas.ucla.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 20, 2003 at 11:49:51 ---------------------------------------------------------------------------
Does anybody have experience to get rid of Newton's rings on the scanned TEM images. I use Minolta Dimage Scan Multi Pro and I'm quite pleased with the quality of the scanned images except Newton's rings sometime appear on them.
Hi Gary, Thank you for the reply. I am not interested in auto fill, our LN2 vendor AirProducts actually does that as a part of their service. The downside is that we forget how much effort goes into that simple task and take it for granted. They do not issue a phase separator, but we need it to transfer LN2 for filling in cold traps.
To all of you who replied, Thank you for numerous tips.
Regards,
Jerzy
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Saturday, October 18, 2003 11:15 AM To: Gazda, Jerzy Cc: MSA listserver
If this is for filling an EDS Dewar, check into a product called Safe Fill. It connects via a vacuum hose to a big LN2 Dewar or LN2 source and automatically fills the EDS Dewar when the level gets low. It is totally hands-off. I can get more info for you if you can't find the product.
gary g.
At 02:25 PM 10/16/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 20 16:10:56 2003
Looks like sort of algae. I remember Russians do find a lot of microorganisms buried very deep in pretty old Antarctic ice. Those creatures used to live in the water films on the borders between ice crystals. Some of them used to color Antarctic icebergs in funny colors. Those from the deep are anaerobic of coarse. Yours are looks like escaped ice trap and pretty happy enjoying the freedom. Sergey
At 07:36 PM 10/17/2003, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Steve, I believe the way the phase separator works is this:
What is coming down the delivery tube is a boiling mix of gas and liquid at 40 to 200 psi. If it exits the open tube, there is a large amount of atomization and subsequent evaporation. In other words, most of what comes down the tube never makes it into the container. The phase separator is able to cause a lot of the fine mist to coalesce and gravity pulls the liquid to the bottom end. The gas is free to escape from the upper part, less its liquid load. The yield for the liquid part is much greater with it just running freely into the container (without pressure).
Your example shows how important it can be. I'm assuming that the valve was opened the same mount and the feed rate from the bulk tank was the same. That means that you used less than half as much from the bulk tank using the separator. And it saved you time in the bargain.
Ken Converse owner Quality Images third party SEM service Delta, PA
Stevewerner2001-at-aol.com wrote:
} Mr. Converse, } Being new to this career field (microscopy) and new to } science-as-a-livelyhood I was interested in your reasoning to the } outfitting of the live end of the LN line. I have experienced the same } thing here at my microscopy job: pre-frit it took about 42 minutes to } fill the first empty 35 liter dewar on a 85-87 degree day at (usual?) } pressure. Now with the frit nozzle an mere 18 minutes. The distance to } the bulk tank has not changed. Why does a nozzle, that appeares to be } an arreator, cause the fluid to remain fluid and not phase into a gas? } } I understand that you are a business operater and if you don't have } time or reason to respond, that is OK. Any reply is well appreciated. } Thank you, Steve Werner.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 07:48:16 2003
I have been told by knowledgable folks that coating the (face down side of the) negative with "Kami mounting fluid" will stop the formation of Newton rings. Haven't tried it.
Geoff
by way of MicroscopyListserver wrote:
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-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 08:27:03 2003
My colleague is using fluorescent-labeled antibodies to stain bacteria living on agar plates. He is currently imaging these stains/bacteria by picking up the bacteria and placing them on slides. He says that he often is not getting the best images, most likely from interference and background from the agar. Is there a better way to isolate the bacteria from the agar? Should he use a water immersion objective? I've never worked with bacteria before, so I'm finding it a bit difficult to help him in this situation.
Thanks so much,
Elke Pravda Biostructure Core Facility Forsyth Institute Boston, MA epravda-at-forsyth.org
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 11:05:54 2003
If one is using an LN2 tank at 40 to 200 psi that is a major part of the liquid delivery problem. A tank intended to deliver liquid, as opposed to gas-phase coolant, should be a low pressure tank that maintains the liquid only under about 20 psi, enough to boost it out the nozzle and little more. Most delivery services can provide either low pressure (~20 psi) tanks or high pressure tanks. The typical 110 L size of each is very similar in appearance except for the plumbing and pressure regulation apparatus at the top. The high pressure tanks are preferred by users of temperature cycling equipment where the objective is to maximize cooling throughout a protracted discharge from the tank without delivering liquid into the test chamber. It can be virtually impossible to collect the liquid effectively from a high pressure tank.
John Twilley
qualityimages wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Steve, } I believe the way the phase separator works is this: } } What is coming down the delivery tube is a boiling mix of gas and liquid } at 40 to 200 psi. If it exits the open tube, there is a large amount of } atomization and subsequent evaporation. In other words, most of what } comes down the tube never makes it into the container. The phase } separator is able to cause a lot of the fine mist to coalesce and } gravity pulls the liquid to the bottom end. The gas is free to escape } from the upper part, less its liquid load. The yield for the liquid } part is much greater with it just running freely into the container } (without pressure). } } Your example shows how important it can be. I'm assuming that the valve } was opened the same mount and the feed rate from the bulk tank was the } same. That means that you used less than half as much from the bulk } tank using the separator. And it saved you time in the bargain. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } Stevewerner2001-at-aol.com wrote: } } } Mr. Converse, } } Being new to this career field (microscopy) and new to } } science-as-a-livelyhood I was interested in your reasoning to the } } outfitting of the live end of the LN line. I have experienced the same } } thing here at my microscopy job: pre-frit it took about 42 minutes to } } fill the first empty 35 liter dewar on a 85-87 degree day at (usual?) } } pressure. Now with the frit nozzle an mere 18 minutes. The distance to } } the bulk tank has not changed. Why does a nozzle, that appeares to be } } an arreator, cause the fluid to remain fluid and not phase into a gas? } } } } I understand that you are a business operater and if you don't have } } time or reason to respond, that is OK. Any reply is well appreciated. } } Thank you, Steve Werner.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 14:09:31 2003
I've dug through the documentation I've got but haven't found anything.
Does anyone, have a suggestion for getting it clean inside?
Most of the use now is with ethanol suspended copper nano-particles. And there has been a bit of a build up/discoloration of some of the parts. And with a bit of glass this delicate and expensive I just want to be extra cautious.
Thanks,
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 14:53:43 2003
We have a JEOL 1200EX that uses freon in the HV tank.
Yesterday a campus enforcer raided the lab and confiscated our supply of Freon 12 used to replenish the HV tank and just about took me to jail for having it.
I understand and sympathize with the efforts to contain this stuff, but you would think it was heroin or enriched uranium the way this guy acted. He wants it inventoried, under lock and key, and will keep it 'for safe keeping' at the central campus facilities site. If we need it, we can call him and he will arrange for a technician to come to the lab and maybe top off the tank.
No amount of explaining that we hardly ever use it or that when we need to use it we are careful or that my independent service provider doesn't carry this stuff around in his car would satisfy him. We don't have any gross leaks and have only kept the tank around because it came with the microscope and there are directions for checking the HV tank etc. You know, just in case we need it and it is a part of the instrument.
Has anyone had a similar problem? How did you resolve it? Are there any alternatives to this scenario?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 14:57:14 2003
A few decades ago I made many 3.25x4 inch lantern slides (pre-35mm slide presentations). To elimintate Newton Ring formation between the glass slide and the glass cover, we put a frame of paper between them. I do not know the debth of the focus plane in your scanner but if it is not necessary to have the negative in direct contact with the scanner plate, you can easily cut a rectangle out of a piece of paper or thin cardboard, slightly smaller than your negative, put the negative on top and try it out. Perhaps a top frame is also be needed. If this is true a file folder can be used. Inexpensive and not at all messy.
The scanner that I currently use can accomodate a plastic negative holder that was made for it. It must be at least 1mm from the glass plate to the negative surface and the image is still in focus.
Pat Stranen Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 15:22:02 2003
Dear Michael, I don't know if anyone answered your questions, so here is what I know. 1. the Ovonics company originally marketed the layered pseudocrystals with the big d-spacing for the efficient diffraction of light-element x-rays. A search of the Web found an Ovonics company, but I don't know if they are the same. You might contact Oxford Instruments, particularly Joe Carr, their WDX specialist, to see if they are still around, because everyone uses these crystals for light elements. 2. My P-10 gas is delivered with a two-stage regulator set for ten PSI. The original tank I got when the spectrometer was installed in 1987 is still going, over half full. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Michael Cheatham" {mmcheath-at-mailbox.syr.edu} To: {microscopy-at-ns.microscopy.com} Sent: Thursday, October 16, 2003 10:02 AM
A few years ago we had the tank of a JEOL 2000 converted from Freon to SF6 for the same reason. I don't recall the cost but it wasn't way over the top. I will check. Perhaps you will need to do this, too.
cheers, John
******** John S. Vetrano Sr. Research Scientist Materials Structure and Performance Group Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
} ---------- } From: Jon Krupp } Sent: Tuesday, October 21, 2003 12:53 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: [Microscopy] Freon alternatives? } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi: } } We have a JEOL 1200EX that uses freon in the HV tank. } } Yesterday a campus enforcer raided the lab and confiscated our supply of } Freon 12 used to replenish the HV tank and just about took me to jail for } having it. } } I understand and sympathize with the efforts to contain this stuff, but you } would think it was heroin or enriched uranium the way this guy acted. He } wants it inventoried, under lock and key, and will keep it 'for safe } keeping' at the central campus facilities site. If we need it, we can call } him and he will arrange for a technician to come to the lab and maybe top } off the tank. } } No amount of explaining that we hardly ever use it or that when we need to } use it we are careful or that my independent service provider doesn't carry } this stuff around in his car would satisfy him. We don't have any gross } leaks and have only kept the tank around because it came with the } microscope and there are directions for checking the HV tank etc. You know, } just in case we need it and it is a part of the instrument. } } Has anyone had a similar problem? How did you resolve it? Are there any } alternatives to this scenario? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 20:54:59 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (martin.hohmann-marriott-at-asu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 21, 2003 at 18:17:41 ---------------------------------------------------------------------------
Email: martin.hohmann-marriott-at-asu.edu Name: Martin Hohmann-Marriott
Organization: Arizona State University
Title-Subject: [Microscopy] Fixation & embedding with DMSO
Question: Dear hard-working microscopists,
Unfortunatelly, alcohols and acetone dissolve a biological structure that I am interested in visualizing. Therefore I am looking for alternative solvents.
-Is it possible to fix cells in DMSO? -Does DMSO interacct with OsO4? -Does it interact with glutaraldehyde? -Is there a resin that mixes with DMSO?
Jean-Baptiste Comiti has a web page in French on scanning light photography using equipment he built. The only thing more stunning than his images is the equipment he built to get them.
Babel Fish does a passable translation http://babelfish.altavista.com/babelfish/tr enter http://comiti.com in translate a web page and chose French to English. All computer translations have some awkward phasing and part that incorrect but you should be able to follow this one OK.
Thanks to Ted Clarke for bringing this to my attention.
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 01:52:26 2003
1) Replace R-12 with SF6, if this is OK with JEOL.
2) Have any refrigeration service (large one or a single contractor) to store your R-12 for you, and refill HT tank when necessary. They charge from $30 to $80 per hour.
3) Get EPA license for handling refrigerants yourself. It did cost me half-a-day and $80 to take a test. Test is easy. Such license allows purchasing, handling, storing, and using refrigerants, including severely restricted chlorocarbons such as R-12 and others, in all USA. You will be required to have proper equipment, which is simple and inexpensive for refills (refrigeration service valves and hoses). Main rule- do not vent R-12 on purpose. Leaks are OK- same as with old household refrigerator or AC (air conditioner). Just refill alone is easy. But, ask your AC service people to pump out freon when and if HT tank needs repair- this will require more expensive equipment and more work. Make sure that your organization is still part of USA and does not have different rules. I am serious :-)
More important point is that R-12 is not manufactured for a number of years. You can still buy it (recycled), but last time I checked it was close to $1,100 per pound (eleven hundred). Perhaps SF6 is the best way to go.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
This message is made of 100% recycled electrons.
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----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, October 21, 2003 3:53 PM
Jonathan,
I work on automotive air conditioning as a hobby. Perhaps the easiest way to resolve this problem is as Vitaly suggested, to certify one of the cognizant personnel in the lab for use of r12. Though I haven't certified myself, I heard it's quite quick and painless to do online and costs only $20. Click on (http://www.epatest.com/) to get started. Once you have your license, you're certified to own and use r12 refrigerant. This will take the teeth out of the enforcers. For actual service that involves removing refrigerant, you may want to contract a refrigeration specialist.
I heard the price of r12 has actually come down to $20 per 12 oz. can. This is because a lot of pre-1995 cars that use r12 are being retired from service. The used freon supply is increasing and demand for use is decreasing. Freon is available on eBay (with proof of a license).
Be careful with conversions. I'm not familiar with SF6 refrigerant. But if you convert, make sure the lubricant is compatible. If it isn't, the entire refrigeration system will need to be flushed with solvent during the conversion and recharged with the proper type and amount of lubricant. Your cheapest and least compromising option is probably to stay with r12 in your refrigerator.
A good source of information on refrigeration can be found on: http://www.aircondition.com/wwwboard/
Stu Smalinskas Senior Metallurgist SKF North American Technical Center Plymouth, Michigan (734) 414-6862 stu.smalinskas-at-skf.com
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Jonathan wrote:
We have a JEOL 1200EX that uses freon in the HV tank.
Yesterday a campus enforcer raided the lab and confiscated our supply of Freon 12 used to replenish the HV tank and just about took me to jail for having it.
I understand and sympathize with the efforts to contain this stuff, but you would think it was heroin or enriched uranium the way this guy acted. He wants it inventoried, under lock and key, and will keep it 'for safe keeping' at the central campus facilities site. If we need it, we can call him and he will arrange for a technician to come to the lab and maybe top off the tank.
No amount of explaining that we hardly ever use it or that when we need to use it we are careful or that my independent service provider doesn't carry this stuff around in his car would satisfy him. We don't have any gross leaks and have only kept the tank around because it came with the microscope and there are directions for checking the HV tank etc. You know, just in case we need it and it is a part of the instrument.
Has anyone had a similar problem? How did you resolve it? Are there any alternatives to this scenario?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 08:36:08 2003
I was going to suggest "chemical" dehydration with 2,2 dimethoxypropane but then I found out that it yeilds methanol and acetone which you want to avoid. How about dioxane? Or could you just skip solvents and dehydrate by critical point drying or freeze drying, then embed in whatever resin you choose?
Geoff
by way of MicroscopyListServer wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (martin.hohmann-marriott-at-asu.edu) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Tuesday, October 21, 2003 at 18:17:41 } --------------------------------------------------------------------------- } } } Email: martin.hohmann-marriott-at-asu.edu } Name: Martin Hohmann-Marriott } } Organization: Arizona State University } } Title-Subject: [Microscopy] Fixation & embedding with DMSO } } Question: Dear hard-working microscopists, } } Unfortunatelly, alcohols and acetone dissolve a biological structure } that I am interested in visualizing. Therefore I am looking for } alternative solvents. } } -Is it possible to fix cells in DMSO? } -Does DMSO interacct with OsO4? } -Does it interact with glutaraldehyde? } -Is there a resin that mixes with DMSO? } } Thank you for your help } Cheers, } } Martin Hohmann-Marriott } } --------------------------------------------------------------------------- } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 10:16:13 2003
Recently I have been asked to run EDS on scraps and chips of steel alloys. The surfaces are too rough and irregular for quantification but I would like to run standards of different stainless steels to get a ball park idea of relative element responce. Is anyone aware of a source of inexpensive steel reference standards. I don't need NIST traceablity, I just need to know the ASTM alloy type (316 or what ever).
Thanks!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 11:17:19 2003
Freon is used in the tank as an insulator not as a refrigerant.
SF6 is the only (costly) alternative. On a JEOL 1200EX it is not just a case of removing the Freon and replacing it with SF6. It is necessary to get JEOL in to replace components in the HT tank in order to make it compatible with SF6.
Regards
Alan
At 06:21 AM 10/22/2003 -0700, Kestutis Smalinskas wrote:
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Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Please contact me off line if you are interested in a 1980s vintage Philips 430 TEM with one single tilt holder. The price has been reduced considerably. The buyer must pay shipping and handling charges. Full payment is required prior to shipping from ASU.
Please contact me today (I won't be here Thursday and Friday) or next week by e-mail or phone.
If you have specific questions regarding payment, please call Patricia Chase, Surplus Property Manager at 480-965-1850 or e-mail: patricia.chase-at-asu.edu.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Any steel supplied by a reputable metal supply company will come with a certified mill test report (CMTR) containing composition data. You might be able to work out a deal with a friendly neighborhood steel supplier to obtain small samples of several different bars and alloys with a CMTR for each little sample. They might be able to provide you with short remnants from larger bars, or if they are really friendly, let you cut off a small piece that would not diminish the value of a full bar. This could allow you to get several "standards" at minimal cost.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 13:14:24 2003
You should be able to get a pretty good idea of the composition even without flat pieces and probably without SS references. We have one client that is sending us samples in almost every form: chucks, pieces, shavings, drillings, scrapings, etc. They are trying to get an idea of what materials are present throughout their food processing plant so that they can narrow down the source should a piece find its way into their food stream. We find that they have a lot of stainless steel of about 18% Cr content. Sometimes we find a piece with 20% Cr, but I am reluctant to get too excited about the change in content. It might well be a real difference, but I am not sure what tolerances are allowed on alloy compositions.
Since most of the alloying elements are at moderate energy 5-10 kV, the intensities are not terribly affected by geometry. Still, I try to find a flat and level place on the sample to minimize the effects. If elements are being determined from lower energy lines (1-3 kV), then I have to be somewhat suspicious of the results. Intensities can vary significantly based on the local geometry. Fortunately, many alloys are not specified too tightly, e.g., 0.4-0.8% of an element. It may be enough to determine the presence of an element.
You can try some experiments to determine the sensitivity to geometry. You might put a rod in your SEM perpendicular to your x-ray detector axis and move the location of the analysis to see how the results change with tilt. For that matter, if you can tilt your stage both ways with respect to the detector, you could perform the same exercise with a polished, flat sample and know the geometry exactly. You might also try some shavings with roughness and change the size of the raster used to scan the sample and see what effect that has on the results.
As you develop familiarity with what a "normal" spectrum and background looks like (e.g., position of the background hump) you should be able to recognize what should be good spectra from your samples. That should help improve your results. Of course, flattening out the samples as well as you can before analysis will also help.
Warren
At 11:02 AM 10/22/2003 -0400, you wrote:
} Recently I have been asked to run EDS on scraps and chips of steel alloys. } The surfaces are too rough and irregular for quantification but I would } like to run standards of different stainless steels to get a ball park idea } of relative element responce. Is anyone aware of a source of inexpensive } steel reference standards. I don't need NIST traceablity, I just need to } know the ASTM alloy type (316 or what ever). } } Thanks!! } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
} Unfortunatelly, alcohols and acetone dissolve a biological structure that } I am interested in visualizing. Therefore I am looking for alternative } solvents. } } -Is it possible to fix cells in DMSO?=====} DMSO will not chemically fix } -Does DMSO interact with OsO4?=====} perhaps no, check Merk Index for } compatibility. } -Does it interact with glutaraldehyde?======} see above } -Is there a resin that mixes with DMSO?====} perhaps no
Dear Martin I don't understand your point. How do you know that alcohol and acetone dissolve your structure? Did you chemically fix your sample before? If so, it quite unlikely that your "structure" will be "dissolved". I am surprise: DMSO is such good solvent, it should "dissolve" even better than ethanol or acetone. Nevertheless, you may try some water-compatible embedding media to avoid organics. Some acrylates may do the job. Durcupan also comes to my mind (old stuff). Another choice is to use "freeze-substitution" - it's shown that at low temperature acetone has much smaller "dissolving" ability... Good luck, Sergey
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 14:28:35 2003
I installed digital locks on all my Lab doors, so nobody without PIN could enter (including the head of Department). The idea is that EM "presence" risk to your health, so you have to be trained and certified (by me) in order to have access to that "hazardous, nasty" place... So, everyone who want to enter my Lab should be "certified" by me. To made it more "professional" they actually have to sign a special form... I also established the strict rule that everyone who want to see my rooms should set appointment with me. My previous experience with Russian bureaucracy helps me so much here in US. The bottom line here - to do everything legally and in agreement with local laws. Local policies are not a law by the way. Sergey
At 06:21 AM 10/22/2003, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Thanks for your comments and suggestion for the negative film scanner. By comparing the specifications and prices of scanners your suggested, we decided to modify our present scanner holder. The holder type we are using for TEM films is Nikon FH-869S. We polished the two strips, which were used to fix the film size. The two strips are much easy to be milled by lathe in our machine shop. Then we got the space, which is a little bit larger than the film size. Right now, the films are fixed just by two sides upper holders with 4 hooks, however, the films are as stable as before. If anyone who is using this kind of holder has the similar problem as we have, please try this way to modify your holder. Thanks all again.
Regards, Qi Zhang
*************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu qizhang-at-email.unc.edu
--- Zhang Qi {qzhangtj-at-yahoo.com} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear colleagues, } } We are in great need of a negative film scanner used } to scan HRTEM films with the size of 3.25x4 inches. } Although we have a Nikon super coolscan 8000 film } scanner, the film holder is kind of small. It is } annoying to cut the films every time when we scan } them. If you use scanner to scan the electron } microscopy films, please give me your suggestion for } the vendor and model. Actually, this question has } been } discussed before, so any new comment? Thanks. } } Regards, } Qi Zhang } } *************************************************** } Qi Zhang, Ph.D } 164 Phillips Hall, CB#3255 } Department of Physics and Astronomy } the University of North Carolina at Chapel Hill } Chapel Hill, NC 27599 } Tel: 919-843-2407 } Fax: 919-962-0480 } E-mail: qizhang-at-physics.unc.edu } qizhang-at-email.unc.edu } } } __________________________________ } Do you Yahoo!? } The New Yahoo! Shopping - with improved product } search } http://shopping.yahoo.com }
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 15:09:22 2003
Dear Frank, When I am trying to identify alloys, I use the listings in the American Society of Metals Handbook. They list the composition ranges for most of the alloys known. There is a desk addition (the full Handbook is 16 or more volumes) that covers the basics. You might go to the ASM web site and see if they sell a book or CD that will give you the basic alloy compositions. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {Frank.Karl-at-degussa.com} To: {microscopy-at-msa.microscopy.com} Sent: Wednesday, October 22, 2003 8:02 AM
I wonder if the new replacement for R-12 will work ( I think it is 134). It is compatible with the same seals in refrigeration systems. The only problem is compatibility with the lubricants, but that is not a problem here. When I needed to stop using R-114, I called DuPont and spoke with an engineer. I explained what my system was doing with the R-114, and he helped me find an alternative in the new "SUVA" refrigerants. The specification of interest here would be the dielectric properties and the composition of the seals. There are still rules on the handling of it, but it is not looked at as if it is going cause the end of the universe, which would, of course, find us all at Milliways.
Sorry, I strayed a bit. If it were me, I would call DuPont and talk to an engineer there. If you switch to something not on the EPA hit list, the campus enforcers should be more willing to work with you. You will, of course need a qualified contractor to remove the R-12 in an approved manner. You may be allowed to do the top-offs yourself, afterwards. They sell the new refrigerant in auto parts stores for home use.
Regards, Darrell
Alan Nicholls {nicholls-at-uic.edu To: microscopy-at-sparc5.microscopy.com } cc: Subject: [Microscopy] Re: Re: Freon alternatives? 10/22/2003 12:25 PM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Jonathon
Freon is used in the tank as an insulator not as a refrigerant.
SF6 is the only (costly) alternative. On a JEOL 1200EX it is not just a case of removing the Freon and replacing it with SF6. It is necessary to get JEOL in to replace components in the HT tank in order to make it compatible with SF6.
Regards
Alan
At 06:21 AM 10/22/2003 -0700, Kestutis Smalinskas wrote:
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Jonathan, } } I work on automotive air conditioning as a hobby. } Perhaps the easiest way to resolve this problem is as } Vitaly suggested, to certify one of the cognizant } personnel in the lab for use of r12. Though I haven't } certified myself, I heard it's quite quick and } painless to do online and costs only $20. Click on } (http://www.epatest.com/) to get started. Once you } have your license, you're certified to own and use r12 } refrigerant. This will take the teeth out of the } enforcers. For actual service that involves removing } refrigerant, you may want to contract a refrigeration } specialist. } } I heard the price of r12 has actually come down to $20 } per 12 oz. can. This is because a lot of pre-1995 } cars that use r12 are being retired from service. The } used freon supply is increasing and demand for use is } decreasing. Freon is available on eBay (with proof of } a license). } } Be careful with conversions. I'm not familiar with } SF6 refrigerant. But if you convert, make sure the } lubricant is compatible. If it isn't, the entire } refrigeration system will need to be flushed with } solvent during the conversion and recharged with the } proper type and amount of lubricant. Your cheapest } and least compromising option is probably to stay with } r12 in your refrigerator. } } A good source of information on refrigeration can be } found on: http://www.aircondition.com/wwwboard/ } } Stu Smalinskas } Senior Metallurgist } SKF North American Technical Center } Plymouth, Michigan } (734) 414-6862 } stu.smalinskas-at-skf.com } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } Jonathan wrote: } } We have a JEOL 1200EX that uses freon in the HV tank. } } Yesterday a campus enforcer raided the lab and } confiscated our supply of } Freon 12 used to replenish the HV tank and just about } took me to jail for } having it. } } I understand and sympathize with the efforts to } contain this stuff, but you } would think it was heroin or enriched uranium the way } this guy acted. He } wants it inventoried, under lock and key, and will } keep it 'for safe } keeping' at the central campus facilities site. If we } need it, we can call } him and he will arrange for a technician to come to } the lab and maybe top } off the tank. } } No amount of explaining that we hardly ever use it or } that when we need to } use it we are careful or that my independent service } provider doesn't carry } this stuff around in his car would satisfy him. We } don't have any gross } leaks and have only kept the tank around because it } came with the } microscope and there are directions for checking the } HV tank etc. You know, } just in case we need it and it is a part of the } instrument. } } Has anyone had a similar problem? How did you resolve } it? Are there any } alternatives to this scenario? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } __________________________________ } Do you Yahoo!? } The New Yahoo! Shopping - with improved product search } http://shopping.yahoo.com
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com ----- Forwarded by Robert Fowler/TCU/TDK-US on 10/23/2003 07:44 AM -----
Robert Fowler To: Microscopy-at-sparc5.microscopy.com 10/22/2003 cc: 02:01 PM Subject: [Microscopy] Help with Nikon 990 Threaded Adapter
Listers My Nikon 990 threaded adapter (on the camera end) has seen its better days after repeated threading and unthreading (I know shame on me for not dedicating....budget). It now has no more threads to attach to the MDC/c-mount adapter. Does anyone out there have a solution to this problem? Is there a threaded adapter available from Nikon or elsewhere? Any solution is better than no solution. Thank you
BTW Sorry if this is double posted had some trouble with my subscription
Also I talked to many people at Nikon before someone informed me this was not a replacement part so ALL comments are welcome
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com
THIS TRANSMISSION IS INTENDED FOR THE SOLE USE OF THE INDIVIDUAL AND ENTITY TO WHOM IT IS ADDRESSED AND MAY CONTAIN PRIVILEGED AND/OR CONFIDENTIAL INFORMATION.
If you are not the intended recipient, be advised that any use, dissemination, distribution or duplication of this transmission is strictly prohibited. If you received this transmission in error, please notify the sender immediately by electronic reply to this transmission or by phone (847-803-6100). Thank you.
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 07:48:17 2003
DMSO isn't a fixative, but can be added to improve fixative penetration. It doesn't react with glutaraldehyde and is commonly used in combination with it. Use caution with this combination so as to not fix yourself, too. It is my understanding that DMSO does, however, react badly with osmium tetroxide forming a tar-like substance in and around your samples.
Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 13:30:37 2003
My supervisor has just requested our travel and training needs for the upcoming year. I am new to TEM and would like to attend some biological type seminars. Does anyone have any suggestions or recommendations for future meetings?
-- Sue Tyler Biologist Cooperative Oxford Laboratory Center for Coastal Environmental Health Biomolecular Research at Charleston ( CCHEBR) USDOC/NOAA/NOS/NCCOS 904 S. Morris St. Oxford, Maryland 21654-9724 410-226-5193 Fax: 410- 226-5925 Sue.Tyler-at-noaa.gov
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 14:18:14 2003
Dear Martin How do you know DMSO is safe? Also, if your structures are soluble in alcohol and acetone, there is a good chance they will be dissolved or damaged by the resins themselves at 20oC unless they are chemically fixed. Sergey is right - you should consider using freeze-substitution and low temperatures for the entire process - dehydration, infiltration with resin and polymerization. Power of solvents is dramatically altered by temperature. e.g. fats and waxes soluble in chloroform at +20oC become insoluble at -20oC.
But none of us can give you really good advice unless you tell us what your structure is, or at least its composition.
Chris
Dr. Chris Jeffree
} ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, October 22, 2003 8:10 PM } Subject: [Microscopy] Re: via-WWW: Fixation & embedding with DMSO } } } } } } } } -------------------------------------------------------------------- } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ----------- } } } } } } } Unfortunatelly, alcohols and acetone dissolve a biological } structure that } } } I am interested in visualizing. Therefore I am looking for } alternative } } } solvents. } } } } } } -Is it possible to fix cells in DMSO?=====} DMSO will not } chemically fix } } } -Does DMSO interact with OsO4?=====} perhaps no, check Merk Index } for } } } compatibility. } } } -Does it interact with glutaraldehyde?======} see above } } } -Is there a resin that mixes with DMSO?====} perhaps no } } } } Dear Martin } } I don't understand your point. How do you know that alcohol and } acetone } } dissolve your structure? Did you chemically fix your sample before? } If } } so, it quite unlikely that your "structure" will be "dissolved". I } am } } surprise: DMSO is such good solvent, it should "dissolve" even } better than } } ethanol or acetone. Nevertheless, you may try some water-compatible } } embedding media to avoid organics. Some acrylates may do the job. } } Durcupan also comes to my mind (old stuff). Another choice is } to use } } "freeze-substitution" - it's shown that at low temperature acetone } has much } } smaller "dissolving" ability... Good luck, Sergey } } } } }
------- End of forwarded message ------- ========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 14:19:58 2003
I don't know if our adapter would help you (it is a 28mm threaded adapter as well). But, if you would like to order one and give it a shot, you can do so with a 30 day no-hassle return policy.
What makes our adapters better? http://www.mvia.com/Coolpix/clpxadpt.htm#What_makes_Our_Adapters_Better
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Robert.Fowler-at-tdktca.com wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Robert Fowler } Quality Assurance Failure Analysis Technician } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.315 } Fax: (770) 487-1460 } email: rfowler-at-tdktca.com } www.component.tdk.com } ----- Forwarded by Robert Fowler/TCU/TDK-US on 10/23/2003 07:44 AM ----- } } Robert Fowler } To: Microscopy-at-sparc5.microscopy.com } 10/22/2003 cc: } 02:01 PM Subject: [Microscopy] Help with Nikon 990 Threaded Adapter } } } } Listers } My Nikon 990 threaded adapter (on the camera end) has seen its better days } after repeated threading and unthreading (I know shame on me for not } dedicating....budget). It now has no more threads to attach to the } MDC/c-mount adapter. Does anyone out there have a solution to this problem? } Is there a threaded adapter available from Nikon or elsewhere? Any } solution is better than no solution. Thank you } } BTW Sorry if this is double posted had some trouble with my subscription } } Also I talked to many people at Nikon before someone informed me this was } not a replacement part so ALL comments are welcome } } Robert Fowler } Quality Assurance Failure Analysis Technician } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.315 } Fax: (770) 487-1460 } email: rfowler-at-tdktca.com } www.component.tdk.com } } THIS TRANSMISSION IS INTENDED FOR THE SOLE USE OF THE INDIVIDUAL AND ENTITY } TO WHOM IT IS ADDRESSED AND MAY CONTAIN PRIVILEGED AND/OR CONFIDENTIAL } INFORMATION. } } If you are not the intended recipient, be advised that any use, } dissemination, distribution or duplication of this transmission is strictly } prohibited. If you received this transmission in error, please notify the } sender immediately by electronic reply to this transmission or by phone } (847-803-6100). Thank you.
--
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 17:22:02 2003
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: {Robert.Fowler-at-tdktca.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 23, 2003 7:44 AM
Thanks for the response.
the info I seek is specific to STEM on/in an SEM. Major SEM makers offer a STEM option. Some tough the BF and DF feature. I just don't understand how a dark field STEM image is acquired on a SEM which has a STEM stage option.
gary g.
At 06:56 PM 10/23/2003, you wrote: } Gary, } } The detectors are the same, the difference is in geometry. Bright field is } a circle, and dark field is a ring around the circle. In } a properly aligned STEM all that is required to switch from BF to DF is to } switch the sensors. Both BF and DF signals are present } all time, it matters what sensor you are acquiring from. I am not sure } what you mean by SEM. This sensor arrangement will produce } results identical for both sensors in SEM. Did you mean TEM? Sensors, BTW, } are same (well, almost) as for SEM backscattered } electrons detectors. I can look up something and fax it to you if you } wish. Several pages. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } (678)467-0012 mobile } www.sia-cam.com } } This message is made of 100% recycled electrons. } } This address can not receive messages larger than 15 kb without prior } notification. } } ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} } To: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} } Sent: Thursday, October 23, 2003 6:31 PM } Subject: [Microscopy] Darkfield STEM } } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------- } } } } Hi all: } } } } Is there a reference source that explains how } } a BF/DF STEM detector works on a SEM? In } } particular, how does it do DF? } } } } tnx, } } gary g. } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 24 02:25:35 2003
http://www.microscopy-uk.net/mag/artaug01/vrcoolpix.htm and http://www.microscopy-uk.net/mag/artsep01/vrcoolpix2.htm
It's a way to mount a coolpix or any other digital camera on a microscope, using the tripod thread insted of the objectif thread. You'll need perheps to modify your microscope interface. Of coarse you need a workshop ! It works well, particulary if you can have a centring of your interface inside (or outside) the "late thred" of the camera objectif.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Sorry folks, but I can't seem to post the list these days, please respond directly to me: Richard Edelmann {edelmare-at-MUOHIO.EDU} and not the sender of this message
------- Forwarded message follows ------- } From: Richard Edelmann {edelmare-at-MUOHIO.EDU} To: emsabbs
Gary
The geometry, what is and how it is collected is exactly the same as STEM in TEM.
The BF/DF detectors are after the specimen and collect electrons that have been transmitted through the (thin) specimen. The BF detector collects electrons close to the optical axis which have not suffered a scattering interaction with the specimen (direct beam). The Annular DF detector, which is a ring scintillator with a hole in the center, simultaneously collects electrons that have been forward scattered from the specimen in to a particular angular range (typical 10mrad to maybe 300mrad). Close to the direct beam these are diffracted and inelastically scattered electrons, further out these are elastically scattered electrons. Assuming the dark field image collects most of the scattered electrons the image will look like the negative of the bright field image.
The technique is briefly mentioned in "Scanning Electron Microscopy and X-ray Microanalysis" by Goldstein et al. and also more extensively in "Transmission Electron Microscopy" by Williams and Carter
Alan
At 07:10 PM 10/23/2003 -0700, Gary Gaugler wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
There is a (relatively) cheaper substitute for Durcupan called Aquembed, available from Ladd. I used to use it routinely for processing alcohol-sensitive material into Epon and it worked very well, though it extends the processing times considerably. I never tried it after DMSO, but I was always careful to wash all traces of fixative and buffers out before going to the first dilution of Aquembed.
Lesley Weston.
on 22/10/2003 12:10 PM, Sergey Ryazantsev at sryazant-at-ucla.edu wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } } } Unfortunatelly, alcohols and acetone dissolve a biological structure that } } I am interested in visualizing. Therefore I am looking for alternative } } solvents. } } } } -Is it possible to fix cells in DMSO?=====} DMSO will not chemically fix } } -Does DMSO interact with OsO4?=====} perhaps no, check Merk Index for } } compatibility. } } -Does it interact with glutaraldehyde?======} see above } } -Is there a resin that mixes with DMSO?====} perhaps no } } Dear Martin } I don't understand your point. How do you know that alcohol and acetone } dissolve your structure? Did you chemically fix your sample before? If } so, it quite unlikely that your "structure" will be "dissolved". I am } surprise: DMSO is such good solvent, it should "dissolve" even better than } ethanol or acetone. Nevertheless, you may try some water-compatible } embedding media to avoid organics. Some acrylates may do the } job. Durcupan also comes to my mind (old stuff). Another choice is to use } "freeze-substitution" - it's shown that at low temperature acetone has much } smaller "dissolving" ability... Good luck, Sergey } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 25 09:32:53 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tpepper-at-iastate.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 23, 2003 at 13:29:48 ---------------------------------------------------------------------------
Question: Greetings: I had a client come by today with a bunch of slides made from paraffin sections and some cryo sections of various rat parts that had lost their hematoxylin counter-staining (Gill's formula). Any clues as to what is causing/caused this? They were stored in dark in slide boxes, some slides have sections with stain and some without. It is showing up on slides from this last year and is inconsistent. Very strange! Thanks, Tracey Pepper
Can you change the angular acceptance as you would by changing the camera length on a TEM/STEM ? On a TEM/STEM you can have ADF, HAADF and "medium" or "low" angle ADF, which can be useful (i.e., for strain imaging). I don't know if you can have the same range of options on a SEM/STEM and what the angle(s) are and if they completing eliminate diffraction contrast. Anyone know?
----- Original Message ----- } From: "Alan Nicholls" {nicholls-at-uic.edu} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Friday, October 24, 2003 7:47 AM
Call for Nomination of Individuals to be considered for Major Awards by the Microscopy Society of America.
Awards:
Distinguished Scientist Awards:
These Awards recognize preeminent senior scientists from both the Biological and Physical disciplines who have a long-standing record of achievement during their career in the field of microscopy or microanalysis.
Burton Medal:
The Burton Medal was initiated to honor the distinguished contributions to the field of microscopy and microanalysis of a scientist who is less than 40 years of age on January 1st of the award year.
Optical Imaging Association-MSA Outstanding Young Investigator Award:
This Award, initiated in 1999, recognizes the distinguished contributions in the field of optical microscopy made by a scientist who is less than 40 years of age on January 1st of the award year.
Outstanding Technologist Awards:
These Awards honor technologists from both the Biological and Physical Sciences who have made significant contributions such as the development of new techniques which have contributed to the advancement of microscopy and microanalysis.
Morton D. Maser Distinguished Service Award:
This Award was initiated to recognize outstanding volunteer service to the Society as exemplified by Mort Maser, who served the Society for many years with great dedication. This award is made to honor an MSA member who has provided significant volunteer service to the Society over a period of years.
Nomination Requirements:
The Distinguished Scientist, Burton Medal, OIA-MSA Outstanding Young Investigator and Outstanding Technologist Awards Nominations should include:
1) a letter from the primary MSA nominator describing the research accomplishments of the candidate with particular emphasis on the unique technical achievements in the Physical or Biological Sciences; and 2) supplemental letters of support from other members of the scientific community.
The Morton D. Maser Distinguished Service Award Nomination should include:
1) a letter from the primary MSA nominator describing the basis for the nomination; and 2) supplemental letters of support from other members of MSA.
The Deadline for receipt of Awards Nomination Packages is December 15, 2003.
Please contact the MSA Business Office for additional information.
Judy Janes, Administrative Manager Bostrom Corporation 230 E. Ohio Street, Suite 400 Chicago, IL 60611-3265 (800) 538-3672; Fax (312) 644-8557, jjanes-at-MSA.microscopy.org
Thanks,
William T. Gunning, Ph.D. Associate Professor of Pathology Medical College of Ohio Department of Pathology BHSB 140 3035 Arlington Avenue Toledo, Ohio 43614-5806 Phone: 419-383-5256 Fax: 419-383-3066 email: wgunning-at-mco.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 27 07:22:04 2003
Robert, Thales-Optem has couplers for Coolpix and most other compact digital cameras. Our customers have been very successful with them. Check out http://www.thales-optem.com/dca.html versions are available to fit C-mount, eyepiece, and many photoport or phototube mountings. We can assist with configuration if needed.
George
George Laing National Graphic Supply 800.223.7130 x3109 scisales-at-ngscorp.com
} } Listers } My Nikon 990 threaded adapter (on the camera end) has seen its better days } after repeated threading and unthreading (I know shame on me for not } dedicating....budget). It now has no more threads to attach to the } MDC/c-mount adapter. Does anyone out there have a solution to this problem? } Is there a threaded adapter available from Nikon or elsewhere? Any } solution is better than no solution. Thank you }
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 27 10:08:54 2003
Most digital cmaeras I hear of in the consumer market use FireWire (IEEE 1394), but in more professional high-speed applications Cameralink seems to be a better solutions as it has more bandwidth to offer. Does anyone have experience with CameraLink cameras for microsocpy ?
It seems to me that a CameraLink camera and interface are the best choice for the highest speed imaging applications. A CameraLink interface can achieve real-time 30fps 12-bit video at 3.2 Megapixel resolution. In its base configuration, CameraLink can transfer at up to 1.6 Gb/sec. and in full configuration, using 2 cables, the rate is extended up to 4.8 Gb/sec.
With the FireWire cameras I know of you pay a serious speed penalty when capturing at full frame size without binning.
The simple answer is no. The detector is a fixed distance from the specimen and there are no lenses in between. In principle you can change the inner angle by using a different hole size, a mask in front of the detector or a segmented solid state detector made up of many rings - not sure any of these have been used in an SEM, all three have been tried in dedicated 100kV STEMs which also have no post specimen lenses.
Alan
At 08:17 AM 10/25/2003 -0700, Edward Principe wrote: } However, } } Can you change the angular acceptance as you would by changing the camera } length on a TEM/STEM ? } On a TEM/STEM you can have ADF, HAADF and "medium" or "low" angle ADF, which } can be useful (i.e., for strain imaging). } I don't know if you can have the same range of options on a SEM/STEM and } what the angle(s) are and if they completing eliminate diffraction contrast. } Anyone know? } } } ----- Original Message ----- } From: "Alan Nicholls" {nicholls-at-uic.edu} } To: "Gary Gaugler" {gary-at-gaugler.com} } Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} } Sent: Friday, October 24, 2003 7:47 AM } Subject: [Microscopy] Re: Re: Darkfield STEM } } } } } } } } -------------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------------- } ----- } } } } Gary } } } } The geometry, what is and how it is collected is exactly the same as STEM } } in TEM. } } } } The BF/DF detectors are after the specimen and collect electrons that have } } been transmitted through the (thin) specimen. The BF detector collects } } electrons close to the optical axis which have not suffered a scattering } } interaction with the specimen (direct beam). The Annular DF detector, } } which is a ring scintillator with a hole in the center, simultaneously } } collects electrons that have been forward scattered from the specimen in } to } } a particular angular range (typical 10mrad to maybe 300mrad). Close to } the } } direct beam these are diffracted and inelastically scattered electrons, } } further out these are elastically scattered electrons. Assuming the dark } } field image collects most of the scattered electrons the image will look } } like the negative of the bright field image. } } } } The technique is briefly mentioned in "Scanning Electron Microscopy and } } X-ray Microanalysis" by Goldstein et al. and also more extensively in } } "Transmission Electron Microscopy" by Williams and Carter } } } } Alan } } } } At 07:10 PM 10/23/2003 -0700, Gary Gaugler wrote: } } } } } } } } --------------------------------------------------------------------------- } --- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } --------------------------------------------------------------------------- } ---- } } } } } } Thanks for the response. } } } } } } the info I seek is specific to STEM on/in } } } an SEM. Major SEM makers offer a STEM option. } } } Some tough the BF and DF feature. I just don't } } } understand how a dark field STEM image is } } } acquired on a SEM which has a STEM stage option. } } } } } } gary g. } } } } } } } } } At 06:56 PM 10/23/2003, you wrote: } } } } Gary, } } } } } } } } The detectors are the same, the difference is in geometry. Bright field } } } } is a circle, and dark field is a ring around the circle. In } } } } a properly aligned STEM all that is required to switch from BF to DF is } } } } to switch the sensors. Both BF and DF signals are present } } } } all time, it matters what sensor you are acquiring from. I am not sure } } } } what you mean by SEM. This sensor arrangement will produce } } } } results identical for both sensors in SEM. Did you mean TEM? Sensors, } } } } BTW, are same (well, almost) as for SEM backscattered } } } } electrons detectors. I can look up something and fax it to you if you } } } } wish. Several pages. } } } } } } } } Vitaly Feingold } } } } Scientific Instruments and Applications } } } } 2773 Heath Lane, Duluth GA 30096 } } } } (770)232-7785 ph. } } } } (770)232-1791 fax } } } } (678)467-0012 mobile } } } } www.sia-cam.com } } } } } } } } This message is made of 100% recycled electrons. } } } } } } } } This address can not receive messages larger than 15 kb without prior } } } } notification. } } } } } } } } ----- Original Message ----- } } } } From: "Gary Gaugler" {gary-at-gaugler.com} } } } } To: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} } } } } Sent: Thursday, October 23, 2003 6:31 PM } } } } Subject: [Microscopy] Darkfield STEM } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------------- } ----- } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } } To Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } } ------------------------------------------------------------------------- } ------ } } } } } } } } } } Hi all: } } } } } } } } } } Is there a reference source that explains how } } } } } a BF/DF STEM detector works on a SEM? In } } } } } particular, how does it do DF? } } } } } } } } } } tnx, } } } } } gary g. } } } } } } } } } } } } } } } } } } } } } } } } } Alan W Nicholls, PhD } } Director of Research Service Facility (Electron Microscopy) } } Research Resources Center - East (M/C 337) } } Room 100 Science and Engineering South Building } } The University of Illinois at Chicago } } 845 West Taylor St } } Chicago, IL 60607-7058 } } } } Tel: 312 996 1227 } } Fax: 312 996 8091 } } Office: Room 110 } } } } Web site www.rrc.uic.edu } } } } } }
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
I agree with Peter, that the Cameralink currently is the faster interface, and for applications that need this speed it is probably the better one. However, for cameralink you need to add a card to your computer or a PCMCIA card to your laptop, whereas IEEE 1394 is nowadays directly build into the computer and operating system. As far as speed is concerned, Firewire in it's present generation allows up to 400 Mbps, and we will soon see 800 and 1.6 Gbps.
At present, many cameras (especially in the TEM and fluorescence sectors) are limited not by the transfer rate of the interface, but by intensity issues. If you want to transfer 30 frames per second, you must also be able to live with exposure times of 30 msec or less. This may only be possible through larger pixels on the chip (=lower resolution) or binning (again lower resolution).
Another factor is, that for higher transfer rates you also need to read out the chip faster. The so-called "readout noise", however, is proportional to the readout speed. This again may lead to the transfer speed being of less importance.
If you have applications that require the high bandwidth (several cameras, or situations where noise and intensity is not a problem), the cameralink interface provides higher throughput. In other cases, the bandwidth may not be the most important factor. I don't think, there is a general answer for this question.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Peter Van Osta [mailto:pvosta-at-unionbio-eu.com] Sent: Monday, October 27, 2003 9:14 AM To: Confocal Microscopy List; MSA
Hi,
Most digital cmaeras I hear of in the consumer market use FireWire (IEEE 1394), but in more professional high-speed applications Cameralink seems to be a better solutions as it has more bandwidth to offer. Does anyone have experience with CameraLink cameras for microsocpy ?
It seems to me that a CameraLink camera and interface are the best choice for the highest speed imaging applications. A CameraLink interface can achieve real-time 30fps 12-bit video at 3.2 Megapixel resolution. In its base configuration, CameraLink can transfer at up to 1.6 Gb/sec. and in full configuration, using 2 cables, the rate is extended up to 4.8 Gb/sec.
With the FireWire cameras I know of you pay a serious speed penalty when capturing at full frame size without binning.
I've searched the list-server archives as much as I have patience for, and the 'library' here is a collection of just about every biological book on EM but none seem to document a procedure for making lacey grids.
Before I make the recommendation to purchase grids from one of the EM supply houses, I would like to make sure that making these isn't something that can be done easily and routinely.
The intended application is not for biological use. If anyone has a protocol they can share it would be much appreciated.
Thank you,
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 27 17:58:05 2003
Lacey films are extremely holey films containing numerous holes so that specimens can be suspended over the thin, web-like extensions of plastic or carbon-coated plastic. We describe a method in our textbook (page 95, 2nd edition) using glycerine suspended in Formvar.
Here is another method, described by F.S. Sjostrand in 1956 (Proceedings of First Europena Regional Conference on electron Microscopy, Stockholm, p. 120). This is also described in detail on pages 294-297 of his textbook: Sjostrand, F.S. 1967. Electron Microscopy of Cells and Tissues. Academic Press, 462 pages.
1. Dip microscope slide into solution of 2% Formvar (or Butvar) in ethylene dichloride. 2. IMMEDIATELY transfer the slide to a beaker containing a saturated atmosphere of ethylene dichloride (ED) for 20 min. (Make this by lining a beaker with filter paper and soaking the paper with the ED. Keep tightly covered.) 3. After 20 min, remove cover of ED chamber and blow in a stream of air saturated with water vapor. (Do this by blowing air through a flask of water at about 70 degrees C.) 4. Float the formvar net on a water surface. About the upper 2/3 of the slide has the lacey part of the film. 5. Coat with carbon for strength, if desired.
JB
} I've searched the list-server archives as much as I have patience for, } and the 'library' here is a collection of just about every biological } book on EM but none seem to document a procedure for making lacey grids. } } Before I make the recommendation to purchase grids from one of the EM } supply houses, I would like to make sure that making these isn't } something that can be done easily and routinely. } } The intended application is not for biological use. If anyone has a } protocol they can share it would be much appreciated. } } Thank you, } } Geoff Williams } Microscopy Facility Supervisor -- ############################################################## 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 ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 27 21:41:09 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (timothy-at-che.utexas.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 27, 2003 at 08:09:50 ---------------------------------------------------------------------------
Question: We are in initial stages of purchasing a cryogenic microtome and would like to gain some insight on the quality and performance of commercial microtomes. The brand of microtomes that we are currently looking at are RMC, Microstar, and Leica. Of these brands, which one would you recommend or not recommend and why? The microtome will be primarily used to cut polymeric samples.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stefan.Gunnarsson-at-ebc.uu.se) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 27, 2003 at 09:19:51 ---------------------------------------------------------------------------
Email: Stefan.Gunnarsson-at-ebc.uu.se Name: Stefan Gunnarsson
We are in the process of buying a FEGSEM with EDS and are thinking about an EDS-system with an LN2-free detector. Both EDAX and Noran claim to have such detectors that compare in performance (regarding resolution and lifetime) with the LN2-cooled ones. If anyone has experience of these things, I would be very happy for a bit of input and opinions on this. (There are several reasons why we don't want LN2: safety, workload, availability etc.)
thanks for any replies,
Stefan
...................................................................................................................... Dr Stefan Gunnarsson Evolutionary Biology Centre Microscopy and Imaging Unit Norbyvgen 18A SE75236 Uppsala, Sweden Tel & Fax: +46 - 18471 2638 ......................................................................................................................
We here in Botswana have a "in between" detector from EDAX on our ESEM. We are very happy with it. You fill it with LN2 when you need it. Wait for ~30 min to stabilize and use it. Keep LN2 for roughly two days. No forced 365 day a year forced filling even on Christmas we can be free (Was it not for the TEM!) I personally feel uncomfortable to let a crystal to room temp since the "fear of God" was put in me if it does happen. Battling to break the bond on the law. Otherwise it is great. Have no experience with LN2 free detrectors. Just worht having a look at the ressolution and light element sensitivity.
hope it helps
-----Original Message----- } From: by way of MicroscopyListserver [mailto:Stefan.Gunnarsson-at-ebc.uu.se] Sent: Tuesday, October 28, 2003 5:48 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stefan.Gunnarsson-at-ebc.uu.se) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 27, 2003 at 09:19:51 ---------------------------------------------------------------------------
Email: Stefan.Gunnarsson-at-ebc.uu.se Name: Stefan Gunnarsson
We are in the process of buying a FEGSEM with EDS and are thinking about an EDS-system with an LN2-free detector. Both EDAX and Noran claim to have such detectors that compare in performance (regarding resolution and lifetime) with the LN2-cooled ones. If anyone has experience of these things, I would be very happy for a bit of input and opinions on this. (There are several reasons why we don't want LN2: safety, workload, availability etc.)
thanks for any replies,
Stefan
..................................................................................................................... Dr Stefan Gunnarsson Evolutionary Biology Centre Microscopy and Imaging Unit Norbyv"gen 18A SE75236 Uppsala, Sweden Tel & Fax: +46 - 18471 2638 .....................................................................................................................
Colleagues, (1) has anybody successfully adapted the NIKON COOLPIX 5400 to a microscope via a C-mount adapter? (2)by use of the c-mount adapter as available for the Coolpix 990 / 995 / 4500 ?? (3)are you content with the results ? Please respond short and informally, at best off-line, to peter.heimann-at-uni-bielefeld.de Thanks, Peter
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 02:39:59 2003
Dear all, Not quite microscopy directly, but is anyone using software to collate and organise references to the literature, which can also export to Word format or that sort of thing. Any hints would be greatly appreciated.
Thanks
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 06:24:55 2003
a general problem with digital cameras and microscope adapters is the lens diameter of the camera and the lens diameter of the microscope eye piece/c-mount. If you buy a digital camera from a microscope manufacturer with c- mount adapter, you get a modified eye piece with a c-mount thread and a holder for the camera (which places the lens of the camera in front of the eye piece); Leica sold that a few month ago for app. 2000 with a Canon S40. For good pictures you need a camera with a small front lens, and an eye piece with a large lens; and still you have to use the zoom. Otherwise you get circular pictures; surrounded with a black frame. Before I bought the Canon, I thought of the Nikon 5400 (b/c of the wide field lens), but then decided for the Canon (because it comes with a nice software "remote capture" to operate the camera with a PC). I think the Nikon 5400 is not the best choice for taking pictures with a microscope, the Nikon 4500 is much better. The Canon G3 is as bad as the 5400, but I bought it not just for microscopy, but also for taking pictures with a ring flash e.g.
:-) Torsten
} } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Colleagues, } (1) has anybody successfully adapted the NIKON COOLPIX 5400 to a } microscope via a C-mount adapter? (2)by use of the c-mount adapter as } available for the Coolpix 990 / 995 / 4500 ?? (3)are you content with } the results ? Please respond short and informally, at best off-line, } to peter.heimann-at-uni-bielefeld.de Thanks, Peter } } }
I like Reference Manager, b/c it has an internal pubmed search. What I don't like is, that these programs only support MS Word directly, and not other formats (anymore), like Lotus Wordpro/Wordperfect/OpenOffice.
:-) Torsten
} --------- } } Dear all, } Not quite microscopy directly, but is anyone using software to collate } and organise references to the literature, which can also export to } Word format or that sort of thing. Any hints would be greatly } appreciated. } } Thanks } } --
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 06:43:39 2003
Dear all, Can anyone explain to me the difference between "Endnote" and "Reference Manager"? They both seem to be made by the same company, cost about the same, but I couldn't extract a comparison of what the two packages do differently from their websites.
Thanks again
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 07:47:17 2003
Or, given the amount of effort involved, you could just buy some. I am sure EBS, SPI, Fullam and Pella would be glad to supply some at reasonable cost. If you have never made them before I would have thought that buying was the almost surefire way of getting what you want.
On Monday, October 27, 2003, at 07:03 PM, John J. Bozzola wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Geoff, } } Lacey films are extremely holey films containing numerous holes so } that specimens can be suspended over the thin, web-like extensions of } plastic or carbon-coated plastic. We describe a method in our textbook } (page 95, 2nd edition) using glycerine suspended in Formvar. } } Here is another method, described by F.S. Sjostrand in 1956 } (Proceedings of First Europena Regional Conference on electron } Microscopy, Stockholm, p. 120). This is also described in detail on } pages 294-297 of his textbook: Sjostrand, F.S. 1967. Electron } Microscopy of Cells and Tissues. Academic Press, 462 pages. } } 1. Dip microscope slide into solution of 2% Formvar (or Butvar) in } ethylene dichloride. } 2. IMMEDIATELY transfer the slide to a beaker containing a saturated } atmosphere of ethylene dichloride (ED) for 20 min. (Make this by } lining a beaker with filter paper and soaking the paper with the ED. } Keep tightly covered.) } 3. After 20 min, remove cover of ED chamber and blow in a stream of } air saturated with water vapor. (Do this by blowing air through a } flask of water at about 70 degrees C.) } 4. Float the formvar net on a water surface. About the upper 2/3 of } the slide has the lacey part of the film. } 5. Coat with carbon for strength, if desired. } } JB } } } } } I've searched the list-server archives as much as I have patience for, } } and the 'library' here is a collection of just about every biological } } book on EM but none seem to document a procedure for making lacey } } grids. } } } } Before I make the recommendation to purchase grids from one of the EM } } supply houses, I would like to make sure that making these isn't } } something that can be done easily and routinely. } } } } The intended application is not for biological use. If anyone has a } } protocol they can share it would be much appreciated. } } } } Thank you, } } } } Geoff Williams } } Microscopy Facility Supervisor } -- } ############################################################## } 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 } ############################################################## }
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 08:14:49 2003
} } Dear all, } Not quite microscopy directly, but is anyone using software } to collate and } organise references to the literature, which can also export to Word } format or that sort of thing. Any hints would be greatly appreciated. } } Thanks } } -- } Ian MacLaren } Technische Universität Darmstadt } Material- und Geowissenschaften } Petersenstr. 23 } 64287 Darmstadt } Germany } http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Datei en/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 08:31:11 2003
Microscopy Today, June 1997, Bruce Wagner, "Home Made Holey Film Grids".
Phil
} Dear list, } } I've searched the list-server archives as much as I have patience for, } and the 'library' here is a collection of just about every biological } book on EM but none seem to document a procedure for making lacey grids. } } Before I make the recommendation to purchase grids from one of the EM } supply houses, I would like to make sure that making these isn't } something that can be done easily and routinely. } } The intended application is not for biological use. If anyone has a } protocol they can share it would be much appreciated. } } Thank you, } } Geoff Williams } Microscopy Facility Supervisor } } CMU Biology Department Microscopy Facility web page. } http://www.cst.cmich.edu/centers/microscopy/
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 08:44:45 2003
You can find recipes the the "Tips & Tricks" resource at http://www.biotech.ufl.edu/EM/tips/tem.html
in the section titled "holey grid recipie" (sic) The process is not too difficult especially if you don't mind a bit of variability in the hole size, etc.
Thanks to Greg Erdos, et al, for maintaining the "Tips & Tricks" resource!
Cheers, Henk Colijn
At 03:18 PM 10/27/2003 -0500, Geoff Williams wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 09:17:34 2003
You might want to try the program, Endnotes. I think that it is just what you want.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: Tuesday, October 28, 2003 3:45 AM To: Microscopy Listserver
Dear all, Not quite microscopy directly, but is anyone using software to collate and organise references to the literature, which can also export to Word format or that sort of thing. Any hints would be greatly appreciated.
Thanks
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 09:19:36 2003
I use Biblioscape. It's cheaper than some of the others, with all the functions. A few years ago, I researched the various options and decided on Biblioscape. It includes a built-in web browser and MS Word and Wordperfect integration. There is actually a freeware version of the software (Biblioexpress). Here's the web site. http://www.biblioscape.com/ Shannan
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all, Not quite microscopy directly, but is anyone using software to collate and organise references to the literature, which can also export to Word format or that sort of thing. Any hints would be greatly appreciated.
Thanks
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
Shannan Little Research Technician/Technicien de recherche Electron Microscopy and Image Analysis / Microscopie électronique et Analyse d'images Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 403-317-3446 Facsimile/Télécopieur: 403-382-3156 P.O. Box 3000 / CP 3000 Lethbridge, Alberta T1J 4B1 littlesm-at-em.agr.ca http://res2.agr.ca/lethbridge/emia/index_e.htm
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 09:33:56 2003
I can't answer your question but I will make a related comment. One of my grad students badgered me into getting Endnote. I have had similar packages in the past and not thought they were worth the effort since I don't write so many papers that it is a huge burden to enter the references in my hand. Most of the citations were in previous papers and I can cut and paste. And one can use Medline or Pubmed to search and organize the latest references on a topic so I didn't think I would use it. But I have found it very useful. I have begun writing summary notes on every paper I read and storing them in the Comments section on Endnotes. This means I can search for all the papers on a topic and avoid having to pull them and re-read them to get the important facts again. I find that I can write a shorthand summary of the key experimental details and findings in a paragraph. I can note whether I thought the data was impressive or worthless. This has proven to save me a lot of time. In addition, students can log into this file, see my views on the paper and enter their own comments. If I point out a flaw or insightful finding, the students and postdocs can have that info as they review the paper. In summary, I have found Endnote a useful tool. I have no financial interest in this software.
At 01:48 PM 10/28/2003 +0100, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Thank you for all the replies... In going through the emails this morning I realized now that maybe the question I posed should have been re-phrased: "I've made holey films before, what is different or is there a difference making lacey carbon films..."
Specifically I've taught/made useable holey films for astigmatism and focus exercises in the TEM classes, but those are small uniform and not terribly numerous and completely unlike images found in some of the references to "lacey carbon films," ie: http://www.personal.rdg.ac.uk/~scsharip/contam.pdf -or- http://www.2spi.com/catalog/grids/cusctgrd.html (i have no reason to display the SPI site other than for example purposes)
For those that are interested: To review the recommendations, there is a significant showing of "just buy them." Many comments recommend breathing on the formvar solution before it dries, or to pass the wet formvar coated slide over boiling water. The second seems to be the same way I've made regular holey films just with more suspend glycerol in the solution, or soap and water in the formvar solution. The final technique that seems promising is the Kuga and Brown method as described by Henk Colijn in the 'tips and tricks' section.
Again thank you all for your suggestions and patience.
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 10:43:00 2003
I installed a new 50W HBO Hg bulb into my Axiophot's lamp housing. It aligned and worked fine for 4 hrs but the next time I went to re-fire it, the power supply just kept clicking and the bulb wouldn't light. I assumed the bulb mounting had loosened during the first usage so I removed it and re-installed it three times with the same result. I gave up on this bulb but your comments on this secondary problem are welcome. The real problem I would like advice on is the second bulb. It fired up just fine but I have had great difficulty aligning it. I get a good typical primary filament image but can't seem to bring the mirror image into focus. If i close down the fluorescence lamp aperature all the way, I get a small dot for the mirror image and a much brighter band for the real filament image (with brighter tips at the top and bottom of the band). The real image is more like I am used to seeing but I usually get a better mirror image. If I open the fluorescence lamp aperature all the way (where I normally leave it during alignment), I get a diffuse round circle of light for the mirror image. I have tried moving the mirror alignment screws for up down and left right and they are working. I went the full distance of both without much improvement. The focus screw for the mirror goes from all clockwise to all counter clockwise with minor changes in the focus. I did the best alignment I could and get a decent illumination of my sample. But I am wondering if any one can explain why I am not getting a good mirror image or if it is dangerous not to align it better - i was taught that if the two filament images overlap it can result in overheating of the bulb. Thanks for any advice on this. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I'm looking for a pdf or word-document of the following qrticle. Can anyone help me out? Thanks!
Histochemistry. 1985;82(3):205-8. Cowen T, Haven AJ, Burnstock G. Pontamine sky blue: a counterstain for background autofluorescence in fluorescence and immunofluorescence histochemistry.
Sven Terclavers
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 12:22:02 2003
Friday Harbor Labs, a marine biological station in the San Juan Islands affiliated with the University of Washington, has an immediate need for a functional transmission electron microscope for biological work.
As the labs' Philips 300 is currently nonfunctional, any TEM work must be done at the UW campus in Seattle - a ferry boat ride and two hour drive away. We have a core group who would like to pursue TEM projects year round and additional researchers who come to the labs in the summertime.
We would like to find a scope that has been maintained under a service agreement.
Please contact:
Paulette Brunner Center for Cell Dynamics Friday Harbor Labs Friday Harbor, Washington 98250 (206)616-0895 pbrunner-at-u.washington.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 12:46:53 2003
Johns Hopkins University still has an opening for an Electron Microscopy Technician in the Department of Pathology. If anyone is interested or knows anyone, pleace contact me.
Thanks
Lois Anderson Johns Hopkins University Dept. of Pathology Laboratory Manager Electron Microscopy/Immunofluorescence 600 N. Wolfe Street/Pathology 709 A Baltimore, MD 21287 (410) 955-2861/fax (410) 614-7110 landers-at-jhmi.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 13:02:58 2003
Dear Geoff, Many years ago a grad student on my TEM wanted holey carbon films and decided to make his own. We make our usual carbon films by casting colldion dissolved in amyl acetate on water, adding grids, scooping up the film+grids on filter paper, then drying the paper, carbon-coating the collodion film on the grids and then dissolving the collodion for 48 hours in chloroform. To make the holey films, he mixed the collodion liquid, which is the plastic carried in amyl acetate, with one or two drops of Triton X-100, which is a surfactant. I'm not sure if he diluted the collodion with more amyl acetate. He shook the bottle with collodion/amyl acetate/ Triton X-100 until it was foamy, then cast his films from that on water. He said it worked better than the other methods that he tried. You may be able to make the films lacey by shaking harder or longer. I'm sorry I cannot be more specific, but it was long ago and I didn't make them myself. Good luck. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Geoff Williams" {willi1gl-at-cmich.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, October 27, 2003 12:18 PM
Dear list,
I've searched the list-server archives as much as I have patience for, and the 'library' here is a collection of just about every biological book on EM but none seem to document a procedure for making lacey grids.
Before I make the recommendation to purchase grids from one of the EM supply houses, I would like to make sure that making these isn't something that can be done easily and routinely.
The intended application is not for biological use. If anyone has a protocol they can share it would be much appreciated.
Thank you,
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 13:22:09 2003
Cowen T, Haven AJ, Burnstock G. Pontamine sky blue: a counterstain for background autofluorescence in fluorescence and immunofluorescence histochemistry. Histochemistry. 1985;82(3):205-8.
The stain pontamine sky blue (PSB) has been shown to reduce background autofluorescence in catecholamine fluorescence and immunofluorescence histochemical preparations. Using PSB as a counterstain on whole-mount stretch preparations of human mesenteric blood vessels, a medium dense noradrenergic nerve plexus is clearly revealed, which previously had been only partially visible because of background autofluorescence. Image analysis of nerve densities in whole-mount stretch preparations of guinea-pig arteries containing noradrenergic, substance P-, and vasoactive intestinal polypeptide (VIP)-positive nerve plexuses shows that PSB staining does not alter the specific neuronal fluorescence and that it improves image definition.
If you subscribe to Loansome Doc, you can order a pdf copy for a price.
Gary Gill
-----Original Message----- } From: Sven Terclavers [mailto:Sven.Terclavers-at-med.kuleuven.ac.be] Sent: Tuesday, October 28, 2003 11:55 AM To: Microscopy-at-msa.microscopy.com; histonet-at-lists.utsouthwestern.edu
Hi all,
I'm looking for a pdf or word-document of the following qrticle. Can anyone help me out? Thanks!
Histochemistry. 1985;82(3):205-8. Cowen T, Haven AJ, Burnstock G. Pontamine sky blue: a counterstain for background autofluorescence in fluorescence and immunofluorescence histochemistry.
Sven Terclavers
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 13:28:25 2003
Reference manager, I think they are up to version 10 or 11 by now.
Geoff
Ian MacLaren wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Dear all, } Not quite microscopy directly, but is anyone using software to collate } and organise references to the literature, which can also export to } Word format or that sort of thing. Any hints would be greatly } appreciated. } } Thanks }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 14:21:27 2003
I want to get a string going about whom many consider the Greatest Microscopist, and research scientist of the 20th Century. After all He and Carl Zeiss worked together pre 1920. And everyone of you have utilized Dr. Rife's science even to this day.
Check this out.
http://www.rife.de/mscope/mscope2.htm
Let's have a sane discussion about why all you geniuses are not following in his footsteps.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 15:50:55 2003
For the secondary problem, it might not be a problem with the lamp or lamp-house, but with the starter (where you switch on the light). It also happened to me once, I got the same problems, and it turned out that the electrical supply-box, the 'starter', did not function correct anymore (just like a neon-lamp, if the small white starter here doesn't work anymore, it also keeps on clicking and flashing, but will not (or just after a while) start burning.
Your first problem might be a problem of the screw for the lens in fron of the lamphouse. Even though you can turn it complete, does the lens fully moves? The same for the mirror, the screw might work, but does the mirror actually moves? you can always try adding a little drop of oil (WD-40).
And indeed, be careful, I was taught the same: if the two filaments overlap, your lamp gets heated too much and can explode.
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
--
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 20:33:25 2003
On Thursday, October 23, 2003, at 11:33 AM, Sue Tyler wrote:
} My supervisor has just requested our travel and training needs for the } upcoming year. I am new to TEM and would like to attend some biological } type seminars. Does anyone have any suggestions or recommendations for } future meetings? } Dear Sue, I have found the M&M meetings to be excellent. I'm sure that every session will have several biological TEM sections--your main problem will be choosing among them. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 29 05:12:09 2003
} Dear all, } Not quite microscopy directly, but is anyone using software to collate } and organise references to the literature, which can also export to } Word format or that sort of thing. Any hints would be greatly } appreciated. } } Thanks }
I like Reference Manager, b/c it has a nice internal pubmed search and database function. The others have it, too, but I did not test it. (But you get trialware for all of them). What I don't like is, that these programs only support MS Word directly, and not other formats (anymore), like Lotus Wordpro/Wordperfect11/OpenOffice. Refman, Procite, and Endnote are all from the same company (isiresearchsoft), I don't know why they sell different ones and don't fuse them to a single product. Probably b/c there are "traditionalists" out there, and two out of three where originally for Apple OS?
Isiresearchsoft compares the three products:
http://thomsonisiresearchsoft.com/compare/
Some other links: http://ist.uwaterloo.ca/ew/biblio/index.html http://www.cse.bris.ac.uk/~ccmjs/rmeval.htm (quite old)
I attended my first M& M meeting this summer and did not find much to offer in the way of biological applications. Of course every years program is different so I would certainly continue to consider it as a viable option. MSA however has been an invaluable asset and I recently learned of another society ( The Ultrastructural Pathology Society) that I am going to check out. Here is the website www.sup.ultrakohl.com Hope that helps.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On Thursday, October 23, 2003, at 11:33 AM, Sue Tyler wrote:
} My supervisor has just requested our travel and training needs for the } upcoming year. I am new to TEM and would like to attend some biological } type seminars. Does anyone have any suggestions or recommendations for } future meetings? } Dear Sue, I have found the M&M meetings to be excellent. I'm sure that every session will have several biological TEM sections--your main problem will be choosing among them. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 29 10:44:44 2003
M&M is an excellent meeting for all - well most! - forms of microscopy. While TEM probably comprises about half the meeting, I'll guarantee that you will learn many useful things from the presentations and posters, not to mention the first-rate trade show of new instruments. Note that it is primarily an instrumentation/techniques oriented meeting that is useful to physical and biological participants alike.
The Ultrastructural Pathology meeting is a very good and much smaller, specialized event and is sharply focused on specific, and generally clinical, research issues. They sometimes have one or two technique papers but no trade show. It is often held in other countries.
Peter Ingram
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
If you want a "sane discussion" about this man's work how about providing some real evidence of what he has accomplished, that is, where his work has been published. Claims made on a website are just that, claims made on a website. When I went on to the homepage and read about how the FDA and the AMA are conspiring to keep his work out of the medical manistream I got a bit skeptical, but that's just me.
Geoff
Bruce Grosso wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 29 12:33:49 2003
http://www.sumeria.net/tech/rife2.html The Smithsonian Report
http://www.navi.net/~rsc/seidel.htm#rife Journal of the Franklin Institute Volume 237(2):103-130 (1944) Click on the Rife line at the beginning
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------------- ----- } } } Down here in the Southern Hemisphere it's October 30. } } It's not April 1 up there yet, is it? } } cheers } } rtch } } } } } } } From: "Bruce Grosso" {bgrosso-at-AssetRecovery.Net} } To: {Microscopy-at-msa.microscopy.com} } Subject: [Microscopy] Royal Raymond Rife } Date sent: Tue, 28 Oct 2003 14:26:28 -0800 } } } } } } } ---------------------------------------------------------------------- } } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } --------- } } } } I want to get a string going about whom many consider the Greatest } } Microscopist, and research scientist of the 20th Century. After all } } He and Carl Zeiss worked together pre 1920. And everyone of you have } } utilized Dr. Rife's science even to this day. } } } } Check this out. } } } } http://www.rife.de/mscope/mscope2.htm } } } } Let's have a sane discussion about why all you geniuses are not } } following in his footsteps. } } } } } } } } -- } Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 } Microanalyst Fax : 64 9 3737435 } Department of Geology email : r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand } }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 29 14:16:58 2003
After reviewing Dr. Rife's accomplishments one observation is possible. It was probable the inability to fritter time on the internet which allowed him to accomplish these improbable accomplishments.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238 ----- Forwarded by Frank Karl/AKR/Degussa-Huels/US on 10/29/2003 03:16 PM -----
Geoff McAuliffe To: Bruce Grosso {bgrosso-at-AssetRecovery.Net} {mcauliff-at-umd cc: Microscopy-at-msa.microscopy.com nj.edu} Subject: [Microscopy] Re: Royal Raymond Rife
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Bruce:
If you want a "sane discussion" about this man's work how about providing some real evidence of what he has accomplished, that is, where his work has been published. Claims made on a website are just that, claims made on a website. When I went on to the homepage and read about how the FDA and the AMA are conspiring to keep his work out of the medical manistream I got a bit skeptical, but that's just me.
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } I want to get a string going about whom many consider the Greatest } Microscopist, and research scientist of the 20th Century. After all He and } Carl Zeiss worked together pre 1920. And everyone of you have utilized Dr. } Rife's science even to this day. } } Check this out. } } http://www.rife.de/mscope/mscope2.htm } } Let's have a sane discussion about why all you geniuses are not following in } his footsteps. } } } } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 29 14:47:01 2003
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, October 29, 2003 1:42 PM To: Microscopy-at-msa.microscopy.com
Sorry to disappoint you all, but I really don't think M&M is the best place to go if one's interest is TEM applied to biological questions. I agree there were a few good talks at this year's meeting in that particular field, but too few to justify the expenditure and the time (5-6 days). There are some very good TEM courses organized around the country that would be more suitable for someone who is simply looking for some basic training in Electron Microscopy of biological systems. including immuno- cytochemistry. One such course is the one organized by Elaine Humphrey and Kent McDonald at the University of British Columbia in June 2004. Courses like this are frequently advertised on the listserver. You could also check the websites of the affiliated local societies listed on the MSA website. These societies organize several meetings per year, some in the form of mini-symposiums. Best,
Marc
On Wednesday, October 29, 2003, at 11:50 AM, Peter Ingram wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Sue: } } M&M is an excellent meeting for all - well most! - forms of } microscopy. While TEM probably comprises about half the meeting, I'll } guarantee that you will learn many useful things from the } presentations and posters, not to mention the first-rate trade show of } new instruments. Note that it is primarily an } instrumentation/techniques oriented meeting that is useful to physical } and biological participants alike. } } The Ultrastructural Pathology meeting is a very good and much } smaller, specialized event and is sharply focused on specific, and } generally clinical, research issues. They sometimes have one or two } technique papers but no trade show. It is often held in other } countries. } } Peter Ingram } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } --------- } } } } } } On Thursday, October 23, 2003, at 11:33 AM, Sue Tyler wrote: } } } } } My supervisor has just requested our travel and training needs for } } } the } } } upcoming year. I am new to TEM and would like to attend some } } } biological } } } type seminars. Does anyone have any suggestions or recommendations } } } for } } } future meetings? } } } } } Dear Sue, } } I have found the M&M meetings to be excellent. I'm sure that every } } session will have several biological TEM sections--your main problem } } will be choosing among them. } } Yours, } } Bill Tivol } } EM Scientist and Manager } } Cryo-Electron Microscopy Facility } } Broad Center, Mail Code 114-96 } } California Institute of Technology } } Pasadena CA 91125 } } (626) 395-8833 } } tivol-at-caltech.edu } } } -- } Peter Ingram } Sr. Physicist } Adj. Professor of Pathology, } Duke University Medical Center } Box 90319 } LaSalle Street Extension } DURHAM NC USA 27708-0319 } } Tel: (919) 660-2695 } Fax: (919) 660-2671 } e-mail: p.ingram-at-cellbio.duke.edu } http://152.3.54.107/AEM_LAB.html } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 29 16:32:43 2003
I guess I was warned by a friend, fairly enough, that most of you would be dismissive but also that there would be an open mind or two as to the possibility that he (Rife) was right and the dismissive are wrong. I always find that to be true quite often. You overlook the man's accomplishments, his positions he held, his papers, his patents, his relationship with Carl Zeiss, and the Army of doctors, scientists, researchers and allies he had and just sweep all those people away and focus on the smallest (if there really is any) negative and exploit that as reason enough to totally dismiss all the positive documented accomplishments this man has to his credit. and pooh pooh it all with snide commentary as if you have anything to lose by keeping an open mind for just a little while and just for giggles digging a little deeper to see if there is a probability rather than an improbability.
I am not trying to sell Rife to anyone, I was just hoping for a "sane" discussion, with real scientists, between one another of yourselves but I see with some of you that is not possible.
It is too easy to crack wisely and dismiss, than to briefly study and discuss the what if and say instead; impossible.
The one great thing about doing the impossible is there is no competition.
I am just hanging around this site, waiting to sell a microscope and thought I would try to get to know few of you. Not start WWIII.
I do know that the ones of you who are curious and are looking at the "What if it is true?" are going to be the ones who say nothing so they can ponder that rather than imperiously dismiss it as balderdash.
You know the mind is like a parachute! It only works when it is open.
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists ----- Original Message ----- } From: {Frank.Karl-at-degussa.com} To: {microscopy-at-msa.microscopy.com} Sent: Wednesday, October 29, 2003 12:05 PM
The copy of the journal The Scientist that arrived in my mailbox this afternoon contains a comparison of the major players in the reference manager software arena (http://www.the-scientist.com/yr2003/oct/lcprofile1_031020.html); the review seems a bit cursory, but might be of use to those newly-entering the morass of citation management.
|| Aaron Barnes The Evergreen State College Olympia, WA USA
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: Tuesday, October 28, 2003 12:45 AM To: Microscopy Listserver
If that is the way you see my approach, then you have my sincerest and my most humble apologies and many times over and again in spades.
I just wanted to get to know a few of you guys and gals better as I wait to sell my scope. (for what it is worth it is a Zeiss LSM-510 META confocal).
I have to wait for the wheels of academic bureaucracy to turn and I was trying to make some new acquaintances in case I get another scope or useful piece of research equipment to sell one day.
I wish I had the vernacular to engage in a discussion about the differences between light frequencies and cellular wall mortal oscillatory rates as compared to the features of the light microscope he built and how in using it he had the intention of finding:
#1) Cancer was a viral pathogen #2) Each specimen emitted it's own distinct light signature, and this was not limited to the first specimens but after years of study he discovered it applied to virtually every pathogen and living cell. #3) That by discovering it light frequency he could emit the matching sonic (for lack of a better more educated word) frequency thus doing a Crystal glass style shattering of the cell wall and destroying the Pathogen without affecting the surrounding tissue in any way as they had different mortal oscillatory rates.
AND what if it is true? Doesn't anyone want to try it out to see if it is or not? I mean we already do it on gall stones and crystal glass. What is the harm on living tissue samples? What could it hurt to just take a look see?
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists ----- Original Message ----- } From: "Webster, Paul" {PWebster-at-mail.hei.org} To: {microscopy-at-msa.microscopy.com} ; "'Bruce Grosso'" {bgrosso-at-AssetRecovery.Net} Sent: Wednesday, October 29, 2003 5:25 PM
Hi all,
Our Hitachi H-600 TEM is failing to provide an HV reading at any kV (ie 25, 50, 75, 100). As the microscope is 20 years old I believe the HT cable is the problem. Do you agree? What's the best plan of attack from here? Apart from the HT cable and a dirty gun chamber, are there any other potential causes of this failure? I have never attempted to isolate the HT cable. The vacuum system is fine. I've turned the microscope completely on and off a few times and have vented all compartments to air and back to high vacuum but this has had no effect.
Two days ago I reassembled the rear cosmetic guard that encases the upper vacuum system and the HT cable. Could a slight knock to the cable be enough to finish it off? The microscope was working yesterday, though. I've tried gently wiggling the cable but that gave no response.
The microscope has not been hissing and the HV has appeared quite stable. The only sign that something was not quite right was that some mornings the HV would fail to read at 25kV (but would always read at 50, 75 and 100kV).
In the light of problems encountered by Sarah Ellis (refer to archives) a few months ago I am tempted to clean the gun chamber first (however I doubt if this will fix anything).
Any advice would be gratefully accepted before I contact our institution's electrical engineers.
John Brealey EM Unit The Queen Elizabeth Hospital Institute of Medical & Veterinary Science Adelaide Australia (08) 8222 6612 john.brealey-at-imvs.sa.gov.au
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 29 22:31:48 2003
Geoff Williams asked how to make lacey carbon coated grids. There may be many methods; a number of methods may be found in textbooks. Here's how we do it. The problem with instructions like these is that the devil is in the details, and times, temperatures and concentrations all vary with the season.
Take a glass slide. Clean it thoroughly. Dip it into a solution of Formvar in ethylene dichloride. Remove the slide from the Formvar solution and allow it to dry briefly; our current time is about one second, but there's a "feel" for this step. Insert the slide into the "steam chamber" for several seconds, remove the slide and allow it to dry thoroughly. "Score" the film at the edges of the slide using a fresh razor blade and float the film off the slide onto freshly filtered, deionized water.
The "steam chamber" which we use consists of a beaker of water, sitting on a hot plate and covered with a Maxwell House coffee can which has a hole cut in the side about 4 cm square. We do not know if other brands of coffee cans will work as well. The temperature of the water in the beaker is maintained so that there is visible condensation on the side of the beaker, but the water is not actively boiling.
When you have floated the film onto the water, look at it. If it looks clear, it is probably good. If it looks cloudy, or if wrinkles can be seen, you might as well discard it now. Lay a pattern of the grids of your choice onto the film, and pick the film and the grids up using a piece of filter paper. Allow the film, the grids and the filter paper to dry thoroughly. What you now have is grids which have a damaged Formvar film. The grids are made "lacey" by exposing them to the ethylene dichloride vapors above ethylene dichloride-soaked filter paper in a covered Petri dish for several seconds. The exact time of exposure is determined using a stop watch and inspecting sample grids in the TEM; there is no substitute for this inspection step and the feedback of checking sample grids as you process each batch.
The lacey grids can now be carbon coated. After carbon coating, the Formvar film is removed by placing the grids on chloroform-soaked filter paper overnight. Example grids are examined in the TEM to assure that the film is beam stable and free from artifacts. The finished grids are inspected under a low power light microscope and packaged for shipment.
At each step, it is necessary for the specific laboratory to determine the precise concentration, time, temperature, etc. which is required for each step; there is no universal recipe, and the recipe for our laboratory varies from day to day. There is no substitute for inspection of sample grids at each step of the process using a TEM; you simply can't see what you need to see by light microscopy. The cornerstone of our quality assurance program for lacey carbon and other custom coated grids is inspecting example grids from each batch in our own TEM. If the grids do not meet our quality standards, the customer never sees them. This may explain why we have virtually no returns of coated grids from customers.
And it helps to have been doing the process for several decades. This is a skilled art, and the only substitute for going through the learning curve yourself is to ask someone who has "been there" before you to make the grids you need.
Disclaimer: SPI Supplies sells all of the materials necessary to do this except the TEM itself. We also sell lacey carbon coated grids. We have an obvious interest in promoting this technology.
Andy
Andrew W. Blackwood, Ph.D. Vice President, Technical SPI Supplies P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 X108 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 30 09:02:26 2003
Thank you for sharing your instructions. We make them a bit differently here but what works for one may not always work for another. I think that sharing your instructions was particularly generous in that you obviously make these grids for sale. However, I suspect that those that typically buy them will continue to do so and those that struggle to make them themselves will just have a bit easier time of it.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 10/30/03 7:43 AM, "Blackwood, Andrew" {ablackwood-at-2spi.com} wrote:
} Geoff Williams asked how to make lacey carbon coated grids. There may be } many methods; a number of methods may be found in textbooks. Here's how we } do it. The problem with instructions like these is that the devil is in the } details, and times, temperatures and concentrations all vary with the } season. } } Take a glass slide. Clean it thoroughly. Dip it into a solution of Formvar } in ethylene dichloride. Remove the slide from the Formvar solution and allow } it to dry briefly; our current time is about one second, but there's a } "feel" for this step. Insert the slide into the "steam chamber" for several } seconds, remove the slide and allow it to dry thoroughly. "Score" the film } at the edges of the slide using a fresh razor blade and float the film off } the slide onto freshly filtered, deionized water. } } The "steam chamber" which we use consists of a beaker of water, sitting on a } hot plate and covered with a Maxwell House coffee can which has a hole cut } in the side about 4 cm square. We do not know if other brands of coffee cans } will work as well. The temperature of the water in the beaker is maintained } so that there is visible condensation on the side of the beaker, but the } water is not actively boiling. } } When you have floated the film onto the water, look at it. If it looks } clear, it is probably good. If it looks cloudy, or if wrinkles can be seen, } you might as well discard it now. Lay a pattern of the grids of your choice } onto the film, and pick the film and the grids up using a piece of filter } paper. Allow the film, the grids and the filter paper to dry thoroughly. } What you now have is grids which have a damaged Formvar film. The grids are } made "lacey" by exposing them to the ethylene dichloride vapors above } ethylene dichloride-soaked filter paper in a covered Petri dish for several } seconds. The exact time of exposure is determined using a stop watch and } inspecting sample grids in the TEM; there is no substitute for this } inspection step and the feedback of checking sample grids as you process } each batch. } } The lacey grids can now be carbon coated. After carbon coating, the Formvar } film is removed by placing the grids on chloroform-soaked filter paper } overnight. Example grids are examined in the TEM to assure that the film is } beam stable and free from artifacts. The finished grids are inspected under } a low power light microscope and packaged for shipment. } } At each step, it is necessary for the specific laboratory to determine the } precise concentration, time, temperature, etc. which is required for each } step; there is no universal recipe, and the recipe for our laboratory varies } from day to day. There is no substitute for inspection of sample grids at } each step of the process using a TEM; you simply can't see what you need to } see by light microscopy. The cornerstone of our quality assurance program } for lacey carbon and other custom coated grids is inspecting example grids } from each batch in our own TEM. If the grids do not meet our quality } standards, the customer never sees them. This may explain why we have } virtually no returns of coated grids from customers. } } And it helps to have been doing the process for several decades. This is a } skilled art, and the only substitute for going through the learning curve } yourself is to ask someone who has "been there" before you to make the grids } you need. } } Disclaimer: SPI Supplies sells all of the materials necessary to do this } except the TEM itself. We also sell lacey carbon coated grids. We have an } obvious interest in promoting this technology. } } Andy } } Andrew W. Blackwood, Ph.D. } Vice President, Technical } SPI Supplies } P.O. Box 656 } West Chester, PA 19381-0656 } Ph: 1 610 436 5400 X108 } FAX: 1 610 436 5755 } e-mail: ablackwood-at-2spi.com } WWW: http://www.2spi.com }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 30 11:39:51 2003
One of our lab members was looking for bimetallic grids: Cu on one side and Ni on the other. She couldn't remember the supplier, but remembered using them -- within the last couple of years. I've gone on the web and checked all the usual suspects to no avail. Has any one heard of such grids, and knows who supplies them? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 30 11:45:01 2003
It's very interesting to read all about how the SPI grids we purchase are made...takes me back to my student days! You're absolutely correct that there is an art associated with each step, which varies from lab to lab. That's why it is far more cost effective for my lab to purchase the SPI lacey film products rather than attempt to make them ourselves any more. Of course, we don't have an unlimited supply of graduate students either... ;-)
Larry
Disclaimer: I have no connection whatsoever with SPI Co., and am pleased to pay full price for any product that I purchase from them.
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 30 15:13:21 2003
Dear Phil, I have seen Cu-Pd grids, which Marivac sells, and they look copper-coloured on one side and grey on the other, but I have never seen Cu-Ni.
----- Original Message ----- } From: "Philip Oshel" {peoshel-at-wisc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 30, 2003 9:45 AM
Hi:
Forgive me if this is too simple. Some guys from another lab just came by and asked if I had any quartz coverslips. They are doing Raman microscopy with infrared/red light and they say the ordinary glass coverslips are killing their signal.
I volunteered to check around for them, this is my first stop since, of course, I couldn't think of anyplace else where I could tap directly into the hearts and minds of the world's greatest and most renowned microscopists.
Is there an easy solution to their problem? Inquiring minds want to know. I will check around elsewhere, but I always think of this list whenever questions like this come up.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 30 15:53:37 2003
There are a number of reasons why the HT will not switch on.
1. The Vacuum safety trip is not working preventing the HT system operating. 2. The electronics for HT generation are faulty 3. Not so sure that a duff HT cable would give such a problem.
Without a circuit diagram it is difficult for me to tell you how to overcome (1) other than to say I have shorted out the trip in the past in order to get a customer going.
Number (2) could be a lack of a standard supply but always ask "what did I do last?" i.e. prior to the fault. Did you knock one of the connections to the tank or on the microscope end of the system; check their tightness?
Regarding (3) how to check out the cable problems
1. Clean the top of the tank so that there will be no danger of dirt falling into the oil when you open it up. 2. Remove the high voltage cable from the tank and wrap it in clean aluminium foil. Cover the tank opening with foil. 3. Now switch on the HT 4. Still nothing then you have a supply or connection problem, the circuitry is at fault. 5. All is well (I doubt in your case) you have a cable problem 6. Make contact with a local x-ray or high voltage company (nothing to do with EM but will be able to handle a 125kV cable) who should be able to use your cable ends and make a new cable at a fraction of the Hitachi costs, or any one else for that matter!
Hope this helps. I am away for the next two weeks but come back to me if you need more help?
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, October 30, 2003 3:54 AM
Hi John
There are a number of reasons why the HT will not switch on.
1. The Vacuum safety trip is not working preventing the HT system operating. 2. The electronics for HT generation are faulty 3. Not so sure that a duff HT cable would give such a problem.
Without a circuit diagram it is difficult for me to tell you how to overcome (1) other than to say I have shorted out the trip in the past in order to get a customer going.
Number (2) could be a lack of a standard supply but always ask "what did I do last?" i.e. prior to the fault. Did you knock one of the connections to the tank or on the microscope end of the system; check their tightness?
Regarding (3) how to check out the cable problems
1. Clean the top of the tank so that there will be no danger of dirt falling into the oil when you open it up. 2. Remove the high voltage cable from the tank and wrap it in clean aluminium foil. Cover the tank opening with foil. 3. Now switch on the HT 4. Still nothing then you have a supply or connection problem, the circuitry is at fault. 5. All is well (I doubt in your case) you have a cable problem 6. Make contact with a local x-ray or high voltage company (nothing to do with EM but will be able to handle a 125kV cable) who should be able to use your cable ends and make a new cable at a fraction of the Hitachi costs, or any one else for that matter!
Hope this helps. I am away for the next two weeks but come back to me if you need more help?
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, October 30, 2003 3:54 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (scanning-at-fams.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 30, 2003 at 14:20:26 ---------------------------------------------------------------------------
Organization: SCANNING, The Journal of Scanning Microscopies
Title-Subject: [Microscopy] SCANNING 2004 - Washington, D.C. April 27-29
Question: Dear Microscopists and Friends:
The 15th annual scientific meeting on scanning microscopies, sponsored by the Foundation for Advances in Medicine and Science (FAMS, Inc.), promises to be an exciting and informative scientific meeting on the scanning microscopies. We hope you will join us there to commemorate the first century of scanning electron microscopy and the centennial anniversary of Sir Charles Oatley, pioneer of scanning electron microscopy.
Details of the Meeting:
What: SCANNING 2004 When: Tuesday, April 27, Wednesday, April 28, Thursday, April 29 Where: Hotel Washington, Washington, D.C. Contact: (201) 818-1010 (Tel) (201) 818-0086 (Fax)
Please visit us at www.scanning.org to download the Call for Papers and for the latest information pertaining the the sessions, short courses, hotel information, and sponsorship opportunities.
If that type of bulb is elongated with a terminal at each end, reverse the polarity. i.e. Flip it around and reinsert it. The starter electrode needs to be at the + anode terminal to work. Bob.
Bob Carter 2000 Bayshore Road Lopez Island, WA 98261
----- Original Message ----- } From: "Tom Phillips" {phillipst-at-missouri.edu} } To: {Microscopy-at-msa.microscopy.com} } Sent: Tuesday, October 28, 2003 8:50 AM } Subject: [Microscopy] Hg bulb alignment } } } } } } } } -------------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------------- } ----- } } } } I installed a new 50W HBO Hg bulb into my Axiophot's lamp housing. It } } aligned and worked fine for 4 hrs but the next time I went to re-fire it, } } the power supply just kept clicking and the bulb wouldn't light. I } assumed } } the bulb mounting had loosened during the first usage so I removed it and } } re-installed it three times with the same result. I gave up on this bulb } } but your comments on this secondary problem are welcome. The real problem } } I would like advice on is the second bulb. It fired up just fine but I } } have had great difficulty aligning it. I get a good typical primary } } filament image but can't seem to bring the mirror image into focus. If i } } close down the fluorescence lamp aperature all the way, I get a small dot } } for the mirror image and a much brighter band for the real filament image } } (with brighter tips at the top and bottom of the band). The real image is } } more like I am used to seeing but I usually get a better mirror image. If } I } } open the fluorescence lamp aperature all the way (where I normally leave } it } } during alignment), I get a diffuse round circle of light for the mirror } } image. I have tried moving the mirror alignment screws for up down and } } left right and they are working. I went the full distance of both without } } much improvement. The focus screw for the mirror goes from all clockwise } } to all counter clockwise with minor changes in the focus. I did the best } } alignment I could and get a decent illumination of my sample. But I am } } wondering if any one can explain why I am not getting a good mirror image } } or if it is dangerous not to align it better - i was taught that if the } two } } filament images overlap it can result in overheating of the bulb. Thanks } } for any advice on this. Tom } } } } Thomas E. Phillips, PhD } } Professor of Biological Sciences } } Director, Molecular Cytology Core } } 3 Tucker Hall } } University of Missouri } } Columbia, MO 65211-7400 } } } } 573-882-4712 (office) } } 573-882-0123 (fax) } } PhillipsT-at-missouri.edu } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 02:33:13 2003
} Hello Peter, } } you are perfectly right with your assessment of FireWire cameras. What you } have neglected is the benefit of IEEE1394 being a widespread standard } interface on Apple Macintosh as well as Microsoft Windows computer } platforms. On current computers you do not need expensive framegrabber } boards (e.g. for use with CameraLink) because they are already equipped with } FireWire. This greatly simplifies the usage of one camera in an environment } with more than one microscope wokstation. Furthermore, direct digitization } in the camera's head also minimizes noise that may be introduced to your } signal while beeing transmitted to the framegrabber. So the trade-off is } flexibility vs. speed. } } What do you think? } } Best regards, } } Achim } } } =============================================== } Achim Zimmermann, MSc (Phys) } Product Manager Microscope Cameras } } JENOPTIK Laser, Optik, Systeme GmbH } Business Unit Sensor Systems - Digital Cameras } Göschwitzer Str. 25 } D-07745 Jena } Germany } } Tel.: +49 36 41 65 - 21 39 (office) } Tel.: +49 17 33 99 35 45 (mobile) } Fax: +49 36 41 65 - 21 44 } } eMail: achim.zimmermann-at-jenoptik.com } Web: www.progres-camera.com - www.jenoptik.com } =============================================== } } } } } -----Original Message----- } } From: Peter Van Osta [mailto:pvosta-at-unionbio-eu.com] } } Sent: Monday, October 27, 2003 5:14 PM } } To: Confocal Microscopy List; MSA } } Subject: [Microscopy] Firewire or CameraLink ? } } } } } } } } } } -------------------------------------------------------------- } } ---------------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 02:41:53 2003
We are currently testing our new system in which we use a 75 W Xenon Arc (Carl Zeiss). We have developed a remote control system through which we can control the the power (unipolar switches) from within our software.
Although we got the message that the Xenon Arc is properly shielded it frequently brings other electronic components in the system in disarray and this leads to malfunction of the entire system.
We have tried netfilters and grounding but up to now we have not been able to protect our other equipment from periodic failure due to the spikes in the electricity net and/or the RF generated by the the Xenon Arc.
We take care to schedule the powerup of the Xenon Arc before all other equipment and wait for 15 sec. before proceeding with the powerup sequence. Powerdown is in reverse order and the Xenon Arc is powered down as the last device.
Quartz slides and coverslips are available in various sizes / thicknesses and shapes from Agar Scientific http://www.agarscientific.com ========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 07:42:59 2003
Try SPI (www.spi2.com). I know that Chuck Garber understands this problem well.
Thanks, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 01:23 PM 10/30/03 -0800, Jon Krupp wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 09:13:32 2003
Jonathan Krupp wrote: ====================================================== Hi:
Forgive me if this is too simple. Some guys from another lab just came by and asked if I had any quartz coverslips. They are doing Raman microscopy with infrared/red light and they say the ordinary glass coverslips are killing their signal.
I volunteered to check around for them, this is my first stop since, of course, I couldn't think of anyplace else where I could tap directly into the hearts and minds of the world's greatest and most renowned microscopists
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 11:10:27 2003
Please forgive this attempt to contact Gordon Couger who posted a message regarding scanning light photomacrography on the 21st of October. I have tried several times to email him directly and my messages bounce - they say couger.com is not a local host, not a gateway (whatever that means). Anyway, if I can get a valid email address for Gordon, I will take this off the list. Again, my apologies for dealing with this on the list.
Regards, Bill Sharp
William P. Sharp Arizona State University School of Life Sciences, box 4501 Tempe, AZ 85287-4501 Phone - (480)-965-3210 Fax - (480)-965-6899
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 12:30:17 2003
Listers, I would like to get some insight on anybody who is currently using the fluorescence illumination system from the EXFO company's X-Cite 120 (Exfo.com) particularly on the following aspects. Ease of use, User friendliness, stability of the lamp intensity, life of the lamp and the liquid light guide (influence of its liquid filled model, diameter and the claim on 'less heat more light'), and the cost to efficiency ratio or any relevant information on this product. Although some of this information already I am aware of from the manufacturer, but would like to hear from somebody who is currently using it and having hands on experience. Thanks in advance Shiv
Mayandi Sivaguru, Ph.D., Associate Director Molecular Cytology Core Facility Molecular Biology Program 2, Tucker Hall University of Missouri Columbia, MO 65211-7400 sivagurum-at-missouri.edu
Voice: 1-573-882-4895 Fax: 1-573-882-0123
www.biotech.missouri.edu/mcc/
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 13:44:26 2003
Does anyone know the recommended floor space suitable for an electron microscopy lab? I'm interested in the requirements for both research and also hospital labs space requirements - or at least the recommended square footage requirements.
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 14:50:45 2003
I'm assuming you mean for the microscope itself. Sorry for the U.S. units but we have a 2010F that fits well into a lab with 366 sq. ft., and a Tecnai 30 that fits into a lab with 317 sq. ft. That includes room for sample loading areas (two areas in each case), sample storage and storage of misc. parts and extras in a cabinet.
Our sample prep rooms are a similar size to the microscope rooms.
I hope this helps.
cheers, John
******** John S. Vetrano Sr. Research Scientist Materials Structure and Performance Group Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
} ---------- } From: Garry Burgess } Sent: Friday, October 31, 2003 11:49 AM } To: MSA } Subject: [Microscopy] Space Requirements for EM lab } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Does anyone know the recommended floor space suitable for an electron } microscopy lab? I'm interested in the requirements for both research and } also hospital labs space requirements - or at least the recommended square } footage requirements. } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 18:30:31 2003
Jonathan Krupp wrote: ====================================================== Hi:
Forgive me if this is too simple. Some guys from another lab just came by and asked if I had any quartz coverslips. They are doing Raman microscopy with infrared/red light and they say the ordinary glass coverslips are killing their signal.
I volunteered to check around for them, this is my first stop since, of course, I couldn't think of anyplace else where I could tap directly into the hearts and minds of the world's greatest and most renowned microscopists
From MicroscopyL-request-at-ns.microscopy.com Sat Nov 1 08:23:39 2003
You can not use glass for this kind of application and SPI Supplies has offered a full range of quartz slides and cover slips designed to solve this problem on URL http://www.2spi.com/catalog/ltmic/quartz.shtml
They are generally available from stock for overnight shipment worldwide.
One should note that, like mica, if you don't know the grade you are purchasing, there is no way to tell what you actually do have. So from the above page, you can see that we feature GE 124 quartz in the fabrication of our quartz slides and cover slips. And full transmittance data is disclosed. See URL http://www.2spi.com/catalog/ltmic/quartz-slides-coverslips.html
Whoever you do purchase from, if not from SPI, make sure you know what quartz you are getting and its characteristics.
One further comment: For the near infrared, MgO might be a better alternative. See URL http://www.2spi.com/catalog/submat/magnesium-oxide.shtml and to the graph showing the transmittance spectrum for MgO.
The absorption curves are given for both materials so one can match product selection with objectives in terms of what part of the spectrum they need their greatest transparency.
Disclaimer: SPI Supplies has offered a full range of quartz slides and cover slips to those working in the UV and near infrared range for some number of years. We over standard shapes and sizes but can also produce custom shapes and sizes.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Sat Nov 1 14:07:49 2003
I would like to know the origin of terms of lacey carbon and holey carbon films. When I was in Japan, most people use microgrid to refer carbon films having many holes. I have not yet investigated who developed such carbon films. However, I think that Fukami, Adachi, Sakata, and Sjostrand.
Papers written by Prof. Akira Fukami are: Fukami and Adachi (1965); J. Electron Microscopy, 14, 112-118. Fukami, Adachi, Katoh (1966) The proceedings of 6th International Congress for Electron Microscopy, Vol. 1, 263-264.
Later, he wrote Specimen Preparation Techniques for Electron Microscopy in 1967 (Japan Electron Optics Laboratory Co., LTD.), based on the above two papers.
Prof. Fukami used a term Micro Plastic Grid in his 1967 paper. I do not know who changes it to microgrid, but Japanese Electron Microscopy Dictionary (1986) edited by Hashimoto and Ogawa uses microgrid. Some Japanese textbooks I used in Japan also used "microgrod" or "micro grid".
I think that the preparation method of holey and lacey carbon films is the same as that of microgrid. Japanese prefer to use "microgrid", but most researchers outside Japan use "holey or lacey carbon films". I would like to know historical reason international researchers use holey or lacey carbon films instead of microgrid.
I use holey or lacey carbon in my papers because most people do not know microgrid and the venders use such terms. However, if microgrid" is not strange English, I think that people should use microgrid, assuming that people basically follow Fukamis method to prepare such films.
Thank you,
Hiromi Konishi, Ph.D. Dept. of Earth and Planetary Sciences The University of New Mexico
From MicroscopyL-request-at-ns.microscopy.com Sun Nov 2 14:41:40 2003
Here is the table of contents for the November/December 2003 issue of Microscopy Today. This issue is mailed with the Call for Papers for the 2004 Microscopy and Microanalysis meeting
New Subscriptions via http://www.microscopy-today.com only, please New subscriptions will close on Tuesday 11 November for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting. Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA. MSA members anywhere have free subscriptions. Non-MSA, non-North American subscriptions have been reduced from $80 or $110US to $35US. Additional details in the magazine or on our web site.
THIS WILL BE THE LAST ISSUE NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED OR NOT-CURRENTLY-SUBSCRIBED INDIVIDUALS WILL RECEIVE!
Table of Contents:
Zaluzec The Scanning Confocal Electron Microscope Marko, et al. Correlative Electron Tomography And Elemental Microanalysis In Biology: A Preview Sedgewick Creating Pseudocolored Images in Photoshop Steigerwald Ultra Low Voltage BSE Imaging Fasolka, et al. Techniques for Combinatorial and High-Throughput Microscopy Part 2: Automated Optical Microscopy Platform for Thin h Mustoe Microscopy of Silicified Wood Gerrity, et al. Microwave Processing in Diagnostic Electron Microscopy Shribak and Oldenbourg A Polarizing Microscope for Mapping Birefringent Objects in 3D Space Casavan and Gaidoukevitch Colocalization of Fluorescent Probes Using Image-ProR Plus v. 5.0 Munroe Electron Microscopy in Australia Tengowski Converting Right-Left Stereo Pairs Into Colored Pairs For Electronic Presentation Ahlstrand Cellulose Acetate Replication of Plant Surfaces for SEM Ellis Safe Handling of Embedding Media Schooley Microscopes as Gifts MSA Council Microscopy Society of America Position on Ethical Digital Imaging
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Sun Nov 2 14:45:19 2003
I have 120 square feet for a JXA-840A EPMA, including sample loading and storage of spares, etc and boy, let me tell you, it's not enough!
I wish I had been more assertive.............
cheers
rtch
} From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} To: MSA {microscopy-at-msa.microscopy.com}
Listers,
Here is the table of contents for the November/December 2003 issue of Microscopy Today. This issue is mailed with the Call for Papers for the 2004 Microscopy and Microanalysis meeting
New Subscriptions via http://www.microscopy-today.com only, please New subscriptions will close on Tuesday 11 November for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting.
Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA.
MSA members anywhere have free subscriptions.
Non-MSA, non-North American subscriptions have been reduced from $80 or $110US to $35US. Additional details in the magazine or on our web site.
THIS WILL BE THE LAST ISSUE NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED OR NOT-CURRENTLY-SUBSCRIBED INDIVIDUALS WILL RECEIVE!
Table of Contents: November/December 2003
Zaluzec-The Scanning Confocal Electron Microscope Marko, et al.-Correlative Electron Tomography And Elemental Microanalysis In Biology: A Preview Sedgewick-Creating Pseudocolored Images in Photoshop Steigerwald-Ultra Low Voltage BSE Imaging Fasolka, et al.-Techniques for Combinatorial and High-Throughput Microscopy Part 2: Automated Optical Microscopy Platform for Thin h Mustoe-Microscopy of Silicified Wood Gerrity, et al.-Microwave Processing in Diagnostic Electron Microscopy Shribak and Oldenbourg-A Polarizing Microscope for Mapping Birefringent Objects in 3D Space Casavan and Gaidoukevitch-Colocalization of Fluorescent Probes Using Image-ProR Plus v. 5.0 Munroe-Electron Microscopy in Australia Tengowski-Converting Right-Left Stereo Pairs Into Colored Pairs For Electronic Presentation Ahlstrand-Cellulose Acetate Replication of Plant Surfaces for SEM Ellis-Safe Handling of Embedding Media Schooley-Microscopes as Gifts MSA Council-Microscopy Society of America Position on Ethical Digital Imaging
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 3 00:59:31 2003
Dear Garry Depends what you are referring to. This question is currently wrecking my sleep. I need more space! We have a multiuser facility in the University of Botswana and we are training students (sadly the academics are not interested in learning) to operate the scopes. There is a possibility of a new building somewhere in the distant future and for the planning phase I had to give some input. I will give real distances rather that square meter since I found the real distances (Shape) of the room is important. For the TEM we currently have 4.63m X 3.25m which seems adequate as long as there is not more that 3 people in the room at the same time. Two people work fine but it does get crowded with 3. Often it is the operator + client sometimes the supervisor as well. We do not store samples in the room and except for loading space the rest is dedicated to the scope (Technai 12). The ESEM room we have is a bit to small. We need more space on the sides to walk to the bac where the pumps and electronics are (XL 30 ESEM). We can not fit a cryo, the room is just to small. 3.25m X 4.02 currently a 4x4m room will be my minimum requirement for an SEM. The confocal room is 2.08 X 2.27 it just does not work! 3 X 3 (Just for the confocal) will work. (Again there will be no spare space for "clients"to be involved. All sample prep must take place outside the microscope rooms. The biological sample prep area is 3.25 X 3m. It work fine for a max of 2 people in the area at the same time. The CPD and Sputter coater fit. All chemicals must be stored at another location. We would like to have a microwave in there as well but the room is to small. Thus my suggestion is 4X5m room for biological sample prep. and the same size for materials science prep. 4x3 for light microscopy. assuming 2 stereo and 1 transmitted light as well as one inverted light microscope. Space for the PC for each scope to store images on and the high end scopes are computer driven. Still you need storage space and a general work area gass bottle storage, UPS storage (we have 3 9kvA units)and most EM units have a small collection of books. To get this amount of space in a country is not an easy task. Then there is the long term planning of adding/expanding? Just hope this is useful. Remember these are my personal views.
-----Original Message----- } From: Garry Burgess [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Friday, October 31, 2003 9:50 PM To: MSA
Does anyone know the recommended floor space suitable for an electron microscopy lab? I'm interested in the requirements for both research and also hospital labs space requirements - or at least the recommended square footage requirements.
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 3 07:03:49 2003
We used to buy these from Ernest F. Fullam and SPI.
Fullam's shows a Rh and a Pd 'back coated' Cu grid in the old red catalog.
SPI Inc. also sells them, 2020P.
I am sure other EM suppliers do also.
Paul
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Thursday, October 30, 2003 12:46 PM To: Microscopy-at-sparc5.microscopy.com
Micromavens,
One of our lab members was looking for bimetallic grids: Cu on one side and Ni on the other. She couldn't remember the supplier, but remembered using them -- within the last couple of years. I've gone on the web and checked all the usual suspects to no avail. Has any one heard of such grids, and knows who supplies them? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 3 07:35:58 2003
Dear all, Is there a standard reference book on TEM of semiconductors? A grad student here wants to look for info on this topic and a good text would be a very helpful starting place.
Please copy your answers to sigmund-at-hf.tu-darmstadt.de
Thanks
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 3 08:53:10 2003
Garry Try to get your hands on a copy of "Practical Methods in Electron Microscopy, Vol.4: Design of the Electron Microcope Laboratory" by Ronald H Alderson. It is part of the series edited by Audrey M. Glauert. Even thought it is over 25 years old, it is full of useful information on this topic. 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
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 3 08:53:51 2003
Hi, Anyone out there using a JEOL 6300 and using an inexpensive frame grabber to capture the video output? If so can you provide me with any info on manufacturer, model number, and performance?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 3 12:16:30 2003
Background Information: We have a JEOL 6460LV which was installed about 6 months ago. Recently we have noticed a significant degradation in image quality. We have had field service come in and clean the column and the apertures twice. The response they gave us on both occasions was that this is normal for this instrument.
Normal use and operation is HV and low KV, {2. When LV mode is used, the samples are clean and LV is only used to get rid of charging. "Dirty" samples are never placed into the chamber. Primary use of the instrument is polymer imaging.
The Question: What is the expected frequency of final aperture cleaning, given the above stated operating conditions? (This one is seriously starting to go downhill after about 1.5 months of operation). Previous experience with a JEOL 5900 was that after 6 months there was no significant performance loss. The apertures of other microscopes, i.e., AMR, ISI, TOPCON, were changed when LaB6 filaments were changed. That is after ~4 to 6 months of continuous use. Thanks Nancy Simons,
Molecular Interventions Boston Scientific Corp. 1 Boston Scientific Place A4 Natick, MA 01760-1537
tel. 508-650-8603 fax 508-647-2329
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 3 12:27:26 2003
you can pretty much use any frame grabber that understands the video protocol the machine is using (I think it's RS-170, but I'm not sure. Check the manuals) to acquire images from the SEM. There are many drawbacks using this technique, though:
1) Your images will be relatively low resolution (640x480), due to the limitations of video signals. 2) It is possible that you can only acquire the images recorded at video speed. They are typically noisy compared to slow scan images. 3) You will have no calibration information, unless you can also acquire the data bar and calibrate the images that way. 4) Your images will be 8-bit images (256 levels of gray).
Item 2 you can improve through frame averaging, but the frame grabber and software must support that.
If you need more than that, you need to consider SEM interfaces, such as our ADDA II (there are other manufacturers as well). These devices can acquire slow scan signals at a higher resolution (up to 4Kx4K), and they are usually better than 8 bit (12-bit in the case of the ADDA).
If you need more information, please contact me off-line.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Michael Cheatham [mailto:mmcheath-at-mailbox.syr.edu] Sent: Monday, November 03, 2003 08:03 To: microscopy-at-ns.microscopy.com
Hi, Anyone out there using a JEOL 6300 and using an inexpensive frame grabber to capture the video output? If so can you provide me with any info on manufacturer, model number, and performance?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 3 18:56:54 2003
You may also want to consider utility rooms or corridors for noisy pumps and chillers if space allows. We had an opportunity to setup a lab with the ability to place utility rooms adjacent to the microscope rooms and it makes for a much enjoyable place to work. For our Philips CM200 we also have the power supply in the utility corridor, mechanical pumps are not a problem because they don't run all the time. Our Hitachi S-4700 mechanical pumps do run all the time and I'm glad they are not in the same room. I would prefer smaller rooms with a utility corridor over a large room.
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
I couldn't agree more about Ronald Alderson's book - I have found it very useful on several occasions in the past. There have been some changes over the years such as digital imaging so you may or may not need a darkroom for instance.
One final source of information about lab space that is the microscope manufacturer's data at the back of their brochures/literature. This will give you the basic minimum for the instrument and then you would need to factor in people and preparation space. A difficult one is always allowing for occasional groups of visitors - in my case I am limited to groups of about 10 people because there are several small rooms in the lab.
I know it's not high-tech but it's sometimes useful to sit down with a piece of graph paper and some cut-outs of the instruments going into the lab space. This is very useful for highlighting narrow access points (eg access to the backs of instruments for serving).
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Leona Cohen-Gould {lcgould-at-med.cornell.edu}
Less than 1 year old, Zeiss has agreed to service it on a per call basis. (But not install it) , I have someone to install it, and people who can teach operational in house software maintenance.
We have recently moved into a new building that includes a spacious EM facility. I agree with David about the utility room. Not only are the noises of the pumps and chillers out of the room (The EM users get to sit in a wonderfully quiet space), but we have less problem with temperature fluctuations in the EM room, because all that happens on the other side of the wall. Lastly, this keeps the dirty work--oil changes etc --away from the scope. We also have placed the LN2 tank in this room. If you have the space and flexibility, I fully recommend it.
Our engineers recommendeded at least 3' on each side and the back of the console to allow for access for service. (In our old building, there was precious little space for even the thinnest engineers). The amount of space on the operator side probably depends on how you use the space. We went from 3 feet of space (you could not fully open the door while a user was seated) to 10 ft of space between the console and the wall. This has left plenty of room for door sweeps, a small work table, and room for my class of 12 to sit and watch a demonstration.
Don
On Mon, 3 Nov 2003, David R Hull wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } You may also want to consider utility rooms or corridors for noisy pumps and chillers if space allows. We had an opportunity to setup a lab with the ability to place utility rooms adjacent to the microscope rooms and it makes for a much enjoyable place to work. For our Philips CM200 we also have the power supply in the utility corridor, mechanical pumps are not a problem because they don't run all the time. Our Hitachi S-4700 mechanical pumps do run all the time and I'm glad they are not in the same room. I would prefer smaller rooms with a utility corridor over a large room. } } -- } David R. Hull } NASA Glenn Research Center at Lewis Field } Advanced Metallics Branch } Mail Stop 49-1 } 21000 Brookpark Road } Cleveland, OH 44135 } } (216) 433-3281 } fax (216)977- 7132 } david.r.hull-at-nasa.gov } http://www.lerc.nasa.gov/WWW/AdvMet/ASGWEB2000/asghome.html } } }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 4 08:15:23 2003
All Some kind soul has presented me with samples of debris which are very small (I would guesstimate {10 microns). These are stuck to magic tape and then onto paper. What I now need to do to prepare for EDX analysis is to get them out of the adhesive of the tape... On ordinary tape, the adhesive, I have found, can be dissolved using chloroform (acetone just makes a sticky goo). The Scotch Magic Tape appears different though and I am left with a sticky goo again. Question: Anyone any idea how I can dissolve this adhesive?
Regards Adrian Jenkinson
__________________________________ Do you Yahoo!? Protect your identity with Yahoo! Mail AddressGuard http://antispam.yahoo.com/whatsnewfree
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 4 08:54:30 2003
All, Let me add couple cents to the discussion. We have moved to a new lab in January. The lab was designed into a old FAB space so we did not worry about vibrations etc. When we were designing the facility we went ahead and used the manufacturers brochures as a minimum requirements. After hearing our opinions, the interior designer forced the rooms to be exactly the dimensions provided by FEI for their scopes. Lesson 1: plan and fight for as much room as possible within your budget.
Our two TEMs were disjoined into separate rooms. JEM2010 fits nicely and there is plenty of space to move around. Our mechanical pumps and chillers were relocated to separate utility rooms (great idea!!!), power supplies needed to stay in microscope rooms due to cabling distance. Works great!
Our CM300 on the other hand was shoehorned into Technai F30 minimum room. The mechanicals of the microscopes are essentially the same, so it looked as a good fit in view of future upgrade. Besides, we could not get the original spec from FEI for the CM.
Lesson 2: What went wrong? The two scopes have different power supply, chiller, and pump requirements, we again were able to re-locate mechanical pump and chillers to utility room (great), but the power supplies need to stay in the main room (boo). Now we need to put up with fan noise coming out of the main PS (we used to live with it in the old lab, but now we actually notice it). Annoying but not detrimental to daily operation. Moreover, we did not realize that FEI made minimum requirement for the height of the room different. In order to disassemble the FEG, the whole assembly needs to be lifted on a crane supplied with the microscope. With Techani room, we are 1" short of the height. Now each time FEI works on the gun, we need to remove soft panels from the ceiling and essentially open up a large AC plenum that is above the whole lab. This dumps a lot of particulate and dust into the room since this bypasses the filters. That's a NO-NO when working on a ultra high vacuum systems ! and something to take into account when designing air flows. I get ribbed for it each time FEI service comes in.
Our SEM, FIB, and SIMS tools were relocated into new rooms doubling up. The used to be in a large ball-room setting in the old lab. This tremendously helps in facilitation of work and nap times. Great idea, stay away from ballrooms even with curtains around tools. One thing to take into consideration is the selection of tools that are put together. We tried to pair a daily workhorse with a tool that was specialized and used only on occasion. This helps to reduce interference between operators or service.
Hope this helps.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 512 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-amd.com ******************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 4 15:28:08 2003
During my exploration of adhesives to make some "poor man's" grid glue, I only used chloroform. Bozzola discusses the use of ethylene dichloride as your solvent. I believe you can trial-and-error with most any organic solvent. I found that with enough time, the backing on any of the tapes we had in the lab became a "sticky goo" (is that a scientific term?) in the chloroform.
Good luck and let us know what you come up with!
Chris
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 4 16:10:46 2003
This meeting is sponsored by the Midwest Microscopy & Microanalysis Society (MMMS) in association with the ASM Chicago Chapter and will be held at Harper College in Palatine, Illinois on Friday November 14th. Pre-registration is open through Friday November 7th. For a registration form please contact Robb Mierzwa (see below).
For further information including hotel accommodations please contact either:
Robert Mierzwa (MMMS President - Elect): Tel: 920-803-8945, email: mierzwa-at-jeol.com Arvid Casler (MMMS Program Coordinator): Tel: 847-674-7700, email: arvid_casler-at-fmo.com Jim DiOrio (MMMS President): Tel: 847-270-4676, email: jim_diorio-at-baxter.com
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 4 18:10:11 2003
} We also have placed the LN2 tank in this room. If you have } the space and flexibility, I fully recommend it. }
Always dreaming up the worst scenarios and I apologise in advance for that. However:
How big and how well ventilated should a separate "utility room" be if an LN2 tank is being stored in there? How big is the LN2 tank? Also, where are the LN2 transfers from that larger vessel into smaller hand-held dewars being done? If you are storing a large LN2 tank that lets out a "lot" of nitrogen into that room over time, and it is a room that is normally closed up nearly all the time because "nobody goes in there that much", then could there be a slight risk of too much air (oxygen) getting displaced? Maybe this could be a potential problem if there is no formal ventilation in that room, or if the room is only serviced as part of a closed A/C system.
There have been a few postings on this list in past years relating tales of air displacement by evaporating LN2 in confined spaces. Perhaps this is also something that should be considered when thinking about where to put stuff associated with running EMs these days.
Just my paranoid thought for the day!
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/
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 4 18:10:45 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (anjeanette.ormonde-at-unilever.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 4, 2003 at 14:44:59 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] MListserver: Dismantling TEM
Question: We have a Philips 400 TEM that we are no longer going to be keeping. Right now it is under vacuum and everything is working well. However, we do not use the system enough to justify the costs of keeping it (we let the service contract expire in Dec. 2002 but even beyond that depreciation costs come out of our budget). Anyhow, I was not able to find anyone who wanted the system (free to a good home!) so I am (sadly) faced with dismantling and discarding the system. However, I have no experience with this. Are there components we can separate out and either keep or give to others? What happens with unwanted pieces? Can most things go out with regular garbage? Can anything be recycled? Thanks in advance for any advice or tips you can offer.
Adrian Jenkinson wrote: ======================================================= Some kind soul has presented me with samples of debris which are very small (I would guesstimate {10 microns). These are stuck to magic tape and then onto paper. What I now need to do to prepare for EDX analysis is to get them out of the adhesive of the tape... On ordinary tape, the adhesive, I have found, can be dissolved using chloroform (acetone just makes a sticky goo). The Scotch Magic Tape appears different though and I am left with a sticky goo again. Question: Anyone any idea how I can dissolve this adhesive? ======================================================= The fact that you don't worry about the chloroform either dissolving or in some other way altering the particles, would suggest that the particles are either metal or ceramic, neither of which will be affected by an oxygen plasma.
Therefore, I would suggest putting the particles, goo and all into a glass petri dish and placing the entire dish with gooey particles into a plasma etcher, such as the SPI Plasma Prep II plasma etcher (see URL http://www.2spi.com/catalog/instruments/etchers1.shtml ).
The oxygen plasma will remove all traces of the organics while leaving the inorganics behind undisturbed and unchanged (from the oxygen plasma).
What is left is a dry residue on the bottom of the glass petri dish.
In our own laboratory, we would take an SEM mount, apply a double sided conductive carbon adhesive disc such as on URL http://www.2spi.com/catalog/spec_prep/cond_adhes-discs.shtml and then either a) sprinkle on the dry ashed residue particles onto the surface of the adhesive or if static or other forces keep the particles from being released from the glass, then use either b) a Zerostat antistatic gun (see URL http://www.zerostat.com ) to cause the particles to release and if that does not work, then c) press the SEM mount and adhesive gently onto the particles on the glass surface which then will be efficiently lifted off and will be attached to the SEM mount, ready for carbon coating and viewing.
If you do not have a plasma etcher, please contact me off line and we can maybe set up a demo for you and at the same time, clean up your particles.
If the collected particles are organic, then of course ,this approach most definitely should not be used.
Disclaimer: SPI Supplies manufactures the Plasma Prep II plasma etcher and supplies the carbon discs and the Zerostat antistatic gun.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 5 07:35:49 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pastasalda-at-aol.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 5, 2003 at 05:52:22 ---------------------------------------------------------------------------
Email: pastasalda-at-aol.com Name: john downes
Organization: J.G.M.Downes. consultants.
Title-Subject: [Microscopy] MListserver:
Question: Retired SEM required for good home in the UK. To be used as part of a training project.
Thanks for your word of warning, especially since I was not explicit about the situation in my facility. Due in part from previous discussion from this group, I "designed in" a concern about LN2. My current utility room is 8 x 15 ft in size. It has one door that opens directly to the corridor--so tanks can be delivered without zig-zagging through the lab-- and another door to the TEM room. All users are instructed to never dispense LN2 with the doors shut.
Your paranoia (caution) is well-placed!
Don On Wed, 5 Nov 2003, Arthur Day wrote:
} } We also have placed the LN2 tank in this room. If you have } } the space and flexibility, I fully recommend it. } } } } Always dreaming up the worst scenarios and I apologise in advance for } that. However: } } How big and how well ventilated should a separate "utility room" be } if an LN2 tank is being stored in there? How big is the LN2 tank? } Also, where are the LN2 transfers from that larger vessel into } smaller hand-held dewars being done? If you are storing a large LN2 } tank that lets out a "lot" of nitrogen into that room over time, and } it is a room that is normally closed up nearly all the time because } "nobody goes in there that much", then could there be a slight risk } of too much air (oxygen) getting displaced? Maybe this could be a } potential problem if there is no formal ventilation in that room, or } if the room is only serviced as part of a closed A/C system. } } There have been a few postings on this list in past years relating } tales of air displacement by evaporating LN2 in confined spaces. } Perhaps this is also something that should be considered when } thinking about where to put stuff associated with running EMs these } days. } } Just my paranoid thought for the day! } } } } } } } 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/ } }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 5 08:47:41 2003
Angie, A few years ago we had a similar situation with an old JEOL TEM. Over several weeks, I dismantaled the column and separated out the various metals for recycling. The take-home-message was that I was able to realize about $35.00 for working through about 10 lunch hours plus carrying MANY pounds of metal to a recycling facility on a Saturday morning. Our facilities department took care of the rest of the scope so I do not know what they needed to do for disposal. Hoping someone else has a better solution, Pat
Patricia Stranen Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu ============================ } Below is the result of your feedback form (NJZFM-ultra-55). } It was submitted by (anjeanette.ormonde-at-unilever.com) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html on } Tuesday, November 4, 2003 at 14:44:59 } ------------------------------------------------------------------- } Email: anjeanette.ormonde-at-unilever.com } Name: Anjeanette Ormonde } Organization: Unilever HPC } Title-Subject: [Microscopy] MListserver: Dismantling TEM } } Question: We have a Philips 400 TEM that we are no longer going to } be keeping. Right now it is under vacuum and everything is working } well. However, we do not use the system enough to justify the costs } of keeping it (we let the service contract expire in Dec. 2002 but } even beyond that depreciation costs come out of our budget). } Anyhow, I was not able to find anyone who wanted the system (free to } a good home!) so I am (sadly) faced with dismantling and discarding } the system. However, I have no experience with this. Are there } components we can separate out and either keep or give to others? } What happens with unwanted pieces? Can most things go out with } regular garbage? Can anything be recycled? Thanks in advance for } any advice or tips you can offer. } } Sincerly, } Angie Ormonde
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 5 09:01:01 2003
Adrian, If your chem lab has hexane, you might try that, or just use gasoline which is mostly a mix of alkanes of varying chain length. Of course, all appropriate precautions need to be taken for a very dense, highly flammable vapor.
Ken Converse owner Quality Images third party SEM service Delta, PA
ady jenkinson wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 5 09:20:00 2003
I'm a 20 year veteran of a clinical (and some research) EM laboratory. I believe that I am underpaid and under appreciated and intend to do something about it. However, I need some hard facts to support my theory. I'm one of those people that don't make a fuss about things and just take what I'm given...the problem is that I think that I've been taken advantage of. Up to 5 years ago, I was the 'junior tech'. When the 'senior tech' left, I took over all the responsibilities and haven't been given any compensation (title or pay wise). To make matters worse, I believe that I'm very underpaid in general for being the only person who can do what I do. I just have to make administration aware of this.
My question is this...would anybody be willing to share what a 20 year clinical specialist would be making in your area??? I'd be happy to tell you what I am making but don't want to publicly announce it in this forum...too many people would laugh at how low it is!
Another question is how are clinical EM technologists classified in your hospital/facility? I do it all...the technical preparation and then investigate the case by taking (hopefully!) representative pictures and then presenting this to the pathologist. Investigating a case seems to put a tech into a different category but how does administration in your facility look at this...any comments?
Any help/ideas are sincerely appreciated.
Unappreciated in Nebraska, Karen Bovard
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 5 10:15:17 2003
Why do you want to put the particles away from the tape ?
Pull the tape away from the paper, cut the zone where the particles are, et fix it on a SEM stub, with carbon paint, carbon tape, or what you normally use. Than sputter some carbon on it, to avoid charges on the tape. It works !
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } All } Some kind soul has presented me with samples of debris } which are very small (I would guesstimate {10 } microns). These are stuck to magic tape and then onto } paper. What I now need to do to prepare for EDX } analysis is to get them out of the adhesive of the } tape... } On ordinary tape, the adhesive, I have found, can be } dissolved using chloroform (acetone just makes a } sticky goo). The Scotch Magic Tape appears different } though and I am left with a sticky goo again. } Question: Anyone any idea how I can dissolve this } adhesive? } } Regards } Adrian Jenkinson } } __________________________________ } Do you Yahoo!? } Protect your identity with Yahoo! Mail AddressGuard } http://antispam.yahoo.com/whatsnewfree }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 5 12:28:21 2003
Have you looked into donating it to a local college, community college, high school or museum? I've usually found that somewhere can be found, often a college that someone in the company attended in the past. A growing destination (more so for SEMs than TEMs, because of their more intuitive imaging and interface) is a local high school. Also gives you or others a chance to donate a little of your time integrating it into a curriculum, instructing the teachers and perhaps demonstrating to the kids. Especially nice if you happen to have a student there.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Tuesday, November 04, 2003 6:20 PM, by way of Nestor J. Zaluzec [SMTP:anjeanette.ormonde-at-unilever.com] wrote: } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (anjeanette.ormonde-at-unilever.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 4, 2003 at 14:44:59 } --------------------------------------------------------------------------- } } Email: anjeanette.ormonde-at-unilever.com } Name: Anjeanette Ormonde } } Organization: Unilever HPC } } Title-Subject: [Microscopy] MListserver: Dismantling TEM } } Question: We have a Philips 400 TEM that we are no longer going to be keeping. Right now it is under vacuum and everything is working well. However, we do not use the system enough to justify the costs of keeping it (we let the service contract expire in Dec. 2002 but even beyond that depreciation costs come out of our budget). Anyhow, I was not able to find anyone who wanted the system (free to a good home!) so I am (sadly) faced with dismantling and discarding the system. However, I have no experience with this. Are there components we can separate out and either keep or give to others? What happens with unwanted pieces? Can most things go out with regular garbage? Can anything be recycled? Thanks in advance for any advice or tips you can offer. } } Sincerly, } Angie Ormonde } } --------------------------------------------------------------------------- } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 5 13:23:07 2003
On Wednesday, November 5, 2003, at 07:14 AM, Pat Connelly wrote:
Dear Angie and Pat, Our old TEM in Albany NY found a good home in one of the local high schools. We thought that it was a much better solution than recycling it as scrap. Unilever could even write off the donation, so the company would likely be happy to cooperate with any arrangement you might make. We found a school in the Albany area (population ~ 0.5M), so it is likely you can find one near you. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
} Angie, } A few years ago we had a similar situation with an old JEOL TEM. Over } several weeks, I dismantaled the column and separated out the various } metals for recycling. The take-home-message was that I was able to } realize about $35.00 for working through about 10 lunch hours plus } carrying MANY pounds of metal to a recycling facility on a Saturday } morning. Our facilities department took care of the rest of the scope } so I do not know what they needed to do for disposal. } Hoping someone else has a better solution, } Pat } } Patricia Stranen Connelly } The University of Pennsylvania } Department of Biology } Philadelphia, PA 19104-6018 } 215-898-7145 } psconnel-at-sas.upenn.edu } ============================ } } Below is the result of your feedback form (NJZFM-ultra-55). } } It was submitted by (anjeanette.ormonde-at-unilever.com) from } } http://microscopy.com/MicroscopyListserver/MLFormMail.html on } } Tuesday, November 4, 2003 at 14:44:59 } } ------------------------------------------------------------------- } } Email: anjeanette.ormonde-at-unilever.com } } Name: Anjeanette Ormonde } } Organization: Unilever HPC } } Title-Subject: [Microscopy] MListserver: Dismantling TEM } } } } Question: We have a Philips 400 TEM that we are no longer going to be } } keeping. Right now it is under vacuum and everything is working } } well. However, we do not use the system enough to justify the costs } } of keeping it (we let the service contract expire in Dec. 2002 but } } even beyond that depreciation costs come out of our budget). Anyhow, } } I was not able to find anyone who wanted the system (free to a good } } home!) so I am (sadly) faced with dismantling and discarding the } } system. However, I have no experience with this. Are there } } components we can separate out and either keep or give to others? } } What happens with unwanted pieces? Can most things go out with } } regular garbage? Can anything be recycled? Thanks in advance for } } any advice or tips you can offer. } } } } Sincerly, } } Angie Ormonde
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 5 18:11:40 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mccaulak-at-wfu.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 5, 2003 at 13:46:45 ---------------------------------------------------------------------------
Email: mccaulak-at-wfu.edu Name: Anita McCauley
Organization: Wake Forest University
Title-Subject: [Microscopy] lymphocyte preparation for the SEM
Question: I am in need of information on how to prepare CD4 and CD8 lymphocytes for viewing on the SEM. I would appreciate any protocols, references, tricks of the trade, etc...
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dwaraka-at-casimir.ece.uic.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 5, 2003 at 16:12:42 ---------------------------------------------------------------------------
This question is regarding a Digital Instruments Nanoscope III AFM/STM. I wanted to know if it was ok to leave the piezo scanner plugged in "before" switching on/off the Nanoscope controller. Initially I was of the opinion that the scanner had to be unplugged before switching on/off the controller.
I would like to transfer the license of MacTempas, software, and hardware key with a reasonable price. The transfer does not violate the license agreement. Please contact me at: hkonishi-at-unm.edu
I also handle Japanese university or Japanese NSF funds (KOHI BARAI in Japanese)through third company.
Thank you,
Hiromi Konishi, Ph.D Dept. of Earth and Planetary Sciences The University of New Mexico
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 6 02:10:25 2003
We are using the Bal-Tec CDP 030 critical point dryer. A collegue, who's working on another type of machine just told me that the specimen chamber of the CPD 030 is filling up too slowly. This may lead the the material being exposed to surface tension forces, hence deforming the specimen. Is it normal that the specimen chamber on the CPD 030 is filling up slowly or could it be that the problem lies in the fact that our liquid CO2 cylinder is placed in a safety cabinet and that the tubes leading to the CPD are too narrow (outside diameter 3mm)?
Dear folks liquid nitrogen always scares me a little. You need to be very careful about room size, ventilation and potential displaced air.
Three things you should consider above all else are: 1. you can't smell or in any other way detect oxygen depletion. 2. you will be comatose long before all of the oxygen is depleted and sleepy and less aware long before that. 3. the amount of air displaced by one litre of liquid nitrogen is 830 litres.
Add a few more points to this. If you're unconscious in a room and someone tries to rescue you, they will almost certainly suffer the same fate and so on. Further if cleaners, maintenance or security ever go into that room how will they be protected?
The worst possible scenario should always be considered in a risk assessment over something as insidious as this. How big a volume of liquid nitrogen and how much air will it displace. Remember dewars can lose their insulation (we've had 2 or 3 go over 20 years) so that would release all of the nitrogen in a few hours.
You must have warning signs and should seriously consider an oxygen depletion monitor in any area where there is a serious risk. If your room is not ventilated then should you store liquid nitrogen in there under any circumstances?
There have been several well publicised tragedies in the past.
Malcolm
PS paranoia is only when you think everyone's out to get you - in this case 'paranoids' have a much better long term chance of survival.
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Donald Lovett {lovett-at-TCNJ.EDU}
LAST CALL FOR REGISTRANTS
31st Scottish Microscpy Symposium Wednesday 12th November 2003
University of Dundee, West Park Centre 319 Perth Rd, Dundee, Scotland
Registration deadline extended to 7/11/03 - see http://www.abdn.ac.uk/emunit/smg2003.htm for contact details.
Scientific Programme and Trade Exhibition
Programme
09.30-10.0 Registration and coffee
Morning Session
Chair William Maxwell
10.00-10.45 Photochemical internalisation: From microscopy towards treatment Kristian Berg, Norwegian Radium Hospital, Oslo
10.45- 11.15 Overview and Recent Advances in SPM for the Life Sciences' Mike Conroy, Applications Scientist, Veeco Instruments Ltd, Cambridge, UK
11.15-12.00 High pressure freezing - the ultimate approach for immunolabelling? Paul Monaghan, Institute of Animal Health
12.00-13.45 Lunch, Trade Exhibition and Posters
Afternoon Session
Chair / Laurence Tetley
13.45-14.30. A comparison of cryo-SEM with other preparation techniques for the study of plant cell walls. Kim Findlay, John Innes Centre, Norwich 14.30-14.45 Tobacco mosaic virus-movement protein function investigated by FRAP Kathryn M Wright, Cell-cell communications programme, Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA. 14.45-15.00 In situ hybridisation to study gene expression patterns in nematode infected root tissue. Jane Wishart, Alison Paterson, Hui Liu, Vivian Blok. Scottish Crop Research Institute, Dundee Refreshments/Trade exhibition
Chair / Eric Lachowski
15.30-16.15 Mapping surface properties at micro/nanometre levels - by a multi-function Tribological Probe Microscope (TPM) Xianping Liu, School of Engineering, University of Warwick
16.15-16.30 Prize presentations and End of Meeting announcements
This Symposium is CPD accredited by the IBMS (1 Credit).
Exhibitors :
Veeco Instruments Ltd Microscopy Supplies and Consultants Ltd, Improvisionl Leica Microsystems (UK) Ltd, Oxford Instruments Analytical TAAB Laboratories Equipment Ltd Royal Microscopical Society ISS Group Services Ltd Agar Scientific Ltd Hitachi High-Technologies Corporation Princeton Gamma Tech (UK) Ltd Imaging Associates Ltd Emitech Ltd LEO Electron Microscopy Ltd Gatan Olympus Optical Co. (UK) Ltd Quorum Technologies
Dr Laurence Tetley Division of Infection & Immunity, IBLS, Integrated Microscopy Facility Joseph Black Building University of Glasgow Glasgow G12 8QQ
l.tetley-at-bio.gla.ac.uk tel 0141 330 4431 FAX 0141 330 3516
I've looked at neutrophils and macrophages in the SEM using the following protocol: Have the investigator plate the cells onto(round) coverslips that have been treated with poly-l-lysine or some other ECM-type molecule) Wash then fix with buffered glutaraldehyde (2-4% is usual, you can use 1%). They should be well adhered to the glass. After that, you can use a pretty standard SEM prep (post-fix, dehydrate, CPD, sputter). I use round coverslips because they fit into a holder I have for my CPD. I load them into the holder after the glut. fix and then do the rest of the processing. Its easier than having many wells or dishes with coverslips, and it minimizes the chances of having your samples flip over and you don't know which side is up. 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
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 6 09:01:23 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (useitzer-at-fz-borstel.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, November 6, 2003 at 08:49:52 ---------------------------------------------------------------------------
Email: useitzer-at-fz-borstel.de Name: Ulrike
Organization: Research Center Borstel, Germany
Title-Subject: [Microscopy] MListserver:
Question: Hello! I don't know if this is a stupid question, but here goes: I am interested in imaging DNA/RNA using acridine orange and fluorescence microscopy. Is there any way of fixing and preserving the staining without changing the DNA/RNA signals and is there a possibility to use anti-fading reagents for AO? I would be most grateful for any help! Thank you and good bye Ulrike
Are these cells spread or in suspension? If the former, leave them on their substrate, if the latter, filter them down onto a 0.2 or 0.45 micron membrane filter. The kind with neat, round holes, not a torturous-path filter. Use a syringe filter holder, and keep the fliters in the holder -- use the syringe to change the solutions. Fix in 1 or 1.25% glutaraldehyde + 1% tannic acid (Mallinckrodt 1764 seems to work best) in whatever buffer you're using for the cells *minus* any serum proteins or other peptides. The tannic acid reduces or prevents holes in membranes. Fix for 1 (maybe 2 hrs) hour at room temp or overnight at 4 deg C. OsO4 is usually not needed, although if desired, you can try 1% OsO4 + 1% tannic acid for 1 hr at room temp. Wash in buffer and dehydrate starting with 30% EtOH through 3 X 100% *dry!* EtOH, with 5 to 10 min. steps. Usually 5 minutes for cells. Critical point dry using 4 or 5 5 min. soaks and 2 or 3 min purges (depending on your CPD's specimen holders, and what nooks and crannies they have for trapping EtOH/lqCO2). After the EtOH series, you can also try drying with HMDS (hexamethyldisilizane). There are various recipes for this on the U Florida Tips & Tricks page: http://www.biotech.ufl.edu/EM/tips/index.html I haven't done lymphocytes with HMDS, though, just CPD, so I can't say how well HMDS works for this purpose.
Phil
} Email: mccaulak-at-wfu.edu } Name: Anita McCauley } } Organization: Wake Forest University } } Title-Subject: [Microscopy] lymphocyte preparation for the SEM } } Question: I am in need of information on how to prepare CD4 and CD8 } lymphocytes for viewing on the SEM. I would appreciate any } protocols, references, tricks of the trade, etc... } } Thanks } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 6 09:22:08 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (couryhouse-at-aol.com) from on Thursday, November 6, 2003 at 09:07:36 ---------------------------------------------------------------------------
Email: couryhouse-at-aol.com Name: ed sharpe archivist for smecc
Organization: southwest museum of engineering, communications and computation
Title-Subject: [Microscopy] Museum needs several micro manipulators
Question: For several projects and demonstrations we need several micro manipulators Please respond off list and let us know what you can share....
Ed Sharpe archivist for SMECC
see the museum's web site at www.smecc.org or when in Arizona stop in and visit....
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (couryhouse-at-aol.com) from on Thursday, November 6, 2003 at 09:09:44 ---------------------------------------------------------------------------
Email: couryhouse-at-aol.com Name: ed sharpe archivist for smecc
Organization: southwest museum of engineering, communications and computation
Title-Subject: [Microscopy] Museum needs RCA EMT electron microscope parts and docs....
Question: The museum has an RCA EMT electron microscope we are restoring.... need some parts and more documentation
to see what one of these looks like go to the museum's page at... http://www.smecc.org/rca_emt_tabletop.htm
There are some nice images you can download of the brochure, the large ones are really large so a hi speed connection is preferable!
Identification of Man-Made Textile Fibers using PLM.
December 5, 2003
class=Section2} Step by Step Procedure for the Identification of Man-Made Textile Fibers using PLM.
The day will begin with a brief overview of a personalized approach to modern textile fiber identification using PLM and then move on to a full workshop in which the participants will be given unknown fabric samples throughout the day to identify. As a beginning review, each participant, together with the group, will look at the natural fibers such as cotton,linen, silk (cultivated and tussah), and wool to be sure we can eliminate them from our unknown pool. The workshop will then concentrate on the most common man-made fiber types found in contemporary and historic textiles, such as viscose, cuprammonium rayon, acetate, triacetate, polyester, acrylic, nylon, olefins, and even elastomerics, Beginning with sample taking and mounting techniques, this day will cover the most effective techniques to use in the identification of unknown man-made fibers: refractive index, the 530nm red tint plate, and the Michel Levy chart. Although the day is about how far one can go with fiber identification using PLM alone, a discussion of corroborative solvent and burning tests will also be covered. The day will end with a "final exam" for which each student will be given a mystery fabric of mixed fiber types to identify. The instruction will be supplemented by printed materials to take home.
The Workshop Instructor is Denyse Montegut, the Chair of the Museum Studies Department in the School of Graduate Studies at the Fashion Institute of Technology where she has taught conservation science courses since 1991. She received her B.A. in art history and mathematics from Brooklyn College, and holds an M.A. in art history and a certificate in conservation from the Institute of Fine Arts, NYU. She is currently a Ph.D. candidate in art conservation research at the University of Delaware. Denyse also has a small private conservation and consulting practice.
WHEN: Friday December 5, 2003, from 10 A.M. to 5 P.M.
Because the fumes of the resin are so objectionable, is it permissible to do the actual embedding step out of the cold chamber (-50) in the fume hood or must the specimens remain at the low temperature suggested for the entire infiltration series? We have figured out the logistics of changing our specimens from lower to higher concentrations of resin while maintaining temperature discipline, but the act of teasing out individual specimens, placing them in embedding molds or gel-caps and filling them with resin seems problematical if some warming isn't tolerated. Any advice or guidance would be much appreciated.
As always, thanks to Nestor for keeping up with us and the web world and thanks to those who share their hard won expertise.
Regards, Bill Sharp
William P. Sharp Arizona State University School of Life Sciences, box 4501 Tempe, AZ 85287-4501 Phone - (480)-965-3210 Fax - (480)-965-6899
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 6 13:21:25 2003
Hi all, I would like to purchase a good modern histology text to use as a general reference for this facility. I would like one that is sufficiently up-to-date to include SEM, TEM and ICC (Light & EM).
I would appreciate recommendations.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 6 15:10:29 2003
A workshop on electron micro-diffraction and direct methods for crystalline structure determination using electron diffraction/imaging will be held at the National Center for Electron Microscopy (NCEM), Lawrence Berkeley National Laboratory, University of California, Berkeley from April 18 through April 23, 2004. For further details, which will be posted as they are finalized, see http://ncem.lbl.gov/workshop.htm.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm2-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 6 16:03:03 2003
If you know the room can have an atmosphere that won't support life you pull some one out by holding your breath and working in short sprints. You should first get some one to back you up and tie a line to your leg so he can drag you out without having to go in himself. And your effort should be to get a rope on the person that is down and get out of the room and drag them out.
Spend some time thinking before you go in that room because as soon as you blood oxygen saturation drops you are through thinking. You can only act on plans made before hand.
With all the safety warriors at every turn if there is a room that has a liquid nitrogen container large enough to pose any risk at all without a oxygen monitor you should roast your safety officer over a bed of low coals for ignoring real risks and enforcing petty ones.
Gordon Gordon Couger red-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. : : Dear folks : liquid nitrogen always scares me a little. You need to be very careful about room size, ventilation and potential displaced air. : : Three things you should consider above all else are: : 1. you can't smell or in any other way detect oxygen depletion. : 2. you will be comatose long before all of the oxygen is depleted and sleepy and less aware long before that. : 3. the amount of air displaced by one litre of liquid nitrogen is 830 litres. : : Add a few more points to this. If you're unconscious in a room and someone tries to rescue you, they will almost certainly suffer the same fate and so on. Further if cleaners, maintenance or security ever go into that room how will they be protected? : : The worst possible scenario should always be considered in a risk assessment over something as insidious as this. How big a volume of liquid nitrogen and how much air will it displace. Remember dewars can lose their insulation (we've had 2 or 3 go over 20 years) so that would release all of the nitrogen in a few hours. : : You must have warning signs and should seriously consider an oxygen depletion monitor in any area where there is a serious risk. If your room is not ventilated then should you store liquid nitrogen in there under any circumstances? : : There have been several well publicised tragedies in the past. : : Malcolm : : PS paranoia is only when you think everyone's out to get you - in this case 'paranoids' have a much better long term chance of survival. : : Malcolm Haswell : e.m. unit : School of Health, Natural and Social Sciences : Fleming Building : University of Sunderland : Tyne & Wear : SR1 3SD : UK : e-mail: malcolm.haswell-at-sunderland.ac.uk : : : : ----- Original Message -----
: } : } : } Thanks for your word of warning, especially since I was not : } explicit about : } the situation in my facility. Due in part from previous : } discussion from : } this group, I "designed in" a concern about LN2. My current : } utility room : } is 8 x 15 ft in size. It has one door that opens directly to the : } corridor--so tanks can be delivered without zig-zagging through : } the lab-- : } and another door to the TEM room. All users are instructed to never : } dispense LN2 with the doors shut. : } : } Your paranoia (caution) is well-placed! : } : } Don : } On Wed, 5 Nov 2003, Arthur Day wrote: : } : } } } We also have placed the LN2 tank in this room. If you have : } } } the space and flexibility, I fully recommend it. : } } } : } } : } } Always dreaming up the worst scenarios and I apologise in : } advance for : } } that. However: : } } : } } How big and how well ventilated should a separate "utility room" be : } } if an LN2 tank is being stored in there? How big is the LN2 tank? : } } Also, where are the LN2 transfers from that larger vessel into : } } smaller hand-held dewars being done? If you are storing a large LN2 : } } tank that lets out a "lot" of nitrogen into that room over time, and : } } it is a room that is normally closed up nearly all the time because : } } "nobody goes in there that much", then could there be a slight risk : } } of too much air (oxygen) getting displaced? Maybe this could be a : } } potential problem if there is no formal ventilation in that : } room, or : } } if the room is only serviced as part of a closed A/C system. : } } : } } There have been a few postings on this list in past years relating : } } tales of air displacement by evaporating LN2 in confined spaces. : } } Perhaps this is also something that should be considered when : } } thinking about where to put stuff associated with running EMs these : } } days. : } } : } } Just my paranoid thought for the day! : } } : } } : } } : } } : } } : } } : } } 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/ : } } : } } : } : } ____________________________________________________________________ __ : } Donald L. Lovett e-mail: lovett-at-tcnj.edu : } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 : } P.O. Box 7718 fax: (609) 637-5118 : } The College of New Jersey : } Ewing, NJ 08628-0718
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 6 16:41:42 2003
Fawcett's The Cell has the best B&W TEM and SEM photos around but little text. The old Bloom and Fawcett "A Textbood of Histology" (later editions were only with Fawcett as the author but still carried Bloom and Fawcett above the title) has the most authoritative text and excellent B&W images but lack the pizazz of modern histology texts. Most of my students prefer Wheater's "Functional Histology" which is a short but probably sufficient text with lots of excellent LM's and a more limited number of EM's. Berman's Color Atlas of Basic Histology is also quite good. It has minimal text but large LM images. Good luck. Tom
At 02:30 PM 11/6/2003 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cachuca-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, November 6, 2003 at 16:49:34 ---------------------------------------------------------------------------
Organization: UNAM, (National Autonomous University of Mexico)
Education: Undergraduate College
Location: Mexico City, Mexico
Question: I would like to know several questions: 1)Are vacuoles exclusive of plant cells? 2)Are lisosomes exclusive of animal cells? 3)If there exists a plant or/and animal cell model (accepted in the scientific community)which shows the proportionate sizes of the ultrastructures in each prototype and is it based on reconstructions from electron microscope observations. If so, I would like to know where it has been published. Thank you very much
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ldemp-at-mse.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 7, 2003 at 07:05:34 ---------------------------------------------------------------------------
Organization: Major Analytical Instrumentation Center, University of Florida
Title-Subject: [Microscopy] Florida Society for Microscopy Symposium
Question: Florida Chapter of the AVS Science and Technology Society Florida Society for Microscopy Florida Section of the American Ceramic Society 2004 Annual Joint Symposium
March 8-12, 2004 University of Central Florida Student Union Orlando, Florida
Web: www.flavs.org
You are invited to attend the 2004 Annual Joint Symposium of the Florida Chapter of the AVS Science and Technology Society (FL AVS), the Florida Society for Microscopy (FSM) and the Florida Section of the American Ceramic Society (Fl ACerS). This symposium is sponsored by the Florida Chapter of the AVS Science and Technology Society and will be held jointly with the Florida Microscopy Society and the Florida Section of the American Ceramic Society at the University of Central Florida, Student Union Building in Orlando from March 8-12, 2004. The scientific program consists of a keynote lecture, technical sessions, a tutorial session on MEMS, a poster session and competition for students, an equipment exhibit, an AVS national short course program, a Latin-American school of electron microscopy (LASEM) program, and a science workshop for high and middle school teachers. A reception will be held on Monday evening, March 8, for all symposium participants and attendees. There is no registration fee for either the symposium or equipment exhibit but pre-registration is encouraged. On-line registration will be available by the end of December on the Florida AVS website.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (baskin-at-bio.umass.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 7, 2003 at 12:05:59 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] postdocs in microscopy
Question: Microscopists, Please bring this to the attention of qualified candidates...
Postdoctoral positions in Computational Biology
Two postdoctoral positions are available to join a research project in the quantification of deformable motion in biology, with emphasis on cell motility. The positions are supported by an NIH-funded collaboration between Tobias I. Baskin (a biologist at Umass Amherst) and K. Palaniappan (a computer scientist at University of Missouri, Columbia). One position will be at Columbia, the other at Amherst. Baskin and Palaniappan have developed new software for quantifying the spatial distribution of velocity within a growing plant organ (a root). The software is called RootflowRT and the biological application is described by van der Weele et al (2003 Plant Physiology, 32:1138-1148). The software implements a novel algorithm for quantifying deformable motion that combines structure-tensor and robust-matching approaches. The current project is to enhance and validate RootFlowRT, apply software engineering principles to the current code base, explore new computational algorithms, and extend the robust-tensor approach to other kinds of biological objects, in particular motile animal cells and embryos. The position at Missouri is primarily computational (with bio-imaging opportunities possible based on candidate interest). The successful candidate for this position will have a strong background in any area of image processing. The position at Amherst will involve imaging different kinds of biological object as well as modifying the software. The successful candidate will have experience with both imaging in biology as well as computer programming, preferably in the area of image processing.
Those interested in the computational position (Missouri) should contact Dr Palaniappan (email: palani-at-cecs.missouri.edu), and can find further information from his web page: http://www.cs.missouri.edu/facultypages/palani.html and the multimedia communications lab page: http://meru.cecs.missouri.edu/ .
Those interested in the biological computation position (Amherst) should contact Dr Baskin (email: baskin-at-bio.umass.edu), and can find further information from his web page: http://www.bio.umass.edu/biology/baskin/ and the page for RootflowRT http://meru.rnet.missouri.edu/mvl/bio_motion.
We encourage applications from anyone regardless of skin color, religion, sex, sexual orientation, or nationality.
Thank you all very much for the procedures. I was going to wait to report on the quality until the negatives I've got are scanned and formatted for a web page, but I've got the negatives scanned and no time to put the following in a web page yet.
On the 28th (maybe the date even makes a difference with formvar films :wink:), I set up to do a few different methods. And now having done my different trials, I can say what I think works for me.
} From the suggestions here, I set about to do two different methods, 'breath' and 'steam' and of course a 'control.' The common procedure for all: -0.5% Formvar (too thick, commenting further down) in Ethylene Dichloride -Fisher Superfrost Slides, fresh out of the box, and wiped vigorously with dry fisher lens paper. -'Cleaned' slide was dipped into a dip-miser containing the fomvar solution -It was pulled out, letting the excess wash down (this process is all about watching and observing the formation of the film - too slow, too fast - lumpy draining...) -Treatment: ----Breath: As one suggestion said fog it up but DO NOT breath in, and the proximity of the wet ED coated slide to my mouth was disconcerting but I carried on in the name of 'education' (and with great caution NOT to breath in). So I fogged it up... the formvar on the slide immeadiately turned a bit white, foggy. The slide was then allowed to dry completely
----Steam: I put an Erlenmeyer flask on a hot plate and had it boiling prior to starting any of the previous steps. The still wet slide was wafted above the mouth of the flask in the steam... (I made 6 slides for this test). The slides were affected very differently with different times in the steam, and what I found later was that different areas of the same slide had different results. After steaming the slides were allowed to dry.
----Control: Dip and dry.
-Collection of the film was standard: cut sides of film with a razor blade, float off in a 9" straight sided bowl in clean water at a 45 degree angle, breath on it before floating and after each cut on the sides, but only after eating a taco salad and spinning around three times on the chair chanting "formvar is my friend..." (sorry, couldn't help myself).
-Grids were placed dull side down on the film -Grids and film were picked up with parafilm covered slides (my preferred method - but after the next step and reading Andrew's procedure I might chance it for making lacey films). -Grids were removed and placed on filter paper in a 110 degree C oven for 3 minutes (or not) -Imaged on the TEM
Results: The control grids were not heated and yielded a thick film (with one or two small holes-it was an older batch of formvar). But in general a standard film. The Breath grids were covered with lacey patterns but very few true holes, most of the lace was just thicker areas of formvar. The Steam grids, both heated and un-heated, were lacey. Hole size was a bit variable, but they were true holes, and from what I can tell very suitably for our purposes here. Many of the holes had stragler filaments of formvar, and heating them for the full 12 minutes at 110 didn't appreciably change the holes or the filaments... The next batch I will add the ED dish vapor variable as outlined by Andrew Blackwood, to see if that changes anything. (images will be posted)
To be honest I was rather surprised how well they turned out. And as luck might have it they might not be the same the next time. But as specific as this is and as much sample as can be loaded on a single grid, the time spent checking the grids at each stage is well spent. One observation I have is that the film didn't look like the clean 'control' film. I might find that as I move on to the sample loading phase that the holes in the lace are too big.
Another observation noted was the thickness of the film and the supports. They should be acceptable, esp since they are for looking at samples suspended over the holes, but the film is thick, and the next batch I try (to verify and refine my technique) I will do with both a 0.5% and a 0.25% formvar/ED solution, and evaluate that variable. The un-coated film is, as to be expected, increasingly unstable as the intensity of the beam goes up.
After carbon coating even the thinnest filaments are stable. Even at crossover. And the films are clean and free from evaporated debris as reported in one or two papers.
Finally (11/7/03) I managed to get a sample of the carbon "nanotube" (no nanotubes - just an amorphous carbon sample unfortunately) and was very successful in imaging the sample.
I also had the great pleasure to switch from the old formula 4489 film to the new stuff in process of collecting before carbon and after carbon coating. This film is aweful - simply horrible. Even constant hand agitation yielded poor results (better than just agitating ever 30 seconds as we had with the old formula). This is incredibly frustrating (the new film that is).
And of course as with a few things in the EM world, explaining and duplicating the lacey carbon procedure in a different place under a different phase of the moon in a different drainage basin might be impossible... or more seriously as Larry Allard said "there is an art associated with each step."
(and I will get the procedure and images on the cmich facility web page soon)
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: Blackwood, Andrew [mailto:ablackwood-at-2spi.com] } Sent: Thursday, October 30, 2003 7:43 AM } To: microscopy-at-sparc5.microscopy.com } Subject: [Microscopy] Making Lacey Carbon Grids }
} 29 October 2003 } } Geoff Williams asked how to make lacey carbon coated grids. There may be } many methods; a number of methods may be found in textbooks. Here's how we } do it. The problem with instructions like these is that the devil is in } the } details, and times, temperatures and concentrations all vary with the } season.
snip
} And it helps to have been doing the process for several decades. This is a } skilled art, and the only substitute for going through the learning curve } yourself is to ask someone who has "been there" before you to make the } grids } you need. } } Andy } } Andrew W. Blackwood, Ph.D. } Vice President, Technical } SPI Supplies } P.O. Box 656 } West Chester, PA 19381-0656 } Ph: 1 610 436 5400 X108 } FAX: 1 610 436 5755 } e-mail: ablackwood-at-2spi.com } WWW: http://www.2spi.com }
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 7 14:35:21 2003
I don't think there is one book that covers everything you need but there are several good texts available. "Histology, a text and atlas", 4th edition, by Ross, Kaye and Pawlina is just out, looks very nice. I liked the third edition muchly. One of the few books with a good balance of text and atlas. "Wheater's Functional Histology", 4th edition, by Young and Heath, is good but is more atlas than text. Earlier editions are also good. "Color Textbook of Histology", 2nd edition, by Gartner and Hiatt is good but is lacking in the atlas department. There is a seperate atlas by the same authors. For EM only (mostly TEM) "Cell and Tissue Ultrastructure" by Cross and Mercer is excellent.
Geoff
Debby Sherman wrote:
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-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 7 15:45:49 2003
Dear Listers, We would like to set up reflected-light Kohler illumination on an optical bench using a half-silvered mirror to show our students how an epi-fluorescence microscope works. We've managed the transmitted-light set-up and it was very time-consuming.
If any of you have been down this path before and can help us save time from re-inventing the wheel, by giving us an idea of distances, which lens focal lengths and what diameter lenses work, I'd really appreciate hearing from you.
Cheers, Jeremy
Jeremy Sanderson.
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From MicroscopyL-request-at-ns.microscopy.com Fri Nov 7 16:00:56 2003
I've looked at a lot of TB & BCG bacteria. Could you tell me more about this:
"Rife was the first worker known to have isolated and photographed the tuberculosis virus."
-mc
} Hi Bruce: } If you want a "sane discussion" about this man's work how about } providing some real evidence of what he has accomplished, that is, where } his work has been published. Claims made on a website are just that, } claims made on a website. When I went on to the homepage and read about } how the FDA and the AMA are conspiring to keep his work out of the } medical manistream I got a bit skeptical, but that's just me. } Geoff } } Bruce Grosso wrote: } } } Check this out. } } } } http://www.rife.de/mscope/mscope2.htm } } } } Let's have a sane discussion about why all you geniuses are not following in } } his footsteps.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 7 16:28:12 2003
Geoff, You left out a critical detail. Do you spin clockwise or counter-clockwise? Thanks for the advice. Kim
On 7-Nov-03, at 11:44 AM, Geoff Williams wrote:
} -Collection of the film was standard: cut sides of film with a razor } blade, float off in a 9" straight sided bowl in clean water at a 45 } degree angle, breath on it before floating and after each cut on the } sides, but only after eating a taco salad and spinning around three } times on the chair chanting "formvar is my friend..." }
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 7 18:36:52 2003
Of all the replies I received you are the first person who understands that Rife's work has less to do with Microbiology or cancer research as it has to do with the general search for the "Mortal Oscillatory Rate" of virtually ALL pathogens.
He didn't set out to "cure cancer" that was never his goal. His goal was to see what if any effects his wavelength theory applied to cell structure. Not unlike breaking a crystal glass with sound wave, in fact exactly the same principle. That was it in a nut shell. BUT, in order to do that he had to first invent the device to View the living specimen.
Then; POOF! The universal microscope was invented and then continually upgraded for several generations. Now everyone focuses on the biological properties of Rife's work. Everyone is barking up the wrong tree. Everyone tears Rife's work apart based on his work as a biologist, which he was not, the biologists came to him after the fact.
I believe he shattered the shattered the glass and now everyone is looking at his flawed biological science when that isn't even the subject and never has been.
You are right! Someone needs to explore Rife's work from Rife's perspective. Someone needs to prove you can shatter the glass, not the biology, the biology will come all by itself and without any help. It will just happen in cadence with the real science.
Thank you for being so insightful. That makes you one of those geniuses I am always talking about on the list.
Thanks you also helped me encapsulate Rife into less than a paragraph for future discussions should the opportunity ever arise again. Who knows maybe one day Rife will be right up there where he belongs with Tesla and others.
From MicroscopyL-request-at-ns.microscopy.com Sat Nov 8 01:17:34 2003
I know a lot of people in this forum are biting their cheeks, or simply deleting any messages in this particular thread. But, in my usual fashion, I'm more than happy to jump into an area that I confess to have little knowledge of.
I would just like to add my two cents worth, since this captured my attention - and my interest comes down to one simple fact. If you (Bruce) want to argue some miracle cure, you can find a more suitable soapbox than the microscopy listserver.
There are plenty of venues out there for this avenue of thought. Whether or not Rife managed to produce an optical instrument capable of resolutions exceeding current instruments is a valid topic here. I, personally, haven't seen any recent serious work proposing that, but would love to see a peer reviewed research paper on it. I suspect that there are more than a few individuals on this listserver that would love to find this all true, as the commercial aspects alone would be nirvana, not to mention the accolades to the researcher. However, this stinks of an early 1900's Pons and Fleischmann - bad and incomplete science brought to media attention to aggrandize the researcher and attract money to an endeavor that was never really viable.
I'm sure that you will want to respond to this, so while you're at it, you might want to provide us with the particular credentials that entitle you to rant to such a wide variety of professionals who have spent their careers in microscopy. This isn't a particularly large field, and the real experts in optical microscopy are getting fewer, but we all share an interest in the sub-atomic particle interactions at the heart of this problem.
Your desire to see Rife elevated to the heights of scientific minds such as Tesla tends to put your rantings in perspective. Aside from his influence in Westinghouse's decision to promote the use of AC currents in electrical power distribution, I really can't think of a single contribution of Tesla that has resulted, after all of the ensuing years, in any practical application. Of course, I'm a Troglodyte, and don't have idea of the vision of the future that you have. But please don't clog the microscopy listserver to enlighten me - email me directly. I'll be more than happy to engage you for a while.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Friday, November 07, 2003 8:45 PM, Bruce Grosso [SMTP:bgrosso-at-AssetRecovery.Net] wrote: } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Michael: } } Of all the replies I received you are the first person who understands that } Rife's work has less to do with Microbiology or cancer research as it has to } do with the general search for the "Mortal Oscillatory Rate" of virtually } ALL pathogens. } } He didn't set out to "cure cancer" that was never his goal. His goal was to } see what if any effects his wavelength theory applied to cell structure. Not } unlike breaking a crystal glass with sound wave, in fact exactly the same } principle. That was it in a nut shell. BUT, in order to do that he had to } first invent the device to View the living specimen. } } Then; POOF! The universal microscope was invented and then continually } upgraded for several generations. Now everyone focuses on the biological } properties of Rife's work. Everyone is barking up the wrong tree. Everyone } tears Rife's work apart based on his work as a biologist, which he was not, } the biologists came to him after the fact. } } I believe he shattered the shattered the glass and now everyone is looking } at his flawed biological science when that isn't even the subject and never } has been. } } You are right! Someone needs to explore Rife's work from Rife's perspective. } Someone needs to prove you can shatter the glass, not the biology, the } biology will come all by itself and without any help. It will just happen } in cadence } with the real science. } } Thank you for being so insightful. That makes you one of those geniuses I } am always talking about on the list. } } Thanks you also helped me encapsulate Rife into less than a paragraph for } future discussions should the opportunity ever arise again. Who knows maybe } one day Rife will be right up there where he belongs with Tesla and others. } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Nov 8 08:16:53 2003
Allen, Are you going to let the rest of us know if there is some REAL information out there? Given that resolution is given as magnification and TB is a virus, I have my doubts that there will be anything to report. It's been a long time since I was a microbiology major, but I've heard of E. coli, can anyone confirm the existence of B. coli?
If it's on a web site, it must be true........
Ken Converse owner Quality Images third party SEM service Delta, PA
Allen Sampson wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sat Nov 8 09:22:37 2003
I've lately been seeing an increase in REJECTED mail questions from subscribers. As you know I have tightened the noose on the Junk/Spam mail filters significantly in the last few months.
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The most common problem I am seeing now is that many subscribers have over the years changed their Email addresses subtly.
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From MicroscopyL-request-at-ns.microscopy.com Sat Nov 8 09:43:43 2003
I sent this already to Allen but in light of Ken's comment I feel I am obligated to share these few thoughts with everyone.
First off I am not on a Rife soapbox. Second, I had already dropped the discussion idea as out of the dozens of mostly derisive replies only a few, not always agreeable people, kept their heads and dissected what they knew about Rife's' work in intelligent and respectful tones, Thirdly, I am not an educated person, I have always meant "You geniuses" in the most flattering tones, or at least tried to on account of the fact that to me you all "ARE" geniuses compared to me.
What I know about Rife's work began after reading, and then on several occasions, rereading the book by Barry Lynes.
Several lengthily phone conversations with Lynes and two personal meetings with one of Rife's living coworkers in his home in San Diego in the late 1980's with him at that time in his 80's., Touring his garage with him, seeing what I saw in the way of documents and equipment and listening to what he had to say intensely. It wasn't until those two meetings that it dawned on me that Rife actually did produce the microscope, I viewed a few of the remaining original 35mm through the lens photos with him, and bits and pieces of what could only be described as detailed documentation of something I could not make heads nor tails of, for want of the academic background to comprehend, but could not imagine that this man, who stood before me, so late in years was clinging to a lie so close to the end of his life.
It also occurred to me begged the question: What is so unfathomable about the theory of cellular wall destruction by waveform? It is already a widely accepted fact that sound wave can kill you given the right frequencies.
Rife never contended that, what he set out to do was to isolate each individual cells particular, individual "Mortal Oscillatory Rate" and for each individual pathogen, be it bacterial or viral.
What is so hard to accept? That he concocted this microscope and lied relentlessly about the results he was reporting, given the man's credentials and well documented personal achievements?
I am more prone to believe his lab was vandalized and parts of the microscope were pilfered than to think someone of his already renowned stature fabricated his claim of magnifications for which he was well qualified to create according to academic records as outlined in the book, duly footnoted, cross referenced and included in the bibliography.
If you haven't read the book there is no point in defending or being offensive towards something you haven't read. I lost my copies (I had six altogether) years and years ago.
And yes! Now after all this, I do plan on acquiring another copy so I can brush up on my rebuttals to repudiations.
Lastly is Barry Lynes himself who I have had the pleasure of several lengthily telephone conversations with and he in no way appears to be the type who would idealize something or someone and instead seems to be a thorough and competent, objective reporter, who has been caught up in a shit storm of his own making but quite unintentionally.
He has no illusions of Rife. Does not idolize Rife and reported the facts as he acquired them with no slant or favoritism in most depictions of his dissertation on Rife's life and work. He respects Rife but does not idolize him.
So what am I then therefore to think? Uneducated by and large except in the most cursory way towards each of these sciences, yet tantalized by the sheer scope and magnitude of the drug companies strangle hold on an industry who sheer might and force of power is nothing less than awesome, combined with the BILLIONS of dollars in cancer research monies and yet the plague still grows exponentially with no end in sight.
For myself I chose to believe in the adage. "Keep It Simple Stupid" which is what I think Rife did. He never set out to cure cancer. That was never his intent. He set out to shatter the crystal glass and I am convinced to whatever degree; he did it, he actually did it. Even with the possibility that his achievements could possibly have been over stated in the broadest sense by others I am convinced he succeeded to some extent and THAT is what needs to be recreated.
But alas, it is so much easier to criticize than scrutinize, or deride than to investigate. It is so much simpler to hold the party line and shout "Impossible" instead of conceding that vandalism and arson destroyed the man's work in an age where backup was not as easy as the click of a mouse. Hell there weren't even copy machines back then.
Given Rife's penchant for detailed reporting of the simplest and most benign movement of the scopes adjustments a (few surviving pages of which I saw myself but made no sense to me) just the simple replication of his work by a clerk would have been nothing less than a monumental undertaking in it's own right.
You say you have some knowledge of this man's work, But do you have any knowledge of the man? Have you ever tried to get into his head? Read about his life from an objective reported viewpoint and try to see into his mind and what he was thinking all those long 10, 15, 20 or 30 hour sessions sitting at the scope recording his every turn of the dial, pencil in hand every time he touched a screw or adjusted a component, this way and that. And then documenting that failure after another and another until he found what he was looking for, all or in part.
Like a movie is an encapsulation of a book, it doesn't all fit into 1 hour and 10 minutes, a book is an encapsulation of a life, it doesn't all fit into a few hundred pages.
No I think you geniuses are the ones who need to get real, not me, not Rife. If you don't think he did it. Prove it. Recreate his every footstep and then tell me he was wrong. Especially when the underlying theory is just all too simple in it's concept and all too complicated in it's implementation.
I can say, yes here is how we can get to the moon and back, but what it took to actually do that was a huge undertaking. I believe so it is too with Rife's work, his theory and I believe he did the work and took on the monumental task and left his foot print in the sand. Now someone needs to go back and with today's technology retrace his footsteps and prove he was wrong or right.
That is what gets me. No one is willing to prove him wrong, yet everyone wants to accuse him of something I just don't think the man was capable of.
I got into his head and I wish I had your education and an extra 20 years left because I would be willing then to undertake the task. I am just a salesman. You guys are the geniuses.
I am convinced someone in your fields of expertise will take up the mantle one day. It is just a matter of time.
Cheers and God Bless
Bruce
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 775-213-9028 FAX bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists ----- Original Message ----- } From: "qualityimages" {qualityimages-at-netrax.net} To: {ars-at-sem.com} ; "Microscopy" {Microscopy-at-Sparc5.Microscopy.Com} Sent: Saturday, November 08, 2003 6:24 AM
Colleagues.....
I see no discussion of microscopy in this thread and note that it is beginning to breakdown into off-topic issues unrelated to the subject of the Listserver.
Further postings should be taken off-line, between those that are interested in continuing.
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Sun Nov 9 09:24:10 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dougbaldwin-at-mindspring.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, November 8, 2003 at 22:54:42 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] Macro-Nikkor Lenses and Nikon Multiphot Macroscope
Question: Dear Listers,
I'm shooting the circuitry of microchips such as a Pentium-type processor. The chip faces range in size from 1mm to 20mm. My 6 Mb Fuji S2 digital camera with 60mm Micro-Nikkor lens don't resolve enough detail and resolution on the larger chip faces (for one important client) and the large chips are too big for a microscope to capture in one shot.
I'm thinking of shooting them with a MACRO-Nikkor, either the 65mm f4.5 or 120mm f6.3 very high resolution lens on a 4x5" camera or a Nikon Multiphot MACROscope which was specifically designed for macro photography in the 1x-40x range.
The Mulitphot system was last made in the mid 1980's by Nikon. They were not standard camera store items so they don't show up in current used camera inventories. I can't find either the lenses or the Multiphot for sale anywhere.
I'm hoping one of you microscopists out there might have one or both of the lenses or even the whole Microphot system (including the 19mm f2.8 and 35mm f4.5 Macro lenses) sitting in a closet unused and lonely, just waiting to be put back into service shooting hi-tech photos of microchips in the 21st century.
There are other macro lenses that might work as well. Those would be the Zeiss Luminars or the Leitz Photars. Does anyone have any of these lenses?
I'm thinking of purchasing an older Nikon Optiphot microscope with coaxial lighting (necessary for reflective microchip faces) to shoot the small 1-5mm chips. My budget is limited and thought this might be an appropriate microscope for this part of the project. Any feedback?
I would appreciate any thoughts on this subject and especially a Nikon Multiphot system that might be for sale.
You may well have already rejected this on some other grounds, but why don't you get someone with a SEM to image them for you?
Or use a lower-magnification optical microscope.
cheers
rtch
Date sent: Sun, 9 Nov 2003 09:35:52 -0600 To: microscopy-at-ns.microscopy.com } From: dougbaldwin-at-mindspring.com (by way of MicroscopyListserver)
This is done either of two ways, depending on the final resolution desired. The simplest and the method with least overall resolution is a stereo zoom.
The highest quality method is to take multiple shots of small portions of the die and stitch them together. This method can produce up to 60"x60" full color 300dpi prints. Or, they can be PDF for on-line viewing. This method uses a metallurgical microscope with motorized stage.
I don't think your main problem is resolution, per se. Rather, it is the field of view that you can obtain with a corresponding ability to resolve whatever degree you need.
gary g.
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America To } Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (dougbaldwin-at-mindspring.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Saturday, November 8, 2003 at 22:54:42 } --------------------------------------------------------------------------- } Email: dougbaldwin-at-mindspring.com Name: Doug Baldwin Title-Subject: [Microscopy] } Macro-Nikkor Lenses and Nikon Multiphot Macroscope Question: } Dear Listers, I'm shooting the circuitry of microchips such as a } Pentium-type processor. The chip faces range in size from 1mm to 20mm. My } 6 Mb Fuji S2 digital camera with 60mm Micro-Nikkor lens don't resolve } enough detail and resolution on the larger chip faces (for one important } client) and the large chips are too big for a microscope to capture in one } shot. I'm thinking of shooting them with a MACRO-Nikkor, either the 65mm } f4.5 or 120mm f6.3 very high resolution lens on a 4x5" camera or a Nikon } Multiphot MACROscope which was specifically designed for macro photography } in the 1x-40x range. The Mulitphot system was last made in the mid 1980's } by Nikon. They were not standard camera store items so they don't show up } in current used camera inventories. I can't find either the lenses or the } Multiphot for sale anywhere. I'm hoping one of you microscopists out there } might have one or both of the lenses or even the whole Microphot system } (including the 19mm f2.8 and 35mm f4.5 Macro lenses) sitting in a closet } unused and lonely, just waiting to be put back into service shooting } hi-tech photos of microchips in the 21st century. There are other macro } lenses that might work as well. Those would be the Zeiss Luminars or the } Leitz Photars. Does anyone have any of these lenses? I'm thinking of } purchasing an older Nikon Optiphot microscope with coaxial lighting } (necessary for reflective microchip faces) to shoot the small 1-5mm chips. } My budget is limited and thought this might be an appropriate microscope } for this part of the project. Any feedback? I would appreciate any } thoughts on this subject and especially a Nikon Multiphot system that } might be for sale. Thanks in advance. Doug Baldwin Baldwin Photography } Scottsdale, Arizona dougbaldwin-at-mindspring.com } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 10 17:34:59 2003
This is kind of a fishing expedition. We are thinking of adding a 120KV cryoTEM to our lab and would like to get any helpful hints from more experienced users.
I know practically nothing about this area so anything would be helpful to bring me up to speed. Not only instrument selection questions, but things like service, parts, etc.
One issue that has come up, in addition to the choice of instrument, is the question of cameras. I have read lots about the problems with the new formulations of TEM film and can't avoid noticing the rise in popularity of digital cameras. Anyone have experience that would help guide us in the right direction? Film or digital, 1K, 2K, or 4K? Same questions about service, availability, parts, etc.
Any comments would be welcome.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 10 22:19:50 2003
Hi all, Does anyone have a method to jazz up the uranyl acetate followed by lead citrate post-staining procedure? I am working with some tissue that just doesn't seem to want to stain...it looks very dull in the scope. I'm using a saturated aqueous UA then Reynold's lead citrate. Should I try UA in 50% ethanol rather than aqueous? Any suggestions would be greatly appreciated, Beth -- ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I need to modify or replace our standard fixation protocol to handle pedicels of legume pods for preparation of wax sections for light microscopy. These are about 1 mm in diameter and combine woody (phloem fibers and xylem) and soft tissues (cortex, phloem, cambium, pith) and the tissues readily distort or tear during sectioning. Currently we use an age old protocol consisting of standard fixation in FAA or parafomaldehyde, then ethanol series dehydration through TBA and into Paraplast. Clearly this is not good enough. Any suggestions for an improved protocol that would produce undamaged sections would be greatly appreciated.
Greg
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 04:13:00 2003
On Mon, 10 Nov 2003 23:30:09 -0500 Beth Richardson {beth-at-plantbio.uga.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi all, } Does anyone have a method to jazz up the uranyl acetate followed by } lead citrate post-staining procedure? } I am working with some tissue that just doesn't seem to want to } stain...it looks very dull in the scope. } I'm using a saturated aqueous UA then Reynold's lead citrate. } Should I try UA in 50% ethanol rather than aqueous? } Any suggestions would be greatly appreciated, } Beth } -- } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } *************************************************************************** } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 04:53:09 2003
Dear Beth! You will find very good results with UA 2% in methanol (100º) , then the usual lead citrate. (8 minutes for UA, whash in 70º,50º and 30º methanol then double destilated water and them , prior wash dd water, the lead citrate(10 minutes), sometime there are some variation related with the hardness and the tipe of the resin used for inclusion. We get very good results in plant tissues with the above technique. Francisco
Prof. Francisco Freire FRMS Head Service Electron Microscopy Service University of Las Palmas de Gran Canaria Fac. Cs. Salud P.O. Box 550 Las Palmas Canary Islands Spain
Beth Richardson wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi all, } Does anyone have a method to jazz up the uranyl acetate followed by } lead citrate post-staining procedure? } I am working with some tissue that just doesn't seem to want to } stain...it looks very dull in the scope. } I'm using a saturated aqueous UA then Reynold's lead citrate. } Should I try UA in 50% ethanol rather than aqueous? } Any suggestions would be greatly appreciated, } Beth
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 06:40:08 2003
Years ago, we used uranyl acetate dissolved in anhydrous methanol. As I remember, you can dissolve something like 25% of the UA. We would dip the grids in the the methanol solution for a few seconds and then rinse in methanol. The only drawback was that it seemed to etch the plastic and make the sections weak. The stain was a bit grainy as well. However, in emergencies, it might be useful.
Date sent: Mon, 10 Nov 2003 23:30:09 -0500 To: microscopy-at-msa.microscopy.com } From: Beth Richardson {beth-at-plantbio.uga.edu}
Beth, I use 70% ethanol with UA and it appears to yield better contrast than the aqueous solution. Good luck, Mary Gail Engle
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -------------------------------------------------------------------------------
Hi all, Does anyone have a method to jazz up the uranyl acetate followed by lead citrate post-staining procedure? I am working with some tissue that just doesn't seem to want to stain...it looks very dull in the scope. I'm using a saturated aqueous UA then Reynold's lead citrate. Should I try UA in 50% ethanol rather than aqueous? Any suggestions would be greatly appreciated, Beth -- ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Looking at getting a color laser printer for general quick and dirty printing. Particularly looking for feed back from anyone one either the Minolta QMS Magicolor 3100 or the Lexmark C752n. We have excellent experience with our Lexmark B&W printers. And I like the example prints from the Miniolta I pick up at M&M (Thank you Vitally). Now I'm look for more opinions Good the bad and the ugly.
Yes, we have a number of ink jet printers, excellent quality but slow, looking for some thing faster, basically to serve for walk away prints as we do more and more digital.
Thank you!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 08:00:10 2003
Jon It is perfectly understandable that you should want a digital camera on your new TEM. Anyone purchasing today would want one for a variety of reasons, but I am alarmed that you should be driven to this choice by scare stories about film quality. In my experience these are completely unjustified, and I worry that this sort of of talk will unnecessarily accelerate the demise of EM film. Despite enormous advances in digital camera performance the film image still sets the standard to beat for sheer image quality. Kodak electron image films are every bit as good as they have ever been. They are a complete piece of cake to process, and it mystifies me that anyone with darkroom experience could have serious difficulty with this.The process is quick, easy and in our hands completely reliable. The most difficult bit is to find a rack that holds the film securely. After that the film practically processes itself, no voodoo required*. Best wishes with your purchase Chris
p.s. I have to concede that the results are improved if the D19 is matured six months buried in the gizzard of a ritually-slaughtered chicken. Anyway it's a good excuse for a lab party.
On 10 Nov 03, at 15:41, Jon Krupp wrote:
} } } -------------------------------------------------------------------- } -- -------- The Microscopy ListServer -- Sponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } -- --------- } } Greetings: } } This is kind of a fishing expedition. We are thinking of adding a } 120KV cryoTEM to our lab and would like to get any helpful hints } from more experienced users. } } I know practically nothing about this area so anything would be } helpful to bring me up to speed. Not only instrument selection } questions, but things like service, parts, etc. } } One issue that has come up, in addition to the choice of instrument, } is the question of cameras. I have read lots about the problems with } the new formulations of TEM film and can't avoid noticing the rise } in popularity of digital cameras. Anyone have experience that would } help guide us in the right direction? Film or digital, 1K, 2K, or } 4K? Same questions about service, availability, parts, etc. } } Any comments would be welcome. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 08:02:16 2003
I have a user with gold-coated samples after doing ion probe analyses, and now he needs to do more electron microprobe work, both imaging and quantitative analyses. The gold coats (I'm not certain about thickness) need to be removed. We cannot, however, polish the gold off because he needs to preserve the pits left from the ion probe. As a result, we are looking for chemical means to remove the gold coats. Any suggestions are welcome (particularly those known to work and with details about concentrations, temperatures, etc.).
Thanks, Ellery
--------------- Ellery E. Frahm Research Scientist/Manager Electron Microprobe Laboratory Department of Geology & Geophysics University of Minnesota - Twin Cities Lab Website: http://probelab.geo.umn.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 09:12:02 2003
I have been using the Minolta QMS Magicolor 3100 for about 1 year, and am extremely pleased with it. Good image quality (both resolution and color fidelity), fast results, low cost. The dye-sub printer is now rarely used and the ink-jet has been completely retired.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 09:58:20 2003
You didn't mention the tissue type nor the resin, but if its Spurr's epoxy, that can make staining of embedded tissues more dificult, especially with Reynold's lead, I think. Or maybe the tissue type just doesn't pick up much stain.
I don't usually use Reynold's stain here, but prefer to use the Sato triple lead stain (lead citrate, lead nitrate, lead acetate), which I've been using for many years. The way I make it up, its VERY stable and gives excellent staining, usually with no lead ppt contamination, tho if its older than 2 months, I may use a 0.2 micron filter as a precautionary measure. A 100 ml batch typically lasts 3-4 months. If you want the protocol for making Sato lead, lemme know off-line.
Before the Sato lead, I stain with 3% aqueous UA for about 20 minutes, room temp. But you may need to cut the UA with methanol and/or elevate staining temperature as others have suggested on this thread for your special case.
Good luck, of course!
Gib
} Hi all, } Does anyone have a method to jazz up the uranyl acetate followed by } lead citrate post-staining procedure? } I am working with some tissue that just doesn't seem to want to } stain...it looks very dull in the scope. } I'm using a saturated aqueous UA then Reynold's lead citrate. } Should I try UA in 50% ethanol rather than aqueous? } Any suggestions would be greatly appreciated, } Beth
-- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 10:42:55 2003
Richard, We are getting very satisfactory results from our recently purchased HP 5500 color laser. It was purchased with image printing in mind but also has to provide color printing for the rest of the facility. Turned out to be a very good compromise and is reasonably quick. Sorry I cannot compare it to the others you mentioned, but thought you like another data point.
No $ connection or interest, just another user.
Kevin Battjes Impact Analytical Voice 989-832-5555, ext 556 Michigan Molecular Institute Fax 989-832-5560 1910 W. St Andrews Road e-mail: battjes-at-mmi.org Midland MI 48640 battjes-at-impactanalytical.com
Visit us at PittCon, March 8 - 11, 2004, booth 2306
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOHIO.EDU] Sent: Tuesday, November 11, 2003 8:56 AM To: microscopy-at-MSA.Microscopy.com
Looking at getting a color laser printer for general quick and dirty printing. Particularly looking for feed back from anyone one either the Minolta QMS Magicolor 3100 or the Lexmark C752n. We have excellent experience with our Lexmark B&W printers. And I like the example prints from the Miniolta I
pick up at M&M (Thank you Vitally). Now I'm look for more opinions Good the
bad and the ugly.
Yes, we have a number of ink jet printers, excellent quality but slow, looking for some thing faster, basically to serve for walk away prints as we do more and more digital.
Thank you!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 10:55:41 2003
Beth, If you are using an epon type embedding, then yes the saturated UA in 50% ethanol would be a great help. Some cautions in working with it (besides the radioactive chemical safety ones) are: 1. to really keep it in the dark, even while in the staining dish 2. make it in small batches (5 ml in a small vial works well) because it looses its "umph" after just a few days 3. no need to filter the stain if you let it settle for about 15 minutes after shaking and the stain is taken gently from near the top of the vial so as not to disturb the bottom crystals.
A second place for lack of contrast in staining is to leave the lead stain on too long. The metal will be leached back into the solution.
The samples I work up are usually block stained in 0.5-1.0% aqueous UA in the refrigerator overnight after fixation. My section staining proceedure is to place a dry grid onto a drop of UA stain in a petri dish on parafilm for 8 to 10 min. The lid of the dish is covered with cardboard and opaque tape. Water rinse several times and put the grid into a large drop of water also on parafilm in another dish and leave the grid there until all grids are washed. Transfer right out of the water drop into a drop of lead stain in a third covered dish for about 1-2 min. then wash well before drying it down.
Although I have used Reynold's Lead stain in the past, the lead stain that I am using now is Lead Citrate as described by Aly Famy in Proceedings of the 25th Annual EMSA Meeting - 1967 - 50ml cooled, boiled, distilled water + 1 dry pellet of sodium hydroxide, after it is disolved add 0.2g (reference states 0.25g) lead citrate. I use a plastic (PP) 50ml centrifuge tube so it can be discarded later and store the staining solution in several syringes stuck into a large stopper to keep out the air and keep it in a cabinet to limit light exposure.
Patricia Stranen Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu } ---------------------------------------------------------------------- } The Microscopy ListServer --Sponsor:The Microscopy Society of America } } Does anyone have a method to jazz up the uranyl acetate followed by } lead citrate post-staining procedure? } I am working with some tissue that just doesn't seem to want to stain... } it looks very dull in the scope. } I'm using a saturated aqueous UA then Reynold's lead citrate. } Should I try UA in 50% ethanol rather than aqueous? } Any suggestions would be greatly appreciated, } } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 11:01:36 2003
i've used ethanolic UA for almost 35 years, with very good results. the procedure calls for saturated UA in 50% ethanol, stir for 5 minutes, stain for 10 minutes in reduced light. depending on how much stain you need at a time, you can make it up in as little as 5ml preps. takes just under 200mg in 5mls to get a saturated preparation. i make it up fresh and filter it through a 0.2micropore filter just before use. keeps ok in glass for several days, i've never tried to keep it longer. it keeps very poorly in disposable syringes, only 2-4 hrs - the rubber on the end of the plunger reacts with the solution.
the reference for the procedure is: Stempak,JG; Ward,RT (1964): An improved staining method for electron microscopy. J. Cell Biol. 22, 697-701.
paul hazelton
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 11:14:24 2003
Well, first off the usual sort of advice would be to extend some of the processing times involved, especially the embedding series of dilute and pure Paraplast. I think we are talking times on the order of days for some of those steps, right? - you are probably doing that already. Then, of course, make sure you have a good sharp knife, and have convinced yourself that the microtome is not the problem.
But if you are going to be doing wax embedding of plant tissues for LM on an ongoing basis, you might want to consider throwing a completely different approach at the problem, which is microwave oven processing. In the MW oven, you can do all the steps from fixation of plant tissues to final infiltration in 100% Paraplast in about 5 hours. A book on microwave processing techniques for both LM and EM is available from Ted Pella,Inc.:
Microwave Techniques and Protocols, A New Book on Microwave Processing Edited by Richard T. Giberson and Richard S. Demaree, Humana Press
Chapter 15 is entitled: Microwave Paraffin Techniques for Botanical Tissues.
See also: Schichnes D, Nemson J, Rusin, SE (1998) Microwave protocols for paraffin microtechnique and in situ localization in plants. Microscopy & Microanalysis, 4:491-496.
Here at the CBS Imaging Center, we have taken these two sources, modified the protocols a bit for various reasons and to fit our MW processor and its accessories. We have processed alfalfa nodules for paraffin sectioning, and barley leaf infected with fungus, leaf with viral infections, other fungal specimens in epoxy resin for TEM. Again, in about 5 hours time, including cured blocks in the eposy resin for TEM case.
That approach won't solve your present problems of the moment, but its an approach well worth looking into for many kinds of biological samlple prep for LM and EM.
Laboratory microwave ovens specially designed for this kind of processing are available from vendors such as Ted Pella., Inc., and Electron Microscopy Sciences, maybe others. By the way, I have no financial interst in these two companies. If you want any more info on MW processing of plant materials, contact me off line.
Good luck!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra
} I need to modify or replace our standard fixation protocol to handle pedicels } of legume pods for preparation of wax sections } for light microscopy. These are about 1 mm in diameter and combine woody } (phloem fibers and xylem) and soft tissues } (cortex, phloem, cambium, pith) and the tissues readily distort or tear during } sectioning. Currently we use an age old protocol } consisting of standard fixation in FAA or parafomaldehyde, then ethanol series } dehydration through TBA and into Paraplast. } Clearly this is not good enough. Any suggestions for an improved protocol that } would produce undamaged sections would } be greatly appreciated. } } Greg
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 12:12:27 2003
Try this article from the Sept.-Oct. 2002 Microscopy Today: Peter Tomic Method for Metallization Stripping of Gold Interconnected Semiconductors Using an Aqueous KI Solution The issue is available on-line as a pdf file at http://www.microscopy-today.com/TableofContentsPDF.html
Phil
} Hello all, } } I have a user with gold-coated samples after doing ion probe analyses, and } now he needs to do more electron microprobe work, both imaging and } quantitative analyses. The gold coats (I'm not certain about thickness) } need to be removed. We cannot, however, polish the gold off because he } needs to preserve the pits left from the ion probe. As a result, we are } looking for chemical means to remove the gold coats. Any suggestions are } welcome (particularly those known to work and with details about } concentrations, temperatures, etc.). } } Thanks, } Ellery } } --------------- } Ellery E. Frahm } Research Scientist/Manager } Electron Microprobe Laboratory } Department of Geology & Geophysics } University of Minnesota - Twin Cities } Lab Website: http://probelab.geo.umn.edu
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 12:35:42 2003
I just wanted to thank everyone who replied - there are too many of you to thank individually. The most popular method for improved staining was: UA in 50% ethanol or methanol. Various times (5-20 min) Add 1 drop of acetic acid/per 10mls of solution. Store in an amber bottle. Stain should last a year or longer. Follow UA staining with Reynolds lead citrate.
I appreciate the help! Thanks again, Beth -- ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
The first way is to use a sputter coater in the Argon ETCH mode. Use low power and a vacuum of 50mT. The key to using this method is to not have enough energy to zap the devices yet have enough ionic bombardment to clean off the coating. Gradual etching can be examined for effectiveness by SEM imaging at moderate (10KV) energy for charging. Once you get charging, odds are that the coating is gone. I typically use 50A-80A of Au/Pd or Pt.
The second method is to use a Gatan ion mill with Argon. This too is at low power and 50mT or default. Use 2KEV, 40-50uA and 20 minutes, 10RPM, 10 degree rock. You might try building up to 20 minutes in 5 minute increments. This method will not bash metal but it will affect oxide if done too long or too "hot."
gary g.
At 06:13 AM 11/11/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 18:02:23 2003
I am in the process of surplusing our old TN5500. It is available, either as a complete unit or individual components. We have all original monitors, cables, boards, documentation, software, printer,etc., and even the Tracor Northern Training slide show (a collectors item!). The TN5500 was last used in 1996 and was retired in a completely healthy condition. It has since been used as a table top for our new automation system.
The property is offered for sale "As-Is, Where-Is". The University of Washington makes no guaranty, warranty, or representation expressed or implied as to the condition of the property or its fitness for any use or purpose. Seller confirms that it has clear title to the property. The buyer will be responsible for the cost of packing and transporting the equipment from the UW campus.
Cost is negotiable, but will be real cheap. You must pay for shipping but I can do the packaging if the items are small.
************************************************ ....amphiboles do violence to history... T. Feininger, 2001. (taken out of context) ****************************
Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Earth and Space Sciences ph.206-543-8393 Mail Stop 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 20:52:21 2003
} Dear list friends: } } I am trying to apply AC current to AFM to do some measurement, if teh AFM we have is DI nanoscope III. We do nothave any conductive module yet. But is that possible we can apply AC to AFM? } } Also how can I obtain more information about biological samples preparation for AFM measurement? } } I do appreciate your kindness. } } Chris } } } Life is like a box of hand grenades, you don't know what will blow you to kingdom come.
===== Life is like a box of hand grenades, you don't know what will blow you to kingdom come.
Mr.Yung-fou Chen E-mail: yung_fou-at-yahoo.com
__________________________________ Do you Yahoo!? Protect your identity with Yahoo! Mail AddressGuard http://antispam.yahoo.com/whatsnewfree
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 21:57:25 2003
I have been looking for a printer for publication-quality color figures and high-resolution grayscale images. I have been considering the Magicolor 3100, but was put off by some of the reviews I read -- for example, http://www.pcmag.com/article2/0,4149,908304,00.asp
Michael A. O'Keefe Materials Sciences Division LBNL
Richard Edelmann wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Looking at getting a color laser printer for general quick and dirty printing. } Particularly looking for feed back from anyone one either the Minolta QMS } Magicolor 3100 or the Lexmark C752n. We have excellent experience with } our Lexmark B&W printers. And I like the example prints from the Miniolta I } pick up at M&M (Thank you Vitally). Now I'm look for more opinions Good the } bad and the ugly. } } Yes, we have a number of ink jet printers, excellent quality but slow, } looking for some thing faster, basically to serve for walk away prints as we do } more and more digital. } } Thank you! } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 350 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } http://www.emf.muohio.edu } } "RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 11 23:29:53 2003
Dear all: An investigator of my facility is interested in looking at the cross-section of a peptide nanotubes on a TEM. These tubes have a diameter of 50-60 nm, and the wall of tube is about 4 nm. We would like to start with conventional aldehyde fixation and epoxy resin embedding, however, the sample is very sensitive to pH, and anything above pH 4 would cause disassembling of the peptide tubes. Can anyone recommend an alternative EM fixative that works well at low pH, and explain how this fixative works? Thank you very much.
Hong
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 02:14:03 2003
There was a thread on this in January 2002 The following are representative of the non-polishing methods suggested. Chris ================================================= We routinely use a potassium iodide mixture off the shelf made by Acton Industries, Pennsylvania, USA, on GaAs semiconductors. It's called "GE-6". I imagine for "gold etch." The pure Au metal is ~ 3.5 uM and is removed cleanly by this material. The only caveat is that it attacks exposed GaAs very quickly.
If anyone needs details, please contact me directly.
Peter Tomic Group Leader Failure Analysis & Analytical Services Anadigics, Inc. Warren, New Jersey ============================== Chris,
The solution is
65g iodine 113g potassium iodide dissolved in 100ml cold water.
Dissolve the KI first for a saturated solution, followed by the iodine!
Happy stripping!!
Best Wishes Barry ************************ Barry Lamb Manager, Materials Evaluation Centre C-MAC Engineering Ltd web: {http://www.cmacengineering.com/} e-mail: blamb-at-har.cmac.com tel: +44 (0)1279 403671 (ESN 742-3671) ============================== Chris,
I've used 1:1 Nitric(70%):HCl mixture for gold coat and gold wire removal from semiconductor samples. Although I don't know the etch rate, it will etch off a moderate gold coat in about 20-30 seconds.
Frank Martini ZiLOG Nampa, Idaho ============================= Chris:
A similar question arose on another listserver, although it was removing a 2 mciron gold metallizton from GaAs. The responses from there were as follows:
Response 1 "Greetings!
It is possible that a cut or buffered aqua regia might do the trick. I have had some success with the recipe below on Au metallization at M2 and above on GaAs dice. The interlayer dielectric was silicon dioxide, so there was not a lot of seepage into the die substrate layer - this, more than the recipe, might have kept the damage down. Assuming SiO2 as the ILD on all layers of your part, this might still work:
50 ml deionized water
30 ml HCl
20 ml HNO3
Swirl or agitate the part in the solution for five minutes. Inspect and repeat once if needed.
One minute rinse in deionized water, followed by 30 seconds rinse in acetone. Dry under heat lamp to minimize surface staining.
As always, try this on a practice part first in case this does not buffer the reaction with GaAs enough.
Good Luck !
Regards,
Carl Nail National Semiconductor Carl.Nail-at-nsc.com"
Response 2 "A solution of potassium iodide and iodine in water is a decent gold etch. I have no idea how it would affect GaAs, but I suspect it would be less destructive than Aqua Regia.
Alan Street alan-at-irsi.com"
I hope this helps!
David ================================ Chris; I have had very good luck using a solution of potassium ferrocyanide. Hope this helps.
Jon McGovern Manager, Microscopy and Imaging Facility University of Calgary =================================== We use sodium cyanide solution to remove gold.
Robert Champaign Raytheon Failure Analysis Lab ===================================
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 02:52:30 2003
A short paper outlining some chemical methods for dissolving gold can be accessed at http://fbe.erciyes.edu.tr/Turkce/eufbedergisi/ DER99/119-127.pdf =================================================== Gold will dissolve into a solution if the solution has an oxidizer (like nitric acid, H2O2, ozone, sodium bromate, etc.) and a chlorine, bromine, iodine, cyanide, thiourea, thiocyanate, or thiosulfate source. This gives a large number of combinations of substances that will dissolve gold. The most famous is aqua regia (muratic acid and nitric acid). Joshua Gulick http://www.ormus.ws/~pyramid/dissolve_gold.htm =================================================== MacArthur was not in fact the first person to realize that cyanide could dissolve gold. That discovery had been made over one hundred years before, in 1783, by the Swedish chemist Scheele who was the first person to prepare the gas hydrogen cyanide. (The companion story of the history of cyanides will be the subject of next week's article.). By the mid)nineteenth century, solutions of gold in cyanide solutions were being used to electroplate gold on to other metals. This process also is used almost unchanged today. Thus we see that MacArthur was not the first person to observe that gold dissolves in cyanide solutions. His contribution to technology, which was recognized in his patent, was to realize that the chemical reaction could be applied economically to the recovery of gold from low)grade ores. As is often the case with technology, cyanide was being used to dissolve gold long before the scientific details of the process were understood. As already mentioned, electroplating of gold was a commercial process by 1850, and gold extraction from ores using MacArthur's patent began in 1889, yet the chemistry of the process was not completely understood until the 1950's. Michael Faraday (1857) was the first person to recognize that oxygen is needed in order for gold to dissolve in a cyanide solution. Neither oxygen alone nor cyanide alone does the job; both substances are needed. An especially puzzling phenomenon is that extremely low concentrations of cyanide (e.g. 0.01 per cent) are sufficient to dissolve gold, and that the rate of dissolution does not seem to be affected by the concentration of the cyanide solution. Eventually it was shown that the limiting factor under typical conditions is how fast the oxygen of the air dissolves in the cyanide solution. A modern view point of the process is that the gold is corroded in a manner not unlike that of iron rusting. extract from THE SCIENCE CORNER by Nigel Bunce and Jim Hunt College of Physical Science University of Guelph http://helios.physics.uoguelph.ca/summer/scor/articles/scor174.htm =================================================== In 1856, M. Faraday prepared the first colloidal gold dispersion in water. To repeat the experiment the reader might reduce gold chloride in water with sodium citrate (which he did with phosphorous). After a short time, a blue coloration will appear and then a ruby-red gold dispersion. =================================================== finally, if all else fails..... Collect a good amount of boys' urine, let it putrefy for some time, and distill the first and subtle spirit over like brandy. Set the filtrate in digestion for 8 days and distill again as before. Keep the spirit but boil the left-overs of both distillations quite dry in a kettle. Calcine it in a potter's furnace, extract from it its fixed salt with rain water, knead it under potter's clay, and distill it like common spirit of salt. You will obtain a yellow, sharp spirit, rather heavy in weight. Rectify it to remove all phlegma, then pour on it by drops the first- prepared volatile spirit. It will effervesce strongly, so that you will be surprised to find so many opposites together in one subject. A white substance will precipitate. Let it settle, pour the phlegma off from it, dry the rest, put it in a curcurbit and sublimate it with a strong fire. A beautiful bright sublimate will rise into the alembic. Remove it and keep it, as it is good for many things. Take one part of it, add to it 3 parts of spirit of salt, digest this together and distill it. Now you will have a wonderful menstruum for dissolving not only gold but all the other metals and minerals. http://www.levity.com/alchemy/agric_06.html ========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 06:21:30 2003
} I have been using the Minolta QMS Magicolor 3100 for about } 1 year, and am extremely pleased with it. Good image quality } (both resolution and color fidelity), fast results, low cost. } The dye-sub printer is now rarely used and the ink-jet } has been completely retired.
Because I'll soon be looking into a high throughput printer as well, this is good info to have. My only question, in the context of grayscale SEM micrographs, is how well the color lasers print good detail in neutral gray(?) That is, I imagine most of them, if not all, achieve better detail when allowed to dither all colors for achieving gray. Is the result neutral? ... and, does the neutrality hold up over time?
Why do one-step immuno-EM? We routinely do GFP immuno-localization using anti-GFP antibody followed by protein A-gold. We get great results. Directly coupling an antibody to gold is never a good idea as it usually decreases the affinity and the complex is not stable very long. There might be some sources of GFP antibody-gold conjugates out there, but I wouldn't really trust them. We have obtained good results using anti-GFP antibody from Molecular Probes. We purchase our protein A-gold from a lab at the University of Utrecht in the Netherlands, but there are many other sources for this reagent.
Marc
On Tuesday, November 11, 2003, at 04:35 PM, Richard Edelmann wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } } ------- Forwarded message follows ------- } Subject: [Microscopy] gold anti-GFP } Send reply to: kissjz-at-muohio.edu } } From: "JOHN KISS" {kissjz-at-muohio.edu} } Date sent: Fri, 31 Oct 2003 14:23:42 -0500 } } Question for listserv; } } There are lots of papers where people use indirect immunofluroescence } to } localize a GFP fusion protein. Does anyone know of a company that } sells an } anti-GFP antibody directly coupled to collodial gold so one can do a } 1-step } labelling for EM level localization? } } } } John Z. Kiss, Ph.D. } Professor } Dept. Botany, Miami Univ. } Oxford OH 45056, USA } tel./Fax: 513-529-5428/-4243 } http://www.cas.muohio.edu/botany/bot/jzk.html } } } ------- End of forwarded message ------- } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 08:35:17 2003
Can you make the fixative (glutaraldehyde) in the same buffer the nanotubes are in now? I think a bigger problem might be getting the nanotubes orientated so you can cut them in cross section.
Geoff
Hong Yi wrote:
} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Dear all: } An investigator of my facility is interested in looking at the } cross-section of a peptide nanotubes on a TEM. These tubes have a } diameter of 50-60 nm, and the wall of tube is about 4 nm. We would } like to start with conventional aldehyde fixation and epoxy resin } embedding, however, the sample is very sensitive to pH, and anything } above pH 4 would cause disassembling of the peptide tubes. Can anyone } recommend an alternative EM fixative that works well at low pH, and } explain how this fixative works? Thank you very much. } } Hong } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 08:44:44 2003
We are running a mostly automated Olympus IX81 with IPLab Spectrum 3.6.1. Fortunately, Scanalytics does provide control plugins for the IX81 (for instance, "Set filter to Dapi"), however, we cannot query the status of the microscope from IPLab. For instance, the software cannot ask, "Which filter is the microscope set at?" and make the answer available to a script. Has anybody solved this problem yet? Thanks.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 08:53:11 2003
In a message dated 11/12/03 8:40:16 AM, michael-at-shaffer.net writes:
} Because I'll soon be looking into a high throughput printer as well, this is } good info to have. My only question, in the context of grayscale SEM } micrographs, is how well the color lasers print good detail in neutral } gray(?) That is, I imagine most of them, if not all, achieve better detail } when allowed to dither all colors for achieving gray. Is the result } neutral? ... and, does the neutrality hold up over time?
That's a good question and raises several points worth considering: 1. As far as the neutrality of the laser print, when given a monochrome image the Minolta (and in my experience, all other) laser printers use only the black toner so there is no consumption of the color toners (in the Minolta, the CMYK toners are all separate and you can replace K individually as needed) and of course there is no color whatever in the print, and they are smart enough to print with a monochrome halftone pattern rather than leaving space for the color rosettes that are used for full color printing. I've just printed an 8x10 test image consisting of a grey scale ramp, and it shows no banding or "steps" in the grey scale. I would judge that the lightest 3-4% is pure white (no visible toner on the paper) and the darkest 5% or so is "plugged" (i.e., no white showing at all), which is typical of halftone printing. The default halftone pattern used in the Minolta is apparently about 140 cells per inch (by inspection of the print), but the printer is Postscript controlled so this can be set as desired (frankly, I've just left it with default settings and have no complaints). 2. The laser prints have a very long life (i.e., I haven't seen any evidence yet of any deterioration, regardless of some pretty sloppy storage conditions, exposure to light, etc. The same cannot be said for most ink jet prints. The dye based inks fade badly in strong light or humidity, and the "archival" pigment based inks don't have as good tonal resolution and exhibit strong m etamerism (different colors under different lighting conditions). 3. Neither a color laser printer nor a color ink jet printer is the optimum solution for printing only monochrome images. By using an ink jet mechanism with custom inks having multiple shades of grey, Jon Cone (check out piezography.com) has come up with a monochrome printing system that has more tonal range and control than even traditional darkroom printing. But it is slow, and costly per copy. And there are other monochrome-only solutions that use essentially photographic methods (using laser light to expose the "print" which is then developed). These are also slow and costly per copy. For making display images that would rival Ansel Adams prints when hung on the wall, I would not hesitate to go with Piezography. But for routine output I find the halftone printing from the Minolta to rival the quality of a decent offset press, the kind used to print quality books. It is fast, cheap per copy (after an initial investment of nearly $2k), and runs happily on a network with a mixture of Mac, Unix and Windows computers. 4. I have a set of sample (color, but probably now I will include a few monochrome examples as well) images that I carry around printed on dye sub, ink jet and laser printers. Viewed side by side from a normal viewing difference, there are very few differences. I would judge that the dye sub shows slightly more saturation and somewhat richer colors, but not by much. Unless you hold them up very close to see the individual dots in the ink jet, the halftone pattern in the laser output, and the inherent softness in the dye sub, you would probably not be able to guess which was which. And for some of us older folks, it takes a hand lens to really see those details.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 09:00:20 2003
Dear Listers I've found two ex-Leo/Phillips people founded a company to provide SEM & TEM, LM service and support centre at Queens- New York City, they've started by the freelancing services earlier. They've fixed our facility SEM S-435 and dismantled and moved the TEM-900 to another facility with Gemini filter in a very efficient and cost-effective way. We've always faced problem with the companies in their after sales support and Maintenance contract. Hope their service would be very beneficial for many of our fellows who can get many EM related problems solved by them in a inexpensive way. You can contact them in the e-mail id dc-at-infosoftusa.net Regards } From Dr. John Kurtz's Lab Dave NYU ----- Original Message ----- } From: "Greg Barclay" {gbarclay-at-trinidad.net} To: {} Sent: Tuesday, November 11, 2003 2:24 PM
Michael brought up a good point about printing gray images. Sometimes I need to print both a color image and a gray SEM image on the same page. Gray is the hardest color to reproduce. Nearly every color printer I've used uses color ink to reproduce a gray image on a "mixed image" page, resulting in a particualr hue to the gray image. One exception is the Lexmark Optra SC 1275. It can distinguish between a color image and a gray image on the same page, using color ink for the color image and black ink for the gray image and text, even though all are on the same page.
I can't recommend this particular printer because it has a bad reputation of breaking down a lot, plus color saturation was so-so. But I cherished this abovementioned peculiar feature of the printer, and recommend anybody to look for this feature on their future printer purchase.
Stu Smalinskas Metallurgist SKF USA Plymouth, Michigan (734) 414-6862
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
michael-at-schaffer.net wrote:
Because I'll soon be looking into a high throughput printer as well, this is good info to have. My only question, in the context of grayscale SEM micrographs, is how well the color lasers print good detail in neutral gray(?) That is, I imagine most of them, if not all, achieve better detail when allowed to dither all colors for achieving gray. Is the result neutral? .. and, does the neutrality hold up over time?
} I have been looking for a printer for publication-quality color } figures and high-resolution grayscale images. } I have been considering the Magicolor 3100, but } was put off by some of the reviews I read -- for example, } http://www.pcmag.com/article2/0,4149,908304,00.asp
Interesting reading, and this review certainy contrasts with a similar review of a less expensive Magicolor printer, the Minolta-QMS 2350 EN ... http://www.pcmag.com/article2/0,4149,1217172,00.asp ... which gets kudos instead of kriticism for its "photographic" quality and color saturation. Logical questions not answered in the review would be with respect to the expense of the toner cartridges.
} Richard Edelmann wrote: } } } } Looking at getting a color laser printer for general quick and dirty } printing. } } Particularly looking for feed back from anyone one either the } Minolta QMS } } Magicolor 3100 or the Lexmark C752n. We have excellent experience with } } our Lexmark B&W printers. And I like the example prints from } the Miniolta } I } } pick up at M&M (Thank you Vitally). Now I'm look for more opinions Good } the } } bad and the ugly. } } } } Yes, we have a number of ink jet printers, excellent quality but } slow, } } looking for some thing faster, basically to serve for walk away } prints as } we do } } more and more digital. } } } } Thank you! } } } } Richard E. Edelmann, Ph.D. } } Electron Microscopy Facility Supervisor } } 350 Pearson Hall } } Miami University, Oxford, OH 45056 } } Ph: 513.529.5712 Fax: 513.529.4243 } } E-mail: edelmare-at-muohio.edu } } http://www.emf.muohio.edu } } } } "RAM disk is NOT an installation procedure." } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 10:21:27 2003
Hi Jonathan We have a Hitachi H-7600 with a Gatan cryo stage and an AMT camera.
We really like the H-7600 and have had very good service from Hitachi. We have adapted the transfer technique for putting grids into the Gatan cryo holder and it is now pretty slick. So we are happy with the cryo holder. My suggestion is from experience that you should try and get a second transfer rod straight away. What I like about the Hitachi-Gatan system is that the liquid nitrogen does not pour out when you put the transfer rod in. This happens with the FEI system but I have no experience with the Joel system.
We have a 1985 Zeiss 10C without digital, just film. Since we got the digital camera, no-one has used the Zeiss for taking pictures. It seems to be used mostly for checking grids and such which makes it extremely useful in that the Hitachi is very often booked when you just want to check your grid! For those odd occasions when someone wants higher resolution, the Hitachi has a film camera too.
We have a 1Kx1K digital camera and it is not enough for an 8x10. A 2Kx2K would now be the basic camera I would want but if I could afford it, I would want the 4Kx4K. One of the nice things about having a digital camera is that you can turn the beam down so much that you cannot see anything on the fluorescent screen, but the camera can pick up the signal and give a good image. This is important if you have a section which blows up just as the beam hits it.
Spare parts to specifically ask for when you are buying are spare Wehnelt assembly, spare objective aperture strip, spare condenser aperture strip, and a box of 10 filaments. These may not seem much but when the filament blows, having a spare system ready to go means the TEM is down only a short time. Same with particulary the objective aperture. It can take a week or more to get a spare but if you have one ready to go, you can replace it in a very short time.
I can't think of anything else at the moment Elaine
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 10:39:46 2003
} In a message dated 11/12/03 8:40:16 AM, michael-shaffer.net writes: } } } ..., in the context of grayscale SEM } } micrographs, is how well the color lasers print good detail } } in neutralgray(?) ... } } That's a good question and raises several points worth considering: } 1. As far as the neutrality of the laser print, when } given a monochrome image the Minolta (and in my experience, } all other) laser printers use only the black toner ...
That's certainly different from the way inkjets work. Leastwise, if you send the printer neutral RGB data, they'll use all colors. It's my experience with Epsons, this use of all inks is impossible to control, whereas other printers can be told to print "don't use color". For clarification, can you confirm the QMS printer drivers will discriminate neutrality even if RGB data is sent?
} ... I've just printed an 8x10 test image consisting } of a grey scale ramp, and it shows no banding or } "steps" in the grey scale. I would judge that the } lightest 3-4% is pure white (no visible toner on the paper) } and the darkest 5% or so is "plugged" (i.e., no white } showing at all)
Although Photoshop "printer transfer functions" are implied to work with CMYk EPS files only, I find they will work with RGB data ... and could possibly remedy the loss of detail for either end of the histogram (depending on the printer).
This question is regarding a Digital Instruments Nanoscope III AFM/STM. I wanted to know if it was ok to leave the piezo scanner plugged in "before" switching on/off the Nanoscope controller. Initially I was of the opinion that the scanner had to be unplugged before switching on/off the controller.
Does it really matter
thank you dwarakanath
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 11:10:56 2003
I'm looking for information on any procedure using Davidson's Fixative for eyes for both Light and Electron Microscopy
Any information would be greatly appreciated
Thanks,
Pedro
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 11:33:28 2003
Hong, It sounds like this is a sample that would benefit from high pressure or plunge freezing followed by freeze substitution. That way you would minimize collapse and pH would not be a critical factor. Even adding fixative to the freeze substitution solution wouldn't matter since you would not be buffering it.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
On 11/12/03 12:41 AM, "Hong Yi" {hyi-at-emory.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Dear all: } An investigator of my facility is interested in looking at the } cross-section of a peptide nanotubes on a TEM. These tubes have a } diameter of 50-60 nm, and the wall of tube is about 4 nm. We would } like to start with conventional aldehyde fixation and epoxy resin } embedding, however, the sample is very sensitive to pH, and anything } above pH 4 would cause disassembling of the peptide tubes. Can anyone } recommend an alternative EM fixative that works well at low pH, and } explain how this fixative works? Thank you very much. } } Hong } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 12:30:47 2003
In a message dated 11/12/03 1:19:42 PM, michael-at-shaffer.net writes:
} That's certainly different from the way inkjets work. Leastwise, if you } send the printer neutral RGB data, they'll use all colors. It's my } experience with Epsons, this use of all inks is impossible to control, } whereas other printers can be told to print "don't use color". For } clarification, can you confirm the QMS printer drivers will discriminate } neutrality even if RGB data is sent?
Yes. Even if the document has black text with both color and monochrome image insets, the printer correctly uses no color toner except in the color image region. This is indeed a much better solution than ink jets, which must use colored inks as well as black in order to get enough paper coverage to generate a "black" appearance. Laser printers are capable of complete area coverage with any of the toner colors, although for printing color images they normally lay down the colors using a halftone grid pattern.
} Although Photoshop "printer transfer functions" are implied to work with } CMYk EPS files only, I find they will work with RGB data ... and could } possibly remedy the loss of detail for either end of the histogram } (depending on the printer).
Setting up a printer curve that skips the very ends is easy, and solves the problem as you note. I printed my test sheet intentionally without that correction. By the way, the printer drivers for the Minolta (and all other laser printers I've worked with) expect an RGB image not CMYK. They perform the conversion themselves. If you do print a CMYK image, it first gets converted back to RGB! The Photoshop CMYK conversion is appropriate for making color separations for offset web press, but is not useful for laser (or ink jet) printers.
Also, to respond to a point raised in a private email, I have no financial interest in Minolta, or in Piezographics. Just a very satisfied user.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 18:26:49 2003
I've been using the HP Color LaserJet 4550N for several years. Excellent results and long life for the toner. It prints with CMYK so greyscale is done with just the black toner. Grey, black and color on one page all render perfectly. Image quality is excellent. The printer is 600dpi and produces outstanding color prints. The newer version is the 5500. With the network option, any computer on the LAN can get to the printer. Using DAVE, a Mac can also print to the color laser using TCP/IP.
toner cartridges cost about $175 each. But they last a long time and only need to be replaced individually when empty.
gary g.
At 08:05 PM 11/11/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 20:36:54 2003
I have used these old protocols with success on woody materials. I am not sure how long you processed your tissues, but plant material always requires greatly extended times. Also we softened the embedded material in Gifford's solution. The difficulty is always trying to get the harder tissue soft enough so it doesn't tear the softer tissue during sectioning.
The details we used are as follows: Fixation for 12 to 48 hours (you might also try Navashin's fixative which contains chromic acid) Infiltrate through TBA series (Johansen series) for 2 hours each step 3 changes of pure TBA over 24 hours 3 changes of Paraplast over 24 hours Embed
Soften the blocks by soaking overnight to 7 days in water at up to 38°C If this doesn't work try 2 days to 2 weeks in Gifford's (room temp in fume hood): 20 ml glacial acetic acid 80 ml 60% ethanol 5 ml glycerine You will have to try the times with your own tissue. If you leave it too long the soft tissue will become macerated. If this fails then you may have to go with a resin embedding.
Good luck,
Kim On 11-Nov-03, at 12:54 AM, Greg Barclay wrote: } } I need to modify or replace our standard fixation protocol to handle } pedicels of legume pods for preparation of wax sections } for light microscopy. These are about 1 mm in diameter and combine } woody (phloem fibers and xylem) and soft tissues } (cortex, phloem, cambium, pith) and the tissues readily distort or } tear during sectioning. Currently we use an age old protocol } consisting of standard fixation in FAA or parafomaldehyde, then } ethanol series dehydration through TBA and into Paraplast. } Clearly this is not good enough. Any suggestions for an improved } protocol that would produce undamaged sections would } be greatly appreciated. } } Greg } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 12 22:13:45 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (steven.young-at-sri.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 12, 2003 at 18:39:57 ---------------------------------------------------------------------------
Email: steven.young-at-sri.com Name: Steve Young
Organization: SRI International
Title-Subject: [Microscopy] MListserver:Vintage Cambridge SEM available
Question: We have a Cambridge Mark IIA SEM that was purchased new in 1968 that has been up and running until retired recently. This instrument is 90% vacuum tube electronics and was completely overhauled 10 years ago. It is clean and reasonably reliable. We need to find a good home for it, preferably a museum or a private collector. I can't stand to see it end up in a dumpster. It includes a spare stage and many options including all manuals and documentation. The only cost would be packing and shipping from Menlo Park, CA. 94025
Any other ideas? I've tried several local museums.
The point is that we need to connect Link AN 10000 computer with IBM PC. We tried to use XMODEM program. But it was useless in spite of the program works well with other applications. May be the reason is not up-to-date programs (1986-87). So we need your advice. Thanks. Best regards, Sadova
http://www.ipm.sci-nnov.ru/
mailto:sadova-at-ipm.sci-nnov.ru
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 13 08:23:23 2003
Hi, has anybody longer experience with diamond knives from Drukker (The Netherlands / Europe)? Can You recommend them? Have you ever had re-sharpened a knive? Thanks for a short informal and offline reply to
peter.heimann-at-uni-bielefeld.de
Peter Heimann
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 13 10:37:05 2003
I currently using a Gatan GIF for EELS on a Tecani thermal field emitter. I was told that extraction voltage can affect energy resolution to the spectrometer. If so how may I go about optimizing extraction voltage for optimizing Gif for high resolution PEELS?
Thanks
Thurston Herricks
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 13 10:39:49 2003
This may be a broad inquiry, but at this point my knowledge on the topic is limited and thus my need for help from the group:
Our lab is considering expanding some capabilities, and we are investigating the possibility of establishing a hot-lab for various analyses on low level radioactive samples. Theoretically, this could potentially include a SEM, which leads me to contact everyone.
Since all of my experience is with strictly non-radioactive materials, what are some of the key considerations for such an undertaking? Beyond shielding and safe handling (which other people here will help assure), with specific regards to a SEM, imaging and analytical capabilities, what (if any) are some things to become aware of, investigate, and learn about as we get into this issue?
TIA!
If desired, I can be contacted off list.
Chris Holp FirstEnergy Corp. HolpC-at-firstenergycorp.com 440-604-9704
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From MicroscopyL-request-at-ns.microscopy.com Thu Nov 13 12:14:10 2003
Microscopy Society of America 2004 Undergraduate Research Scholarship Program
With this year's call for applications, the MSA Undergraduate Research Scholarship Program begins its 16th year providing funding for undergraduate research. To date over 81 projects covering a wide range of topics in the physical and biological sciences have received support through this program. Over the years, nearly all of the scholarship recipients have maintained a strong interest in imaging sciences and have gone on to graduate school, professional school, teaching, or industry positions.
The program, which is funded by MSA and by matching funds from MSA Sustaining Members, is able to support over 50% of applicants. The maximal award for the Undergraduate Scholarships is $3,000 and helps to provide student stipends, supply costs, and limited travel expenses associated with the research. Additional support in the form of instrument use time, equipment purchases, etc., is generally provided by the student's supervisor and/or through the sponsoring institution. Abstracts reporting the research results, are prepared by scholarship awardees, and published in "Microscopy and Microanalysis".
The program actively seeks sources of matching funds in order to maintain the favorable levels of support both in terms of the number of projects supported and the level of support for each. We are extremely grateful for the matching support provided by MSA sustaining members and individuals. Their support over the years has enabled the program to increase both the number of awards and the maximum amount of each award. For 2003 we are particularly grateful for support provided by Gatan and JEOL.
The MSA Undergraduate Research Scholarship Program is currently soliciting applications from students interested in conducting a research project which involves the use of any microscopy technique. Students should be sponsored by a member of MSA. The maximal award is $3000. The application deadline is Dec 31, 2003. Application forms and instructions are available on the MSA web site, www.msa.microscopy.com. Applications can also be obtained from the MSA Business Office, businessoffice-at-msa.microscopy.com or call toll free at (800) 538-3672. If you have any questions or require additional information regarding the program please contact:
Dr. Ralph Albrecht, University of Wisconsin 1675 Observatory Drive, Madison, WI 53706 (608) 263-3952 or 263-4162; (608) 262-5157 FAX; albrecht-at-ansci.wisc.edu
2003 awardees Renee Lopez-Smith. Microscopic study of fertilization in the fern Ceratopteris: the role of actin in gamete fusion. Southern Illinois University, Advisor: Karen Renzaglia Brigit O'Donnell. Effect of vein pattern alteration on movement of turnip vein clearing virus. New Mexico State University, Advisor: Dr. Souuumitra Ghoshroy Julie Takacs. Inhibition of bacterial invasion mechanisms using anti gC1q-R monoclonal antibody carbon nanotube probes. Franklin and Marshall College, Advisor: Barbara Panessa-Warren Aurelie Thuaire. Characterization of nanocomposites using electron microscopy. McGill University, Advisor: Raynald Gauvin
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 13 13:18:32 2003
Dear Chris, I suggest you contact Woody White of GE (I think). He has a lot of experience running SEMs analysing radioactive materials. His web site is: home.att.net/~woody.white/ and his e-mail address is on the site. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {HolpC-at-firstenergycorp.com} To: {Microscopy-at-msa.microscopy.com} Sent: Thursday, November 13, 2003 8:44 AM
Thurston,
Using a GIF, we measured the zero-loss energy spread from our CM300FEG as a function of extraction voltage from 4kV down to 1.5kV. The measured energy spread dropped from 0.93eV FWHH at 4kV to 0.65eV at 1.5kV extraction voltage. Backing out the contribution from the GIF gave us a true beam spread of 0.85eV FWHH at 4kV and 0.53eV FWHH at 1.5kV -- see figure 2a of Estimation of the Electron Beam Energy Spread for TEM Information Limit, Michael A. OKeefe, Peter C. Tiemeijer and Maxim V. Sidorov, Microscopy & Microanalysis 8 (2002) suppl 2: 480-481.
We wanted to reduce the energy spread to improve our HRTEM information limit, but found that imaging below 3kV extraction voltage was difficult due to the low beam current. For PEELS I guess you also want a low spread, but I don't know how low a beam current you can tolerate.
Mike O'Keefe
thurston e herricks wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello, } } I currently using a Gatan GIF for EELS on a Tecani thermal field emitter. } I was told that extraction voltage can affect energy resolution to the } spectrometer. If so how may I go about optimizing extraction voltage for } optimizing Gif for high resolution PEELS? } } Thanks } } Thurston Herricks
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 13 17:43:53 2003
We routinely analyse filtrate samples in SEM, specifically for morphology of particles and composition. Unfortunately due to the continuous changes in manufacturers and suppliers merging etc, we no longer have an appropriate source of filter for our needs.
This is the type of filter we used to use. { {...OLE_Obj...} } a Support 0.8um gelman filter
since the merger gelman/pall now produce the following
{ {...OLE_Obj...} } which is unsuitable for our needs. the small round nodules can be easily confused with particles - sends our engineers quite a panic!
we are currently using this one below: { {...OLE_Obj...} } a GN-4 0.8um filter
but this is not ideal as there still are small nodules on the filter surface.
The filter needs to be caustic resistant to high pH, 0.8-0.2um pore size, low background for EDS/XRD analysis.
Can anyone recommend a brand or supplier who could help?
thanks for your assistance.
Samantha Taylor TDG Experimental Officer Alcoa of Australia XRD, Microscopy and Thermal Analysis * Phone:(08) 9410 3588 * Fax: (08) 9410 3167 * Email: Samantha.Taylor-at-Alcoa.com.au
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 01:51:13 2003
The Institute for Transuranium Elements in Karlsruhe, Germany, has experience through many years with electron microscopy (both TEM and SEM) of radioactive materials. Their SEM webpage has the address
Joergen Bilde-Soerensen Materials Research Dept. Risoe National Laboratory Roskilde, Denmark.
-----Original Message----- } From: HolpC-at-firstenergycorp.com [mailto:HolpC-at-firstenergycorp.com] Sent: 13. november 2003 17:45 To: Microscopy-at-msa.microscopy.com
Probably a very basic question, but can anyone tell me whether araldite resin actually infiltrates cells in tissue blocks? I've small (1 mm2) para fixed tissue blocks, which are processed into araldite resin. Does the resin cross cell (and intracellular) membranes?
Thanks.
G McGovern
Veterinary Laboratories Agency (VLA)
This email and any attachments is intended for the named recipient only. Its unauthorised use, disclosure, storage or copying is not permitted. If you have received it in error, please destroy all copies and inform the sender. Whilst this email and associated attachments will have been checked for known viruses whilst within VLA systems we can accept no responsibility once it has left our systems. Communications on VLA's computer systems may be monitored and/or recorded to secure the effective operation of the system and for other lawful purposes.
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 07:52:50 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mikko.uittamo-at-m-real.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 14, 2003 at 02:50:05 ---------------------------------------------------------------------------
Email: mikko.uittamo-at-m-real.com Name: Mikko Uittamo
Title-Subject: [Microscopy] Link Isis Autobeam problem
Question: Hi
I dont know if this is the right place for this kind of a question, but bare with me. If you know any better place to post this please inform me.
We have a Oxford link isis software in our microscope and the problem is that when usin autobeam software and opening jobs from network drive, software stops responding for couple of minutes and then it works ok. This happens almost on every job. We just updated the system to w2k and that was when problems started.
Could the problem be with the software or with the new operating system.
Any embedding material must infiltrate all of the cells otherwise you will have a hole where the cell was. Araldite is very viscous and may not best choice for the tissue+fixative combo you are using. On the other hand, it is easier to cut and stain than some other resins. Are you using osmium post-fixation? If not, tissues will be harder to infiltrate (osmium makes membranes more permeable to subsequent processing). If you are not using osmium are you using one or more Araldite:clearing agent steps?
Geoff
McGovern, Gillian wrote:
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-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 10:17:43 2003
Yes, the resin will penetrate right into the cells. In fact, if it doesn't adequately infiltrate right into the cells, then you will be unable to section the tissue ultra-thin, and it will turn into garbage right off your knife.
For this reason, it is important to allow some time for the plastic to penetrate the tissue, usually as a mixture of resin/propylene oxide, until the tissue is adequately infiltrated.
-----Original Message----- } From: McGovern, Gillian [mailto:g.mcgovern-at-vla.defra.gsi.gov.uk] Sent: Friday, November 14, 2003 2:15 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Probably a very basic question, but can anyone tell me whether araldite resin actually infiltrates cells in tissue blocks? I've small (1 mm2) para fixed tissue blocks, which are processed into araldite resin. Does the resin cross cell (and intracellular) membranes?
Thanks.
G McGovern
Veterinary Laboratories Agency (VLA)
This email and any attachments is intended for the named recipient only. Its unauthorised use, disclosure, storage or copying is not permitted. If you have received it in error, please destroy all copies and inform the sender. Whilst this email and associated attachments will have been checked for known viruses whilst within VLA systems we can accept no responsibility once it has left our systems. Communications on VLA's computer systems may be monitored and/or recorded to secure the effective operation of the system and for other lawful purposes.
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 11:02:25 2003
Have you tried the Mykrolis Company (http://www.mykrolis.com/)? They make a polyethylene membrane filter in various pore sizes. I used to use these to do airborne particles in the wafer fab. The filter media has a smooth surface with a support grid on the backside and particles are easily imaged on it.
"Taylor, Samantha" wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear All } } We routinely analyse filtrate samples in SEM, specifically for morphology of } particles and composition. } Unfortunately due to the continuous changes in manufacturers and suppliers } merging etc, we no longer have an appropriate source of filter for our } needs. } } This is the type of filter we used to use. } { {...OLE_Obj...} } a Support 0.8um gelman filter } } since the merger gelman/pall now produce the following } } { {...OLE_Obj...} } } which is unsuitable for our needs. the small round nodules can be easily } confused with particles - sends our engineers quite a panic! } } we are currently using this one below: } { {...OLE_Obj...} } a GN-4 0.8um filter } } but this is not ideal as there still are small nodules on the filter } surface. } } The filter needs to be caustic resistant to high pH, 0.8-0.2um pore size, } low background for EDS/XRD analysis. } } Can anyone recommend a brand or supplier who could help? } } thanks for your assistance. } } Samantha Taylor } TDG Experimental Officer } Alcoa of Australia } XRD, Microscopy and Thermal Analysis } * Phone:(08) 9410 3588 } * Fax: (08) 9410 3167 } * Email: Samantha.Taylor-at-Alcoa.com.au
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 14:32:40 2003
We are using JEM-2010F microscope. The smallest field limiting aperture which JEOL commercially provides is 5 micron. But it is still a little large when we want to get electron diffraction patterns for some special materials. However, we do not want to use NBD mode because of the serious radiation damage. So we are trying to get the information about companies who can make smaller field limiting aperture (~ 1 micron). If you know who can make it, please give me their contact information. Thanks.
Best regards, Qi Zhang ***************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu qizhang-at-email.unc.edu
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From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 14:39:58 2003
I have a project from a faculty member who is studying a protein, the monomers of which form a ring. I'm using 1% uranyl acetate as the negative stain. This was the method used in the background information they brought to me. Does there exist an even finer grained negative stain? I'm looking for a way to improve the visual quality of the material. Also the operating conditions of my TEM (a Philips 410LS with tungsten filament) are 80Kv and 100Kv (the limit of my TEM), aperture sizes are condensor 200micron, objective 20micron, spotsize 2, zero degree tilt. I'm trying to photograph in the range of 240,000X to 500,000X (the upper limit of my TEM). I would appreciate any suggestions to improve the image of the samples either by way of specimen preparation or operating condition of the instrument. Would metal shadowing be worth trying? Thanks in advance for your help.
Tom Bargar Core Electron Microscopy Laboratory UNMC 986395 Nebraska Medical Center Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 15:15:39 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wwiggins-at-carolinas.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 14, 2003 at 10:29:12 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] MListserver: Ilford Service
Question: Mary, I was told that Serco Equipment Services, 800-922-0192, handles maintenance and service contracts in the States for Ilford (an Italian company). We've contracted with them for our 2150. Alternatively, you can try Hunt's Photo & Video, 800-924-8682, in Melrose, MA to see if they can help. Winston
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor Cannon Electron Microscopy Lab Carolinas Medical Center P.O. Box 32861; Charlotte, NC 28232-2861 Ship to: 1000 Blythe Blvd; Charlotte, NC 28203 Ofc:704-355-1267 ; Lab:704-355-7220 ; Fax 704-355-7648 E-mail: Winston.Wiggins-at-CarolinasHealthCare.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Hi, everyone,
Does anyone have the name of a service person for a tabletop Ilford 2150 RC paper printer in the Boston area? Thanks in advance,
Mary McKee MGH Renal Unit Charlestown, MA 02129 (617)726-3696
We have had problems with Cy5 and Alexa 647 where the dye fluoresces in the green channel. Basically, we were told that the molecules cleave and one of the products fluoresces in the green channel.
We have been using Bodipy 650/665 phalloidin with even a worse problem. [One of our users wanted to use this as a FRET acceptor; more on this later.] The staining looked really bright through a narrow band Cy3 filter set, which, according to the published spectra, it shouldn't emit at all. After bleaching with 637 nm light, the signal in the Cy3 channel looked even brighter, the result we'd like to get if there were Cy3 to Bodipy FRET. However, this led me to think that maybe exposure of the Bodipy to 637 light was not so much bleaching it as modifying the molecule such that it absorbed and emitted at shorted wavelengths. Perhaps the Bodipy 650 behaves similar to the shorter Bodipy 581 which "shifts to green fluorescence upon peroxidation, a feature that has been exploited for ratiometric measurements of lipid oxidation in live cells"?
Anybody know if this is correct? We'd like to find the most stable dyes in the infra-red channel in addition to Alexa 633.
And as for the FRET, I recommended not using it regardless because "These reactive dyes contain an additional seven-atom aminohexanoyl spacer ("X") between the fluorophore and the succinimidyl ester group. This spacer helps to separate the fluorophore from its point of attachment, potentially reducing the interaction of the fluorophore with the biomolecule to which it is conjugated and making it more accessible to secondary detection reagents such as anti-dye antibodies."
Thanks.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 17:40:05 2003
At 12:38 PM 11/14/2003 -0800, you wrote: } We are using JEM-2010F microscope. The smallest field } limiting aperture which JEOL commercially provides is } 5 micron. But it is still a little large when we want } to get electron diffraction patterns for some special } materials. However, we do not want to use NBD mode } because of the serious radiation damage.
You can almost always _throw away_ beam current in probe mode if that's what you want to do. Use the smallest condenser aperture, select the smallest possible probe size, and select the smallest possible convergence angle. If that doesn't get you a small enough current, turn down the electrostatic gun lenses starting with A2. I have produced probes ~5 nm in diameter with relatively small convergence angles ( {0.2 mrad) and ~ 1 pA of beam current on a 2010F. And of course, if you are trying to illuminate a larger area, you can reduce the current density by defocusing the focused probe with C2, the brightness control.
There is also some danger with trying to obtain selected area diffraction patterns from very small regions of a sample - spherical aberration of the objective lens means that the higher order reflections in the pattern don't come from the same part of the sample as the lower order reflections (see Williams and Carter, p. 186-187 for an explanation).
Best wishes, Paul Voyles
Paul Voyles Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 20:35:28 2003
Well not really - some of the new inkjets have three or four black cartridges - and if you are printing mostly black and white. i.e. grey scale images, the ink sets fro Cone or Quad Black provide a very wide gamut of true grey scale printing by having a range of black to grey inks.
Bill
At 01:40 PM 11/12/2003, DrJohnRuss-at-aol.com wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Nov 14 21:51:06 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rpowell-at-nanoprobes.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 14, 2003 at 21:18:28 ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell at Nanoprobes
Organization: Nanoprobes, Incorporated
Title-Subject: [Microscopy] Re: Achieving a fine grain negative stain - Vendor reply
Question: Hello Tom:
I can't advise on the microscope, but at Nanoprobes we do offer a vanadate-based negative stain, NanoVan, which several users find gives a very fine grain structure - check the paper by Gregori et al ( J. Biol. Chem., 272, 58-62 (1997)) for an example.
Reprint: http://www.jbc.org/cgi/reprint/272/1/58.pdf Reprint of our 1994 MSA abstract: http://www.nanoprobes.com/MSANV.html
Since it is based on a lower Z element, the staining is lighter. We also offer a tungstate-based analog, Nano-W, for denser staining. Details are on our web site:
http://www.nanoprobes.com/Nstain.html
Hope this helps,
Rick Powell Nanoprobes, Incorporated
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If so, even with a 5 um SA aperture, you potentially have significant errors in your diffraction pattern. The spherical aberation of the objective lens means that in an SA pattern, different reflections come from slightly different areas. This causes displacement errors from ~0.1 nm for low order reflections up to several um for higher order reflections.
In addition, the focus of the objective lens and the vertical position of the SA aperture are critical. Errors in either can result in additional displacement errors, to as much as 20 um or more if the objective is badly out of focus and/or the SA aperture is in the wrong plane.
With a 5 um SA aperture, the projected diameter on your specimen is likely to be ~250 nm to 500 nm. So, even if you have been very careful in setting up the microscope, it is probable that any but the lowest order reflections actually arise from points outside the area selected!
To be confident that you are getting diffraction from the area you want, you must use a technique which limits the illuminated area using the TEM illumunation optics. In the case where you have a very beam-sensitive specimen, I would suggest that the Riecke method for nanodiffraction is most appropriate. In this case, you use a small (~5 um to 10 um) condenser aperture and set up a parallel or near-parallel illumination condition at the specimen (not a focused probe, so beam damage is minimised). This results in diffraction patterns which look very similar to SAD patterns but do not suffer from the displacement errors of SAD. The technique typically allows the selection of areas down to ~100 nm, depending on the demagnification between the C2 aperture and the specimen.
See, for example, David Williams "Practical Analytical Electron Microscopy in Materials Science" for a more detailed discussion of this whole subject.
-- Larry Stoter JEOL UK
From MicroscopyL-request-at-ns.microscopy.com Sat Nov 15 14:41:32 2003
Ellery E. Frahm wrote: =============================================================== I have a user with gold-coated samples after doing ion probe analyses, and now he needs to do more electron microprobe work, both imaging and quantitative analyses. The gold coats (I'm not certain about thickness) need to be removed. We cannot, however, polish the gold off because he needs to preserve the pits left from the ion probe. As a result, we are looking for chemical means to remove the gold coats. Any suggestions are welcome (particularly those known to work and with details about concentrations, temperatures, etc.). =============================================================== Something we learned quite by accident some number of years ago: Approximately ten to fifteen minutes in an (isotropically created) oxygen plasma (for example, in an SPI Plasma Prep II plasma etcher) would remove a 10-15 nm sputtered gold coating (at 100 watts power). We never understood the chemistry of how this happens but it does work. It is essentially a "room temperature" process.
Of course your sample has to be something that won't be etched by an oxygen plasma so if it is a metal, or ceramic, or anything else that is inorganic, you are OK, but not if it is organic. We made our original observations on a clay coated photocopy paper, and the oxygen did not seem to be changing the clay particles (but it did of course remove the acrylic binder holding the clay particles to the paper substrate).
There are advantages to using a "dry process" as opposed to a "wet chemical" approach, at least in some instances.
Chuck
Disclaimer: SPI Supplies is the manufacturer of the Plasma Prep II plasma etcher which is described on URL http://www.2spi.com/catalog/instruments/etchers1.shtml
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Sun Nov 16 19:17:12 2003
On Friday, November 14, 2003, at 12:45 PM, tbargar-at-unmc.edu wrote:
} Also the } operating conditions of my TEM (a Philips 410LS with tungsten } filament) are } 80Kv and 100Kv (the limit of my TEM), aperture sizes are condensor } 200micron, objective 20micron, spotsize 2, zero degree tilt. I'm } trying to } photograph in the range of 240,000X to 500,000X (the upper limit of my } TEM). I would appreciate any suggestions to improve the image of the } samples either by way of specimen preparation or operating condition } of the } instrument.
Dear Tom, Since Rick Powell covered the stain half of your question, I thought I'd take a stab at the other half. The negative stain consists of clumps of material typically ~1 - 2 nm in diameter, so the magnification you're using results in blobs of stain ~1/4 -1 mm on your detector. If you lower the mag, you will have smaller, but still visible clumps of stain, and spreading the beam can reduce effects from heating and radiation damage; additionally, you will get a larger field of view. Even with a W filament, you are concentrating a large number of electrons on a very small sample area and few electrons outside this area, so you can cause motion even of very heavy atoms. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 17 07:39:58 2003
I am looking for a good TEM photo of a Canal of Hering, or cholangiole from the liver. Also, I would like some information on how to maximize one's chances of finding this structure under the beam.
Many thanks, Peggy Harger-Allen EM Lab, VAMC, Indianapolis
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 17 08:17:04 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Donnarae48-at-AOL.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, November 16, 2003 at 14:43:02 ---------------------------------------------------------------------------
Email: Donnarae48-at-AOL.com Name: Donna Morrow
Organization: Moreau Catholic High School
Education: 9-12th Grade High School
Location: Hayward, California
Question: My biology class did a lab using the microscopes. We are able to magnify up to 400 magnification. We used a scraping from our cheeks and dye to make a slide. We were asked to draw what we saw and lable at least 2 things. I am not sure what I saw. At 400 magnification how clear would an epithelial cell be and would I have been able to see a nucleus. All I saw were blue circles and light blue background and very small round things scattered around. I am wondering if the larger dark blue circles were a nucleus and the smaller specks the cell membranes even though they did not seem to be uniform. The teacher said my slide looked good but I am not sure what I was viewing. Also do you know of any other sight that is available to help me with my lab work with the microscope. We have been trying to find a sight that would show me what an epithelial cell would look like at 400 magnification and I keep running in to brick walls. Thank you for your help
I have a Philips CM-12 TEM. Your operationg conditions seem OK to me for the mag range you indicate, except for the spot size. Drop it down to spot size 4-6 and see what you get for particle resolution. Take a photo at your spot 2 setting, then shoot same particle at lower spot sizes for comparison. I have photographed carbon nanoparticles at those mags and the smaller spot size definitely improved the resolution of the details. Your exposure times will increase, so must have stable samlple. You can also kick up the emission level of the electron gun, or even set tungsten filament tip a little closer to Wehnelt cap (but not so clsoe that you get arcing) to increase the brightness at those smaller spot sizes.
Also, just a reminder to use your cold trap. After a half-hour cooldown, you will reduce any beam induced contamination of the particles you are looking at.
Let me know if you get any improvement - or not - with smaller spot sizes. Good luck!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra
..snip!...
} Also the } operating conditions of my TEM (a Philips 410LS with tungsten filament) are } 80Kv and 100Kv (the limit of my TEM), aperture sizes are condensor } 200micron, objective 20micron, spotsize 2, zero degree tilt. I'm trying to } photograph in the range of 240,000X to 500,000X (the upper limit of my } TEM). I would appreciate any suggestions to improve the image of the } samples either by way of specimen preparation or operating condition of the } instrument.
} Tom Bargar } Core Electron Microscopy Laboratory } UNMC } 986395 Nebraska Medical Center } Omaha, NE 68198-6395 } 402-559-7347 } tbargar-at-unmc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 17 11:04:17 2003
We can supply gold foil apertures down to 2 micron. We have other size limitations though, so please contact me directly with the OD and thickness of the blank.
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Tel: 413 786-9322 Fax: 413 789-2786 mnesta-at-ebsciences.com www.ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: Paul Voyles [mailto:voyles-at-engr.wisc.edu] Sent: Friday, November 14, 2003 6:46 PM To: microscopy-at-msa.microscopy.com
At 12:38 PM 11/14/2003 -0800, you wrote: } We are using JEM-2010F microscope. The smallest field } limiting aperture which JEOL commercially provides is } 5 micron. But it is still a little large when we want } to get electron diffraction patterns for some special } materials. However, we do not want to use NBD mode } because of the serious radiation damage.
You can almost always _throw away_ beam current in probe mode if that's what you want to do. Use the smallest condenser aperture, select the smallest possible probe size, and select the smallest possible convergence angle. If that doesn't get you a small enough current, turn down the electrostatic gun lenses starting with A2. I have produced probes ~5 nm in diameter with relatively small convergence angles ( {0.2 mrad) and ~ 1 pA of beam current on a 2010F. And of course, if you are trying to illuminate a larger area, you can reduce the current density by defocusing the focused probe with C2, the brightness control.
There is also some danger with trying to obtain selected area diffraction patterns from very small regions of a sample - spherical aberration of the objective lens means that the higher order reflections in the pattern don't come from the same part of the sample as the lower order reflections (see Williams and Carter, p. 186-187 for an explanation).
Best wishes, Paul Voyles
Paul Voyles Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 18 00:11:58 2003
Hi all, I just realised that I've been replying to myself so I'll post my emails here in a new post. Therefore, my initial email is at the bottom. Any advice gratefully accepted.
John Brealey __________________________________________________________________
Hello again,
Still no electron beam. We believe the HV Stabilzer board (new capacitors) and the cable are OK. Gun vacuum is 10 (-5) Torr. Column and camera are 10 (-2) Torr. Gun and camera green lights are on. Can anyone advise on interlocks within the system? We think there are at least four... the gun valve, specimen airlock, camera shutter and camera valve. Something in the system is preventing the filament from switching on. Is there anything else that will prevent the filament turning on? We have tried several filament setups.
Thanks again,
John Brealey
-----Original Message----- } From: John Brealey [mailto:john.brealey-at-imvs.sa.gov.au] Sent: Tuesday, 11 November 2003 01:15 PM To: John Brealey
Hi all,
Our Hitachi H-600 TEM is failing to provide an HV reading at any kV (ie 25, 50, 75, 100). As the microscope is 20 years old I believe the HT cable is the problem. Do you agree? What's the best plan of attack from here? Apart from the HT cable and a dirty gun chamber, are there any other potential causes of this failure? I have never attempted to isolate the HT cable. The vacuum system is fine. I've turned the microscope completely on and off a few times and have vented all compartments to air and back to high vacuum but this has had no effect.
Two days ago I reassembled the rear cosmetic guard that encases the upper vacuum system and the HT cable. Could a slight knock to the cable be enough to finish it off? The microscope was working yesterday, though. I've tried gently wiggling the cable but that gave no response.
The microscope has not been hissing and the HV has appeared quite stable. The only sign that something was not quite right was that some mornings the HV would fail to read at 25kV (but would always read at 50, 75 and 100kV).
In the light of problems encountered by Sarah Ellis (refer to archives) a few months ago I am tempted to clean the gun chamber first (however I doubt if this will fix anything).
Any advice would be gratefully accepted before I contact our institution's electrical engineers.
John Brealey EM Unit The Queen Elizabeth Hospital Institute of Medical & Veterinary Science Adelaide Australia (08) 8222 6612 john.brealey-at-imvs.sa.gov.au
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 18 05:06:17 2003
you mentioned in your e-mail that several interlocks may affect the operation of the filament. But you didn't say whether you had any way of monitoring them.
I use an H7000 so it may not have the same interlocks but camera shutter and camera valve have no effect on the filament. The camera can be closed or at air and the filament will still run and the shutter should not affect the filament (I assume you mean the exposure shutter). If the electron gun was closed or interlock had failed you wouldn't get the high voltage operating either. So this means that if it is one of these interlocks it is most likely to be the specimen airlock.
In the H7000 you must have a proper specimen rod actually in the column to be able to turn the filament on normally. I wonder if there is some simple logic attached to the airlock interlock operation which has gone out of sequence and so maybe the specimen rod should be in a different position from normal. It's worth a try if you haven't already checked it.
Also we have a couple of plug in leads just beside the specimen airlock. The important one is HK-CN4 on our machine - if it is loose or detached then the filament will not come on because presumably this carries the signal from the airlock mechanism that there is a specimen in the microscope. I know this because it happened to us once and it's so easily knocked when you're working near the airlock in the dark.
I am assuming in all this that you mean you've got a high voltage but no beam from the filament.
Good luck,
Malcolm
Malcolm Haswell e.m. unit Chemispec School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK tel no: 0191 515 2872 / 3468 e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: John Brealey {john.brealey-at-imvs.sa.gov.au}
Dear all, I asked about this once before but got no replies. Is there any introductory book or nice review paper on the TEM of semiconductors? A student needs to learn about basic features such as 60° threading dislocations and such things but at least in recent publications everything is taken as if one already understands the basics, which we don't. Please give us a hint about where to start.
Thanks
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 18 12:03:53 2003
With the H600 as with many other instruments you may find that you cannot obtain a beam until the specimen rod has tripped the microswitch within the specimen exchange unit? Try putting the empty rod into the microscope? Also a green light means that the system has received an instruction - not that it has completed an action. Another common problem is that the gun airlock may jam closed - you will have an emission current in this case but no beam. The figures that you give for column and camera suggest backing pump pressure which is not enough to allow the system to operate correctly - when you run from air do you hear valves switching as the system moves from backing pump to diffusion pump? To check a valve opens place a stethoscope against the valve or rest a screw driver blade against the valve with your ear on the handle end! You should hear the action if its working? Anther reason for no filament is a failure of the diodes in the filament circuit within the HT tank but this is a rare problem.
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Tuesday, November 18, 2003 6:21 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (weazzi-at-unity.ncsu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 17, 2003 at 11:45:51 ---------------------------------------------------------------------------
Email: weazzi-at-unity.ncsu.edu Name: Wassim Azzi
Organization: NCSU
Title-Subject: [Microscopy] MListserver:
Question: I would please like to get information about E-Beam damage of samples in the SEM. I am primarily looking for recent publications and advancements. Any inofrmation provided will be greatly appreciaed. Regards. Wassim Azzi
Tom: Another negative stain to consider is uranyl formate. It is a bit fussy about staying in solution so you need to make it up just prior to use.
Mr. Donald Gantz Dept. Physiology and Biophysics Center for Advanced Biomedical Research Boston University School of Medicine 715 Albany Street Boston, MA 02118 email: {mailto:gantz-at-bu.edu} gantz-at-bu.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 18 14:57:17 2003
The 37th Annual Fall Symposium of the New England Society for Microscopy (NESM) will be held on Tuesday, December 9th at Gordon College in Wenham, MA. This is the second year that Gordon College has hosted this meeting.
The symposium will begin at 12Noon with registration and at 1 pm, the Technical Sessions (with talks from both the biological and materials sciences) will begin with a short coffee break between the sessions. Following the talks, the Annual Business meeting will convene at 5:00 pm with the election of new officers as one of the main agenda items. A dinner will follow with Prof. Paul Goldberg of the Archaeology Department of Boston University as the after-dinner speaker.
For details re: registration, fees, program (speakers and titles), directions to Gordon College etc., please visit NESM's website: http://prism.mit.edu:8083. Click on "Current Newsletter".
We hope to see many of you there!
Peggy Sherwood Corresponding Secretary, NESM
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 18 18:50:43 2003
Thankyou to all respondents for your replies. The microscope is in pieces at the moment so I can't check your suggestions.
To clarify... yes, we have fixed the HV but now there is absolutely no response from the filament. Our technicians have checked the circuitry of the HV Stabilzer board and believe all is well there. Voltages change as we turn the filament knob and the bias knob however there is no movement in the HV / beam current meter. I have run the microscope for 30 minutes at 100kV with filament and bias on maximum, then, leaking the gun chamber to air, have checked the filament shield and it was stone cold. The filament assembly can only be inserted one way due to the presence of a set screw in the gun assembly. We have turned the filament clockwise (as stated in the manual) until we feel the resistance of the filament pins against the gun contacts. We have tried several clean filament assemblies. All moveable apertures are out. Specimen holder in or out doesn't make any difference. We have removed the gun cable twice from the HV tank. Our technicians are happy with the circuitry of the cable and believe it is making good contact in the HV tank.
I may have been incorrect with the vacuum units on the vacuum gauge. The meter is at 10(-5) for the gun chamber (Penning gauge) and 10 (-2) for the column and camera chambers (Pirani gauge). These readings are the same as they always have been.
Currently we are checking the gun valve. Could this be stuck shut even though the vacuum system thinks it is open? When the camera valve opens I can feel its movement, so that appears to be operating. The camera system appears to be working normally when I take a dummy exposure, ie, negative plates are feeding normally.
I will try your suggestions when the microscope is reassembled.
Thanks again, John Brealey
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 05:50:47 2003
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: 18. november 2003 16:40 To: Microscopy Listserver
Dear All,
We are interested in purchasing a new ESEM. The ESEM will mostly be used for biological and some material sciences. I would like to hear your suggestions regarding which ESEM I should choose (manufacturer and model of the ESEM) and any particular reasons. Thank you in advance.
Ping Li
-- Ping Li, Ph.D. Director, Scientific Imaging Suite Department of Biology Dalhousie University Halifax, NS B3H 4J1 Canada
We are considering using a cheaper, Au target instead of a Au/Pd target, for our sputter coater. With our FEI XL30 SEM we should not notice any significant difference, is that correct?
I still wanted to know if the Au coating may be of lesser quality, and that this may show up in the future, examining the same samples with a better SEM or FESEM. In the long term, can we obtain a good quality preparation by re-coating with Au/Pd a sample that was previously coated with Au?
I will very much appreciate any hint on this.
Best regards,
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 07:41:24 2003
Listers I have a file in JPEG format. I need to reduce the file size. Currently the file size is 22 Meg and too large to import into my application. Is there a function in Paintshop that will perform a file size reduction? Thank you
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.249 Fax: (770) 487-1460 email: robert.fowler-at-tdktca.com www.component.tdk.com
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 08:34:43 2003
Hello all, I've noticed that ASTM method D:3015-95 "Standard Practice for Microscopical Examination of Pigment Dispersion in Plastic Compounds" has been withdrawn (2001). Can anyone provide an EM method for the qualitative or quantitative assessment of pigment dispersion in polymeric materials.
Thanks, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
} Hello Microscopist, Ladies and gents, } } Let me set the stage, no pun intended. We have a Jeol 2010. I } managed and trained students on this instrument from 1993 to some time } } in 2000. During that time I had to tongue lash, in a nice way, users } for } not activating the ACD cycle at the end of the day. This happened ~ 20 } } times with the only failing being a bellows in the gun valve after ~ } 10,000 cycles. This will make since to you if you have a 2010 with the } } old gun valve design. } I still on occasion use the instrument. Please understand, I need } to } refer to specifics of day. This in not intended to be a assault on } Jeol. I am the culprit. } I logged onto the instrument a few days after an new filament was } installed. I found the image of the filament to be moving back and } forth, ~1 Hz, ~1mm at x100K.... This I interpreted as an instrument } charging } problem. I continued to survey samples, encountering 2 more technical } annoyances followed by a 4th that brought the session to an end. We } all } know these things happen. } Perhaps more frustrated than I realize, I left the instrument } without activating the ACD and with the HT at 100 kV. My mistake was } found the following morning. } It's been implied that my mistake caused a contamination of the } gun } so severe that 2 additional filaments were contaminated by it as fast } as they were installed. This I } find hard to believe but I suppose stranger things have happened and } what do I know. } I'll not share other thoughts or the opinions of others for now } least I } bias your response. } } Well what do you think? } 1. has anyone had this happen to their instrument? } 2. Has anyone ever heard of a gun contaminating a filament to this } extent?. } 3. other comments? } } } thanks, } Bruce Brinson } Rice U.
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 10:01:32 2003
In a message dated 11/19/03 10:56:36 AM, Paul.Gerroir-at-crt.xerox.com writes:
} I've noticed that ASTM method D:3015-95 "Standard Practice for Microscopical } Examination of Pigment Dispersion in Plastic Compounds" has been withdrawn } (2001). Can anyone provide an EM method for the qualitative or quantitative } assessment of pigment dispersion in polymeric materials.
I'm unfortunately not familiar with the ASTM method and don't have it here in my library, but let me take a chance and jump in with something that may be helpful. Dispersion of pigment is probably ideally random but with some tendency toward clumping or clustering. On the other hand, for appearance purposes, it would be nice if the pigment could be self-avoiding, which simply means the particles would be as uniformly spaced as possible (not sure how you would produce that in this case, but in some products like Dupont Corian countertops you can see that they manage to do it, and nature does it easily - usually by chemical depletion of the region around a particle - but that's another long story).
Anyway, the measurement procedure is to determine the mean nearest neighbor distance from suitable images. Some software packages will do this directly (e.g. Fovea Pro), otherwise you would need to record the coordinates and calculate the distance to neighbors to find the minimum or nearest neighbor for each particle. A subtle refinement, which may be unimportant if you have a lot of small features and large images, is to skip any feature that is nearer to the border than to another particle, because you may not have found its actual nearest neighbor. Once you have the mean nearest neighbor distance, you can characterize the degree of clustering or self-avoidance. A true random distribution would have a mean nearest neighbor distance equal to 0.5 divided by the square root of the number of features per unit area. Notice that the units of that come out in distance, as they should. If the ratio of actual nearest neighbor distance to the "ideal" value calculated from that equation is less than one it indicates clumping or clustering (and the ratio measures the extent). And vice versa if the ratio is greater than one it indicates and measures self-avoidance.
Hope that helps
John Russ
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 10:12:08 2003
It is my understanding that Au/Pd does a better job at coating rougher surfaces because of its smaller grain size. It may depend on what you coat.
Ron L
-----Original Message----- } From: Martin Ramirez [mailto:ramirez-at-amnh.org] Sent: Wednesday, November 19, 2003 8:40 AM To: Microscopy-at-sparc5.microscopy.com
Hi,
We are considering using a cheaper, Au target instead of a Au/Pd target, for our sputter coater. With our FEI XL30 SEM we should not notice any significant difference, is that correct?
I still wanted to know if the Au coating may be of lesser quality, and that this may show up in the future, examining the same samples with a better SEM or FESEM. In the long term, can we obtain a good quality preparation by re-coating with Au/Pd a sample that was previously coated with Au?
I will very much appreciate any hint on this.
Best regards,
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 10:40:35 2003
I had mentioned in my Lacey grid review that the new batch of 4489 film had made its way into rotation here. And after trying a few suggestions, I did finally call Kodak. From some reports (from vendors and users on the list) Kodak wasn't acknowledging a problem, and that assumption contributed to a reluctance to prioritize calling and complaining about the new film.
I finally called them and was surpised. Sure enough! They've got a new procedure for hand developing that is different than the printed package. I tested the method this morning, and had another user test it after me.
And first looks seem to say: Back in business! (phew - no need to design/construct a nitrogen bubbler system)
I've quoted the pertinent parts of the procedure below. I might be behind the ball on this but looking back at the list traffic it seems this particular solution hasn't been announced here. -quote- PROCESSING Develop: KODAK Developer D-19 diluted 1 part developer and 2 parts water can be used for 4 minutes at 20°C (68°F). Hand agitate by quickly inserting the film rack into the developer tank, tapping the rack on the tank bottom to dislodge any air bubbles that may cling to the film surface. Every 10 seconds, rapidly remove the film rack completely from the developer tank and tip the film rack 45° to one side of the tank and then tip the film rack 45° to the other side of the tank, allowing the developer to drain into the tank (complete tipping procedure is only a few seconds.) Rapidly return the film rack into the tank. Repeat this process throughout the development cycle. Development times longer than 4 minutes will result in a rapid increase in background fog.
Rinse: For 1 1/2 minutes in running water at 18 to 21°C (65 to 70°F) with continuous agitation. Do not use conventional stop bath solutions because they may produce a mottled appearance on films used for electron exposures. -quote- Fix as normal....
I hope that helps, and I apologize to those who find this old news.
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 11:14:40 2003
Hello Everyone: I am looking for a something that will selectively stain aromatic or amide functional groups. Sawyer and Grubb list ruthenium tetroxide, silver sulfide, and mercuric trifluoracetate as possible stains for aromatics, but RuO4 and Ag2S are not selective and mercuric trifluoracetate is very toxic (we are trying to limit our usage of Hg anyway).
Does anyone know of anything else that will select an aromatic group (vs. alcohol, ester, ether, and acid groups)?
Another option is to stain the amide groups in my sample. Sawyer and Grubb list Tin chloride as a possible stain, but don't give a method for using it. Does anyone have experience with this stain?
Thank you,
Jessica Cervantes Bend Research, Inc 64550 Research Rd Bend, OR 97701 (541) 382-4100 page (541) 382-0212 x240
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 11:25:14 2003
} } } Listers I have a file in JPEG format. I need to reduce the file size. } Currently the file size is 22 Meg and too large to import into my } application. Is there a function in Paintshop that will perform a file size } reduction? Thank you }
If the image does not import because the number of pixels is too great, you can easily resize. With Jasc Paintshop Pro, from the image menu, choose resize. You will have the choice of choosing a specific pixel size or a percentage change in size. You should make sure that the "Maintain aspect ratio" box is checked so that the aspect ratio of the image is not changed.
A similar function is available with Irfanview - choose Resize/Resample from the Image menu.
Resizing will change the resolution of the image, so keep this in mind.
Hope this helps.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 12:12:42 2003
} Hi, } } We are considering using a cheaper, Au target instead of a } Au/Pd target, } for our sputter coater. With our FEI XL30 SEM we should not } notice any } significant difference, is that correct?
I think you will see difference starting with x20k-50k magnifications. Au/Pd target is definitely better for high magnification work.
} I still wanted to know if the Au coating may be of lesser } quality, and that } this may show up in the future, examining the same samples } with a better } SEM or FESEM. In the long term, can we obtain a good quality } preparation } by re-coating with Au/Pd a sample that was previously coated with Au?
No. Au is forming "grains" which is bigger then Au/Pd "grains". Recoating will not eliminate them.
Vladimir
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 12:26:57 2003
my understanding is that gold coating produces a coarser surface coating because Au alone nucleates on the surface as it is coated and islands of Au limit the image quality in high resolution SEM. The Au/Pd alloy forms smaller more numerous islands and so provides some improvement in resolution over gold. Simply coating a previously gold coated sample with Au/Pd will not improve the situation but should make it worse because the sample will be coated with a thicker layer.
I am surprised that you find much difference in price between Au and Au/Pd targets. In the UK they are either the same price or I am sure that I have seen Au/Pd a bit cheaper.
I think the problem is defining what you mean by high resolution in SEM but normally this should be achieved by a good traditional SEM or certainly will be by a field emission SEM. I'm sorry I have no figures - the information I have just says Au/Pd is noticeably better than Au at high resolution. Instinctively I would guess the difference would be noticeable somewhere between 20k and 40k but I would be interested to know if anyone could quote any real hard figures.
There are now lots of alternatives to Au and Au/Pd which give even better resolution and I have seen some remarkable images taken with very low voltages on a high pressure field emission SEM - but I can only dream ...
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Martin Ramirez {ramirez-at-amnh.org}
Dear Friends,
A user of the lab has asked me to separate urban myth from reality regarding HBO bulbs used for fluorescence microscopy.
His questions are:
What is the difference between a 50W bulb and a 100W bulb? He says he set up a demo, identical except one scope had a 50W bulb, the other a 100W bulb, and he didn't think he could tell much difference.
Do bulbs have a finite life, ie., should they be changed at some maximum hours even if still lighting up and appearing OK? He has heard various advice, like change after 200 hours or 250 hours. Change when the glass gets dark, how dark? Is it true that bulbs with many hours may explode?
He has only used Osram bulbs, are there other brands available and what criteria would one use to select one brand over another?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 14:49:57 2003
The method Geoff outlines below is the only one we have managed to get to work with any consistency with the new formulation. It is similar to the technique we had used previously, the critical difference being do not ever, repeat ever, let the film stand still. Our method for the old formulation processed up to 25 plates in two batches - we would alternately agitate half the films at the time leaving the other standing in developer (for 20 or so seconds before its turn). When we tested this with the new formulation around a third of the sheets were very unevenly developed (even though developer would be moving around the still batch). If we develop in smaller batches using more or less constant agitation we have no serious problems. I an
Ian Hallett HortResearch Mt Albert Research Centre, Private Bag 92 169 Auckland, New Zealand Fax +64 9 815 4201 Telephone +64 9 815 4200 EMail ihallett-at-hortresearch.co.nz
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Dear list,
I had mentioned in my Lacey grid review that the new batch of 4489 film had made its way into rotation here. And after trying a few suggestions, I did finally call Kodak. From some reports (from vendors and users on the list) Kodak wasn't acknowledging a problem, and that assumption contributed to a reluctance to prioritize calling and complaining about the new film.
I finally called them and was surpised. Sure enough! They've got a new procedure for hand developing that is different than the printed package. I tested the method this morning, and had another user test it after me.
And first looks seem to say: Back in business! (phew - no need to design/construct a nitrogen bubbler system)
I've quoted the pertinent parts of the procedure below. I might be behind the ball on this but looking back at the list traffic it seems this particular solution hasn't been announced here. -quote- PROCESSING Develop: KODAK Developer D-19 diluted 1 part developer and 2 parts water can be used for 4 minutes at 20°C (68°F). Hand agitate by quickly inserting the film rack into the developer tank, tapping the rack on the tank bottom to dislodge any air bubbles that may cling to the film surface. Every 10 seconds, rapidly remove the film rack completely from the developer tank and tip the film rack 45° to one side of the tank and then tip the film rack 45° to the other side of the tank, allowing the developer to drain into the tank (complete tipping procedure is only a few seconds.) Rapidly return the film rack into the tank. Repeat this process throughout the development cycle. Development times longer than 4 minutes will result in a rapid increase in background fog.
Rinse: For 1 1/2 minutes in running water at 18 to 21°C (65 to 70°F) with continuous agitation. Do not use conventional stop bath solutions because they may produce a mottled appearance on films used for electron exposures. -quote- Fix as normal....
I hope that helps, and I apologize to those who find this old news.
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
______________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 14:54:14 2003
Hi I'm looking for a supplier of carbon string (fiber) for making carbon films. Any recommendation/suggestion is greatly appreciated.
Gang (Greg) Ning, Ph.D. Director, Electron Microscopy Facility Huck Institute for Life Sciences The Pennsylvania State University 1 South Frear Lab University Park, PA 16802 Phone: 814-863-0994 Fax: 814-863-1357 Email: gxn7-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 16:25:17 2003
There are often contradicting explanations in the Hitachi manuals in relation to filament position in its holder. Some say in line with the two slots some show 90 degs. If in doubt insert a filament, remove the HT cable from the tank and check with a multimeter for continuity across two of the contacts; never done it but it should work?
If the gun valve does not open, even when instructed to do so, most Hitachi TEM will still allow HT and filament ON, its just impossible to find the beam (ask some of my customers!!$$??) Under these conditions you have indications on the emission meter that you have HT and an emission current that responds to filament control change. This is not a dirt problem forget cleaning, it appears not to be a supply problem to the tank, it may still be a vacuum problem in relation to the column from your figures, my bet is the filament supply diodes in the tank; if I remember correctly they are rated at 6 amp?
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, November 19, 2003 1:00 AM
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Martín J. Ramírez wrote: ====================================================== We are considering using a cheaper, Au target instead of a Au/Pd target, for our sputter coater. With our FEI XL30 SEM we should not notice any significant difference, is that correct?
I still wanted to know if the Au coating may be of lesser quality, and that this may show up in the future, examining the same samples with a better SEM or FESEM. In the long term, can we obtain a good quality preparation by re-coating with Au/Pd a sample that was previously coated with Au?
I will very much appreciate any hint on this. ========================================================== You should use caution here.
When you say "cheaper", just why is it cheaper? If it is because you found someone who will sell you the same identical item at a lower profit margin, then you are getting a good deal and you should take it. But you sound worried that this might not be the case, that it might be cheaper because it might be lacking in some way in quality and that would be purity. The higher the purity the more expensive is the precious metal, not because you are getting more of the precious metal, but because of the economics associated with the production step, when comparing for example, 0.999 vs. 0.9999 vs. 0.99999.
So what if you are getting something of inferior quality, how does it affect the final result? The main problem is that while the gold sputters easily (because of its low work function), the impurities stay behind on the cathode, forming a barrier, preventing further sputtering from occurring. We have heard stories that some of these impurities at some point start to come down on the sample producing an indeterminable artifact effect. True the stories are anecdotal, but it does make sense that that could happen.
You can always decide at some time to put on another layer of gold. However , each time you make the total layer thicker, you start covering up the smallest features you are seeing, so you lose detail. If impurity particles are showing down on the same, you are then going to be covering them up and "seeing" them in your recoated sample. That, in our opinion, does not make for good laboratory practice.
So while I admit that is will sound like a very self-serving kind of statement, since SPI Supplies supplies high purity cathodes, the best advice is to stick to suppliers that you know well and who are selling to you cathodes of known high purity, at least 0.9999 and perhaps even 0.99995. If you can find a jeweler who has gold foil of those levels of purity, by all means, if you can save some money, why not? But make sure you know your vendor and that you trust what you are being told about purity.
RE: Au vs Au/Pd and difference in grain size. Can some one cite a reference or two where that has actually been studied and confirmed for sputtered coatings? I know that this was done years ago on vacuum evaporated coatings , but I can't put my finger on a reference where it was done with sputtered coatings.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 16:40:51 2003
We tried constant agitation, but in the bath and just vertical (the holder and solution bath don't allow for anything but up and down agitation). The results were somewhat satisfactory but there seems to be something to the 45º and the tapping, because the film suddenly looked more like the old film when I was finished with it.
I did the 10 second interval as outlined with success. So shaking for 2-3 seconds and resting for 5 or so seconds.
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: Ian Hallett [mailto:IHallett-at-hortresearch.co.nz] } Sent: Wednesday, November 19, 2003 3:51 PM } To: willi1gl-at-cmich.edu; microscopy-at-sparc5.microscopy.com } Subject: [Microscopy] Re: Kodak 4489 (working solution for low budgetlabs) } } } } ------------------------------------------------------------------------ -- } ---- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ -- } ----- } } Geoff and listers } } The method Geoff outlines below is the only one we have managed to get to } work with any consistency with the new formulation. It is similar to the } technique we had used previously, the critical difference being do not } ever, repeat ever, let the film stand still. Our method for the old } formulation processed up to 25 plates in two batches - we would } alternately agitate half the films at the time leaving the other standing } in developer (for 20 or so seconds before its turn). When we tested this } with the new formulation around a third of the sheets were very unevenly } developed (even though developer would be moving around the still batch). } If we develop in smaller batches using more or less constant agitation we } have no serious problems. } I } an } } Ian Hallett } HortResearch } Mt Albert Research Centre, Private Bag 92 169 } Auckland, New Zealand } Fax +64 9 815 4201 } Telephone +64 9 815 4200 } EMail ihallett-at-hortresearch.co.nz } } } } } } "Geoff Williams" {willi1gl-at-cmich.edu} 20/11/2003 5:48:47 } } } } } } ------------------------------------------------------------------------ -- } ---- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ -- } ----- } } Dear list, } } I had mentioned in my Lacey grid review that the new batch of 4489 film } had made its way into rotation here. And after trying a few } suggestions, I did finally call Kodak. From some reports (from vendors } and users on the list) Kodak wasn't acknowledging a problem, and that } assumption contributed to a reluctance to prioritize calling and } complaining about the new film. } } I finally called them and was surpised. Sure enough! They've got a new } procedure for hand developing that is different than the printed } package. I tested the method this morning, and had another user test it } after me. } } And first looks seem to say: Back in business! (phew - no need to } design/construct a nitrogen bubbler system) } } I've quoted the pertinent parts of the procedure below. I might be } behind the ball on this but looking back at the list traffic it seems } this particular solution hasn't been announced here. } -quote- } PROCESSING } Develop: KODAK Developer D-19 diluted 1 part developer and 2 parts water } can be used for 4 minutes at 20°C (68°F). } Hand agitate by quickly inserting the film rack into the developer tank, } tapping the rack on the tank bottom to dislodge any air bubbles that may } cling to the film surface. Every 10 seconds, rapidly remove the film } rack completely from the developer tank and tip the film rack 45° to one } side of the tank and then tip the film rack 45° to the other side of the } tank, allowing the developer to drain into the tank (complete tipping } procedure is only a few seconds.) Rapidly return the film rack into the } tank. Repeat this process throughout the development cycle. Development } times longer than 4 minutes will result in a rapid increase in } background fog. } } Rinse: For 1 1/2 minutes in running water at 18 to 21°C (65 to 70°F) } with continuous agitation. Do not use conventional stop bath solutions } because they may produce a mottled appearance on films used for electron } exposures. } -quote- } Fix as normal.... } } I hope that helps, and I apologize to those who find this old news. } } Geoff Williams } Microscopy Facility Supervisor } } Central Michigan University Biology Department Microscopy Facility } http://www.cst.cmich.edu/centers/microscopy/ } } } } } } } } ______________________________________________________ } The contents of this e-mail are privileged and/or confidential to the } named recipient and are not to be used by any other person and/or } organisation. If you have received this e-mail in error, please notify } the sender and delete all material pertaining to this e-mail. } ______________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 17:04:08 2003
On Wednesday, November 19, 2003, at 10:36 AM, Jon Krupp wrote:
} Do bulbs have a finite life, ie., should they be changed at some } maximum } hours even if still lighting up and appearing OK? He has heard various } advice, like change after 200 hours or 250 hours. Change when the glass } gets dark, how dark? Is it true that bulbs with many hours may explode? } Dear Jon, I think that the bulbs you're talking about are low-pressure, so there is not a serious explosion hazard--more experienced people may correct me here. When I was using a high-pressure Hg lamp for photochemistry, rated for 100 hours, I turned on one that had been in use and was adjusting the air flow valve (to achieve the correct temperature) when there was a loud bang and a shower of quartz dust about 10 cm from my hand. The lamp housing protected me from injury, but there were several dents in the steel, one of the expensive quartz lenses was bruised (it could have been worse) and the reflector was destroyed. The company informed me that that was the usual failure mode. The next time, we set up all the experiments, turned the new lamp on, ran 24 hours a day until we were done--at about 400 hours--turned the lamp off, and carefully removed it, wrapped it in bubble paper, and left it in the back of a hood until the safety people could dispose of it. Temperature cycling is a lot worse for the lamp than running it continually, but for a low-pressure lamp, YMMV. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 17:12:13 2003
Gang (Greg) Ning wrote: ========================================================== I'm looking for a supplier of carbon string (fiber) for making carbon films. Any recommendation/suggestion is greatly appreciated. ========================================================= Both carbon string and fiber is available from the main suppliers of EM consumables such as SPI Supplies. You can see the range of diameters offered on URL http://www.2spi.com/catalog/spec_prep/carbon-fiber.shtml
I would urge a word of caution: If you are contemplating using the string in a mechanical pumped vacuum system for the production of TEM support films , you will be disapointed with the result. You really do need a diffusion pumped system or better such as in a vacuum evaporator. And in that instance, carbon rods will give a better result than the string.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 19 22:02:31 2003
Depending on which XL-30 you have, there may or may not be a difference. If you are using the thermal SFEG, it will definitely make a difference. With SFEG or non-SFEG systems, if resolution is good enough at high mag (} 75KX), you can see the Au coating as a web-like structure on the surface. If you coat over this with Au/Pd or Pt, it will just make the structure more obvious. Cold/magnetron coaters with Au/Pd targets do a very good job laying down an ultra fine film. Pt works well too. I use a Denton Desk II coater for semiconductor work up to 350KX. Au alone is not satisfactory even at 50KX since the Au structure distorts the image. I have used an Anatech Hummer VII coater before and also got good results. So it would seem to me that a good coater with an Au/Pd target will do a superior job over Au for high resolution, high mag imaging. Pt targets are more costly but are even better.
Use a low current, high vacuum setting. I'm running the Denton at 20mA, 60mT for between 30-40 seconds.
There are basic price point differences between Au/Pd and Pt. Pt is higher cost. However, keep in mind that many places sell targets with different thicknesses. A cheaper thin target will not last as long as a modestly more expensive thicker target. Used on a regular basis, my targets last over two years before needing replacement. The coating is only about 50-70A thick. My work ranges from 100X to 350KX. Au/Pd works for all but the most demanding specimens. If I were to choose one target, it would be Pt. I would use it for every specimen. But some specimens are better analyzed with EDS using Au/Pd, others with Pt. So it depends.
The other factor is to keep coated specimens under vacuum storage when not being used. Invariably, there will be Carbon polymerization that shows up as scan areas. These can be reduced by keeping oil out of the chamber and atmosphere away from the specimen. A small trap in the coater vacuum line helps keep the coater from backstreaming. Probably the ultimate coater is a turbo pumped system. But I have yet to find one that is user friendly enough and cost effective to justify purchasing.
gary g.
At 08:21 AM 11/19/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Nov 20 00:17:59 2003
Our technician Terry Fitton has summarised the current situation on our microscope. Here is his email to me...
Hi John. Thanks for the new emails. I cant get over what a great bucketful of talent you can call on. If we could get help like this on other instruments life would be much easier. I think we should clarify the current situation for others so we dont waste their time and cause confusion. Here is the situation at present as I see it. 1 Outputs from the stabiliser board to the tank are within spec. for HV, filament and bias. Voltages are OK and vary with adjustment of controls on console as required. 2 We have checked filament connections, hv cable continuity and its connections to the tank and filament and all appears OK. 3 Gun airlock was jammed closed and is currently being repaired. 4 We have indication of HV but no apparent filament current. 5 Still have to check vacuum values for column and camera. 6 Still have to check interlocks on specimen airlock and camera airlock. I am interested in Steve Chapman's idea of faulty diodes in the filament circuit inside the tank. Could he be persuaded to elaborate a little as we have no experience of tank insides. Can we do any checking without getting covered in stinking oil. Joel's suggestions have already been covered with all present and correct. That should do for starters. Cheers Terry
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 20 02:38:48 2003
Like Many others we suffered from the change in the formulation of our EM film. We tried many alternative methods of reducing the fogging and by far the best, and the easiest was changing to Ilford PQ universal developer. We don't even have to agitate to get smooth, repeatable results! Ken Blight Senior Scientific Officer Cancer Research UK London
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 20 04:47:46 2003
I am not familiar with the ACD you mention but am very familiar with electron guns so lets have a look at your problem?
Contamination in the gun area may usually be attributed to vacuum levels and the resulting high voltage discharge. Spoil the vacuum and you reduce the insulating effect such that high voltage discharge occurs and this cracks gasses that deposit around the gun chamber as a brown/orange layer. This layer seems to be very absorbent and acts like a gas sponge when you re apply a vacuum. The constant outgassing of course leading to even more contamination of the gun chamber. To get back to normal ALL this contamination must be removed
Within the cathode assembly the filament and its base will also give an indication of poor vacuum or poor quality filaments. I have set this out in detail on our interactive EM maintenance CD but this is a brief resume of what you will see.
1. The filament will have suffered from gas attack resulting in extreme thinning over the complete filament, not just at the tip as you would see in a normal failure. 2. The filament base will also become discoloured taking on the same orange/brown tint as the gun chamber. 3. Poor quality filaments that are made up of a high percentage of rubbish will very rapidly contaminate the cathode assembly as the rubbish evaporates. The filament break is normal but the life and the base will indicate problems; the base will be orange/brown.
I have worked with almost every type/make of EM and from time to time the manufacturers have a batch of filaments that are, through no fault of their own, of poor quality. Short life high contamination are instant indicators. I have probably mentioned before that if you have a good box of filaments NEVER use all of them, save two. In a situation where you believe you have a filament problem you may then go back to your good box and confirm that they run correctly. Pop the remainder of the bad box back to your supplier whom I am sure will provide you with a new box.
Good luck with your problem, is it a poor filament problem have you just opened a new box?
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "Bruce Brinson" {brinson-at-rice.edu} To: {microscopy-at-msa.microscopy.com} Sent: Wednesday, November 19, 2003 6:00 PM
Robert writes ...
} Listers I have a file in JPEG format. I need to reduce the file size. } Currently the file size is 22 Meg and too large to import into my } application. Is there a function in Paintshop that will perform a } file size reduction? Thank you
I cannot speak for Paintshop, but Irfanview should be able to load the JPEG and resave it with a lower quality compression setting. In the context of scientific imagery, I should warn you that resaving as JPEG is a lossy operation.
If you cannot make the file much smaller with Irfanview, you can change the print resolution (pixels/inch), and make the file size more appropriate for your printer. I.E., a 8x11" print (as JPEG) shouldn't be much larger than 3-5Mb (mileage may vary).
We used ILford PQ Universal with Ilford EM film then switched to 4489 and D19. When you develop 4489 with PQ Universal do you dilute 1+9 and develop for 4min, rinse 1min and fix for 4min (in Ilford Hypam)?
Dave
On Thu, 20 Nov 2003 08:47:31 +0000 Ken Blight {Ken.Blight-at-cancer.org.uk} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Like Many others we suffered from the change in the } formulation of our EM film. We tried many alternative methods of } reducing the fogging and by far the best, and the easiest was } changing to Ilford PQ universal developer. We don't even have to } agitate to get smooth, repeatable results! } Ken Blight } Senior Scientific Officer } Cancer Research UK } London } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 20 09:45:00 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (osakhi2003-at-yahoo.co.in) from on Thursday, November 20, 2003 at 08:58:48 ---------------------------------------------------------------------------
Hello Sousan and Steve and all, Let me answer your questions.
We have a LaB6 Filament.
The Anti-Contamination Device (ACD) is a LN2 cooled metal (no charcoal) cryo pump that pretty much surrounds the sample. It accumulates molecules in this vicinity, most water vapor I am sure. When the ACD Heat (regeneration) cycle is initiated, the ion pump is valved off and the diffusion pump valve is opened to pump the molecules that are desorbed as the ACD is warmed up. The ion pump can not keep up with the gas load so if the ACD cycle is not run, the column pressure increases. I am not sure how high it goes but 10-4 Torr is probably not a unreasonable guestamation.
Yes we have a maintenance contract which includes bi-annual service.
Steve Chapman, Thank you, as you have probably seen provided a very detailed response. The obvious interpretation is that I trashed our gun. The explanation of the plasma effect is opposite is that I was thinking, that being that atomic hydrogen and oxygen from the ionized water would lean up existing carbon as in plasma etching with air and carbon cleaning experiments using H and O that I am aware of here at Rice. Ok so I am wrong. 100kV would tend to change things a bit :).
Thank you, Bruce
Steve Chapman wrote:
} Hi } } I am not familiar with the ACD you mention but am very familiar with } electron guns so lets have a look at your problem? } } Contamination in the gun area may usually be attributed to vacuum levels and } the resulting high voltage discharge. Spoil the vacuum and you reduce the } insulating effect such that high voltage discharge occurs and this cracks } gasses that deposit around the gun chamber as a brown/orange layer. This } layer seems to be very absorbent and acts like a gas sponge when you re } apply a vacuum. The constant outgassing of course leading to even more } contamination of the gun chamber. To get back to normal ALL this } contamination must be removed } } Within the cathode assembly the filament and its base will also give an } indication of poor vacuum or poor quality filaments. I have set this out in } detail on our interactive EM maintenance CD but this is a brief resume of } what you will see. } } 1. The filament will have suffered from gas attack resulting in extreme } thinning over the complete filament, not just at the tip as you would see in } a normal failure. } 2. The filament base will also become discoloured taking on the same } orange/brown tint as the gun chamber. } 3. Poor quality filaments that are made up of a high percentage of } rubbish will very rapidly contaminate the cathode assembly as the rubbish } evaporates. The filament break is normal but the life and the base will } indicate problems; the base will be orange/brown. } } I have worked with almost every type/make of EM and from time to time the } manufacturers have a batch of filaments that are, through no fault of their } own, of poor quality. Short life high contamination are instant } indicators. I have probably mentioned before that if you have a good box of } filaments NEVER use all of them, save two. In a situation where you believe } you have a filament problem you may then go back to your good box and } confirm that they run correctly. Pop the remainder of the bad box back to } your supplier whom I am sure will provide you with a new box. } } Good luck with your problem, is it a poor filament problem have you just } opened a new box? } } Steve Chapman } Senior Consultant Protrain } Electron Microscopy Training and Consultancy World Wide } Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 } www.emcourses.com } } ----- Original Message ----- } From: "Bruce Brinson" {brinson-at-rice.edu} } To: {microscopy-at-msa.microscopy.com} } Sent: Wednesday, November 19, 2003 6:00 PM } Subject: [Microscopy] TEM, GUN? } } } } } } } -------------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------------- } ----- } } } } Bruce Brinson wrote: } } } } } Hello Microscopist, Ladies and gents, } } } } } } Let me set the stage, no pun intended. We have a Jeol 2010. I } } } managed and trained students on this instrument from 1993 to some time } } } } } } in 2000. During that time I had to tongue lash, in a nice way, users } } } for } } } not activating the ACD cycle at the end of the day. This happened ~ 20 } } } } } } times with the only failing being a bellows in the gun valve after ~ } } } 10,000 cycles. This will make since to you if you have a 2010 with the } } } } } } old gun valve design. } } } I still on occasion use the instrument. Please understand, I need } } } to } } } refer to specifics of day. This in not intended to be a assault on } } } Jeol. I am the culprit. } } } I logged onto the instrument a few days after an new filament was } } } installed. I found the image of the filament to be moving back and } } } forth, ~1 Hz, ~1mm at x100K.... This I interpreted as an instrument } } } charging } } } problem. I continued to survey samples, encountering 2 more technical } } } annoyances followed by a 4th that brought the session to an end. We } } } all } } } know these things happen. } } } Perhaps more frustrated than I realize, I left the instrument } } } without activating the ACD and with the HT at 100 kV. My mistake was } } } found the following morning. } } } It's been implied that my mistake caused a contamination of the } } } gun } } } so severe that 2 additional filaments were contaminated by it as fast } } } as they were installed. This I } } } find hard to believe but I suppose stranger things have happened and } } } what do I know. } } } I'll not share other thoughts or the opinions of others for now } } } least I } } } bias your response. } } } } } } Well what do you think? } } } 1. has anyone had this happen to their instrument? } } } 2. Has anyone ever heard of a gun contaminating a filament to this } } } extent?. } } } 3. other comments? } } } } } } } } } thanks, } } } Bruce Brinson } } } Rice U. } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 20 10:21:17 2003
I have developed the "new" 4489 with no problems using the same technique I used with the old film. I have never have problems with uneven development.
1. I make my D-19 from raw ingredients using Kodak's recipe. I don't use prepackaged D-19 because I do so little EM these days I wind up throwing most of it away. The prepackaged stuff only keeps 2 years anyway. 2. Use fresh developer! Stuff that has been sitting in a tank without a floating lid goes bad in just 2-3 days, even if it is covered. Even with a floating lid I never use developer that is more than 5 days old. Chemisty is the cheapest part of the process, a few sheets of film costs as much as all of the chemicals in D-19. 3. I agitate with the "tilt" method but more slowly, it takes me 8-10 seconds to remove-tilt left-reimmerse-tilt right-reimmerse. I do this every 30 seconds. 4. Rinse and fix as usual.
Geoff
Ken Blight wrote:
} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Like Many others we suffered from the change in the formulation of } our EM film. We tried many alternative methods of reducing the } fogging and by far the best, and the easiest was changing to Ilford PQ } universal developer. We don't even have to agitate to get smooth, } repeatable results! } Ken Blight } Senior Scientific Officer } Cancer Research UK } London }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 20 11:48:49 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rbremer-at-acpub.duke.du) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, November 20, 2003 at 11:31:41 ---------------------------------------------------------------------------
Email: rbremer-at-acpub.duke.du Name: Ron
Organization: Duke
Education: Graduate College
Location: Durham. NC
Question: Hey,
I work with an Olympus BH2 which for we recently got a blue filter set (filters both excitation and emission). When I view my specimens with my 20x objective I get a very nice signal over the microscopeís black background. If I use any of my other objectives (at least 10-40x), I get a blue haze over the whole field (that is even with no slide/specimen on the stage). I imagine this has to be a problem with the objectives, but since i've not done much blue fluorescence in the past I thouhgt I would check up on it a bit. All the objectives work fine when using red or green IR. Thoughts?
Gram-positive bacteria have relatively simple cell walls with a relatively high peptidoglycan content. Gram-negative bacteria have a more complex cell membrane system with relatively low peptidiglycan content. The crystal violet stain in the first step of Gram staining will bind to the peptidoglycan layer in Gram-positive cells effectively masking membrane counterstaining by safrannin. Gram-negative cells will bind relatively little crystal violet in the thin peptidoglycan layer, the rest being washed out in the decolorizing step. Cell membranes stained by safrannin in the last step will appear red in Gram-negatives. Hope this helps Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 20 12:21:49 2003
In reply to your request for more information on the high voltage tank. Yes you are correct it can be a messy task!
Disconnect all cables and move the tank to an area that allows plenty of space to work. Run the tank onto an expanse of plastic sheet at least nine times the size of the tank, i.e. plenty of floor space covered. The area selected should be as clean as possible with no through way for persons not involved with the task. The tank is built with all the electronics suspended from the top plate. Give the tank outsides a good clean and then remove the bolts holding top plate to tank. If you have a crane it helps but I have carried out this procedure with some willing helpers! Tank fluid ruins shoes so we always work bare footed with trousers rolled up! Get a picture of this?
Provided the HT has been off overnight you should have no residual charge problems. Very slowly lift the tank top allowing the fluid to drain back into the tank. Take your time here. Once clear of the tank and well drained you may place this unit on the protected floor but it must be on a CLEAN surface.
Its a long time since I went into the tank, but from memory if you have the circuit diagrams you should not have a problem finding the appropriate board. I assume you know how to check the diodes, lets hope there lies the fault?
Once fixed make sure all under surfaces are clean before you very very slowly lower the tank top back into place. The air needs to be driven out of the components so very gently rock the top as you lower it into place. Once in place and bolted down once again rock the tank but you may now be a little more violent.
Run the HT up very slowly, stay on the lowest kV for at least half an hour for the tank to warm, convection currents should ensue that no gas is trapped. Lets hope you are able to fix the beast?
Best of luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, November 20, 2003 6:27 AM
It was recently brought to my attention that a few members of the Microbeam Analysis Society (not Microscopy Society of America) did not receive their 2004 renewal membership forms and election ballots sent out a few weeks ago. If you are a current MAS (not MSA) member and did not receive this mailing, please contact me (rosslm-at-missouri.edu) and I will have it sent out to you.
We apologize for this unfortunate problem . Lou Ross MAS Membership Services -- Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211-5120 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.biotech.missouri.edu/emc
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 20 16:20:45 2003
Hi all, I'm giving up on Google: Does anyone know where I can get a cloth dust covers that would fit a RMC 6000XL ultramicrotome? thanks for any help, Beth -- ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
In a message dated 11/20/2003 5:28:29 PM Eastern Standard Time, beth-at-plantbio.uga.edu writes:
} Hi all, } I'm giving up on Google: } Does anyone know where I can get a cloth dust covers that would fit a } RMC 6000XL ultramicrotome? } thanks for any help, } Beth } -- } ************************************************************ } ********** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805
Hi Beth,
I'll check on this tomorrow for you. Don't know if we have any in stock, but I think a dust cover for an MT-7 will work also. I'll let you know what I find.
Best,
Bob Chiovetti
Robert (Bob) Chiovetti, Ph.D. Senior Product Manager Boeckeler Instruments, Inc. 4650 South Butterfield Drive Tucson, AZ 85714 USA Tel. 520-745-0001 Fax 520-745-0004 www.rmcproducts.com www.boeckeler.com bob-at-boeckeler.com
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 21 05:02:36 2003
Hello all, Does anyone know of a stain for araldite resin to stain it more grey or white under the SEM ? I'm investigating little pieces of charcoal with a XL30SFEG and have some problems with the black background caused by the resin.
TIA, Erwin Vermeij Forensic Scientist Microtraces Netherlands Forensic Institute Rijswijk, The Netherlands
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 21 08:29:00 2003
Hi all, I am looking for a new inkjet printer and an considering the Epson Stylus Photo R800 due out in February, 2004. I was also considering using only continuous tone inks since this printer will be for EM prints only.
What I am looking for is a system that will really replace a good darkroom print. I want a true grayscale without the color toning or cast that you get with all inkjet printers that have color cartridges. I also want to maximize the shades of gray to capture fine nuances in images.
Does anyone have personal experience with the Piezography BW ink systems (www.piezography.com)? Which ink tones do you prefer, what paper do you use, and can you give me an idea of cost per 8x10 page? Also how many pages per cartridge set or is it better to bite the bullet and get the continuous feed system?
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 21 09:06:57 2003
If you don't mind "staining" before the fact, I had a similar problem examining coal in epoxy many years ago. Our solution was to dissolve iodaform in the epoxy resin before mixing with the hardener. We could dissolve up to about 18% iodaform by weight by using mild heating to speed the process. I think the iodaform would precipitate out if we tried to push it further. We were trying to do automated analyses and they worked reasonably well at that level of doping. If you just need to find particles by eye, then lower levels should be adequate.
For the record, this was a simple two-component epoxy such as is sold by Leco, Buehler and others. It seemed to reduce the need for hardener. Since it adds mass to the resin, you will need to recalculate the resin to hardener ratio. You should also experiment with the mix before adding your important samples. We lost some due to the severe exotherm of the new mixture.
Depending on the size of your charcoal, you might be able to add some filler to the araldite to help locate the particles. I just received a sample of Ni powder from Buehler intended to make the epoxy conductive. I have not tried it so I don't know what particle sizes are present.
Warren
At 05:06 AM 11/21/2003, you wrote:
} Hello all, } Does anyone know of a stain for araldite resin to stain it } more grey or white under the SEM ? I'm investigating little } pieces of charcoal with a XL30SFEG and have some problems } with the black background caused by the resin. } } } TIA, } Erwin Vermeij } Forensic Scientist Microtraces } Netherlands Forensic Institute } Rijswijk, The Netherlands
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
While evaluating the latest in digital SEM technology, I notice a few manufacturers are still specifying the size of their frame stores, and one even claims acquiring 2560x1920 with a 1280x960 frames store. My 1st question might be how is this accomplished?, ... but my primary question would be, with modern computer interface speed and RAM, why is a frame store even necessary?
tia & cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 21 09:54:58 2003
} } } -------------------------------------------------------------------------- ---- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------------- ----- } } Dear Friends, } } A user of the lab has asked me to separate urban myth from reality } regarding HBO bulbs used for fluorescence microscopy. } } His questions are: } } What is the difference between a 50W bulb and a 100W bulb? He says he set } up a demo, identical except one scope had a 50W bulb, the other a 100W } bulb, and he didn't think he could tell much difference. } } Do bulbs have a finite life, ie., should they be changed at some maximum } hours even if still lighting up and appearing OK? He has heard various } advice, like change after 200 hours or 250 hours. Change when the glass } gets dark, how dark? Is it true that bulbs with many hours may explode?
The HBO 100w lamps put out more light. There are issues of lamp alignment and condensing lens choices for the lamphouse which can have a dramatic influence on the amount of light that reaches the specimen. If your fluorophore is bright you may be able to use the 50 watt lamp and not notice much difference between the two. If there is not much fluorescence you may not be able to see it at all with a 50 watt lamp.
The 100 watt lamp is much more stable then the 50 watt lamp. There is nothing more irritating then using a microscope with the light intensity flickering wildly. The 100 watt lamp is not likely to do this.
The rated life of the lamps is 100 hours for the HBO 50w AC and 200 hours for the HBO 100. Osram says that you can safely exceed the rated life by 20% without danger of bulb explosion. The bulb will explode if run too long, based on several factors. The rated life is determined by the factory to be the point at which the light output has been reduced by 30% from the initial output of a new lamp. A lamp driven by a properly operating power supply should look new when you change it based on maximum allowable hours. How dark it is should not be a criteria. Some people run their lamps much longer then is safe. They may have power supplies which help them do this, they may get lucky... If not, the damage done inside the lamphouse by an exploding lamp will cost $300-500 to repair. You can purchase a lot of lamps for that much money and also avoid the dubious pleasure of breathing poisonous mercury fumes. These lamps have significant amounts of mercury in them, please dispose of them properly.
The lamps cost the same amount, but the life is different. You should be paying under $100 US for them. Shop around. It seems like quite a bargain to get twice the power of the 100w, for half the cost of a 50w (based on lamp life).
We have had the best luck with Osram lightbulbs based on the 3,000 or so we have purchased over the last 20 years, and have found no difference in price which would warrant using another brand. In general we have found no significant differences in performance either.
The lamp must be run for 20 minutes at a minimum every time it is turned on or it may be ruined. This is the only use issue with this type of low wattage mercury lamp. The newer power supplies are different then the old ones some people remember and so stories of what "used" to work are of little help today. The newer power supplies will run every lamp to its rated life if the power supply is working correctly. We see lots of power supply problems which cause short lamp life. HBO power supplies seem to be problematic for some reason. If you purchase a new system and experience short lamp life I would have the supplier of your system install and align a new lamp for you (showing you the proper way to install it). If the lamp still does not reach its rated life, insist that they swap out a known, good power supply for you to try. In my experience that invariably fixes short lamp life problems...
Good luck, David Burton Optical Specialist University of Washington
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 21 13:26:36 2003
Before you buy a printer make sure the piezography (or MIS or Lyson) system will work with it. Several leading printer manufacturers have put proprietary chips on their ink cartridges so you are forced to buy their cartridges. In the past it has taken several months (at least) for the folks a piezography to 'engineer' a way around this. All of the inkjet black and white prints (piezography and others) I have seen have been on non-glossy paper so I don't know how good the final result will be for traditional EM work. From what I read in the photography press, talking to users and what I have seen a exhibts, there are a lot of variables (dye-based or pigment-based ink, paper, calibrating the monitor so what you see is what you get, etc.) to work out before getting photo-quality B&W prints that do not have a color cast. Study prints are another matter. If you are going to make a lot of prints a continuous ink system will save a lot of money in the long run. Have you ever worked out the cost of cartridge ink on a per ounce basis? Shocking! How about sending piezography a jpeg or tiff image and see what they can do for you before spending your money? They could probably answer your cost questions as well. By the way, I get very nice glossy B&W prints from my Epson stylus 1280 if I check the box that allows me to print without color ink. As much as I love a real silver print, inkjet printing is very fast and convenient. Since most journals are moving to digital submission of figures (or have already done so) I don't think the longevity issues with dye-based inks will be a problem.
Geoff
Debby Sherman wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 21 14:16:40 2003
If not a frame store, then what option would be used?
The idea of a frame store, I believe, is that once an image is captured in the store, it can be annotated, measured and then saved to disk. Direct capture does not allow in-SEM annotation.
The other factor to consider is that most of the frame stores are the same aspect ratio of the Polaroid 4x5 film.
gary g.
At 07:39 AM 11/21/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 21 15:26:01 2003
We're having a difficulties imaging CFP expressing live cells with a 405 nm laser because of autofluorescence, unhappiness with light and corrections (or lack of) in the objectives.
Has anybody tried the Pointsource 440 nm laser for CFP or have other suggestions for a laser source ideally in the 430 to 436 nm range?
Thanks.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 21 15:36:37 2003
Warren's suggestion to use iodoform is a good one.
Regarding his comment on commercial conductive embedding media. The problem with the conductive embedding media that I have used and/or read about is that they are have only macroscopic electrical conductivity. This means that the material will conduct a current but, on a microscopic scale, will contain expanses of nonconductive resin that will charge under the electron beam.
My suggestions are to use polymers that contain high loadings (~50 w%) of carbon black or other conductive particle with small particle size. I presented a paper at the 2003 M&M conference (see proceedings) on the subject. The approaches that I have used are (1) carbon black-filled epoxy resins and (2) carbon black-filled thermoplastics. Regarding the choice of medium, the epoxy component of the first material is available to infiltrate into porous samples. The higher viscosity, low crystallinity plastic component of the second preparation is unable to infiltrate into microporous samples and encapsulates the sample rather than infiltrating into it. One advantage of this prep is that a porous sample is not adulterated by the infiltration of an epoxy resin into it. An advantage of the first technique is that epoxy infiltration into the sample may allow microtomy of ultrathin sections for TEM.
Hope that this provides some guidance in finding a better preparation. Feel free to write off-line for more details.
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Warren E Straszheim {wesaia-at-iastate.edu} To: Microscopy-at-msa.microscopy.com cc: Erwin Vermeij {e.vermeij-at-nfi.minjus.nl} Subject: [Microscopy] Re: Stain for araldite resin 11/21/03 09:13 AM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If you don't mind "staining" before the fact, I had a similar problem examining coal in epoxy many years ago. Our solution was to dissolve iodaform in the epoxy resin before mixing with the hardener. We could dissolve up to about 18% iodaform by weight by using mild heating to speed the process. I think the iodaform would precipitate out if we tried to push
it further. We were trying to do automated analyses and they worked reasonably well at that level of doping. If you just need to find particles
by eye, then lower levels should be adequate.
For the record, this was a simple two-component epoxy such as is sold by Leco, Buehler and others. It seemed to reduce the need for hardener. Since it adds mass to the resin, you will need to recalculate the resin to hardener ratio. You should also experiment with the mix before adding your important samples. We lost some due to the severe exotherm of the new mixture.
Depending on the size of your charcoal, you might be able to add some filler to the araldite to help locate the particles. I just received a sample of Ni powder from Buehler intended to make the epoxy conductive. I have not tried it so I don't know what particle sizes are present.
Warren
At 05:06 AM 11/21/2003, you wrote:
} Hello all, } Does anyone know of a stain for araldite resin to stain it } more grey or white under the SEM ? I'm investigating little } pieces of charcoal with a XL30SFEG and have some problems } with the black background caused by the resin. } } } TIA, } Erwin Vermeij } Forensic Scientist Microtraces } Netherlands Forensic Institute } Rijswijk, The Netherlands
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (david.audette-at-sylvania.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 21, 2003 at 15:31:00 ---------------------------------------------------------------------------
Email: david.audette-at-sylvania.com Name: Dave Audette
Organization: OSRAM Sylvania
Title-Subject: [Microscopy] MListserver:
Question: I have an EDS dewar (Kevex) system which is not holding liquid nitrogen as long as it use to and was planning on warming it up and pumping on the inside chamber via the external valve. My question: Is there a, hopefully simple, way to regenerate the zeolite that I have heard is in side this chamber? Or is it not worth that much trouble?
I am also a high school teacher. I am not an expert, I lurk on this list to get ideas. I think I can answer this question if you haven't gotten other answers.
Do you have any more questions about viewing skin cells? They are pretty easy to spot after you figure out what's there! Using Methylene Blue, the nuclei should be stained. You can use Iodine (even Iodine soap works fine), and in this case the nuclei are stained brown.
Skin cells are pancake like, very thin and broad---which makes sense if you think about what they are. They are transparent, which is one reason to use stain.
You could try oblique illumination, if you have the same school microscopes that we have: the disk diaphragm is the circular disk underneath the stage. If not, you have an iris diaphram---better, but the disk works better for us for oblique illumination. You can darken the background to some extent by tweaking the disk to one side of one of the click stops, just a little bit. On our scopes, number 2 works best, on the disk, don't know what you have. The light that passes through the specimen is directed to the side of the objective, so the background is dark; but any light that is refracted or diffracted through certain parts of the specimens towards the objective now gives the appearance of a bright subject against the dark background. This works for amoebas too, and small plankton. It's a good way to see movement in a sample of water.
Good luck. Please don't hesitate to ask any more questions. The people on this list are experts, I'm not, but as a teacher like yourself, who, by the way, is a keen amateur microscopist, I am happy to help you. I use the microscope a lot in my biology classes. The students love it.
Alan Davis Marianas High School Saipan, N. Mariana Islands
On Mon, 17 Nov 2003 14:28:05 -0600 Donnarae48-at-AOL.com (by way of MicroscopyListServer) wrote:
} } } --------------------------------------------------------------------- } --------- The Microscopy ListServer -- Sponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } --------------------------------------------------------------------- } ---------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (Donnarae48-at-AOL.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, } November 16, 2003 at 14:43:02 } --------------------------------------------------------------------- } ------ } } Email: Donnarae48-at-AOL.com } Name: Donna Morrow } } Organization: Moreau Catholic High School } } Education: 9-12th Grade High School } } Location: Hayward, California } } Question: My biology class did a lab using the microscopes. We are } able to magnify up to 400 magnification. We used a scraping from our } cheeks and dye to make a slide. We were asked to draw what we saw } and lable at least 2 things. I am not sure what I saw. At 400 } magnification how clear would an epithelial cell be and would I have } been able to see a nucleus. All I saw were blue circles and light } blue background and very small round things scattered around. I am } wondering if the larger dark blue circles were a nucleus and the } smaller specks the cell membranes even though they did not seem to be } uniform. The teacher said my slide looked good but I am not sure } what I was viewing. Also do you know of any other sight that is } available to help me with my lab work with the microscope. We have } been trying to find a sight that would show me what an epithelial } cell would look like at 400 magnification and I keep running in to } brick walls. Thank you for your help } } --------------------------------------------------------------------- } ------ } }
-- adavis-at-saipan.com 1-670-235-6580 Alan E. Davis, P. O. Box 506164 CK, Saipan, MP 96950, NMI
I have steadily endeavored to keep my mind free, so as to give up any hypothesis, however much beloved -- and I cannot resist forming one on every subject -- as soon as facts are shown to be opposed to it. -- Charles Darwin (1809-1882)
The right to search for truth implies also a duty; one must not conceal any part of what one has recognized to be true. -- Albert Einstein
As we enjoy great advantages from the inventions of others we should be glad of an opportunity to serve others by any invention of ours, and this we should do freely and generously. -- Benjamin Franklin
From MicroscopyL-request-at-ns.microscopy.com Sat Nov 22 19:32:26 2003
Dave, Certainly, in part, it depends on what you're paying for LN2, but if the vacuum has deteriorated, then it will also aggravate any oil condensation on the detector snout. I haven't done this myself, but I've been told that if you put hot water in the dewar and use a hot air gun on the outside, you should be able to drive the water out of the zeolite. Room temperature takes care of the adsorbed gasses. Hopefully, someone who has done this can also respond. Ken Converse owner Quality Images third party SEM service Delta, PA
by way of MicroscopyListserver wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sun Nov 23 11:37:48 2003
During some testing and debugging today I inadvertently sent out a number of test messages, some of you will see a handful of test messages, which were not suppose to reach the world but were supposed to be "trapped" locally.
Please excuse the slip up.
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 24 08:39:07 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (oneild-at-hfx.nrc.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 24, 2003 at 08:26:14 ---------------------------------------------------------------------------
Question: Does anyone have any experience with the - Cressington 108 Auto or the Polaron/Quorum SC7620 "Mini" - Sputter Coaters that they are willing to share? Thanks
David O'Neil Institute for Marine Biosciences National Research Council of Canada 1411 Oxford St. Halifax, NS B3H 3Z1 ph. 902-426-8258 fax 902-426-9413 david.o'neil-at-nrc.ca http://www.imb.nrc.ca/techdev/microscopyfac_e.html
I am looking for protocols on how to prepare mushroom spores for scanning. I am very much interested in the practical side of things, e.g. in what to put the spores to pass them through the different solutions and critical point drying. All tips & tricks are welcome.
We have a researcher who needs to select and tack 2 to 5 micron particles on to a slide in a small spot over a 30 minute period, then put epoxy on the top, polymerize, polish, and analyze with SIMS. We need something that will stay sticky on the slide for the 30 minutes. I am not familiar with the methods for this. We've had nitrocellulose suggested, as well as B72. Would double sided sticky tape work? They must all be in the same plane as much as possible. Any suggestions? Many thanks, Nancy
Nancy Piatczyc Bronfman Science Center Williams College Williamstown, Ma
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 24 16:31:18 2003
Nancy Piatczyc wrote: =================================================== We have a researcher who needs to select and tack 2 to 5 micron particles on to a slide in a small spot over a 30 minute period, then put epoxy on the top, polymerize, polish, and analyze with SIMS. We need something that will stay sticky on the slide for the 30 minutes. I am not familiar with the methods for this. We've had nitrocellulose suggested, as well as B72. Would double sided sticky tape work? They must all be in the same plane as much as possible. Any suggestions? Many thanks, Nancy ===================================================== This sounds like the perfect application for the SPI Tacky Dot Slides, see URL http://www.2spi.com/catalog/new/tacky.shtml
You would use for the mounting apparatus, the Circle Analytical Mount which would align your particles within the two circular areas and then using the Teflon® Block Mount, embed the mounted particles, right on the slide with your peferred resin.
When the resin is cured, it will slide out of the Teflon mount and the particles will be perfectly aligned and parallel with the flat face of the plastic mount, which can then be polished down exactly as you have indicated The final result of that kind of cross-sectional polishing can be seen on URL http://www.2spi.com/catalog/new/tackdot_array.html
For particles in the indicated size range, I would suggest a 15 um size for the Tacky Dot Slides.
Disclaimer: SPI Supplies manufactures Tacky Dot Slide arrays and therefore has a vested interest in seeing them used by more workers.....
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 24 17:39:01 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tgruber-at-phelpsdodge.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 24, 2003 at 13:12:47 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: Accuview 789 Camera for Donation
Question: Dear Listers,
My company has a c. 1996 "Gatan Accuview Camera Model 789" available for donation to a suitable recipient. This is a 1.4 megapixel slow-scan format. It is interfaced via a camera interconnect cable to a Kodak Megaplus Camera Control Unit. The camera controller output port is 37-pin male and the output cable is provided. Camera, cables, and controller were working fine when their use was discontinued, and all will be donated together, in ìas-isî condition. This camera was formerly operated through Gatan software and a framegrabber board installed on a Macintosh computer (not available from us). The camera is c-mount and has a macro zoom lens mounted.
This donation is contingent upon my company being able to realize a charitable tax deduction relative to our depreciated book value of this equipment (c. $8,000). Non-profit institutions qualified to receive such a donation will be considered.
Eligible and interested parties may reply to me (OFF-LINE, PLEASE, at tgruber-at-phelpsdodge.com).
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (em_man_1-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 24, 2003 at 17:16:13 ---------------------------------------------------------------------------
Email: em_man_1-at-hotmail.com Name: William McManus
Organization: J & M Analytical, Inc.
Title-Subject: [Microscopy] [Filtered] looking for EM equipment
Question: After 20 years, the EM facility which I managed at Utah State University has been closed. I am expanding my consulting company and looking for good used equipment. I am particularly interested in aquiring a Zeiss (LEO) 902 TEM. I am also looking for Ultramicrotomes and SEM prep equipment. If anyone has equipment that they would like to sell, please contact me. Bill 435-946-8739
Hi, I'm posting this enquiry for a gentleman who rang me today. He's acquired an LKB Nova Ultratome (circa 1985) through an auction, however, it's missing the accompanying electronics box (ie control box). Does anyone have a spare control box they are willing to part with?
Thanks,
John Brealey EM Unit Queen Elizabeth Hospital Adelaide, South Australia
(08) 8222 6612
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 24 21:39:09 2003
We can writing alkanethiols onto a gold surface, by dip-pen nanolithography. Can we remove or alter the patten (like using an eraser) we had created if the sample is biological such as E-coli, cells or single peptide? If so how can we do that?
I do appreciate your kindness.
Sincerely yours,
Chris
===== Life is like a box of hand grenades, you don't know what will blow you to kingdom come.
Mr.Yung-fou Chen E-mail: yung_fou-at-yahoo.com
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From MicroscopyL-request-at-ns.microscopy.com Tue Nov 25 02:16:40 2003
Thanks for the reply. I figured that this might work, but I also found this protocol:
Small pieces of the lamellae (herbarium material) were used. They were pretreated for 12h in water and ammonia. After a period of 1h in 70% ethanol, they were passed 2 x 30 min. in dimethoxymethane, before being submitted to the process of critical point-drying. The material was coated with gold (with Argon-gas, under 0.05 bar) for 3 min. 30 sec., until a layer, 15 nm in thickness, is covering the material. (The scanning electron microscopy was carried out with a JEOL 5800LV with tension of 25kVolt and a working distance op 7-8 mm.)
Can you tell me why or when one should use this protocol?
Regards,
Renaat
----- Original Message ----- } From: "Rosemary White" {rosemary.white-at-csiro.au} To: "Renaat Dasseville" {Renaat.Dasseville-at-UGent.be} Sent: Monday, November 24, 2003 11:06 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (h.grenfell-at-geomarine.org.nz) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 24, 2003 at 18:21:41 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: Any ideas?
Question: Hi, Can anyone help with info as to what these minute ?siliceous objects are? Please see images at http://homepages.ihug.co.nz/~bw.hayward/SEM%20images/
They are commonly found in estuarine samples that we are studying using foraminifera. Would be nice to know what these spheres are trying to tell us!!
They are not radiolaria and many tell us they are of sponge origin but no detail or images from the literature as yet.
Please note the final date for receipt of Abstracts for presentation at the Australian Conference for Microscopy and Microanalysis 2004 (http://www.deakin.edu.au/events/acmm18/) and for registration at the series of workshops scheduled to precede the conference is :
Wednesday 26, November
Please also note that response to our call for papers has been outstanding! In order for abstracts to be considered for oral presentation we must have received them by close of business on Wednesday.
Abstracts received after this date may well not appear in the official proceedings, but form part of an addendum (if received in sufficient time).
Lynne Lucas Lynne Lucas, Manager, Deakin Event Management Services Deakin University Geelong Victoria 3217 Australia. Phone: 03 5227 8121 International: +61 3 5227 8121 E-mail: luco-at-deakin.edu.au Website: http://www.deakin.edu.au Deakin University CRICOS Provider Code 00113B
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From MicroscopyL-request-at-ns.microscopy.com Tue Nov 25 07:51:19 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tholzheu-at-camet-lab.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 24, 2003 at 22:27:40 ---------------------------------------------------------------------------
Email: tholzheu-at-camet-lab.com Name: Thomas Holzheu
I am probably pushing my luck here, but then you nerver know. We have an EDXRF spectrometer with a kevex 8000 system and an iomega bernoulli box (scsi). Upon booting the DEC computer, the system is accessing the first drive, but is not loading the software. The issue here is, that I want to replace the bernoulli box with a superdisk drive (scsi), but just exchanging the drives is not working. I would appreciate any input and thoughts on this matter.
Hi Renaat, I would recommend not using any chemicals at all.
You can collect the Lactarius basidiospores by making a spore print: Remove the stalk. Place the cap on a piece of paper for several days (dark paper for light spore and vice versa). The spores will be discharged onto the paper. Remove the cap. Put a carbon sticky tab on a SEM stub and lightly touch the stub to the spore print.
Experiment with the spores after you mount them. Place some in a desiccator for a few days before coating them, and others try coating and scoping right away (when they are fresh like that it is harder on the scope - it takes longer to pump down).
Also, the scoping part depends on your scope. If you have a cryo-SEM then by all means use it. If you don't have the cryo attachment then coat the samples and use low kv (if possible) or scope them quickly at 10 or 15 kv so they don't collapse or look toasted.
have fun with fungi, Beth
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-- ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Thomas, Contact tech support for Thermo Kevex x-ray Tel: 831.461.2326 Fax: 831.438.5892 E-mail: chrisc-at-kxr.com Let them know what you are doing and ask them for tech support. Last year I had trouble with my 8000 and they put me in contact with a tech that really knowledgeable about the system. I think he is in Georgia, but they will connect you to him.
Rich L CMD Tempe
-----Original Message----- } From: by way of MicroscopyListserver [mailto:tholzheu-at-camet-lab.com] Sent: Tuesday, November 25, 2003 7:03 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tholzheu-at-camet-lab.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 24, 2003 at 22:27:40 ---------------------------------------------------------------------------
Email: tholzheu-at-camet-lab.com Name: Thomas Holzheu
I am probably pushing my luck here, but then you nerver know. We have an EDXRF spectrometer with a kevex 8000 system and an iomega bernoulli box (scsi). Upon booting the DEC computer, the system is accessing the first drive, but is not loading the software. The issue here is, that I want to replace the bernoulli box with a superdisk drive (scsi), but just exchanging the drives is not working. I would appreciate any input and thoughts on this matter.
The deadline for all nominations for 2004 Microscopy Society of America awards is nearing (December 15, 2003). Please note the following announcement that was previously posted in October:
Call for Nomination of Individuals to be considered for Major Awards by the Microscopy Society of America.
Awards:
Distinguished Scientist Awards:
These Awards recognize preeminent senior scientists from both the Biological and Physical disciplines who have a long-standing record of achievement during their career in the field of microscopy or microanalysis..
Burton Medal:
The Burton Medal was initiated to honor the distinguished contributions to the field of microscopy and microanalysis of a scientist who is less than 40 years of age on January 1st of the award year.
Optical Imaging Association-MSA Outstanding Young Investigator Award:
This Award, initiated in 1999, recognizes the distinguished contributions in the field of optical microscopy made by a scientist who is less than 40 years of age on January 1st of the award year.
Outstanding Technologist Awards:
These Awards honor technologists from both the Biological and Physical Sciences who have made significant contributions such as the development of new techniques which have contributed to the advancement of microscopy and microanalysis.
Morton D. Maser Distinguished Service Award:
This Award was initiated to recognize outstanding volunteer service to the Society as exemplified by Mort Maser, who served the Society for many years with great dedication. This award is made to honor an MSA member who has provided significant volunteer service to the Society over a period of years.
Nomination Requirements:
The Distinguished Scientist, Burton Medal, OIA-MSA Outstanding Young Investigator and Outstanding Technologist Awards Nominations should include:
1) a letter from the primary MSA nominator describing the research accomplishments of the candidate with particular emphasis on the unique technical achievements in the Physical or Biological Sciences; and 2) supplemental letters of support from other members of the scientific community.
The Morton D. Maser Distinguished Service Award Nomination should include:
1) a letter from the primary MSA nominator describing the basis for the nomination; and 2) supplemental letters of support from other members of MSA.
The Deadline for receipt of Awards Nomination Packages is December 15, 2003.
Please contact the MSA Business Office for additional information.
Judy Janes, Administrative Manager Bostrom Corporation 230 E. Ohio Street, Suite 400 Chicago, IL 60611-3265 (800) 538-3672; Fax (312) 644-8557, jjanes-at-MSA.microscopy.org
Thanks,
William T. Gunning, Ph.D. Associate Professor of Pathology Medical College of Ohio Department of Pathology BHSB 140 3035 Arlington Avenue Toledo, Ohio 43614-5806 Phone: 419-383-5256 Fax: 419-383-3066 email: wgunning-at-mco.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 26 03:34:10 2003
I have some questions regarding the quality of microplates for microscopy, mainly 96- and 384 well SBS-standard layout formats (1536 and 6144 layouts would be interesting too of course).
I have been doing some quality tests for certain types of multiwell plates which are very popular for use in "readers" with our 40x 0.75 N.A. objective and a 63x 0.8 objective in brightfield and fluorescence. These objectives have a very good spatial resolution and capture a lot of light for weak fluorescence (I ~ N.A.^4/M^2). However these objectives have a short working distance and as such the quality of the plate bottoms is important.
Our autofocus system only needs about 5 videoframes to gather enough information to find a focus level, so with a 40 millisec. frametime (PAL 25 fps.) and some time for mechanical movement, it does a complete image content based autofocus in 0.3 or 0.4 seconds (300 or 400 milliseconds). But this only is the case, when a focuslevel is found within a certain range, i.e. the travel range needed for 5 frames, which is in principle determined by the Nyquist sampling theorem for Z-slicing.
We also do a complete range check, as we want to focus in those microscopy modes which can cause multiple maxima in a focus algorithm (patented) in which case we focus on the highest peak/score found ( Geusebroek J.M., Cornelissen F., Smeulders A. W. M., and Geerts H., Robust autofocusing in microscopy, Cytometry, 36(1):1-9, 2000 ).
Using an image content based autofocus system, allows us to use very fine Z-level focusing into cells with a varying offset to the multiwell plate bottom.
I found out that when you plot the bottom profile of standard multiwell plates as a 3D landscape, these plates look more than a mountaneous region than having a flat profile.
I also found some microplate types with very thin plastic bottoms and a flat plate bottom, but they are of course a bit more expensive. Glass bottom plates are an alternative in some cases, but I heard that the glue which is used for these plate-bottoms is not compatible with some of the solvents used in farmaceutical research (DMSO, etc), so the bottoms tend to fall off the plate after 24 h. incubation, which would be rather inconvenient for our time-lapse work :-( ? So using good quality plastic bottom plates seems to be one possible solution ?
A possible solution for the variability of the plate bottoms is to do a prescan at lower mag./N.A., either with a plate bottom finding laser-system or a content-based autofocus and model the plate with a mathematical model, but this takes time.
Using Long Distance (L.D.) objectives is another possible solution, but these objectives have a lower N.A. and as such less spatial resolution, which hampers subcellular imaging and analysis. Also the reduced N.A. has a very dramatic efect on the amount of light captured by the objective (I ~ N.A.^4/M^2) and we always do our image capturing in real-time, we just use very sensitive cameras which allow imaging without the need to integrate frames. The image capturing must be done at 40 msec. (PAL 25 fps.), which is important when doing large volumes of multiwell plates in our screenings. Using the 0.75 and 0.8 N.A. objective in combination with the ultrasensitive camera allows us to dilute reagents up to 1000x, which is very good for our assay costs when doing large screens of subcellular phenomena :-)
I would like to know what is the experience of other researchers with the quality of multiwell plates on microscopes or similar (automated) readers ? Does anyone else uses high N.A. objectives on multiwell plates for screening ? As far as I understand the SBS-standards define XY-measures of multiwell plates, but leave the bottom offset and bottom thickness to the manufactureres of multiwell plates ?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ajitsankar.mallik-at-epfl.ch) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, November 26, 2003 at 04:16:42 ---------------------------------------------------------------------------
Email: ajitsankar.mallik-at-epfl.ch Name: Ajit
Organization: EPFL .
Education: Graduate College
Location: Lausanne , Switzerland
Question: Dear Sir / Madam , 1)I want to know the best visualizing methods for Endothelial cells directly on an artery . 2)Could you please let me know the best fluroscent probes for Endothelial cells if i plan to visualize them on confocal laser scanning microscope ?(least back ground fluroscence ) 3) I would like to know the live and dead endothelial cells on an artery by microscopy . Please suggest the best method .
Postdoctoral Fellow Australian Key Centre for Microscopy and Microanalysis, The University of Sydney, Australia. Reference No. A46/004354
Applications are invited for a Postdoctoral Fellow in the Australian Key Centre for Microscopy and Microanalysis to work on the research project, "Mapping electronic structure and material properties with atomic resolution". This project will use electron energy loss spectroscopy (EELS) to map the bonding and electronic structure of InGaN quantum wells and other materials at the atomic scale. We will measure and correlate the local composition, strain and electronic structure variations within the wells in order to understand the optical emission in the system. The development of EELS spectrum imaging tools for the mapping of bonding states in materials in an important aspect of this project.
Essential criteria for successful applicants include: a PhD or equivalent qualifications in a relevant discipline; extensive experience in the operation of FEG TEM/STEM systems; experience in acquisition of EELS and EFTEM data; proven research ability; evidence of research potential; and the ability to work cooperatively with other members of the university community.
Experience in ab initio calculations of electronic structure and EELS spectra, computer programming experience and demonstration and teaching skills would be desirable.
The position is full-time fixed term for one year, subject to the completion of a satisfactory probation period for new appointees. There is the possibility of further offers of employment for up to twelve months, subject to funding and need. For further information, contact Dr Vicki J. Keast on 9351 4534, fax 9351 7682 or e-mail vicki.keast-at-emu.usyd.edu.au
Remuneration Package: $60,535 - $64,981 p.a. (which includes a base salary Level A $51,153 - $54,910 p.a., leave loading and up to 17% employer's contribution to superannuation)
Closing: 18 December 2003 Applications (five copies) should quote the reference no, address the selection criteria, and include a CV, a list of publications, the names, addresses, e-mail, fax and phone number of three confidential referees.
Forward applications to: The Personnel Officer, College of Sciences and Technology, Carslaw Building, (F07), The University of Sydney, NSW, 2006 ------------------------------------------------------------------------------ Dr. Vicki J. Keast Australian Key Centre for Microscopy and Microanalysis Madsen Building F09 University of Sydney Sydney NSW 2006 Ph: + 61 2 9351 4523 Fax: + 61 2 9351 7682 email: v.keast-at-emu.usyd.edu.au
For correspondence regarding the Australian Microscopy and Microanalysis Society, please reply to amms.secretary-at-emu.usyd.edu.au ------------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 26 08:55:42 2003
I am trying to deliver some protein inhibitors into live worm sperm cell. The nice crawling cell has a long lamelipod and a cell body trailing behind. The lamelidod is about 30 micron in length and 10 micron in width. The tip of micropipette I used is about 1 micron in diameter. The injector I have is from Narishige and filled with mineral oil when used.
Every time when I touch the cell with the micropipette, the cells explode. Does anyone on the list have the same experience before?
Best regards,
Long
----------------------------------------------- Miao, Long Dept of Biological Science 334 Bio. Unit1 Florida State University Tallahassee, FL32306-4370 email: lmiao-at-bio.fsu.edu Voice: (850)644-9817 FAX : (850)644-0481 -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 26 11:52:44 2003
Hi, I have an investigator who is carrying out em immunolabeling on isolated dispersed filaments. The labeling is carried out on the grid but the filaments are not visible without negative staining. It would appear that the labeling procedures deposits considerable material on the grid which shows up as a thick layer and this obscures the filaments. Is this to be expected? Is it an inadequate washing procedure after labeling? Any thoughts?
Regards to this great resource as always
Chris
Christopher J. Gilpin Ph.D. Assistant Professor Director Molecular and Cellular Imaging Facility K1.246 Department of Cell Biology University of Texas Southwestern Medical Center 5323 Harry Hines Boulevard Dallas, Texas 75390-9039 +1 214 648 2827 Phone +1 214 648 6408 Fax christopher.gilpin-at-utsouthwestern.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 26 16:00:48 2003
Christopher- The solutions wouldn't ordinarily be dirty enough to deposit anything. I think he/she may be using too concentrated an antibody solution. Also, leaving the antibodies on for too long a time might accomplish the same thing.
Best of luck to you! Carol Heckman (Bowling Green State University)
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 26 19:06:45 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jtd1-at-psu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 26, 2003 at 08:56:20 ---------------------------------------------------------------------------
Email: jtd1-at-psu.edu Name: Tom Doman
Organization: Animal diagnostic Laboratory, Penn State University
Question: We are seeking to purchase a scanner to digitize 31/4" X 4" TEM negatives; the unit will be used as a dedicated film scanner for this format. The budget for the purchase is up to $700. We have selceted a Umax PowerLook 1000. Does anyone have any experinece with this scanner?
Chris In general, it's bad idea to do labelling on the grid. If you don't have very special reasons to do so, do it in vitro before grids. For good results, molar antigen-antibody ratio should be around 1:1, so you need to perform a serial dilution 1:2; 1:4; 1:8; 1:16 and so on to find optimal one. For negative staining, antibody should be homogeneous - IgG fraction preferably. It's also helpful to remove excess of material by gel-filtration etc... In general: for good consistent negative staining you need BIOCHEMICALLY (at least) homogeneous sample. If it's filaments, it should be filaments, not cell lysate. If it's specific IgGs - it should be IgGs, not serum, etc. Sergey
At 12:03 PM 11/26/2003 -0600, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Nov 27 03:12:34 2003
Hello, I use to do immunolabelling followed by negative staining on virus particles and do not have this kind of problem. You can see the procedure I follow in this paper : Spehner, D., R. Drillien, F. Proamer, C. Houssais-Pecheur, M. A. Zanta, M. Geist, K.Dott, and J. M. Balloul. 2000. Enveloped virus is the major virus form produced during productive infection with the modified vaccinia virus Ankara strain. Virology 273:9-15 Don't hesitate if you wish more informations good luck danièle
-----Message d'origine----- De : Christopher Gilpin [mailto:christopher.gilpin-at-utsouthwestern.edu] Envoyé : mercredi 26 novembre 2003 19:04 À : Microscopy-at-MSA.Microscopy.Com Objet : Immunolabeling and negative staining
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Hi, I have an investigator who is carrying out em immunolabeling on isolated dispersed filaments. The labeling is carried out on the grid but the filaments are not visible without negative staining. It would appear that the labeling procedures deposits considerable material on the grid which shows up as a thick layer and this obscures the filaments. Is this to be expected? Is it an inadequate washing procedure after labeling? Any thoughts?
Regards to this great resource as always
Chris
Christopher J. Gilpin Ph.D. Assistant Professor Director Molecular and Cellular Imaging Facility K1.246 Department of Cell Biology University of Texas Southwestern Medical Center 5323 Harry Hines Boulevard Dallas, Texas 75390-9039 +1 214 648 2827 Phone +1 214 648 6408 Fax christopher.gilpin-at-utsouthwestern.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 27 08:52:46 2003
We are looking for a micromanipulator to handle FIB manufactured TEM samples. We have references from Narishige (distributes in the USA by Tritech Research) but we need a europeen distributor.
By the way, advices from people with some experience in this domain are welcome.
Answer directly to Jacques.Werckmann-at-ipcms.u-strasbg.fr
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Dear all, Does anyone know much about low-loss EELS spectra of gold or silver. I am looking for information on bulk plasmon and surface plasmon energies and energy loss spectra of the region to about 30 or 40 eV I already tried the EELS database at CEMES in France (http://www.cemes.fr/~eelsdb/) and searching the literature but didn't find much. This is to try and explain spectra and energy filtered images of gold or silver nanoparticles.
Thanks for any clues you can give.
Best wishes
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 28 02:07:57 2003
have you tried the EELS Atlas by Ahn and Krivanek, published by Gatan in 1983. (Copies available from Gatan or from the Arizona State University. I can give you adresses.)
Old publications: M. Creuzburg (1966), Zeitschrift fuer Physik 196, 433-463 H. Froehlich and H. Pelzer (1955), Proc. of the Physical Society A, 68, 6, 525 -529
Philip
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em
The phrase 'We have always done things this way.' is as much a reason to change as a reason not to. - Dartwill Aquila _______________________________________
----- Original Message ----- } From: "Ian MacLaren" {maclaren-at-tu-darmstadt.de} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Cc: "Rik Brydson" {mtlrmdb-at-leeds.ac.uk} ; "Alan Craven" {a.craven-at-physics.gla.ac.uk} Sent: Thursday, November 27, 2003 4:55 PM
Dear all, Does anyone know much about low-loss EELS spectra of gold or silver. I am looking for information on bulk plasmon and surface plasmon energies and energy loss spectra of the region to about 30 or 40 eV I already tried the EELS database at CEMES in France (http://www.cemes.fr/~eelsdb/) and searching the literature but didn't find much. This is to try and explain spectra and energy filtered images of gold or silver nanoparticles.
Thanks for any clues you can give.
Best wishes
-- Ian MacLaren Technische Universitdt Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.ht ml
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 28 15:49:12 2003
Our company is considering buying an automatized fluorescence microscope (motorized fluorescence filter wheel, bright field illuminator, nosepiece, condenser, and equipped with a motorized stage).
Considering the quality of optics and reliability of motorized pieces (precision, durability), is there a manufacturer that would be recommended?
Thank you.
Marie-Claude Bélanger
_________________________________________________________________ MSN Search, le moteur de recherche qui pense comme vous ! http://fr.ca.search.msn.com/
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 1 02:44:40 2003
I was wondering if there is already something going on to set up a sort of "Human Cytome Project" ? In my opinion the hardware and most of the software seems to be avaialable to set up such a project ? For the cellular level, light-microscopy based reader technology would be very interesting to use ?
Studying and mapping the genome, transcriptome and proteome at the organisational level of the cell for various celltypes and organ models could provide us with a lot of information of what actually goes on in organisms in the spatio-spectro-temporal space ?
I have been thinking (working) about a concept which could provide the basic framework for exploring and managing this cellular level of biological organisation research on a large scale, but I would like to know if there is already some thought/work going on in the direction of setting up an initiative such as a "Human Cytome Project" ?
This is just an idea, so I am really interested to hear if there is something in it, or even if it is not worth while whet I just wrote.
Best regards,
Peter Van Osta
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 1 10:00:01 2003
I just got the C4P and Nov/Dec issue of MT. Thanks for getting the call for nominations inserted for me. I'm concerned about suitable nominations. We had trouble last year and we didn't award all awards last year; looks like a repeat may happen. Suppose that I shouldn't be too concerned just yet as the deadline is still 2 weeks away?
Bill
William T. Gunning, Ph.D. Associate Professor of Pathology Medical College of Ohio Department of Pathology BHSB 140 3035 Arlington Avenue Toledo, Ohio 43614-5806 Phone: 419-383-5256 Fax: 419-383-3066 email: wgunning-at-mco.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 1 14:47:53 2003
I am having difficulty with immunofluorescence staining of DMSO- differentiated HL-60 cells (a promyelocytic cell line induced to differentiate to a neutrophil-like phenotype). The main problem is nonspecific binding of the secondary reagent--in this case a FITC-labeled donkey F(ab')2 anti-chicken IgY causing very high background. Since anti-chicken reagents are in limited availability, I am going to try to get around using a secondary antibody by biotinylating my primary chicken antibody and then using labeled streptavidin as my secondary reagent. However, I have a mouse antibody that I would like to use as well, and my anti-mouse reagent also gives high background. Does anyone have any helpful pointers on blocking nonspecific binding (or specific, undesirable interactions i.e. with the Fc receptors) on these cells? I am currently blocking and doing my staining in a buffer containing 5% nonfat dry milk and 5% horse serum. I have tried adding in human IgG to block Fc receptors, and the secondary-only controls seem a bit lower, but this background is still unacceptably high.
Any help is greatly appreciated!
Thanks, Amy
Amy Sillman, Ph. D. Visiting Postdoctoral Scholar University of California, San Francisco San Francisco, CA 94143 http://www.ucsf.edu/barber/lab1.html
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 1 15:26:29 2003
Has anyone else been the LUCKY recipient of what seems to me to be one of the worst marketing gimmicks I have seen in a long time? I received a blue plush toy in the mail just before Thanksgiving that looks like some kind of exotic worm, although I think it is supposed to be aquatic, anyone else been sent one?
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48" AIM: thejfmjfm
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 1 18:55:59 2003
Is it just "materialization" of the virtual worms attacked us every day? If so, you have to present it to Norton Antivirus team for identification and including into their antiviral software. Sergey
At 01:23 PM 12/1/2003, you wrote:
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I am looking for a vendor to supply specimen stubs for the upper stage of an ISI DS-130 SEM. They are threaded and about 9 mm in diameter. Does anyone know of a source?
Thanks in advance,
Doug
--------------------- Doug Keene Associate Investigator Imaging Center Shriners Hospital for Children 3101 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 1 19:06:35 2003
John; I got one too, and I loved it. My 1-1/2 year old daughter didn't quite know what to make of it though, so she still prefers to yank my pager and cell phone out of my briefcase and play with them. Maybe when she gets a bit older and learns about microscopy...............
John Mardinly Intel
-----Original Message----- } From: John Mansfield [mailto:jfmjfm-at-umich.edu] Sent: Monday, December 01, 2003 1:24 PM To: microscopy-at-microscopy.com
Has anyone else been the LUCKY recipient of what seems to me to be one of the worst marketing gimmicks I have seen in a long time? I received a blue plush toy in the mail just before Thanksgiving that looks like some kind of exotic worm, although I think it is supposed to be aquatic, anyone else been sent one?
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48" AIM: thejfmjfm
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 1 22:52:26 2003
We are searching for a method in vivo labeling or staining of bladder epithelium and/or it can be detectable in tissue samples after bladder resection. I would appreciate any recommendations.
Thanks.
Robert Molchanov M.D.
Dpt.of Urology State Medical Academy Dniepropetrovsk Ukraine Phone/Fax +380 (562) 466563 E-mail: rob_molch-at-yahoo.com
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From MicroscopyL-request-at-ns.microscopy.com Mon Dec 1 22:54:49 2003
We are searching for a method in vivo labeling or staining of bladder epithelium and/or transitional-cell carcinoma of bladder so that it can be detectable in tissue samples after bladder resection. I would appreciate any recommendations.
Thanks.
Robert Molchanov M.D.
Dpt.of Urology State Medical Academy Dniepropetrovsk Ukraine Phone/Fax +380 (562) 466563 E-mail: rob_molch-at-yahoo.com
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From MicroscopyL-request-at-ns.microscopy.com Tue Dec 2 06:50:03 2003
I have done negative staining ( 1% PTA) on a budded virus for TEM and some of the virus species appears white while other, on the same gridd, have a darker appearence. Does anyone have any experience of this phenomenon? Is this depending on if the envelope membrane is lost or not?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (crimp-at-egr.msu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 2, 2003 at 08:08:12 ---------------------------------------------------------------------------
Email: crimp-at-egr.msu.edu Name: Marty Crimp
Organization: Department of Chemical Engineering and Materials Science
Question: The Department of Chemical Engineering and Materials Science at Michigan State University has an immediate opening for a postdoctoral level researcher with experience in both transmission and scanning electron microscopy of materials. This position will involve a combination of lab supervision, training, and research. Applicants should possess both strong communications skills and experience with a broad range of electron microscopy techniques. An earned Ph.D. in materials science or related field is required.
Please send cover letter, resume, and a list of three references to: Prof. Martin A. Crimp, Dept. of Chemical Engineering and Materials Science, Michigan State University, Lansing, MI 48824-1226. MSU is an Affirmative Action/Equal Opportunity Institution. Women and minorities are strongly encouraged to apply. Persons with disabilities have the right to request and receive reasonable accommodation.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (starovoytov-at-biology.rutgers.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, December 1, 2003 at 13:39:26 ---------------------------------------------------------------------------
Question: We have two scopes that we are removing from our facility. Does anyone want a "Hitachi S-450" SEM or "Philips EM-300" TEM? Both scopes are available for the cost to relocate them to your facility. Both are in good, operating condition.
PLease e-mail or call Val Starovoytov at 732-445-5308
That was pretty common in our studies of the Mammary Tumor Virus. Are you seeing osmotic artifacts as well? In our studies of MTV, the virus often appeared as a "head" with a membranous tail. You can check some of the other ways we prepared the virus in: Cancer Research 35:740-749 (1975)
Joel
} } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Dear Microscopy Listserver members, } } I have done negative staining ( 1% PTA) on a budded virus for TEM and } some of the virus species appears white while other, on the same } gridd, have a darker appearence. Does anyone have any experience of } this phenomenon? Is this depending on if the envelope membrane is } lost or not? } } Sincerely, } Pernilla } } ...................................................................... } ..................................................................... } } Pernilla Nevsten, PhD-student } Materials Chemistry } Lund University, Sweden } } } } } }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 2 11:00:49 2003
congratulations, you are about to receive a deluge of comments recommending different stains. the best advice here is to stick with what you are familiar with. unless you wish to make a comparative study of stains. if you wish to do that let me know and i will share my results from previous studies. the bottom line is that most stains are inherently similar in results. some are more grainy, some sublimate, some are reputed to cause damage to the particles. but if you are happy with what you have, stick with it.
the first question is have you stabilized and inactivated the particles in any way. we use a final concentration of 0.1% glutaraldehyde. as we all know, it is a cross-linking fixative, and therefore, may cause clumping of the material. in our case, we find clumping is not really a problem in clinical samples. also, at a concentration of 0.1% the antigenic sites are retained sufficiently to allow immunoEM procedures to work. alternatively, if you are concerned with clumping you may wish to treat the particles with 2% paraformaldehyde.
the second question is did you adjust the pH of the PTA. it is usually used at a pH of 7.0. if the pH is adjusted with KOH the stain is reported to cause disruption of membranes, whereas adjustment with NaOH will not. of course, fixation should protect against this disruption. if the appearance you see is due to disruption of the virion membranes then the disrupted particles should release the nucleocapsid, which should then be recognizable on its own.
a more likely explanation for the appearance you are seeing is the presence and absence of nucleic acid in the particle. as we all remember, negative staining works by embedding the target particle. good stains will penetrate well into the interstices of the particles. cryoEM studies suggest that most of these small spaces are actually channels which may penetrate ccmpletely into the particle. where there is nucleic acid present, it fills the interior of the particle and prevents excess stain from building up. were there is no nucleic acid present, the stain will fill the inside of the particle. the stain will then deflect the beam in the internal areas, providing for a dark appearance. remember, in negative stain, dark=nothing, white=components.
this appearance is observed with many viruses after gradient purification. in the case of cubic viruses purified on CsCl, the genome defective particles band above the genome complete particles, and are frequently refered to as 'top component'.
if this is not a sufficient explanation, i would be willing to look at a few micrograps if you wish to forward digitized images.
hope this helps
Paul R. Hazelton Department of Medical Microbiology and Infectious Diseases University of Manitoba, Faculty of Medicine 531 Basic Medical Sciences 730 William Avenue Winnipeg, MB Canada, R3E 0W3 phone: Lab: 204-789-3313 Fax: 204-789-3926 Cell: 204-781-1502 Pager: 204-931-9354 e-mail: paul_hazelton-at-umanitoba.ca
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 2 11:54:11 2003
Pernilla, What you are probably seeing is both positive (dark virus) and negative (light virus) staining. True negative staining gives a light particle surrounded by stain which, due to the scattering of electrons, appears darker. You will often also see some darker areas of the particle that have trapped some stain.
Positive staining is when the particle itself has absorbed the stain and appears dark while the background is light. Usually there is very little detail in positively stained particles.
Positive staining often occurs when the amount of sample material is very low. In this case the hydrophobic nature of the film surface results in the stain just rolling off. You can see this happening if you put a droplet of stain on the grid and, when you go to wick it off with filter paper, it all comes off leaving an apparently dry grid behind.
You can reduce this problem by glow discharging the grids in a vacuum evaporator just prior to making your samples. This acts to change the charge on the grid surface and make it hydrophilic. This effect does degrade quickly so grids should be used within a few hours. You can glow discharge grids more than once.
The alternative is to use a sample with sufficient concentration of particles so that there is enough material to hold the stain. It is not unusual to find areas of both positive and negative staining on the same grid based on where the sample has accumulated. The nice thing about negative staining is that you have a fairly homogeneous sample so usually can hunt around for just the right sample and stain distribution.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 On 12/2/03 7:52 AM, "Pernilla Nevsten" {pernilla.nevsten-at-materialkemi.lth.se} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Dear Microscopy Listserver members, } } I have done negative staining ( 1% PTA) on a budded virus for TEM and } some of the virus species appears white while other, on the same gridd, } have a darker appearence. Does anyone have any experience of this } phenomenon? Is this depending on if the envelope membrane is lost or not? } } Sincerely, } Pernilla } } .............................................................................. } ............................................................. } } Pernilla Nevsten, PhD-student } Materials Chemistry } Lund University, Sweden } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 2 12:05:12 2003
} Virus will appear differently (some darkly stained, some lightly } stained) for various reasons:
1. Stain penetration: sometimes the integrity of the virion is disrupted (mechanically, chemically) so that the negative stain can penetrate the particle and deposit inside the virion giving it a dark appearance.
2. Defective particles: in most replicating virus there will be a certain percentage of defective (empty) particles. These are generally "leaky" and permit stain to penetrate. If the population contains too many defective (non-infectious) particles, this leads to a phenomenon called defective interference and could eventually lead to the loss of viability of the virion (infectious particles).
3. Variability in staining: negative stains are rarely exclusively negative and there will always be some degree of positive staining taking place with the stain. Uranyl acetate, for example, often reacts intensely with DNA if it is accessible and PTA sometimes stains polysaccharides (external components of membranes).
} Dear Microscopy Listserver members, } } I have done negative staining ( 1% PTA) on a budded virus for TEM } and some of the virus species appears white while other, on the same } gridd, have a darker appearence. Does anyone have any experience of } this phenomenon? Is this depending on if the envelope membrane is } lost or not? } } Sincerely, } Pernilla } } ........................................................................................................................................... } Pernilla Nevsten, PhD-student } Materials Chemistry } Lund University, Sweden
-- ############################################################## 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 ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 2 12:30:05 2003
} Has anyone else been the LUCKY recipient of what seems to me to be one } of the worst marketing gimmicks I have seen in a long time? } I received a blue plush toy in the mail just before Thanksgiving that } looks like some kind of exotic worm, although I think it is supposed } to be aquatic, anyone else been sent one? } Dear John, No plush toys, but I seem to have won an unbelievably large number of lotteries. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 2 14:07:24 2003
In a message dated 12/2/03 2:50:59 PM, tivol-at-caltech.edu writes:
} No plush toys, but I seem to have won an unbelievably large number of } lotteries.
And it has been my incredibly good fortune to be selected as the one honest person to be entrusted with enormous wealth that has found its way by various ill-gotten means into the hands of numerous persons in various third world countries, all of whom apparently wish to transfer it into my personal bank account. Anyone want to share in this windfall?
Of course, I will probably need all that money if I am to take advantage of the great deals on prescription drugs that I can order on-line, or to visit the numerous casinos that want my patronage.
Thank goodness for spam-blockers, which reduce the flood somewhat.
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 2 15:14:41 2003
John as a long standing customer of that company you should be happy to have received their M&M 2003 giveaway ... only 3 months late but then that is pretty good for them! I received one too.
Alan
At 04:23 PM 12/1/2003 -0500, John Mansfield wrote:
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Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Dear Dr. Osta, I remember vaguely reading about cytomics and what I remember is that it related to complete protein profile in relation to functions ( somewhat like functional genomics) also involving microscopy and facs etc but of single cells. Never followed it much. Shashi Singh
--- Peter Van Osta {pvosta-at-unionbio-eu.com} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi, } } I was wondering if there is already something going } on to set up a sort } of "Human Cytome Project" ? In my opinion the } hardware and most of the } software seems to be avaialable to set up such a } project ? For the } cellular level, light-microscopy based reader } technology would be very } interesting to use ? } } Studying and mapping the genome, transcriptome and } proteome at the } organisational level of the cell for various } celltypes and organ models } could provide us with a lot of information of what } actually goes on in } organisms in the spatio-spectro-temporal space ? } } I have been thinking (working) about a concept which } could provide the } basic framework for exploring and managing this } cellular level of } biological organisation research on a large scale, } but I would like to } know if there is already some thought/work going on } in the direction of } setting up an initiative such as a "Human Cytome } Project" ? } } This is just an idea, so I am really interested to } hear if there is } something in it, or even if it is not worth while } whet I just wrote. } } Best regards, } } Peter Van Osta } }
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From MicroscopyL-request-at-ns.microscopy.com Wed Dec 3 07:38:59 2003
We are in a process of purchasing a new Low Vacuum SEM. One of the suppliers offers LV 1-750 Pa, while the other has LV system 1-260 Pa. I have no experience with Low Vacuum (Variable Pressure) systems. We are trying to decide how important would be to have up to 750 Pa vs 260 Pa. We would like to be able to look at wet samples (plant tissue, soils, compost). I would appreciate your comments.
Thank you for your attention. Val Jeliazkov Ph: (902) 893 7859
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 3 12:17:58 2003
I wonder if this plush toy was something I recently saw in the San Francisco Exploratorium's gift shop - it represented the well-publicized SEM image purporting to show extraterrestrial bacteria.
-- *************************************************************** Do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro Journeys in Microspace (Columbia University Press, 1995) http://www.lsc.org/antarctica/front.html
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 3 13:19:46 2003
I would get the higher pressure model. I think, however, you should consider a model which can support 865Pa (6.5 Torr) at 5 degrees C. One could then image water/ fully hydrated specimens over a long period.
Dave
On Wed, 03 Dec 2003 09:37:50 -0400 "Valtcho Jeliazkov (Zheljazkov) Ph.D." {vjeliazkov-at-nsac.ns.ca} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } We are in a process of purchasing a new Low Vacuum } SEM. One of the suppliers offers LV 1-750 Pa, } while the other has LV system 1-260 Pa. } I have no experience with Low Vacuum (Variable } Pressure) systems. We are trying to decide how } important would be to have up to 750 Pa vs 260 } Pa. We would like to be able to look at wet } samples (plant tissue, soils, compost). } I would appreciate your comments. } } Thank you for your attention. } Val Jeliazkov } Ph: (902) 893 7859 } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 3 13:26:26 2003
Not all low vacuum SEMs are suitable for observation of wet specimens. To avoid substantial drying you need a system that can inject water vapor in the specimen chamber at pressures from 650 to 850 Pa. Microscope should have a cooling Peltier stage for bringing specimen temperature close to a dew point (1-5 Celsius).
Pressure 260 Pa is usually adequate for non-conductive specimens observation with BSE detector, but for some specimens charging can occur. The charging can affect EDS analysis, but for qualitative analysis its usually not a problem.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} } We are in a process of purchasing a new Low Vacuum } SEM. One of the suppliers offers LV 1-750 Pa, } while the other has LV system 1-260 Pa. } I have no experience with Low Vacuum (Variable } Pressure) systems. We are trying to decide how } important would be to have up to 750 Pa vs 260 } Pa. We would like to be able to look at wet } samples (plant tissue, soils, compost). } I would appreciate your comments. } } Thank you for your attention. } Val Jeliazkov } Ph: (902) 893 7859 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 3 23:21:54 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jfrosenberg-at-yahoo.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, December 3, 2003 at 13:05:48 ---------------------------------------------------------------------------
Question: I just purchased a used Olympus binocular compound microscope with a 100x objective. Does the Type A Immersion oil need to be stored in any special way such as a light free or amber bottle? What is the shelf life of this oil? Thanks. Ilana & Julie Rosenberg
Here are some links to websites with information about Cytomics, this will provide you with some information and background about what I mean with a "Human Cytome Project" (see below in this email).
I got the idea about using massive parallel "readers" on a "high-throughput" backbone for Cytomics while I visited the Sanger Center a few years ago where I saw a huge room "buzzing" with DNA-sequencing machines.
I wanted to design and develop a cell-screening system, based on a microscopy-based reader which could do for Cytomics what DNA-sequencing machines did for Genomics.
} } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear Dr. Osta, } I remember vaguely reading about cytomics and what I } remember is that it related to complete protein } profile in relation to functions ( somewhat like } functional genomics) also involving microscopy and } facs etc but of single cells. Never followed it much. } Shashi Singh } } } } --- Peter Van Osta {pvosta-at-unionbio-eu.com} wrote: } } } } } } ------------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The } } Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 4 05:05:24 2003
We have been given a small room that we would like to convert to a cryo-sectioning room. I am concerned about the air flow in the room. I naturally want enough air movement to make it safe for the operators to cut in the room, but I don't want the air conditioning to be so strong that it makes sectioning impossible. Is there a standard (Europe ) air flow rate needed for working with Liquid nitrogen? Or has anyone set up such a room that might have some hints or tips for me? Ken Blight Senior Scientific Officer Cancer Research UK London
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 4 09:37:20 2003
How big is the room, and how many instruments will you have in there?
Some tips from my own experience:
- Have them install an oxygen sensor. But make sure to have it installed at a reasonable height (level with the operator's head). The people at Yale would not listen to us and installed the sensors at floor level. We had to disconnect them as the alarm rang every time we would just open the lid of a nitrogen tank!
- Install "air socks". This is like a giant sock stretched on the ceiling that distributes the air evenly over the entire length of the room. Thanks to this, we have virtually no interference with sectioning. From my recollection of the time spent at ICRF, this is an important issue because we ended up taping a plastic film over the air vent in our sectioning room as the air current was too strong! I can have some diagrams sent to you if you need more information about these "air socks".
- Final tip: we had a glass sliding door installed at the entrance of our sectioning room. This is the best idea we ever had! No more strong air current anytime somebody walks in, and the added safety that people inside the room are seen from outside in case of emergency.
Hope this helps. Best wishes
Marc
On Thursday, December 4, 2003, at 06:04 AM, Ken Blight wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } We have been given a small room that we would like to convert to a } cryo-sectioning room. I am concerned about the air flow in the room. I } naturally want enough air movement to make it safe for the operators } to cut in the room, but I don't want the air conditioning to be so } strong that it makes sectioning impossible. Is there a standard } (Europe ) air flow rate needed for working with Liquid nitrogen? Or } has anyone set up such a room that might have some hints or tips for } me? } Ken Blight } Senior Scientific Officer } Cancer Research UK } London } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 4 09:41:00 2003
Regardless of the flow, one should avoid, for all the obvious reasons, having air venting directly onto a microtome or any other piece of precision equipment. To this end, I recommend a flow diverter be placed in front of the vent. This is achieved by suspending from the ceiling a piece of sheet metal that is slightly larger than the vent. Suspend the sheet about 15-20 cm below the vent and parallel to the ceiling. Lightweight chain makes hanging the sheet straightforward.
We routinely use this arrangement wherever needed in our lab.
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Ken Blight {Ken.Blight-at-cancer.or To: microscopy-at-msa.microscopy.com g.uk} cc: Subject: [Microscopy] Cryo-sectioning Room
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have been given a small room that we would like to convert to a cryo-sectioning room. I am concerned about the air flow in the room. I naturally want enough air movement to make it safe for the operators to cut in the room, but I don't want the air conditioning to be so strong that it makes sectioning impossible. Is there a standard (Europe ) air flow rate needed for working with Liquid nitrogen? Or has anyone set up such a room that might have some hints or tips for me? Ken Blight Senior Scientific Officer Cancer Research UK London
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 5 04:31:38 2003
I have two Ion Tech ion mills, with three chambers, which I wish to dispose of. Liquid nitrogen, iodine, and relatively low angle (down to 7 degrees) ion milling is possible, and one of the systems has beam steering. Also spares, sample holders, shields, etc. I need the space for lab re-organisation, and if no-one is interested in them they will go for scrap. Is there anyone out there in the UK still using these machines? Ah, the happy days (and days) I spent trying to align the guns...
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From MicroscopyL-request-at-ns.microscopy.com Fri Dec 5 10:23:49 2003
I'm looking for some advice and I hope someone has some experience with the Olympus VAS II photographic system. As I understand the operation, I can capture and store images as a TGA file. These files take up so much room that only two fit on a 1.4MB floppy. The software has a provision to convert the files to other formats but these formats aren't readable by powerpoint or the other viewers I have available. I can't get the VAS II installed on our network and I can't install a zip drive. (At least that's what I have been told.)
Any advice on how I can store more files on a floppy, or a reader which will read the conversion files would be gratefully appreciated.
Thanks for any advice......
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 5 11:45:34 2003
I am posting this for a graduate student here who is hoping someone has a technique that will help him out. Please reply to the list (in which case I will forward) and/or directly to Greg:
"I'm searching for a suitable LM staining protocol for distinguishing structures in the stigmatic region on the diminutive pistils of Lupinus perennis. I want to differentiate between the cell walls of the papillae and their overlying cuticle and, if possible, also the cell contents and a pool of lipid-rich exudate which accummulates between the cell wall and the cuticle. The protocol would involve stains that at a minimum differentiate cell wall (cellulose with some lignin) from cuticle (cutin) and ideally also lipids and nuclei. Thanks, Greg Shenk University of Connecticut shenk-at-darwin.eeb.uconn.edu "
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 5 13:34:12 2003
The Department of Molecular, Cellular, and Developmental Biology and the Neuroscience Research Institute are sponsoring an advance course on light microscopy. This 5-day workshop will be offered from March 22 through March 26, 2004 and will consist of lectures and laboratory exercises that will run from 9 am to approximately 5 pm each day. The seminar/workshop will be intensive enabling participants to develop theoretical and hands-on expertise with light microscopes. Attendees will interact closely with the instructors while using modern research grade microscopes, cameras, and computers. The seminars and laboratories will cover basic optical theory and how it pertains to increasing contrast (signal to noise ratio) in biological samples. Fundamental techniques such as fluorescence, phase contrast, Nomarski differential interference contrast, and darkfield imaging will be taught and attendees will use microscopes equipped with these optical enhancement accessories. In addition, the theory and practice of electronic image acquisition (analog and digital) will be discussed and attendees will work with low-light cameras, digital image processing computers, and morphometric programs. There are five research grade microscopes, five electronic imaging cameras, two computer workstations, and one confocal microscope. With a maximum enrollment of 10 students, there will be ample opportunity to work with all of the microscopes and cameras. For those so interested, intensive hands-on instruction and guidance on the confocal microscope will be provided.
For a fuller description of the workshop, please check the web address below. Enrollment forms can be completed online and this workshop provides an opportunity to have a working-vacation in Santa Barbara, California.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (burtonpierce-at-cox.net) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, December 5, 2003 at 23:52:48 ---------------------------------------------------------------------------
Question: My question is regarding ocular micrometer calibration. Are OCULAR DIVISION and OCULAR UNIT synonymus terms? Or is OCULAR UNIT defined as distance/OCULAR DIVISION?
T here was a meeting of facility managers at M&M2003 to discuss the formation of a Focused Interest Group dealing with topics of interest related to operating a core multi-user or service microscopy facility. There was overwhelming support for this and a document will be submitted shortly to the MSA Council requesting authorization for the formation of the FIG on Facility Operations.
The mission statement is: To explore issues regarding management and utilization of microscopy, imaging and microanalytical facilities.
Many of you are already on the E-mail list for information about this topic. Please contact me directly if you are not on the list and want to be included, or want to verify that you are on the list. Future communication about the FIG will be sent through the E-mail list and not via the MSA listserve.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 8 04:35:07 2003
The European Microbeam Analysis Society and the Department for Nanostructured Materials at the Jozef Stefan Institute, Ljubljana, Slovenia, are pleased to invite you to participate in the 6th EMAS Regional Workshop "Electron Probe Microanalysis Today - Practical Aspects" to be held in Bled, Slovenia, from 8 to 11 May 2004.
Updated information on the workshop and on-line registration can be found at the EMAS 2004 website: http://emas2004.ijs.si
Please download the First Announcement for the Workshop at: http://emas2004.ijs.si/EMAS2004_First_Announcement.pdf
Dr. Goran Drazic (chairman) Jozef Stefan Institute Department for Nanostructured Materials Jamova 39 SI-1000 Ljubljana Slovenia
EMAS 2004 Secretariat Email: sanja.fidler-at-ijs.si Fax: +386 1 4263 126
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 8 08:11:20 2003
Dear Burton, It is unlikely that 'ocular unit' is defined as a calibrated distance. Generally, 'ocular division' would refer to the smallest interval in the ocular scale and the the 'ocular unit' would refer to 10 of those divisions. Nevertheless, the main thing to keep in mind, is that there are no language police who arrest folks for incorrect or even unconventional usage. So someone could have defined 'ocular unit' in a sepcific way. If there is no information to indicate that in what you are reading, then I think you can reasonably safely assume that no calibration has gone on.
Tobias Baskin } } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (burtonpierce-at-cox.net) from } http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, } December 5, 2003 at 23:52:48 } --------------------------------------------------------------------------- } } Email: burtonpierce-at-cox.net } Name: Burton Pierce } } Organization: San Diego City College } } Education: Undergraduate College } } Location: City, State, Country } } Question: My question is regarding ocular micrometer calibration. } Are OCULAR DIVISION and OCULAR UNIT synonymus terms? Or is OCULAR } UNIT defined as distance/OCULAR DIVISION? } } ---------------------------------------------------------------------------
in view of the obvious facts that: 1. the DNA-sequence and functional knowledge on all biomolecules will not permit the assembly of living cell from their components 2. the analysis of organ tissue or biopsy specimens by advanced array technologies typically provides averaged result for many cells but does not address the discrete reactivity of specific cell populations being decisevely important for particular organ or organismic functions
it seems important and promising to address molecular principles of cell arrangement and interaction in a human cytome project. Such a project could bundle advanced microscopy, image analysis and other multiparameter cytometric techniques as well as single cell or specific cell group oriented functional genomics arraying and include multidimensional bioinformatics. The goal of the project would be to systematically explore multilevel molecular interrelations within cytomes as they occur in complex diseases like cancers, leukemias, cardiovascular disease, diabetes or in the susceptibility to infection including the detailed search for new pharmacological access points.
At 10:14 04.12.03 +0100, Peter Van Osta wrote: } Hi, } } Here are some links to websites with information about Cytomics, this } will provide you with some information and background about what I mean } with a "Human Cytome Project" (see below in this email). } } I got the idea about using massive parallel "readers" on a } "high-throughput" backbone for Cytomics while I visited the Sanger } Center a few years ago where I saw a huge room "buzzing" with } DNA-sequencing machines. } } I wanted to design and develop a cell-screening system, based on a } microscopy-based reader which could do for Cytomics what DNA-sequencing } machines did for Genomics. } } Websites on Cytomics: } } http://www.cytomics.info/ } http://www.biochem.mpg.de/valet/cytomics.html } http://www.biochem.mpg.de/valet/cellpot.html } http://www.ipd.anl.gov/biotech/programs/func-genomics/ } } Best regards, } } Peter Van Osta } } ( http://ourworld.compuserve.com/homepages/pvosta/cvwww.htm ) } }
It is unclear as to whether you are referring to the markings on the binocular body or on the reticle inside the eyepiece.
The markings on the binocular body are the "interpupillary distance": literally, the space from the center of one pupil to the center of the other when you have set the binoculars so that you see one, round field of view. This distance is marked in mm. You can measure anyone's IPD simply by having them look straight ahead and, using a small ruler, measuring the center-to-center distance. If you wear eyeglasses, you have already gone through this exercise with your local optometrist.
The markings on the reticle inserted inside the eyepiece have no units in and of themselves. They must be calibrated against a known (stage micrometer) for each optical set-up you use (primarily, each objective. If you have any intermediate magnifiers, such as the Zeiss Optovar, you also need to calibrate for each objective/intermediate set). This calibration is very easy to do: 1. Using the stage micrometer as a sample, set up Koehler illumination. 2. Choose some arbitrary number of eyepiece units which match markings on the stage micrometer. For example: 25 eyepiece units might match up with 400 microns on the stage micrometer. 3. Construct a conversion factor of microns/eyepiece units: In the example above: 400 microns/25 eyepiece units = 16 microns per eyepiece unit. 4. To measure with a real sample, simply observe how many eyepiece units cover the dimension you wish to measure in the sample, then multiply by your conversions factor.
Do this type of calibration for each objective (or objective/intermediate mag set), write it down, and tape it near the microscope for easy reference. These measurements will not change unless you use something new in the optical path.
You can also take this exercise one step further: To measure objects on your computer monitor, use the stage micrometer as a sample and project the image to the screen. In this case you won't have an eyepiece reticle. To make an equivalent structure for your screen, use an overhead transparency to set up a scale on the screen by marking (magic marker) 10, 50, or 100 microns on your overhead transparency.
Hope this is helpful.
Good hunting! Barbara Foster Microscopy/Microscopy Education, Inc. (MME, Inc.) 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
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At 05:25 AM 12/6/03 -0600, burtonpierce-at-cox.net wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Dec 8 15:05:40 2003
I'd like to get some feedback frome someone who's using a Leica DMRXA2 automatized microscope, with a Marzhauser or a Prior motorized stage. How would you qualify the quality of optics and motorized pieces?
Thank you!
Marie-Claude Belanger
_________________________________________________________________ MSN Search, le moteur de recherche qui pense comme vous ! http://fr.ca.search.msn.com/
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 8 17:37:07 2003
I would appreciate (off-list of course) any feedback from LEO FESEM users. Under consideration is a LEO Supra 55VP, 20nA gun. Any similar models including 1550 high vacuum or other VP SEMs is very appreciated. The application is semiconductor analysis, with RBSE, EDS and EBSD (EDAX).
thanks, gary g.
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 8 20:02:14 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (taylor-at-research.ge.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, December 7, 2003 at 20:40:58 ---------------------------------------------------------------------------
Email: taylor-at-research.ge.com Name: Seth Taylor
Organization: GE Research
Title-Subject: [Microscopy] [Filtered] MListserver: Epoxies for embedding ceramic particles
Question: Hello,
Can anyone out there suggest an epoxy that can be used to embed ceramic particles for TEM analysis? The trick is that I'd like to be able to thin the particle-epoxy sample in an ion-mill, so I'm looking for an epoxy whose hardness (or thinning rate) closely matches that of SiC.
I'd be grateful for any ideas or suggestions you might have.
Question: Hello all - a coworker has brought me something they found in their house that they suspect might be mold. They were wondering if I might be able to let them know what it is (ie is it mold) and then, if possible, figure out what type (genera) the mold is so they can have some idea of what might be involved in cleaning/removing the mold. We don't usually look at mold so any hints/tips of what direction to take would be helpful. The material was collected via tape sampling and that is what I now have. How should I go about looking at it? Is typical SEM imaging an appropriate place to start? Once I have an image is there a good website or book I can use to try and match my sample to the correct genera? Thanks in advance for any advice you can offer.
Question: We are interested in mounting small sections of PVDF protein transfer membrane for light microscopy. Preferably without disruption to any sample resting on the surface of the PVDF. Has anyone done this?
Question: I would like to identify an exogenous protein that contains a his-tag that was added to an in-vitro eye culture system. Is anyone familiar with identifying his-tags in tissue?
You don't need an EM to identify your mold. A brightfield microscope will do. (But if you want to use an EM, you can too.) You can go by morphology to identify it - note the type and shape of spore, arrangement, presence of sac enclosing spores, other structures. If they have a lot of the mold in the house, they can also note cultural characteristics - color, aerial hyphae (hairy, fuzzy, powdery, etc.). If it's difficult to observe them as they're growing in the house, you could take a sample and subculture it on culture media (potato dextrose agar, saboraud dextrose agar, etc.). Usually, you can identify the genus just by cultural and morphological characterization. When it comes to identification down to species level, it varies. With some molds you can get away with just the above tests, with some you'd need to do physiological tests.
University libraries would usually have several guidebooks on ID of filamentous fungi. These books usually provide descriptions and instructions (and lots of pictures) on characterizing spores.
The EPA and CDC webs provide info on dealing with molds in the house/buildings: http://www.epa.gov/iaq/molds/moldresources.html http://www.cdc.gov/nceh/airpollution/mold/moldfacts.htm
Lizette Tuason
anjeanette.ormonde-at-unilever.com (by way of MicroscopyListserver) said: } } Email: anjeanette.ormonde-at-unilever.com } Name: Anjeanette Ormonde } } Title-Subject: [Microscopy] [Filtered] MListserver: Mold Identification } } Question: Hello all - a coworker has brought me something they found in } their house that they suspect might be mold. They were wondering if I } might be able to let them know what it is (ie is it mold) and then, if } possible, figure out what type (genera) the mold is so they can have some } idea of what might be involved in cleaning/removing the mold. We don't } usually look at mold so any hints/tips of what direction to take would be } helpful. The material was collected via tape sampling and that is what I } now have. How should I go about looking at it? Is typical SEM imaging an } appropriate place to start? Once I have an image is there a good website } or book I can use to try and match my sample to the correct genera? } Thanks in advance for any advice you can offer. } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 9 00:36:40 2003
Comment 2: You work for Unilever so the Quality Control Lab there should have a microbiology division. Why don't you just ask them to identify the mold genus for you?
Lizette Tuason
anjeanette.ormonde-at-unilever.com (by way of MicroscopyListserver) said: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Email: anjeanette.ormonde-at-unilever.com } Name: Anjeanette Ormonde } } Title-Subject: [Microscopy] [Filtered] MListserver: Mold Identification } } Question: Hello all - a coworker has brought me something they found in } their house that they suspect might be mold. They were wondering if I } might be able to let them know what it is (ie is it mold) and then, if } possible, figure out what type (genera) the mold is so they can have some } idea of what might be involved in cleaning/removing the mold. We don't } usually look at mold so any hints/tips of what direction to take would be } helpful. The material was collected via tape sampling and that is what I } now have. How should I go about looking at it? Is typical SEM imaging an } appropriate place to start? Once I have an image is there a good website } or book I can use to try and match my sample to the correct genera? } Thanks in advance for any advice you can offer. } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 9 02:47:28 2003
Speaking as an amateur (as usual).... Whilst it is fun to use the SEM I suspect light microscopy would be more useful eg colour and cell wall appearance. I guess you do not need to know the genera to kill it :)
Dave
On Mon, 8 Dec 2003 20:04:27 -0600 by way of MicroscopyListserver {anjeanette.ormonde-at-unilever.com} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Email: anjeanette.ormonde-at-unilever.com } Name: Anjeanette Ormonde } } Title-Subject: [Microscopy] [Filtered] MListserver: Mold Identification } } Question: Hello all - a coworker has brought me something they found in their house that they suspect might be mold. They were wondering if I might be able to let them know what it is (ie is it mold) and then, if possible, figure out what type (genera) the mold is so they can have some idea of what might be involved in cleaning/removing the mold. We don't usually look at mold so any hints/tips of what direction to take would be helpful. The material was collected via tape sampling and that is what I now have. How should I go about looking at it? Is typical SEM imaging an appropriate place to start? Once I have an image is there a good website or book I can use to try and match my sample to the correct genera? Thanks in advance for any advice you can offer. } } --------------------------------------------------------------------------- } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 9 07:06:12 2003
we use the Leica DMRXA2 for upgrading it to a confocal microscope without using a laser. We are the dealer for this specially developed up-grade system. We also developed a motorized filter wheel, so that you can capture confocal images at different wavelengths and you can do colocalization with more than 3 wavelengths because you don't need different lasers, instead you use different filters.
We use the Leica DMRXA2 microscope because we think its optical quality is excellent. The motorized parts are also working well.
If you need more information about the Märzhäuser stage please let me know what you want to know exactly.
If you would like to see confocal images which were captured with the Leica DMRXA2 and a digital camera, please let me know.
MCB} ------------------------------------------------------------------------------ MCB} The Microscopy ListServer -- Sponsor: The Microscopy Society of America MCB} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver MCB} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html MCB} -------------------------------------------------------------------------------
MCB} Dear all,
MCB} I'd like to get some feedback frome someone who's using a Leica DMRXA2 MCB} automatized microscope, with a Marzhauser or a Prior motorized stage. How MCB} would you qualify the quality of optics and motorized pieces?
MCB} Thank you!
MCB} Marie-Claude Belanger
MCB} _________________________________________________________________ MCB} MSN Search, le moteur de recherche qui pense comme vous ! MCB} http://fr.ca.search.msn.com/
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 9 09:07:25 2003
I would like to demonstrate crystallization from a melt to my Earth Science class by projecting a crystallizing melt using a projection microscope. I am not really familiar with the technique and wonder if anyone could offer some suggestions. I would like to do a simple crystallization from a melt, as well as crystallization of a multicomponent melt that would simulate what happens in an intrusive igneous rock. Are there known multicomponent mixtures that I can use? Any suggestions for a simple hot (warm?) stage to control the rate of crystallization? Any help at all is welcome. Thanks.
Henry Barwood Associate Professor of Science, Earth Science Department of Math and Physics MSCX 312G Troy State University Troy, Alabama 36082 hbarwood-at-troyst.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 9 09:14:02 2003
We routinely re-mount epoxy and acrylic resin embedded tissues to improve their orientation by using two part epoxy glue to mount the trimmed down blocks to wooden dowels. I was wondering if anyone has tried a hot glue gun with success for this purpose. Recommendations on the appropriate type of glue stick welcome. I am mainly trying to find a more convenient way of doing this occasional chore. Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
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Hey Johnny!
Yes. It is Technotrade International, 7 Perimeter Rd Manchester, NH 03103 tel 603 622-5011 Ask for Johnny Hagen. (johnny-at-technotradeinc.com)
Call me if you need any other info.
Happy Hols!
Peter
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
Is anybody aware of any standards of proficiency for EM techs? I'm lookinf for something similar to what CAP has for histotechs. Would anybody be interested in paticipating in a survey to develop them?
Lois Anderson Johns Hopkins University Dept. of Pathology Laboratory Manager Electron Microscopy/Immunofluorescence 600 N. Wolfe Street/Pathology 709 A Baltimore, MD 21287 (410) 955-2861/fax (410) 614-7110 landers-at-jhmi.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 9 12:16:55 2003
Email: vodolan-at-biomed.cas.cz Name: Jana Vodolanova
Organization: Institute of Microbiology
Title-Subject: [Microscopy] [Filtered] protein A-gold detection
Question: Dear all, I have used protein A-gold (5 nm) detection in order to quantitate the antigen on the membrane and detect its distribution.
I have found some useful references concerning protein A-gold detection but not many. I would like to ask you, if you know some good ones, to let me know.
Further, it is believed that protein A-gold binds to primery antibody in a 1:1 ratio under saturation conditions. So, does single protein A-gold particle mean that either one antigen or two antigen at the maximum are detected? Has anyone of you different experience?
Thanks a lot.
Jana
******************** Jana Vodolanova, MSc. Laboratory of Molecular Biology of Bacterial Pathogens Institute of Microbiology Czech Academy of Sciences Videnska 1083 Prague 4, 142 20 Czech Republic
I have some information about a room for cryo-ultramicrotomy from Thomas Keil with the Baumeister Lab in Germany: Total volume is ca. 7.4 cubic meters. Air exchange rate is ca. 60 cubic meters per hour. Temperature around 23 deg C, Air humidity around 33 per cent. Over 40 per cent, ice contamination becomes too high.
This last point about humidity is VERY important if you hope to look at frozen hydrated sections. They also built the air exhaust to take air from near the floor.
Larry Ackerman Advanced Imaging Lab/Dr. John Sedat/Dr. David Agard Dept. of Biochemistry & Biophysics University of California San Francisco 600-16th Street, Box 2140, Room S101 San Francisco, CA 94143
415-476-4441 FAX 415-514-4142
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 9 13:36:42 2003
This is an easy one. Use Super Glue or an equivalent cyanoacrylate product. Application is easy, and if you put the blocks back in the oven after gluing, they are ready to work on within minutes. Just make sure the sides of the block face do not have any adhering glue. Super Glue sections easily, but sections will not lie flat because the glue and the resin have different textures.
Ralph Common Electron Microscopist Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824 517-355-7558; fax 517-432-1053 ralph.common-at-ht.msu.edu
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Tuesday, December 09, 2003 10:15 AM To: Microscopy-at-msa.microscopy.com
We routinely re-mount epoxy and acrylic resin embedded tissues to improve their orientation by using two part epoxy glue to mount the trimmed down blocks to wooden dowels. I was wondering if anyone has tried a hot glue gun with success for this purpose. Recommendations on the appropriate type of glue stick welcome. I am mainly trying to find a more convenient way of doing this occasional chore. Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
} Is anybody aware of any standards of proficiency for EM techs? I'm lookinf for something similar to what CAP has for histotechs. Would anybody be interested in paticipating in a survey to develop them?
The Microscopy Society of America established a certification program in 1978. You can navigate to the information from the MSA web site (http://www.msa.microscopy.com) or jump directly to it at http://www.cvmbs.colostate.edu/emcenter/msa/certboard/
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 9 16:51:47 2003
Dear Jana I don't think you could quantitate with any immunochemistry:
- you don't know how many antigenic determinants are available for primary antibody (AB); - you don't know the efficiency of AB binding. If it's polyclonal - they bind differently (different association constant); - you don't know how many gold particles attached to the protein; - as far as I remember, protein-A molecule has 4 unequal IgG binding sites and IgG molecule has two protein-A binding sites (equal).
Sergey
At 10:18 AM 12/9/2003, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
We find most convenient the use of cyanoacrylic cement (Super Glue) and the stubs of old ultratome blocks, filed flat, for the mounting of reoriented resin-embedded samples. Mike Nesson Electron Mocroscopy Facility Oregon State University Corvallis, OR 97331
-- ÐÏࡱá
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 9 17:39:06 2003
I know of a number of one-phase systems, especially things like urea or any of the cholesteric materials, but I can't think of a two phase system off-hand. Would really like to know about that when you find it.
Most importantly: do this under polarized light. It really brings out the crystalline formation. We did this with a group of jr and sr. high teachers at a course about 3 years ago at Miami U (Ohio). We used a very inexpensive S-video camera, fed into a typical video monitor. The teachers went crazy!!!
Hope your demo is equally successful.
Best regards, Barbara Foster Microscopy/Microscopy Education, Inc. (MME, Inc.) 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
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At 09:07 AM 12/9/03 -0600, Henry Barwood wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Dec 10 04:50:55 2003
Protein A-gold was first introduced by Romano and Romano in 1977 (Immunochemistry 14, 711). Slot and colleagues improved the technology as described a.o. in J. Cell Biol. (1981), 90, 553.
Some useful information on quantification of protein A-gold labeling can be found in: Posthuma et al. (1984) J. of Histochem. and Cytochem. 32, 1028 Posthuma et al. (1986) J. of Histochem. and Cytochem. 34. 203
Hope this is of help. Do not hesitate to contact me when you have additional questions.
Regards, Peter
On Tuesday, December 9, 2003, at 07:18 PM, by way of MicroscopyListserver wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } } Email: vodolan-at-biomed.cas.cz } Name: Jana Vodolanova } } Organization: Institute of Microbiology } } Title-Subject: [Microscopy] [Filtered] protein A-gold detection } } Question: Dear all, } I have used protein A-gold (5 nm) detection in order to quantitate the } antigen on the membrane and detect its distribution. } } I have found some useful references concerning protein A-gold } detection but not many. I would like to ask you, if you know some good } ones, to let me know. } } Further, it is believed that protein A-gold binds to primery antibody } in a 1:1 ratio under saturation conditions. So, does single protein } A-gold particle mean that either one antigen or two antigen at the } maximum are detected? Has anyone of you different experience? } } Thanks a lot. } } Jana } } ******************** } Jana Vodolanova, MSc. } Laboratory of Molecular Biology of Bacterial Pathogens } Institute of Microbiology } Czech Academy of Sciences } Videnska 1083 } Prague 4, 142 20 } Czech Republic } } phone: +420 24106 2380,2770 } fax:+420 24106 2152 } e-mail: vodolan-at-biomed.cas.cz } } ----------------------------------------------------------------------- } ---- } } }
----------- Peter van de Plas Aurion ImmunoGold Reagents Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 10 07:30:15 2003
Lois, I imagine that you have already had responses to this question but I thought to weigh in anyway. I am not familiar with the exact requirements for the CAP. However are you aware of the certification for biologists through MSA? This certification is only for TEM and involves both a written test on theory and a practical exam. The practical requires the preparation of 3 different tissues with full documentation on procedure, thick and thin sections and final micrographs. Most of the people who go through the certification process do so because they or their employer wants certification as a minimal measure of proficiency. This is, as you no doubt know, looked on very favorably in the medical world.
For more information on the MSA Certification process, check out the MSA web site at: http://www.msa.microscopy.com/
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 12/9/03 12:18 PM, "Lois Anderson" {landers-at-jhmi.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Is anybody aware of any standards of proficiency for EM techs? I'm lookinf } for something similar to what CAP has for histotechs. Would anybody be } interested in paticipating in a survey to develop them? } } Lois Anderson } Johns Hopkins University } Dept. of Pathology } Laboratory Manager } Electron Microscopy/Immunofluorescence } 600 N. Wolfe Street/Pathology 709 A } Baltimore, MD 21287 } (410) 955-2861/fax (410) 614-7110 } landers-at-jhmi.edu } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 10 08:32:52 2003
A colleague is looking to buy a light source for a binocular microscope and is especially interested in the fibre optic ones with the long light tubes. With some models, though, these light tubes are "floppy" and need some sort of support. He'd like to find one with the rigid (though flexible) light tubes. I suggested he just apply Viagra to the floppy ones, but, no, he wants a new one. Does anybody have a source or at least a make/model no. for such a unit?
Frank Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 10 08:39:38 2003
For Biological EM techs, MSA offers a certification exam. Please see the link on the MSA website. Lee Cohen-Gould, MSA Certification Board -- 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
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 10 08:41:01 2003
Dynamic, fast growing Scanning Probe company seeks a strongly motivated individual capable of delivering the high level of technical applications and sales support we require. The successful applicant will have extensive practical SPM experience. Strong people and presentation skills, both verbal and written, are essential. A bachelors or graduate degree in materials science and/or physics is required. Business experience is a plus.
Duties will include: · Demonstration support for sales (analyzing samples, and demonstrating instruments to potential customers at an Applications Laboratory or at the customers site) · Technical support for our Marketing Department and at tradeshows, and · Installation and customer training.
It is expected that this job may entail approximately 30% travel. Starting date: early 2004.
Geographic Base: Northeastern U.S. to Mid-Atlantic region, or Bay Area.
Compensation will be commensurate with training and experience. Interested individuals should send CV, short sample of writing/training materials, and 3 references to: spm_info-at-earthlink.net
An equal opportunity employer.
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 10 09:23:22 2003
Can anyone advise me about a protocol for combined FISH (for mRNA) and IF (for protein)? Many of the references I have read don't include fluorescence for mRNA detection but I would really like to use our confocal microscope. Besides, a fluorescent signal may be easier to quantify than, say, brightfield alkaline phosphatase product.
As a philosophical issue, apart from amplification of the expression (prehybridization) or of the signal (post hybridization) has in situ hybridization itself changed much in the last few years? In other words, how valid are references to "critical steps" from the early 90's? I wonder because there seem to be more pure methods papers, carefully testing each step, from that time then seen currently.
Thank you for any advice and I look forward to opinions. Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 7Queen 30 Bond St. Toronto, ON M5B 1W8 Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 10 09:27:53 2003
Got this message today. I think the whole list might be interested if such exists } Date: Tue, 09 Dec 2003 08:35:45 -0600 } From: James Jefferson {James.Jefferson-at-hitachi-hhta.com} } Subject: [Microscopy] Ernst Ruska } To: gwe-at-biotech.ufl.edu } Reply-to: James.Jefferson-at-hitachi-hhta.com } X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2911.0) } Importance: Normal } } Greetings, } } a colleague of mine is looking for a documentary on Ernst Ruska he thought } he had seen on PBS, but multiple attempts to locate this special have left } me empty handed. The special dealt with Ernst Ruska winning the Nobel prize } in relationship to Electron Microscopy. } } Any help would be appreciated. } } Sincerely Yours, } James Jefferson } Hitachi High Technology America } Staff Engineer
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 10 14:56:35 2003
Hello all, I have samples of the same material with suspected difference in porosity, size of pores could be from a few nanometers to a few hundred nanometers.
Is it possible to detect porosity with microanalysis (EDS)? How intensity could change because of porosity? Is it possible to calculate the "detection limit" of porosity?
Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
Cheers, and this is a real gift for technology history buffs,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson-at-wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/
-----Original Message----- } From: Greg Erdos [mailto:gwe-at-biotech.ufl.edu] Sent: Wednesday, December 10, 2003 10:29 AM To: Microscopy-at-sparc5.microscopy.com
Got this message today. I think the whole list might be interested if such exists } Date: Tue, 09 Dec 2003 08:35:45 -0600 } From: James Jefferson {James.Jefferson-at-hitachi-hhta.com} } Subject: [Microscopy] Ernst Ruska } To: gwe-at-biotech.ufl.edu } Reply-to: James.Jefferson-at-hitachi-hhta.com } X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2911.0) } Importance: Normal } } Greetings, } } a colleague of mine is looking for a documentary on Ernst Ruska he } thought he had seen on PBS, but multiple attempts to locate this } special have left me empty handed. The special dealt with Ernst Ruska } winning the Nobel prize in relationship to Electron Microscopy. } } Any help would be appreciated. } } Sincerely Yours, } James Jefferson } Hitachi High Technology America } Staff Engineer
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 10 19:50:04 2003
Pay no attention to the less then "corporate" website. They make good products...
----- Original Message ----- } From: "Thomas, Frank" {FThomas-at-nrcan.gc.ca} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, December 10, 2003 6:34 AM
Fostec (now Schott-Fostec) makes a nice line of FO sources and ring lights and bifurcated illuminators. There is also an LBD daylight correction filter available for each bundle.
gary g.
At 06:34 AM 12/10/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Dec 11 00:31:22 2003
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 11 01:52:27 2003
New conference XVII th Physical Metallurgy and Materials Science Conference "ADVANCED MATERIALS & TECHNOLOGIES AMT '2004
} www.AMT-2004.p.lodz.pl, } } or mail to: } Bogdan WENDLER, D. Sc., Ph. D. } Associate Professor } Secreatary AMT-2004 } Head Division Coatings Engineering } Institute of Materials Engineering (IME) } Dept. Mechanical Engineering } Technical University of Lodz } Ul. Stefanowskiego 1 } 90.924 Lodz } POLAND } Tel (+48) 426 312 265 } Mob (+48) 501 292 922 } Fax (+48) 426 366 790 } E-mail: bowe-at-p.lodz.pl } Web site of IME: http://www.p.lodz.pl/IIM (English version
best regards Chris Hübner
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 11 03:31:04 2003
Focus on Microscopy 2004 Philadelphia, USA, April 4 to April 7, 2004
Dear colleagues,
The FOM 2004 website: www.focusonmicroscopy.org is now open for registration, submission of abstracts and accommodation reservation, All further information is available there.
Deadline for the submission of abstracts will be January 30, 2004.
As the next in a series of unique interdisciplinary meetings on advanced multidimensional light microscopy and image processing, the 2004 meeting will pay special attention to the conjunction of multidimensional microscopies with the areas of bioinformatics, bio-nanotechnology and bioengineering.
The 2004 meeting will be hosted by Drexel University, School of Biomedical Engineering, Science and Health Systems. The conference and exhibition will be located at the Sheraton University City Hotel, central on the Philadelphia campus.
Originally the FocusOnMicroscopy FOM2004 international conference would take place in Singapore but various organizational issues and travel concerns made a move to Philadelphia, PA, USA, April 4 to April 7 necessary. Andres Kriete will be the main local organizer in Philadelphia.
You are cordially invited to participate in this conference on behalf the FOM 2004 organizing committee: A. Kriete, Pittsburgh; F. Brakenhoff, Amsterdam; P.C. Cheng, Buffalo; Banu Onaral, Philadelphia. -- +-----------------------------------------+ Prof. Dr. G.J. Brakenhoff University of Amsterdam Institute for Molecular Cell Biology Section Molecular Cytology Kruislaan 316 1098 SM Amsterdam, The Netherlands
Some years ago we used a lightsource from Illumination Technologies with an optic fiber. It was the CF 1000 model with automated filter wheel and it could be used either manualy or by RS-232 control. We used it for an older type of microscope which did not have an automated filter wheel itself. Illumination Technologies also sells fiber optics for their lightsources
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi } } Sorry to hear about your floppiness problem - happens to us all from } time to time. } } Try Schott - they used to do 'swan-neck' optical fibre illumination with } one or multiple heads - good for macrophotography and other applications. } } Find them at } http://www.us.schott.com/english/index.html } Med vänliga hälsningar/With best regards } } Gareth } } ***Come to Stockholm in 2004 to the 26th IFBLS congress*** Take a look } at http://www.vardforbundet.se/ifbls2004 } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-labmed.ki.se } } Tel +46 8 5858 1038 } Fax +46 8 5858 7730 } } Gareth Morgan MPhil MSc FIBMS, } Department of Laboratory Medicine (Labmed), } Karolinska Institutet, } Huddinge Universitetssjukhus, F46 } SE 141 86 Stockholm } Sweden } } OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. } Laboratoriet för klinisk patologi och cytologi. } } NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. } Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 11 07:46:38 2003
If it is simply pores you are interested in e.g. size, distribution, why not just image your sample in the SEM? Maybe you should elaborate a bit further on your sample and what is of interest about these pores?
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] Sent: Wednesday, December 10, 2003 3:58 PM To: Microscopy (E-mail)
Hello all, I have samples of the same material with suspected difference in porosity, size of pores could be from a few nanometers to a few hundred nanometers.
Is it possible to detect porosity with microanalysis (EDS)? How intensity could change because of porosity? Is it possible to calculate the "detection limit" of porosity?
Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
I'd suggest you also contact the Royal Microscopical Society (Oxford, UK). They have extensive certifications for various types of microscopy.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 08:30 AM 12/10/03 -0500, Debby Sherman wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Dec 11 11:18:49 2003
I'm presently investigating the variety of modern EDX detectors, and I note (via advertising) the high-throughput Roentec Xflash detectors now approach the energy resolutions available from most other detectors. However, what remains a mystery is the counts into the spectrum as a function of beam current. That is, for a detector with a 5mm window area, Roentec still boasts of throughputs of 1000k cps. What must the beam current be?
I understand these detectors shouldn't be compared to other types because of their small processing times, but it doesn't seem to me the difference between 1-2% DT and 20-30% DT can make up for the number of counts through a "normal" 10mm detector at normal beam currents (e.g., 2-5nA), and what Roentec claims would lead me to believe(?)
These modern Roentecs are rare in North America, but I'd enjoy hearing from someone with practical experience (on or off list)
Fundamentals of Microanalytical Entomology: A Practical Guide to Detecting and Identifying Filth in Foods by Alan R. Olsen, Thomas H. Sidebottom is one of the references used by FDA to identify mold using LM.
David A. Foran, chemist Food and Drug Administration Kansas City Elemental Analysis Lab DFORAN-at-ORA.FDA.GOV 913-752-2170 913-752-2727 (voice mail) 913-752-2151 (fax)
----Original Message----- } From: tuasonm-at-unbc.ca [mailto:tuasonm-at-unbc.ca] Sent: Tuesday, December 09, 2003 12:41 AM To: microscopy-at-ns.microscopy.com
Hi,
We are looking around for a ultramicrotome to slice our samples for TEM. The kind of samples we work are semiconductors substrates (Si, SiC, GaAs...) with some epitaxial semi layer or poly metal thin films. What is your experience preparing this kind of sample? Can we prepare sample with large area view under TEM? And what about dislocation or other defects introduced in the layer? As you can see, we are in dark. We do prepare our samples using tripod polishing. What motivated us into looking in ultramicrotomy is a new set of samples that are polymer (coated and bulk). The sample is affect by the superglue and the acetone we use in the polishing process. And it also has a low Tg. And if we can also use ultramicrotomy to prepare our routine samples it is a plus. Any light will be appreciated. :)
And we also nail down our search for the ultramicrotome in two makers, Leica and RMC. Any other you can suggest? And finally, how useful cryo is for material scientists?
Lots of questions. :) Thanks.
Carlos
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 11 16:08:22 2003
If you could microtome Si and other semiconductor specimens the microtome manufacturers would all be driving Rolls Royces!
The problem is that Si, and the others, cleave. As soon as the knife touches the Si, the Si cleaves into many small pieces (it doesn't do your knife a lot of good either). The cleavage is all on [111] planes, which is sort of nice, BUT the Si chip surfaces are [001] and the cross sections are [011]. So the shattered bits of Si debris that results will not be oriented to reveal anything about the chip plan-view or its cross sections.
Tripod polishing is good; cleavage, a'la McCafferty in Canada and Scott Walck at PPG is a lot better; and, of course, FIB, with appropriate protection for the polymers is best. If you don't have a FIB, cleave them. Look at the Southbay Tech web site for cleaving kits.
Ron Anderson
-----Original Message----- } From: Carlos Kazuo Inoki [mailto:kazuo-at-csc.albany.edu] Sent: Thursday, December 11, 2003 3:04 PM To: MSA Listserve
Hi,
We are looking around for a ultramicrotome to slice our samples for TEM. The kind of samples we work are semiconductors substrates (Si, SiC, GaAs...) with some epitaxial semi layer or poly metal thin films. What is your experience preparing this kind of sample? Can we prepare sample with large area view under TEM? And what about dislocation or other defects introduced in the layer? As you can see, we are in dark. We do prepare our samples using tripod polishing. What motivated us into looking in ultramicrotomy is a new set of samples that are polymer (coated and bulk). The sample is affect by the superglue and the acetone we use in the polishing process. And it also has a low Tg. And if we can also use ultramicrotomy to prepare our routine samples it is a plus. Any light will be appreciated. :)
And we also nail down our search for the ultramicrotome in two makers, Leica and RMC. Any other you can suggest? And finally, how useful cryo is for material scientists?
Lots of questions. :) Thanks.
Carlos
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 11 18:06:34 2003
Carlos, you are correct in that Leica and RMC are the two major manufacturers of ultramicrotomes. It is also possible to section coatings on semiconductor substrates, and I have seen many examples done by a colleague, Phil Swab, of Unity Semiconductor in California. He has even sectioned diamond coatings on boron nitride on a Si substrate! However, it is a demanding procedure, plus there is no way you will get large areas for TEM. While I am proponent of materials ultramicrotomy in general, every technique has its limits. Ultramicrotomy works best on samples that are relatively soft, where 'relatively soft' can include many alloys and composites, and is especially applicable for ultrathin (~10nm thick) sections of 'soft' nanomaterials. Hard, brittle samples are a challenge. The fragments produced (think of dropping a sheet of glass on the floor) will be fairly defect-free, however, as the sectioning then becomes more of a crack propogation issue than one of shearing.
Have you considered focused ion beam (FIB) sectioning? It is very good for thin sections of your substrate materials, but it can cause ion damage to polymeric (and biological) materials. You could consider sending one sample to someone with a FIB and see what happens.
Finally, have you thought of corresponding with those experienced in tripod polishing to see if there might be alternatives to your glue/solvent issue? Ron Anderson, editor of Microscopy Today, comes to mind.
Best of luck, Tom
Dr. Tom Malis Scientist Advisor Natural Resources Canada Govt. of Canada Phone: 613-995-7358 cell: 613-371-4577 FAX: 613-947-6606 malis-at-nrcan.gc.ca
PS For those of you who have known me as a 'characterization guy', through and through, over the years, yes, I have succumbed to the lure (read Boss's orders) of moving into upper management. I am having a pretty decent time, however, acting as a 'translator' of science issues for the policy folk in the Canadian government.
-----Original Message----- } From: Carlos Kazuo Inoki To: MSA Listserve Sent: 12/11/2003 3:04 PM
Hi,
We are looking around for a ultramicrotome to slice our samples for TEM. The kind of samples we work are semiconductors substrates (Si, SiC, GaAs...) with some epitaxial semi layer or poly metal thin films. What is your experience preparing this kind of sample? Can we prepare sample with large area view under TEM? And what about dislocation or other defects introduced in the layer? As you can see, we are in dark. We do prepare our samples using tripod polishing. What motivated us into looking in ultramicrotomy is a new set of samples that are polymer (coated and bulk). The sample is affect by the superglue and the acetone we use in the polishing process. And it also has a low Tg. And if we can also use ultramicrotomy to prepare our routine samples it is a plus. Any light will be appreciated. :)
And we also nail down our search for the ultramicrotome in two makers, Leica and RMC. Any other you can suggest? And finally, how useful cryo is for material scientists?
Lots of questions. :) Thanks.
Carlos
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 11 19:40:23 2003
Title-Subject: [Microscopy] [Filtered] books on electron microscopy
Question: Dear colleagues:
Could you please recommend me some books on electron microscopy which might have an interesting problems regarding to SEM, TEM and EDS, which might be given to students on undergraduate level for their homework and tests?
Carlos, I think it will be very difficult to get a good cleaved sample of a polymer layer on a semiconductor substrate. In my experience of polyimide and polyamide passivation/dielectric layers, the substrate cleaves while the polymer remains intact, and you have to pull the two pieces apart, tearing the polymer (and, as often as not, pulling it off the substrate). There are similar problems with thick ductile metal layers such as gold - although if you have a good process they shouldn't delaminate. The only way to get a good section is to have ion milling as the final step - either in a FIB or a conventional ion mill. If you can cleave the samples without too much tearing of the layers, you might be able to cleave wedges (as Ron Anderson mentioned), and then ion mill them to get back to virgin material. I have made 'cold' samples of Pb-Sn solder bumps on Si by using superglue as the mounting medium and soaking them off, rather than using the glycophthalate wax I usually use. Metal layers are no problem since they are resistant to solvents, but your polymers are very tricky.. The only thing I can think of is that you may be able to deal with these samples using tripod polishing or similar standard preparation techniques by changing the mounting media. I have used double-sided sticky tape to hold a sample for (gentle!) grinding and polishing one side, but I don't know what you could use to support the sample as you thin it down to only a few microns.
Good luck!
Richard _______________________________ Richard Beanland Analytical Services Bookham Technology plc Caswell, Towcester, Northamptonshire NN12 8EQ UK Tel: +44 (0) 1327 356362 Fax: +44 (0) 1327 356775 http://www.bookham.com
-----Original Message----- } From: Carlos Kazuo Inoki [mailto:kazuo-at-csc.albany.edu] Sent: 11 December 2003 20:04 To: MSA Listserve
Hi,
We are looking around for a ultramicrotome to slice our samples for TEM. The kind of samples we work are semiconductors substrates (Si, SiC, GaAs...) with some epitaxial semi layer or poly metal thin films. What is your experience preparing this kind of sample? Can we prepare sample with large area view under TEM? And what about dislocation or other defects introduced in the layer? As you can see, we are in dark. We do prepare our samples using tripod polishing. What motivated us into looking in ultramicrotomy is a new set of samples that are polymer (coated and bulk). The sample is affect by the superglue and the acetone we use in the polishing process. And it also has a low Tg. And if we can also use ultramicrotomy to prepare our routine samples it is a plus. Any light will be appreciated. :)
And we also nail down our search for the ultramicrotome in two makers, Leica and RMC. Any other you can suggest? And finally, how useful cryo is for material scientists?
Lots of questions. :) Thanks.
Carlos
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
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From MicroscopyL-request-at-ns.microscopy.com Fri Dec 12 07:29:41 2003
It seems appropriate that we, RONTEC, manufacturer of the XFlash detector make some brief comments on-line regarding the subject. Some clarification on the technical facts could be of interest to the community.
First of all, we don't claim 1000 kcps "throughput" but this is the "input" count rate, i.e. at a pulse load of 1000 kcps a certain XFlash type (2001) is still capable of producing reasonable spectra provided a pulse processor with extremely short shaping time constants such as those offered by RONTEC is used.
Certainly, it won't be possible with every sample in every SEM to produce 1000 kcps within a solid angle covered by a 5 sqmm detector area.
It depends of course on the beam current the cathode can generate, the nature of the sample and the actual size of the solid angle which is determined not only by the active area but also by the distance between the sample surface and the detector.
Regarding the solid angle it is important to say that in many cases it's not true that the solid angle covered by a 5 sqmm XFlash is only half as big as the one covered by a 10 sqmm Si(Li). Due to the more compact inner design the distance between the tip of the endcap and the crystal surface is smaller for the XFlash. Because of the variety of SEM models and their different detection geometries, it is not possible to specify a fixed ratio between the size of the solid angles for 5 sqmm XFlash and 10 sqmm Si(Li) but it may even be 1:1 in some cases.
As for the question of how to generate 1000 kcps, there are certainly many situations where all conditions together are suitable to produce such high x-ray intensities. As a practical example, we see 1000 kcps input from a metallic sample with beam currents adjusted to 60...80 nA (tungsten cathode).
In such case the dead time and energy resolution of the XFlash are comparable with what good conventional Si(Li) detectors would show at a pulse load of 100 kcps (60% DT, 250 eV FWHM).
With the same XFlash type, at lower count rates, say 1...10 kcps (using longer shaping times), the energy resolution goes up to 133...135 eV and the dead time goes down to a few percent.
Another XFlash type (3001) provides even better FWHM at lower count rates (below 130 eV) but the maximum pulse load is limited to 700 kcps (275 kcps into the spectrum or map).
Hopefully, this reply will clarify some of the questions.
RONTEC Thomas Schuelein
} From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-MSA.Microscopy.Com}
Email: mauduit-at-cemes.fr Name: bernadette de MAUDUIT
Question: I would know from which society the Desktop Microscopic software and information about it can now be available because the VIRTUAL LABORATORIES seem to no more exist.
Thanks for the feedback regarding ultramicrotomy. It is interesting to know it doesn't work well with semiconductors, specially to produce large area view samples. This is crucial for us, since we do a lot of defect analysis (count dislocations). Someone asked if I was planning to slice SiC with the ultramicrotome (a very hard material). I was just wondering if ultramicrotomy works with the kind of samples we usually work. Buying something just to work with soft material should be a waste of resources for us. Specially because the bulk of our work is done with semicondutors, or using it as substrate for other materials. :)
About using FIB, we did try with our polymer samples (it is an acrylate based material). But it just melts under the beam. Low glass transition temperature (Tg) I imagine. So no heating, at least over 100C. Also dissolved by solvents and superglue. Cleaving may work, except that we really need to image a large area. Like we obtain with tripod polishing. This samples contains some nanoparticles embedded in it, and we want to count the density and also de distribution. So, this is the problem that start us into look in ultramicrotomy. :)
Anyway, it was good to hear some ideas and suggestions from the Wizards of TEM-sample-preparation-archana. :) Thanks again,
Regards,
Carlos
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 12 19:07:04 2003
SEM--human mucosa Hi, I'm looking for biofilms on human mucosa. Is cryostage SEM superior to standard fixations? would my results be equivalent to using OCT resin? at this point my samples are in formalin. Any suggestions to succesfully finding biofilms? appreciaty any input. Thanks, Jose
__________________________________ Do you Yahoo!? Free Pop-Up Blocker - Get it now http://companion.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 13 01:59:42 2003
Dear friends, it is a pleasure for me to announce that is available on line the new special issue of Microscopy Research and Technique on TPE. Please, also start accepting my best wishes for Xmas and for a New Year aimed to Peace. All my bets Alby
------- Special issue content -----------
Volume 63, Issue 1 (1 January 2004) Special Issue: Two-Photon Microscopy - Part II .Issue Edited by Alberto Diaspro. Published on line, 10 Dec 2003
Articles in the Current Issue:
Rapid dissemination of two-photon excitation microscopy prompts new applications (p 1-2) Alberto Diaspro
Antecedents of two-photon excitation laser scanning microscopy (p 3-11) Barry R. Masters, Peter T.C. So
Notes on theory and experimental conditions behind two-photon excitation microscopy (p 12-17) Alessandro Esposito, Federico Federici, Cesare Usai, Fabio Cannone, Giuseppe Chirico, Maddalena Collini, Alberto Diaspro
Practical limits of resolution in confocal and non-linear microscopy (p 18-22) Guy Cox, Colin J.R. Sheppard
Novel diode-pumped infrared tunable laser system for multi-photon microscopy (p 23-26) Nelly Deguil, Eric Mottay, Francois Salin, Philippe Legros, Daniel Choquet
Ultracompact autocorrelator for multiphoton microscopy (p 27-33) F. Quercioli, A. Ghirelli, B. Tiribilli, M. Vassalli
Distance measurement by circular scanning of the excitation beam in the two-photon microscope (p 34-49) Katarina Kis-Petikova, Enrico Gratton
Probing microscopic diffusion by 2-photon flash photolysis: Measurement of isotropic and anisotropic diffusion in lens fiber cells (p 50-57) M.B. Cannell, M.D. Jacobs, P.J. Donaldson, C. Soeller
Fluorescence lifetime imaging by time-correlated single-photon counting (p 58-66) W. Becker, A. Bergmann, M.A. Hink, K. König, K. Benndorf, C. Biskup
Live cell ultraviolet microscopy: A comparison between two- and three-photon excitation (p 67-71) J. Balaji, R. Desai, S. Maiti
Characterization of two-photon excitation fluorescence lifetime imaging microscopy for protein localization (p 72-80) Ye Chen, Ammasi Periasamy
Performances of high numerical aperture water and oil immersion objective in deep-tissue, multi-photon microscopic imaging of excised human skin (p 81-86) Chen-Yuan Dong, Betty Yu, Peter D. Kaplan, Peter T.C. So
....................................................................... ........................ Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480/309 fax 010314218 e-mail: diaspro-at-fisica.unige.it URL: http://www.lambs.it ....................................................................... .....................
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 13 09:04:31 2003
An advanced course on polarized light microscopy which will cover the following topics:
The nature of polarized light
The origin and interpretation of interference colors
Birefringence and crystal orientation, The Indicatrix
Compensation and variable compensators
Interference figures and their interpretation
The workshop will consist of four Consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of polarized light microscopy. The course instructors include Jan Hinsch of Leica, Inc., Mary McCann of McCann Imaging, John Reffner of SensIR and N.Y.M.S. Instructor Don O'Leary.
WHEN: May 1, 8, 15 & 22, 2004, from 10 A.M. to 4 P.M.
COST: $350 for N.Y.M.S. members, $380 for non-members (includes membership). Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: advanced course for those who have completed "The Use of the Microscope" or are experienced in microscopy and familiar with the theory of its use.
HOW: Register using the form below. Limited to the first 12 registrants.
Return form to Don O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
FURTHER INFORMATION:Call D. O'Leary (201)797-8849 e-mail donoleary-at-att.net
Question: Hi- I am trying to get high-quality images from my microscope recorded on film in my 35mm camera. I am working with birefringent crystals at the moment. My 'scope is excellent quality, great optics, images look sharp and clear. THEN, when I put my camera onto my microscope adapter and look thru the camera, the image magnification is greatly increased, focus is nearly impossible, and images are awful. HELP! I also wonder if anything would be different if I was using a trinocular 'scope. Seems I would have the same problem, since the adapter places the camera further away from the object being viewed. Any suggestions would be greatly appreciated!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pwebster-at-hei.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, December 12, 2003 at 19:46:21 ---------------------------------------------------------------------------
Good morning, I was asked recently whether I had heard of this manufacturer (Precision World) out of China that is a OEM (Original Equipment Manufacturer) for Leica, Nikon, Zeiss and sells their product through ebay. Since I hadn't I thought someone out there may have. Has anyone had any experience with this company or their products?
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
I am basing my comments on work on biofilms on urinary catheters. Bacteria are easier to find after conventional preparation. Processing removes the extracelluar polysaccharides exposing the bacteria. If you want to see their real enviroment then cryoSEM is good. One can sublime off the water and extracellular material (mostly water) eventually. ESEM is good for biofilms but harder, (in my experience; tungsten gun) to get a good image an individual bacterium for various reasons.
Dave
On Fri, 12 Dec 2003 17:08:24 -0800 (PST) JOSE SANCLEMENT {jsanclement-at-yahoo.com} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } SEM--human mucosa } Hi, I'm looking for biofilms on human mucosa. Is } cryostage SEM superior to standard fixations? would my } results be equivalent to using OCT resin? at this } point my samples are in formalin. Any suggestions to } succesfully finding biofilms? appreciaty any input. } Thanks, Jose } } __________________________________ } Do you Yahoo!? } Free Pop-Up Blocker - Get it now } http://companion.yahoo.com/ } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 15 14:45:40 2003
Quick survey here for anyone using XE lamps: for how long do you run a lamp/bulb? Osram claims that the 150W XE has an expected lifetime of 3000 hours, which would be nice BUT I don't know that I trust the lamp not to go pop before 3000h. A new lamphouse would be nice, but lab explosions are such a hassle! Could we (fairly) safely run one for 1500-2000 hours?
Our microscope rep is checking into it, but so far has come up with nothing about the 150W XE, so I thought I'd ask the real experts.
Thanks!
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 05:11:35 2003
I'm not sure, but this question may have been on the List a year or two ago....What is the procedure for changing an HG lamp? I understand there is a right way and a wrong way to do this; unfortunately I know neither one....
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 07:37:36 2003
Question: Could you give me a reference, or a rule of thumb, for good laboratory practice regarding microscope slide drying time before presenting the slide to the pathologist?
Question: I was wondering where to get blade holder for holding blades derived from multi-edged razors, as these do very fine thin sections, normal single-edge razors blades are useless. This thing resembles a credit card into which you can insert the razor blade extracted from multi-edge shaving razors. The ends of teh blade are then gripped, similar in fashion to jewellers saw. Do you have any idea where such a blade holder could be found. It is for preparing thin sections of creosotebush(Larrea tridentata)and the twigs are 1mm in diameter, and covered in resin, so normal single-edge razors blades just mash up specimens.
Carolina Biological Supply Thomas Scientific Ward's Scientific Textbooks of botanical microtechnique
Geoff
by way of Ask-A-Microscopist wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } } Email: rmark-at-nmsu.edu } Name: Mark Robertson } } Organization: NMSU } } Education: Graduate College } } Location: New Mexico } } Question: I was wondering where to get blade holder for holding blades } derived from multi-edged razors, as these do very fine thin sections, } normal single-edge razors blades are useless. This thing resembles a } credit card into which you can insert the razor blade extracted from } multi-edge shaving razors. The ends of teh blade are then gripped, } similar in fashion to jewellers saw. } Do you have any idea where such a blade holder could be found. It is } for preparing thin sections of creosotebush(Larrea tridentata)and the } twigs are 1mm in diameter, and covered in resin, so normal single-edge } razors blades just mash up specimens. } } --------------------------------------------------------------------------- } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 15:06:05 2003
Hi Listers, I'm looking for feedback on different contemporary brands/models for: 1. Vacuum evaporators 2. Sputter coaters 3. Critical point dryers Reliability, serves the needs, bang for the buck, etc.... Thanks for any help! Dee
-- *************************************************************** Do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro Journeys in Microspace (Columbia University Press, 1995) http://www.lsc.org/antarctica/front.html
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 17:03:06 2003
Greetings all, } } I have a question regarding the necessary resolution that would be best } for routine operation. Let me state that I work for a renal path } service where most of the images generated are rather low magnifications } (x1000) to upwards of 100K, although the bulk of photography usually } max's out around 30K or so. We are in the market for a new TEM/CCD } system and were looking at the 2K cameras as an option. My question is } the following: } } For a service such as this, is there any advantage between operating a } 2K camera vs one that is higher (3 or 4K)? If there is no real } difference, does anyone see in the future a move towards the higher res } cameras as prices may or may not come down as the technology becomes } more available? We would like to make our purchase based upon not only } our current needs but thinking of what our needs will be 10 yrs down the } road. } } Thanks in advance, } } Mike Ganger } EM Technicial } Dept. of Surgical Pathology } Weill Cornell Medical College } New York, NY 10021 } 212-746-6437
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 17:17:17 2003
} I'm looking for feedback on different contemporary brands/models for: } 1. Vacuum evaporators } 2. Sputter coaters } 3. Critical point dryers } Reliability, serves the needs, bang for the buck, etc.... } Thanks for any help! } Dear Dee, We are happy with the Cressington evaporator we got from Pella; you can also get a sputter coater with the same vacuum parts. We had the same criteria as you, but we have not had the unit long enough to say anything about reliability beyond that it has worked very well since we got it. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 19:43:13 2003
Finally I can report we are back in operation! The problem was in the HV tank. The HV cable was not contacting one of the three leaf spring contacts in the cable housing, therefore, there was an open circuit to the filament. By physically adjusting the position of the leaf spring we now have good contact. Unless there is an incredible co-incidence here, I believe we may have created the second problem (ie, open circuit to filament) when we were trying to find the first problem (faulty integrated circuit on the HV Stabilzer board). Before we found the faulty IC we had removed the HV cable twice from the tank. Doing this probably moved the leaf spring contact.
Thankyou to Geoff Douglas (Meeco Holdings) for his expert assistance in isolating our problem. Also, thankyou to Terry Fitton and Tony Morgan here at the IMVS, Kerry Gascoigne
____________________________ John Brealey Medical Scientist Electron Microscopy Unit The Queen Elizabeth Hospital IMVS - TQEH Pathology Woodville, 5011 South Australia (08) 82226612
Earlier correspondence...
Hello again,
Still no electron beam. We believe the HV Stabilzer board (new capacitors) and the cable are OK. Gun vacuum is 10 (-5) Torr. Column and camera are 10 (-2) Torr. Gun and camera green lights are on. Can anyone advise on interlocks within the system? We think there are at least four... the gun valve, specimen airlock, camera shutter and camera valve. Something in the system is preventing the filament from switching on. Is there anything else that will prevent the filament turning on? We have tried several filament setups.
Thanks again,
John Brealey
-----Original Message----- } From: John Brealey [mailto:john.brealey-at-imvs.sa.gov.au] Sent: Tuesday, 11 November 2003 01:15 PM To: John Brealey
Hi all,
Our Hitachi H-600 TEM is failing to provide an HV reading at any kV (ie 25, 50, 75, 100). As the microscope is 20 years old I believe the HT cable is the problem. Do you agree? What's the best plan of attack from here? Apart from the HT cable and a dirty gun chamber, are there any other potential causes of this failure? I have never attempted to isolate the HT cable. The vacuum system is fine. I've turned the microscope completely on and off a few times and have vented all compartments to air and back to high vacuum but this has had no effect.
Two days ago I reassembled the rear cosmetic guard that encases the upper vacuum system and the HT cable. Could a slight knock to the cable be enough to finish it off? The microscope was working yesterday, though. I've tried gently wiggling the cable but that gave no response.
The microscope has not been hissing and the HV has appeared quite stable. The only sign that something was not quite right was that some mornings the HV would fail to read at 25kV (but would always read at 50, 75 and 100kV).
In the light of problems encountered by Sarah Ellis (refer to archives) a few months ago I am tempted to clean the gun chamber first (however I doubt if this will fix anything).
Any advice would be gratefully accepted before I contact our institution's electrical engineers.
John Brealey EM Unit The Queen Elizabeth Hospital Institute of Medical & Veterinary Science Adelaide Australia (08) 8222 6612 john.brealey-at-imvs.sa.gov.au
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 19:46:12 2003
Finally I can report we are back in operation! The problem was in the HV tank. The HV cable was not contacting one of the three leaf spring contacts in the cable housing, therefore, there was an open circuit to the filament. By physically adjusting the position of the leaf spring we now have good contact. Unless there is an incredible co-incidence here, I believe we may have created the second problem (ie, open circuit to filament) when we were trying to find the first problem (faulty integrated circuit on the HV Stabilzer board). Before we found the faulty IC we had removed the HV cable twice from the tank. Doing this probably moved the leaf spring contact.
Thankyou to Geoff Douglas (Meeco Holdings) for his expert assistance in isolating our problem. Also, thankyou to Terry Fitton and Tony Morgan here at the IMVS, Kerry Gascoigne at Flinders Medical Centre, and all respondents who provided so much help and advice.
____________________________ John Brealey Medical Scientist Electron Microscopy Unit The Queen Elizabeth Hospital IMVS - TQEH Pathology Woodville, 5011 South Australia (08) 82226612
Earlier correspondence...
Hello again,
Still no electron beam. We believe the HV Stabilzer board (new capacitors) and the cable are OK. Gun vacuum is 10 (-5) Torr. Column and camera are 10 (-2) Torr. Gun and camera green lights are on. Can anyone advise on interlocks within the system? We think there are at least four... the gun valve, specimen airlock, camera shutter and camera valve. Something in the system is preventing the filament from switching on. Is there anything else that will prevent the filament turning on? We have tried several filament setups.
Thanks again,
John Brealey
-----Original Message----- } From: John Brealey [mailto:john.brealey-at-imvs.sa.gov.au] Sent: Tuesday, 11 November 2003 01:15 PM To: John Brealey
Hi all,
Our Hitachi H-600 TEM is failing to provide an HV reading at any kV (ie 25, 50, 75, 100). As the microscope is 20 years old I believe the HT cable is the problem. Do you agree? What's the best plan of attack from here? Apart from the HT cable and a dirty gun chamber, are there any other potential causes of this failure? I have never attempted to isolate the HT cable. The vacuum system is fine. I've turned the microscope completely on and off a few times and have vented all compartments to air and back to high vacuum but this has had no effect.
Two days ago I reassembled the rear cosmetic guard that encases the upper vacuum system and the HT cable. Could a slight knock to the cable be enough to finish it off? The microscope was working yesterday, though. I've tried gently wiggling the cable but that gave no response.
The microscope has not been hissing and the HV has appeared quite stable. The only sign that something was not quite right was that some mornings the HV would fail to read at 25kV (but would always read at 50, 75 and 100kV).
In the light of problems encountered by Sarah Ellis (refer to archives) a few months ago I am tempted to clean the gun chamber first (however I doubt if this will fix anything).
Any advice would be gratefully accepted before I contact our institution's electrical engineers.
John Brealey EM Unit The Queen Elizabeth Hospital Institute of Medical & Veterinary Science Adelaide Australia (08) 8222 6612 john.brealey-at-imvs.sa.gov.au
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 20:10:33 2003
We have been quite happy with our Bal-Tec critical point dryer and our Denton sputter coater. Both have been in service for several years now and have survived users of all levels of expertise.
At 06:43 PM 12/16/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 21:15:17 2003
Mike Digital cameras is fast growing area, so you could not predict what will happens 10 years later. In my point of view, EM digital camera's technology is still under developing: major manufacturers offered to us new (technologically new, not only next model) models nearly every year and it's not only about amount of pixels in it. It's about new CCDs with bigger/smaller pixels, faster readout systems, better coupling etc. Another thing, which comes to my mind: modern scientific grade CCD's life is about 5 years, so, perhaps, you need a new camera sooner than in 10 years... As for "resolution" - this issue has been discussed in this forum many, many times (you may need to check our archive). Camera itself does not provide "resolution". Resolution comes from the microscope and camera is just instrument to transfer it into some amount of discrete pixels. How many pixels you need? It depends. Standard resolution in the pictures published by Science or Nature is 72 dpi. So, if you need your picture will be published 1:1 in those magazines - you actually don't nee too much. Seriously speaking, editors commonly ask for 300 dpi resolution for submitted pictures (to be able to edit image if necessary). If your image is 5x5", at 300 dpi you will have nearly exact 2 megapixels. So, it means, 2K camera is OK for such type of job. What about 2K vs 4K cameras? If you have modern microscope controlled by computer, most camera's manufacturers offered software for "digital montage". So, your camera automatically took a couple of images, then assembled them into a single large image. You may find that this option may work to you (may not at low magnification or drift etc). Another thing to consider: as more pixels you have, as slower camera. It means, that you probably will not have a good TV mode on 4K cameras. Returning back to the resolution issue: the beauty of the digital camera is that you could took as many pictures as you want. So, I usually took a few pictures with different magnification. If you need good detail's resolution on your digital image - you need to go to the higher mag. In this terms, you may have image with very good resolution of details even on 1K camera - you need to use higher magnification and price would be the size of the field. Another thing, which I always recommend: ask manufacturers for demo, use your own samples and took pictures by yourself, then compare side-by-side. You may be surprised to see how different cameras may be.
I hope, it helps. Sergey
At 03:14 PM 12/16/2003, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
After some considerable research, I replaced my long standing Anatech Hummer VII sputter coater with a Denton Desk II coater. After about four months of use, I am really happy with it. Very reliable, easy to use. Fast. Compact (Cressington has a large foot print) and easy to use.
I looked at the Cressington 208FE and found the Denton to be preferable. But of course, your criteria may vary. Anyway, I like the Denton.
gary g.
Oh...no financial interest one way or the other.
At 01:17 PM 12/16/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Dec 16 22:29:55 2003
Many thanks for a number of replies. Unfortunately I formulated my question too fuzzy. I am not interested in the measurements of porosity utilizing EDS. What I am interested in is the effect of porosity (or nanoporosity) on EDS measurements. It seems that an increase in porosity should lead to a decrease in the intensity of X-rays, and that this dependence should not be linear. It's only my gut feeling, I do not want to carry out calculations. I am sure all these calculations were done a long time ago, and I'll appreciate any lead to a proper publication and/or a short explanation of the problem.
Thanks,
Vladimir
________________________________
} From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] Sent: Wed 12/10/2003 2:57 PM To: Microscopy (E-mail)
Hello all, I have samples of the same material with suspected difference in porosity, size of pores could be from a few nanometers to a few hundred nanometers.
Is it possible to detect porosity with microanalysis (EDS)? How intensity could change because of porosity? Is it possible to calculate the "detection limit" of porosity?
Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} Subject: [Microscopy] IASTED Newsletter on Modelling and Simulation - Dec 2003 } } IASTED International Newsletter on Modelling and Simulation } December 15, 2003 } } UPCOMING MODELLING AND SIMULATION CONFERENCES } 1. The IASTED International Conference on Applied Simulation and Modelling - ASM 2004 } June 28-30, 2004, Rhodes, Greece } } Important Deadlines: } Submissions Due: Feb. 15, 2004 } Notification of Acceptance: Apr. 1, 2004 } Registration Deadline: May 1, 2004 } } To submit a paper, tutorial or special session or for more information visit our website at http://www.iasted.org/conferences/2004/greece/asm.htm } } 2. The 4th IASTED International Conference on Modelling, Simulation, and Optimization - MSO 2004 } August 16-18, 2004, Kauai, Hawaii, USA } } Important Deadlines: } Submissions Due: Mar. 5, 2004 } Notification of Acceptance: Apr. 15, 2004 } Registration Deadline: June 1, 2004 } To submit a paper, tutorial or special session or for more information visit our website at http://www.iasted.org/conferences/2004/hawaii/mso.htm } } UPCOMING DEADLINES } The submission deadline for the 15th IASTED International Conference on Modelling and Simulation - MS 2004 being held March 1-3, 2004 in Marina Del Rey, CA, USA has passed. Delegates without papers (attendees) are welcome to register until Jan. 15, 2004. } } For registration information visit our website at http://www.iasted.org/conferences/2004/marina/ms.htm } } **Tutorial Announcement** } "Simulation-Based Engineering of Complex Systems Using EXTEND+OpEMCSS" } Presented by Dr. John R. Clymer - California State University, Fullerton, USA } For additional information visit http://www.iasted.org/conferences/2004/marina/ms-tutorial1.htm } } **Special Session** } "Intelligent Reconfigurable Simulation" Organized by Dr. Tudor Niculiu -University "Politehnica" of Bucharest, Romania. For additional information visit http://www.iasted.org/conferences/2004/marina/ms-specialsessions.htm } } } FUTURE CONFERENCES } Mark your calendars! The IASTED International Conference on Environmental Modelling and Simulation - EM&S 2004 will be held Nov. 22-24, 2004 in St. Thomas, Virgin Islands, USA. } } } MODELLING AND SIMULATION JOURNAL FROM ACTA PRESS } The International Journal of Modelling and Simulation - First published in 1981, this journal covers all aspects of modelling, simulation, languages, software, hardware, methodology, numerical and graphical methods, virtual reality, statistical techniques, tutorials, surveys, and applications. It also includes book reviews, conference notices, call for papers, and new publications. } Editor-in-Chief: Prof. A. Houshyar } Frequency: 4 issues per year } 2003 Rate: US$256.00 } Postage & Handling: US$25.00 } ISSN: 0228-6203 (205) } http://www.actapress.com/journals/journals.htm#Modelling } } It pays to be a member! One of the benefits of your IASTED membership is a complimentary subscription to an ACTA Press journal. Become a member today! http://www.iasted.org/member/OnlineMembershipForm.pdf } } } CONFERENCE PROCEEDINGS AVAILABLE } Past conference proceedings in the area of modelling and simulation are available for purchase from ACTA Press - http://www.actapress.com/proceedings/proceedings.htm } } For more information, to be removed from our mailing list, or to join one of the following mail groups: Power, Telecommunication, Bio-Medicine, Signal and Image Processing, Artificial Intelligence, Business, Software Engineering, Education, Databases and Knowledge Engineering, Internet and Applications, Parallel and Distributed Computing, please contact: } IASTED } #80, 4500 - 16th Avenue N.W. } Calgary, Alberta } Canada T3B 0M6 } Tel: 403-288-1195 } Fax: 403-247-6851 } E-mail: calgary-at-iasted.com } Web site: http://www.iasted.org } } ***************************************************** } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 04:03:06 2003
I assume you are talking about area analysis which provides a display of average composition of the scanned area. As you know, dark regions in SEM image are where the e-detector sees less SE/BSE due to a) less generation and/or less survival of them. Reasons for above a) and b) include when the primary beam hits a hole or a pore. Likewise, depending on the size and shape of a pore and the nature of the material, X-rays may not be generated or less may be generated when the primary e-beam has difficulty reaching there or may not survive well (absorbed) in the location of the pore. That is precisely the reason why a reliable quantitative analysis should start with a flat sample surface and why the detector has an important parameter, take-off angle, for quan-routine.
X-ray mapping of a homogeneous but uneven sample should provide some insight into this.
**************************************** Chaoying Ni, PhD The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
On Tue, 16 Dec 2003, Dusevich, Vladimir wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Many thanks for a number of replies. } Unfortunately I formulated my question too fuzzy. I am not interested in the } measurements of porosity utilizing EDS. What I am interested in is the } effect of porosity (or nanoporosity) on EDS measurements. It seems } that an increase in porosity should lead to a decrease in the intensity of X-rays, } and that this dependence should not be linear. It's only my gut feeling, } I do not want to carry out calculations. I am sure all these calculations were } done a long time ago, and I'll appreciate any lead to a proper publication } and/or a short explanation of the problem. } } Thanks, } } Vladimir } } } ________________________________ } } } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] } Sent: Wed 12/10/2003 2:57 PM } To: Microscopy (E-mail) } Subject: [Microscopy] Microanalysis of porous material } } } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello all, } I have samples of the same material with suspected } difference in porosity, size of pores could be from } a few nanometers to a few hundred nanometers. } } Is it possible to detect porosity with microanalysis } (EDS)? How intensity could change because of porosity? } Is it possible to calculate the "detection limit" } of porosity? } } Thank you, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 08:00:42 2003
We have a copy of a classic wall poster called 'Murphy's Immutable Laws of Microanalysis' which was published in 1989 by Kevex. Does anyone out there know if these are still available anywhere.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 (0)1223 334325 Fax: +44 (0)1223 334567 Email: djv23-at-cam.ac.uk
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 08:31:16 2003
A simple way of confirming your proposed relationship of porosity to the EDS values would be to actually measure the porosity of the sample both in terms of total porosity and the size range the porosity occurs in. Nitrogen adsorption is used for porosity less than 30 nm and mercury intrusion for the porosity range of 7 nm to 300 µm. Non-mercury intrusion is used for well characterized materials. Molecular probe techniques are used to characterize size and shape of pores less than 1 nm.
J. Roy Nelson, Ph.D. Material Testing Lab. Pennington, NJ (609) 730-0575
"Dusevich, Vladimir" wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Many thanks for a number of replies. } Unfortunately I formulated my question too fuzzy. I am not interested in the } measurements of porosity utilizing EDS. What I am interested in is the } effect of porosity (or nanoporosity) on EDS measurements. It seems } that an increase in porosity should lead to a decrease in the intensity of X-rays, } and that this dependence should not be linear. It's only my gut feeling, } I do not want to carry out calculations. I am sure all these calculations were } done a long time ago, and I'll appreciate any lead to a proper publication } and/or a short explanation of the problem. } } Thanks, } } Vladimir } } } ________________________________ } } } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] } Sent: Wed 12/10/2003 2:57 PM } To: Microscopy (E-mail) } Subject: [Microscopy] Microanalysis of porous material } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello all, } I have samples of the same material with suspected } difference in porosity, size of pores could be from } a few nanometers to a few hundred nanometers. } } Is it possible to detect porosity with microanalysis } (EDS)? How intensity could change because of porosity? } Is it possible to calculate the "detection limit" } of porosity? } } Thank you, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 10:07:00 2003
Sergey is right, in that the resolution of a TEM CCD camera is not really determined by the number of pixels. Every TEM camera uses a Phosphor or YAG to convert the electrons into light, and this determines the resolution of the camera. This resolution depends on the thickness of the phosphor and the acceleration voltage, but is on the order of a few microns to a few tens of microns. Each camera, regardless of the number of pixels, can be engineered to resolve this, through optical elements or fiber optics. Once you realize this, the advantage of cameras with more pixels becomes clear: field of view. In most cases, field of view can also be enlarged, as Sergey correctly mentions, by acquiring several images and stitch them together. Many software packages allow to do this multiple image alignment procedure automatically (if you have a motor stage), or semi-automatically. And another point, where Sergey is correct is the readout speed. Most "large" CCD chips have a fairly slow readout speed, due to the electric capacity of their pixels. This reduces the readout in many cases to around 1 frame per second or less. These cameras are replacements for film, but you need to work with the binoculars to find an area and focus. Many of these cameras have partial readouts, which help somewhat. Using "smaller" CCDs with smaller pixel sizes allows a much faster read-out. For example, our KeenView, which uses a smaller chip and a tapered fiber-optic to match the phosphor resolution, allows about 12 frames per second in full resolution (1330x1024). This speed is sufficient to also work with the camera for location and focusing. In other words: There is, like anywhere else in live, pros and cons for each camera, and in the end it depends on your application and your budget that determines the best camera for your TEM.
If you want to discuss specifics about your application, please contact me off line.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Tuesday, December 16, 2003 20:26 To: Microscopy-at-sparc5.microscopy.com
Mike Digital cameras is fast growing area, so you could not predict what will happens 10 years later. In my point of view, EM digital camera's technology is still under developing: major manufacturers offered to us new (technologically new, not only next model) models nearly every year and it's not only about amount of pixels in it. It's about new CCDs with bigger/smaller pixels, faster readout systems, better coupling etc. Another thing, which comes to my mind: modern scientific grade CCD's life is about 5 years, so, perhaps, you need a new camera sooner than in 10 years... As for "resolution" - this issue has been discussed in this forum many, many times (you may need to check our archive). Camera itself does not provide "resolution". Resolution comes from the microscope and camera is just instrument to transfer it into some amount of discrete pixels. How many pixels you need? It depends. Standard resolution in the pictures published by Science or Nature is 72 dpi. So, if you need your picture will be published 1:1 in those magazines - you actually don't nee too much. Seriously speaking, editors commonly ask for 300 dpi resolution for submitted pictures (to be able to edit image if necessary). If your image is 5x5", at 300 dpi you will have nearly exact 2 megapixels. So, it means, 2K camera is OK for such type of job. What about 2K vs 4K cameras? If you have modern microscope controlled by computer, most camera's manufacturers offered software for "digital montage". So, your camera automatically took a couple of images, then assembled them into a single large image. You may find that this option may work to you (may not at low magnification or drift etc). Another thing to consider: as more pixels you have, as slower camera. It means, that you probably will not have a good TV mode on 4K cameras. Returning back to the resolution issue: the beauty of the digital camera is that you could took as many pictures as you want. So, I usually took a few pictures with different magnification. If you need good detail's resolution on your digital image - you need to go to the higher mag. In this terms, you may have image with very good resolution of details even on 1K camera - you need to use higher magnification and price would be the size of the field. Another thing, which I always recommend: ask manufacturers for demo, use your own samples and took pictures by yourself, then compare side-by-side. You may be surprised to see how different cameras may be.
I hope, it helps. Sergey
At 03:14 PM 12/16/2003, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I recently got a smaller size version (11 X 17) after alot of pleading, from a Noran serviceman. But I don't know how available they are. Try contacting Noran at www.thermo.com
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
David Vowles {djv23-at-msm.cam.ac.uk} 12/17/03 09:11 AM
To microscopy-at-ns.microscopy.com cc
Subject Microanalysis Poster
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Dear All,
We have a copy of a classic wall poster called 'Murphy's Immutable Laws of Microanalysis' which was published in 1989 by Kevex. Does anyone out there know if these are still available anywhere.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 (0)1223 334325 Fax: +44 (0)1223 334567 Email: djv23-at-cam.ac.uk
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 10:38:33 2003
I shall appreciate advise on how to connect a Nikon Coolpix 995 to a Reichert-Jung MEF3 (inverted) Light Microscope, and yet be able to photograph the whole field of view, or at least close to it, as is viewed and photographed through the Polaroid or 35mm cameras. I tried to replace the original 35mm camera, normally positioned on the down-left-hand side of the MEF3, by a Coolpix 995 equipped with a x0.63 C-mount and a home-made short adapter, the latter used for matching. But with or without the adapter, and regardless of the position of the Coolpix relative to the microscope port, the field of view was much smaller than the one photographed in the 35mm camera (in other words, the aparent magnification was much higher). In order to get a similar field of view I had to use a lower power objective of the MEF3. This limits the minimal attainable magnification. If the optical magnification of the Coolpix is reduced to a minimum, to get a larger field of view, only a partial, circular field is exposed in the center, the rest is blocked out, like looking through a tube.
Has anyone faced the same problem? can a different C-mount and adapter solve it?
Thanx,
Dr. Uri Admon Beer-Sheva Israel
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 10:38:41 2003
I would like to know where I can buy microscope slide storage (slide mailer) for petrographic thin sections. I am looking for storage in which I can put a glass slide (1x2 inch).
I can find the provider of slide mailer for biological glass slide (1x 4 inch ?) at: http://www.omni-optical.com/micro/sm320.htm
I need such case for 1x 2 inch thin sections. Please advise.
Thank you, Hiromi Konishi The University of New Mexico
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 11:38:40 2003
A few years ago, I was able to obtain a smaller version of the large poster from Noran which purchased Kevex sometime back.
Also, there is a so-so copy on my web site which can be downloaded. It was from multiple flatbed scans (with permission) of the larger poster and the match-up is less than perfect. My web address is below.
Regards, Woody ----------------------------- Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: David Vowles [mailto:djv23-at-msm.cam.ac.uk] Sent: Wednesday, December 17, 2003 9:12 AM To: microscopy-at-ns.microscopy.com
Dear All,
We have a copy of a classic wall poster called 'Murphy's Immutable Laws of Microanalysis' which was published in 1989 by Kevex. Does anyone out there know if these are still available anywhere.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 (0)1223 334325 Fax: +44 (0)1223 334567 Email: djv23-at-cam.ac.uk
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 12:50:52 2003
I've never seen a copy anywhere -- not even at eBay. Mine is actually just a touched-up digital image that someone had scanned and posted on the web.
Ellery
David Vowles wrote: } We have a copy of a classic wall poster called 'Murphy's Immutable } Laws of Microanalysis' which was published in 1989 by Kevex. Does } anyone out there know if these are still available anywhere?
--------------- Ellery E. Frahm Research Scientist/Manager Electron Microprobe Laboratory Department of Geology & Geophysics University of Minnesota - Twin Cities Lab Website: http://probelab.geo.umn.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 13:27:42 2003
All: I'm submitting this for Dr. Stan Trauth of Arkansas State University. Please reply to Dr. Trauth off-line: ----------------------------------------------------------------------------------------------------------------------------------
Would you mind sending me just a few names of EM techs or labs that might be willing to share information regarding the cost of operating a TEM or an SEM--hourly or otherwise?
I would sincerely appreciate the assistance.
Thanks,
Stan Trauth strauth-at-astate.edu
Dr. Stan Trauth Department of Biological Sciences Arkansas State University P.O. Box 599 State University, AR 72467-0599 Ph. 870.972.3082 FAX 870.972.2638 http://biology.astate.edu/faculty/strauth/Dr_%20Stanley%20Trauth.htm
Wasn't this discussed some years ago? If memory serves me someone found it posted on the internet. If you are that interested a search of the archives is in order.
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.249 Fax: (770) 487-1460 email: robert.fowler-at-tdktca.com www.component.tdk.com
ekomarnicki-at-Mac Dermid.com To: David Vowles {djv23-at-msm.cam.ac.uk} cc: microscopy-at-ns.microscopy.com 12/17/2003 Subject: [Microscopy] Re: Microanalysis Poster 11:28 AM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I recently got a smaller size version (11 X 17) after alot of pleading, from a Noran serviceman. But I don't know how available they are. Try contacting Noran at www.thermo.com
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
David Vowles {djv23-at-msm.cam.ac.uk} 12/17/03 09:11 AM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear All,
We have a copy of a classic wall poster called 'Murphy's Immutable Laws of Microanalysis' which was published in 1989 by Kevex. Does anyone out there know if these are still available anywhere.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 (0)1223 334325 Fax: +44 (0)1223 334567 Email: djv23-at-cam.ac.uk
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 15:34:14 2003
We certainly are not totally impartial but for reliability, ease of use and longevity you should look at the Ladd 30000 Vacuum Evaporator.
John Arnott
Disclaimer: Ladd Research sells a variety of vacuum equipment and other EM supplies
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Dee Breger" {micro-at-ldeo.columbia.edu} To: {microscopy-at-msa.microscopy.com} Sent: Tuesday, December 16, 2003 4:17 PM
Hiromi,
Try the sample prep companies, especially Buehler. Also, Carolina Biological often carries storage like this. Make sure that they understand that these are petrographic slides, not regular microscope slides.
Good hunting! Barbara Foster Microscopy/Microscopy Education, Inc. (MME, Inc.) 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Optimizing Light Microscopy is still available through MME. For information on ordering single or small quantities, visit our website. For discounts on classroom-sized, call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&
At 08:50 AM 12/17/03 -0800, Hiromi Konishi wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 17:31:02 2003
I am currently trying to resurrect a 1985 vintage Microspec WDX-2A wavelength spectrometer. The system is mounted on a Philips 535 SEM and seems to be working but I don't have any manuals for it. If anyone has a set a manuals laying around they wouldn't mind parting with or copying they can respond by contacting me directly at wim5-at-lehigh.edu
Thanks, Bill
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 17 22:36:02 2003
The 990 and 995 do a good/decent job of microphoto. However, don't be surprised by many types of aberrations and distortion. Get an Optem coupler and set the camera at infinity...wide open. I think that this is about the best that you will get. It should be very close to 1:1 relative to oculars and digicam. A real digital microscopy camera produces dramatically different results. But the CoolPix price is very attractive. It is a good place to start.
gary g.
At 09:39 AM 12/17/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 01:56:56 2003
AU} ------------------------------------------------------------------------------ AU} The Microscopy ListServer -- Sponsor: The Microscopy Society of America AU} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver AU} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html AU} -------------------------------------------------------------------------------
AU} Dear All, and best wishes for a happy New Year,
AU} I shall appreciate advise on how to connect a Nikon Coolpix 995 to a AU} Reichert-Jung MEF3 (inverted) Light Microscope, and yet be able to AU} photograph the whole field of view, or at least close to it, as is viewed AU} and photographed through the Polaroid or 35mm cameras. I tried to replace AU} the original 35mm camera, normally positioned on the down-left-hand side of AU} the MEF3, by a Coolpix 995 equipped with a x0.63 C-mount and a home-made AU} short adapter, the latter used for matching. But with or without the AU} adapter, and regardless of the position of the Coolpix relative to the AU} microscope port, the field of view was much smaller than the one AU} photographed in the 35mm camera (in other words, the aparent magnification AU} was much higher). In order to get a similar field of view I had to use a AU} lower power objective of the MEF3. This limits the minimal attainable AU} magnification. If the optical magnification of the Coolpix is reduced to a AU} minimum, to get a larger field of view, only a partial, circular field is AU} exposed in the center, the rest is blocked out, like looking through a tube.
AU} Has anyone faced the same problem? can a different C-mount and adapter solve AU} it?
AU} Thanx,
AU} Dr. Uri Admon AU} Beer-Sheva AU} Israel
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 02:46:11 2003
} From: Christian Ronse {cronse-at-dpt-info.u-strasbg.fr} } } INTERNATIONAL SYMPOSIUM ON MATHEMATICAL MORPHOLOGY: } 40 YEARS ON } } Monday 18th to Wednesday 20th April 2005, Paris, France } } } This symposium is the seventh of a series of conferences devoted to } mathematical morphology and its applications in image processing. It } will be held at 40 years after mathematical morphology was born } through the collaborative work of Jean Serra and the late Georges } Matheron in the summer of 1964. It is thus a great opportunity to } present the most recent developments in this field, and to assess its } relevance in image processing, signal processing and computer science. } } The theme of the conference will be mathematical morphology in the } broad sense, around the notion that pictures represent geometrical } objects with luminance (or colour) profiles, that can be analysed by } their interactions with other geometrical objects. More specifically, } the following topics are eligible for submissions: } } Morphological theory: } - lattice theory and algebraic models of images and operators } - combinatorial topology and discrete geometry in image processing } - metrics and topologies for shapes and pictures } - PDEs, level set methods and geometrical scale space } - discrete and continuous geometrical measures, integral geometry } - random sets and geometrical probability } - image connectivity and connected operators } - relations of morphology with signal processing and computer science } Morphological image processing: } - geometrical image analysis } - order-statistics image filtering } - geometrical and topographical segmentation } - colour and multi-channel morphology } - morphological pattern recognition } - motion analysis } - texture analysis } - shape analysis } - image coding } - algorithms and data structures for morphology } Applications of morphology in: } - geoscience and remote sensing } - bio-medical imaging } - materials science } - quality control } - document processing } - data analysis } } The scientific program will include invited talks and contributed } papers (no posters). They will appear in the proceedings volume. } This volume will be available at the beginning of the conference. } } This ISMM will be held in honour of Jean Serra, on the occasion of his } 65th birthday. Its venue will follow that of DGCI in Poitiers (from } Wednesday the 13th to Friday the 15th April 2005). } } } INFORMATION / CONFERENCE WEB SITE: } --------------------------------- } } A conference web site (hosted by esiee.fr) will be available shortly, } check http://ams.jrc.it/mdigest/calendar.html } } It will contain information about submission, registration, invited } speakers, accommodation, etc. For additional information, you can also } contact the organising committee. } } } SUBMISSION PROCEDURES: } --------------------- } } Prospective authors are invited to submit a full paper using the } electronic procedure described in the conference web site. The } manuscript file should be in the PDF format. } } In case of problems with the electronic procedure, it is possible to } submit a paper by sending 5 printed copies of the manuscript to any } one of the 3 conference chairs. Email submissions should be avoided. } } The manuscript title header should include the names, institutions and } addresses of the authors, an abstract of up to 200 words, and } keywords. The submission procedure requires providing full postal and } e-mail addresses, phone and fax numbers of the contact author. } } Acceptance of papers is based on appropriateness of the topic and on } quality, novelty, and clarity of exposition. Each paper will be } reviewed by at least two members of the Program Committee (or referees } chosen by them), and their reviews will be returned to the author. } } Accepted papers will appear in the proceedings volume. The final paper } has to be prepared in LaTeX according to the style file provided by } the organisers (see conference web site). The size should be limited } to 10 pages including artwork and references. Authors are strongly } advised to adopt that LaTeX style already for the initial submission. } } } IMPORTANT DATES: } --------------- } } 10 September 2004: Submission of full paper } 12 November 2004: Notification of acceptance } 14 January 2005: Camera-ready full paper } } } CONFERENCE CHAIRS: } ----------------- } } Christian Ronse } LSIIT UMR 7005 CNRS-ULP } Parc d'Innovation, Boulevard Sébastien Brant } BP 10413 } 67412 ILLKIRCH CEDEX } FRANCE } Tel: +33 3 90 24 45 00 } Fax: +33 3 90 24 44 55 } Email: cronse at dpt-info.u-strasbg.fr } } Laurent Najman } Laboratoire A2SI } Groupe ESIEE } Cité Descartes - BP 99 - 2, Bd Blaise Pascal } 93162 NOISY LE GRAND CEDEX } FRANCE } Tel: +33 1 45 92 66 72 } Fax: +33 1 45 92 66 99 } Email: l.najman at esiee.fr } } Etienne Decencière Ferrandière } CMM - Ecole des Mines } 35, rue Saint Honoré } 77305 Fontainebleau CEDEX } Tel: +33 1 64 69 48 09 } Fax: +33 1 64 69 47 07 } Email: Etienne.Decenciere at cmm.ensmp.fr } } } } PROGRAM COMMITTEE: } ----------------- } } Christian Ronse (Head), Université Louis Pasteur, Strasbourg, France } } Junior Barrera, Universidade de São Paulo, Brazil } Gilles Bertrand, ESIEE, Noisy-Le-Grand, France } Isabelle Bloch, ENST, Paris, France } Gunilla Borgefors, Uppsala Universitet, Sweden } Jose Crespo, Universidad Politécnica de Madrid, Spain } Etienne Decencière, Ecole des Mines de Paris, Fontainebleau, France } John Goutsias, Johns Hopkins University, Baltimore, MA, USA. } Frederic Guichard, Vision IQ & DO Labs, Boulogne-Billancourt, France } Henk Heijmans, CWI, Amsterdam, The Netherlands } Dominique Jeulin, Ecole des Mines de Paris, Fontainebleau, France } Renato Keshet, HP Labs, Israel } Ron Kimmel, Technion, Haifa, Israel } Petros Maragos, National Technical University of Athens, Greece } Fernand Meyer, Ecole des Mines de Paris, Fontainebleau, France } Jean-Michel Morel, Ecole Normale Supérieure, Cachan, France } Laurent Najman, ESIEE, Noisy-Le-Grand, France } Ioannis Pitas, Aristotle University of Thessaloniki, Greece } Gerhard Ritter, University of Florida, Gainesville, FL, USA } Jos Roerdink, Rijksuniversiteit Groningen, The Netherlands } Philipe Salembier, Universitat Politecnica de Catalunya, Barcelona,Spain } Michel Schmitt, Ecole des Mines de Paris, Fontainebleau, France } Pierre Soille, EC Joint Research Centre, Ispra, Italy } Hugues Talbot, CSIRO, Sydney, Australia } Rein van den Boomgaard, Universiteit van Amsterdam, The Netherlands } Marc Van Droogenbroeck, Université de Liège, Belgium } Luc Vincent, Soligence Corporation, Palo Alto, CA, USA } Joachim Weickert, Universität Saarland, Saarbrücken, Germany } } } ORGANISING COMMITTEE: } -------------------- } } Laurent Najman, ESIEE, Noisy-Le-Grand, France } Isabelle Bloch, ENST, Paris, France } Petr Dokladal, Ecole des Mines de Paris, Fontainebleau, France } Christian Ronse, Université Louis Pasteur, Strasbourg, France } } } } ------------------------------ } ======================================================= } 3 ICIAR 2004: ANNOUNCEMENT AND CALL FOR PAPERS } ======================================================= } } From: ICIAR 2004 {iciar04-at-fe.up.pt} } } INTERNATIONAL CONFERENCE ON IMAGE ANALYSIS AND RECOGNITION (ICIAR 2004) } September 29 - October 1, 2004, Porto, Portugal } } General Chair } Aurelio Campilho } campilho-at-fe.up.pt } } General Co-Chair } Mohamed Kamel } mkamel-at-uwaterloo.ca } } Paper submission deadline: April 16, 2004 } } ICIAR - International Conference on Image Analysis and Recognition aims to } bring together researchers in the fields of Image Processing, Analysis and } Recognition. It will be organized annually, alternating between Europe and } North America. In 2004, the conference is held in Porto, Portugal. ICIAR - } 2005 will take place in Toronto, Canada. } The conference is a result of discussion between researchers in Portugal and } Canada to encourage collaboration and exchange between the two Countries } with participation of experts and researchers from other Countries. } The conference will address recent advances in theory, methodologies and } applications. The scientific program will include invited speakers and fully } refereed contributions that will be published in the conference proceedings. } } Please find the call for papers and more information at the webpage } http://www.iciar.uwaterloo.ca } } ------------------------------ } } =======================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 08:28:31 2003
Title-Subject: [Microscopy] [Filtered] Nikon Coolpix Digital Camera
Question: I have a couple of general questions for using Nikon Collpix Digital Camera system, since I am very interested in setting up this system for my photographing to both whole mount embryos and immunostained parrafin slides as a replacement of the conventional films The questions are:
1. What is the advantage of using Nikon Coolpix camera but not other possible digital cameras for the microscopes? 2. I was told to choose Coolpix 5000 over 5700, since the 5700 will give rise to blak periphery field due to the naroow field of view. Is this true? How do I find out what size of chip in different Coolpix Cameras?
In the past everyone has been very helpful so once again I am returning to the well of knowledge for advice:
I have a Philips 400 series TEM (used) currently being installed. To my surprise I found out it has a LaB6 filament. It has been suggested that I consider a CeBix filament. In either case it was suggested I use either the Mini Vogel or Denka style.
I'm light / SEM microscopist, TEM is a largely explored country to me. I anticipate using the TEM for particle sizing and polymer phase studies and I don't expect to exceed 100K magnification at 80 to 100 kv. I would like to get the best value in terms of performance, ease of use, working life for the money. Any advise, experience, recommendations on type, filament material, tip configuration, special awareness of problems or advantages would be welcome.
PS: where can I get a passport to TEM land? :-)
Thanks . . . . .
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 10:19:55 2003
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Energy Beam Sciences is the US distributor for the Polaron Range of SEM prep equipment.
The E6700 benchtop evaporator is one option that offers ease of use, along with choice of options to fit any of your sample preparation needs.
Your welcome to contact me by email MDufraine-at-ebsciences.com, or Tel. 1-800-992-9037 X340, to learn more about the Polaron line of evaporators, coaters, and CPD equipment we offer.
Mike Dufraine Energy Beam Sciences, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 10:42:19 2003
This company makes cardboard boxes for petrographic slides The one I have here holds 25 1X2 slides.
Palouse Petro Products 452 Sand Road Pullman, WA 99163-9620 (509) 332-3695 (800) 632-3695 (509) 332-3606 fax
James Talbot
K/T GeoServices, Inc. Bulk and Clay Mineralogy by X-ray Diffraction www.ktgeo.com (940) 597-9076
At 08:50 AM 12/17/03 -0800, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 10:47:37 2003
I've had bulbs crack, and I have changed them because they became so discolored that they would light but not 'show up for work'. I have also had them, finally, not start and then crumble when I begin to remove them. Now these are the mercury bulbs. I had one 150 XE that ran for 200 hours over three years and still seemed to be OK, so I left it alone. I did not follow it after I left it behind.
Unless there is a quantitative reason for swapping out bulbs, that is, they lose intensity because of the discoloration of the glass or they lose power because they have a micro-leak (or whatever it might be), my own practice has been to keep the bulb until I don't get sufficient fluorescence. Anyway, most published operational durations are only indications of how long the manufacturer feels the bulb SHOULD operate at specs, but we all know from home that many 2000 hr bulbs don't last that long. I just let my high intensity bulbs go, thus, I have Hg bulbs for my old Leitz in their original boxes that are a decade old, because my use has been down of late.
I have NEVER had a bulb explode, so I don't have any advice about that phenomenon.
Cheers and Merry Christmas,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson-at-wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/
-----Original Message----- } From: Tamara Howard [mailto:thoward-at-unm.edu] Sent: Monday, December 15, 2003 3:49 PM To: Confocal Microscopy List; Microscopy Server
Quick survey here for anyone using XE lamps: for how long do you run a lamp/bulb? Osram claims that the 150W XE has an expected lifetime of 3000 hours, which would be nice BUT I don't know that I trust the lamp not to go pop before 3000h. A new lamphouse would be nice, but lab explosions are such a hassle! Could we (fairly) safely run one for 1500-2000 hours?
Our microscope rep is checking into it, but so far has come up with nothing about the 150W XE, so I thought I'd ask the real experts.
Thanks!
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 11:13:52 2003
Dear Bill, I have manuals for the WDX-3PC, which was the first PC-computer-controlled Microspec. I believe the spectrometers were similar but the controller was very different. Which is it you are having trouble with? I might suggest you contact Joe Carr of Oxford Instruments (e-mail carr-at-oxford.usa.com). He is their Microspec specialist and might know if there are any old manuals available. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "William J Mushock (by way of MicroscopyListserver)" {wim5-at-lehigh.edu} To: {microscopy-at-ns.microscopy.com} Sent: Wednesday, December 17, 2003 3:42 PM
Dr. Uri Admon,
We have an adapter for the Coolpix for your microscope that will correct this and allow you to image the entire field of view.
You can remove your C-mount and mount directly to the microscope port using our optics that are specifically designed for the Coolpix cameras to give you a large portion of the zoom range of the Coolpix cameras.
If you take a look at the following page you can find the details: http://www.mvia.com/Coolpix/clpxadpt.htm
Specifically for mounting to your microscope, this section should help: http://www.mvia.com/Coolpix/clpxadpt.htm#Leica,_Leitz,_Wild
Thanks And Happy Holidays! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Admon Uri wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear All, and best wishes for a happy New Year, } } I shall appreciate advise on how to connect a Nikon Coolpix 995 to a } Reichert-Jung MEF3 (inverted) Light Microscope, and yet be able to } photograph the whole field of view, or at least close to it, as is viewed } and photographed through the Polaroid or 35mm cameras. I tried to replace } the original 35mm camera, normally positioned on the down-left-hand side of } the MEF3, by a Coolpix 995 equipped with a x0.63 C-mount and a home-made } short adapter, the latter used for matching. But with or without the } adapter, and regardless of the position of the Coolpix relative to the } microscope port, the field of view was much smaller than the one } photographed in the 35mm camera (in other words, the aparent magnification } was much higher). In order to get a similar field of view I had to use a } lower power objective of the MEF3. This limits the minimal attainable } magnification. If the optical magnification of the Coolpix is reduced to a } minimum, to get a larger field of view, only a partial, circular field is } exposed in the center, the rest is blocked out, like looking through a tube. } } Has anyone faced the same problem? can a different C-mount and adapter solve } it? } } Thanx, } } Dr. Uri Admon } Beer-Sheva } Israel
--
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 12:20:53 2003
} ... } The questions are: } } 1. What is the advantage of using Nikon Coolpix camera but } not other possible digital cameras for the microscopes?
Cost, availability of Coolpix adapters, and number of peers.
} 2. I was told to choose Coolpix 5000 over 5700, since the } 5700 will give rise to blak periphery field due to the } naroow field of view. Is this true? How do I find out what } size of chip in different Coolpix Cameras?
Use the "buying guide" comparison presentation available at http://www.dpreview.com ...
Both Coolpix models use the same size CCD, but there may be other considerations. However, if the 5700 is configured with the correct adapter relative to the same C-mount for the 5000, there should be no vignetting. My experience is with a CP5000, so I stand to be corrected. On the other hand, I see no clear advantage for one over the other, so you may as well same your money regarding the difference in cost.
Thanks for all of your responses regarding EM proficiency. Many of you suggested MSA certification and other similar programs which are excellent resources. More specifically, we are a clinical pathology lab with technicians of varying degrees of skill and experience. As the lab is expanding I need standards that will not only determine competency as a em technician but proficiency with regard to quantity and quality of work. Some of you have expressed and interest in developing the same.
If you have suggestions as to how we can collect and evaluate that data I would certainly be willing to hear them. This is something I am committed to working on in the coming year so I will certainly be in touch with those of you that have expressed the same.
Thanks
Lois Anderson Johns Hopkins University Dept. of Pathology Laboratory Manager Electron Microscopy/Immunofluorescence 600 N. Wolfe Street/Pathology 709 A Baltimore, MD 21287 (410) 955-2861/fax (410) 614-7110 landers-at-jhmi.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 12:57:09 2003
Really, any camera will do, even a really cheapie one, if you can tolerate the aberrations & low resolution. See http://www.aecom.yu.edu/aif/gallery/cheapscope/index.htm
At 08:48 PM 12/17/2003 -0800, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 14:26:53 2003
The Coolpix 990/995/4500 cameras were popular due to the fact that their lens design was easily adaptable to a low cost microscope coupler. This undoubtedly contributes to the popularity of the Coolpix cameras. Couplers are now available to fit a wide assortment of cameras from many manufacturers. Thales Optem http://www.thales-optem.com/DigitalCameraCouplers.html has couplers that will fit many still or video cameras, either directly or via low cost adapters.
Nikon USA lists their digital cameras at http://www.nikonusa.com/template.php?cat=1&grp=2 where you can find various models listed as well as PDF specifications, etc.
Canon USA http://www.powershot.com/powershot2/home.html also has a number of compact digital models. One feature of many Canon cameras is the inclusion of software to operate the camera while tethered to a computer.
Some of the Olympus "C" series cameras can be mounted as well. http://www.olympusamerica.com/cpg_section/cpg_digital_cseries.asp
I hope this is helpful.
George
George Laing National Graphic Supply 800.223.7130 x3109 scisales-at-ngscorp.com
1. What is the advantage of using Nikon Coolpix camera but not other possible digital cameras for the microscopes? 2. I was told to choose Coolpix 5000 over 5700, since the 5700 will give rise to blak periphery field due to the naroow field of view. Is this true? How do I find out what size of chip in different Coolpix Cameras?
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 14:57:10 2003
} Title-Subject: [Microscopy] [Filtered] Nikon Coolpix Digital Camera } } Question: I have a couple of general questions for using Nikon } Collpix Digital Camera system, since I am very interested in setting } up this system for my photographing to both whole mount embryos and } immunostained parrafin slides as a replacement of the conventional } films } The questions are: } } 1. What is the advantage of using Nikon Coolpix camera but not other } possible digital cameras for the microscopes?
Availability of adapters specifically designed for a particular camera line.
} 2. I was told to choose Coolpix 5000 over 5700, since the 5700 will } give rise to blak periphery field due to the naroow field of view. Is } this true? How do I find out what size of chip in different Coolpix } Cameras?
It is true. Due to the optics used in the 5700, the optics must me mounted further from the lens and this results in SEVERE vigneeting. The only way to correct this is to use the digital zoom mode of the camera at maximum zoom. This severly degrades images quality. With the 5000, you only need to use optical zoom, which does not degrade imaeg quality.
There is a small write up of this in the Frequently Asked Questions section on our Nikon Coolpix to microscope adaper webpage: http://www.mvia.com/Coolpix/clpxadpt.htm#FAQ
Thanks And Happy Holidays! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
} Any information is greatly appreciated! } } Yingcui Li, Ph.D } } ---------------------------------------------------------------------------
--
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 15:15:07 2003
Given that you admit to being new to the TEM biz, I'd suggest you carefully box up and shelve your LaB6 filament for awhile, and buy a box of tungsten fialments from FEI/Philips or an EM vendor who sells same for your 400. I think that operating with tungsten would make your (new) life on a TEM easier while you are getting used to it and learning things such as gun alignments, column alignments, filament saturation, spot size control, vacuum system operation. These things are just a bit more complicated on a TEM than an SEM, I think.
Later, when you have your "sea legs", or you discover that you really need the brighter and more coherent beam that LaB6 gives for your work, put in the LaB6 and build on your experience using that. But make sure you get good operating info on LaB6, how to saturate it, prevent oversaturation, whether to leave it on or slightly desaturated when not in use, etc. It takes a bit more care to operate properly, but you can look into that while you are using the tungsten. Do a search in the Microscopy archive to pull up past discussions on this subject.
I have an FEI/Philips CM-12, so I'm extrapolating back to your machine here, but whichever filament type you use, check your Modes/Configuration menus and select either "tungsten" or "Lab6", as that tells the vacuum system which one you are using, as vacuum requirements are more stringent for LaB6 operation.
Just my 2-pence worth, Good Luck!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra
} I have a Philips 400 series TEM (used) currently being installed. To my } surprise I found out it has a LaB6 filament. It has been suggested that I } consider a CeBix filament. In either case it was suggested I use either } the Mini Vogel or Denka style. } } I'm light / SEM microscopist, TEM is a largely explored country to me. I } anticipate using the TEM for particle sizing and polymer phase studies and } I don't expect to exceed 100K magnification at 80 to 100 kv. I would like } to get the best value in terms of performance, ease of use, working life } for the money. Any advise, experience, recommendations on type, filament } material, tip configuration, special awareness of problems or advantages } would be welcome. } } PS: where can I get a passport to TEM land? :-) } } Thanks . . . . . } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 15:16:32 2003
The problem that you are having is because the detector of a digital camera is much smaller than the size of the 35mm film that was used in the original. As a result, the image that once covered an entire film, is now much too big for the chip on the microscope. I went to my local optometrist and got a set of concave lenses, and inserted them in the light path. After trial and error, we found one that worked for our system. Nikon makes an adapter for its microscopes, but charges about $1000 for it. I don't know if that would work with your Reichert-Jung. }
} AU} } ---------------------------------------------------------------------- } -------- AU} The Microscopy ListServer -- Sponsor: The Microscopy } Society of America AU} To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver AU} On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html AU} } ---------------------------------------------------------------------- } --------- } } AU} Dear All, and best wishes for a happy New Year, } } AU} I shall appreciate advise on how to connect a Nikon Coolpix 995 to } a AU} Reichert-Jung MEF3 (inverted) Light Microscope, and yet be able } to AU} photograph the whole field of view, or at least close to it, as } is viewed AU} and photographed through the Polaroid or 35mm cameras. I } tried to replace AU} the original 35mm camera, normally positioned on } the down-left-hand side of AU} the MEF3, by a Coolpix 995 equipped } with a x0.63 C-mount and a home-made AU} short adapter, the latter } used for matching. But with or without the AU} adapter, and regardless } of the position of the Coolpix relative to the AU} microscope port, } the field of view was much smaller than the one AU} photographed in } the 35mm camera (in other words, the aparent magnification AU} was } much higher). In order to get a similar field of view I had to use a } AU} lower power objective of the MEF3. This limits the minimal } attainable AU} magnification. If the optical magnification of the } Coolpix is reduced to a AU} minimum, to get a larger field of view, } only a partial, circular field is AU} exposed in the center, the rest } is blocked out, like looking through a tube. } } AU} Has anyone faced the same problem? can a different C-mount and } adapter solve AU} it? } } AU} Thanx, } } AU} Dr. Uri Admon } AU} Beer-Sheva } AU} Israel } }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 16:33:11 2003
Dee - I reply privately because I don't want to get involved in vendor promotion issues on the list. But here's my experience. Evaporator - Edwards Auto306 - had it 10 years. Excellent machine. Very clean vacuum, easy operation. Reliable machine. I believe that the cost is higher than their competition - but in my 30 years of EM lab work - this was the only evaporator that wasn't troublesome and didn't contaminate delicate samples, especially good at corona treatment. Sputter coater - Edwards scancoat 6. Good machine, coats well. I bought it because I also needed corona (etch) treatment capability (new lab set up when company split) and it was the cheapest with that capability. Had to do 2 fixes in the 2 years I've had it - but they were not major. If you don't need etching capability, you have a wider set of choices. Don't like the "automatic" coating operation setup - modified the machine adding an additional argon bleed valve to suit my preferred coating method.
Good luck - and happy holidays.
Dave Calvert Voridian Division Eastman Chemical Co. P.O. Box 1972 Kingsport, Tennessee Voice: 423-229-4943 Fax: 423-224-7550
-----Original Message----- } From: Mike Dufraine [mailto:MDufraine-at-ebsciences.com] Sent: Thursday, December 18, 2003 11:30 AM To: Dee Breger Cc: microscopy-at-msa.microscopy.com
Dee Breger wrote:
} ----------------------------------------------------------------------- ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Energy Beam Sciences is the US distributor for the Polaron Range of SEM prep equipment.
The E6700 benchtop evaporator is one option that offers ease of use, along with choice of options to fit any of your sample preparation needs.
Your welcome to contact me by email MDufraine-at-ebsciences.com, or Tel. 1-800-992-9037 X340, to learn more about the Polaron line of evaporators, coaters, and CPD equipment we offer.
Mike Dufraine Energy Beam Sciences, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 18 20:37:51 2003
Dear Colleques My congradulations with comig Holidays Season and Happy New Year! Accroding Russian tradition I wish, everyone will be healthy and happy in coming year! I also wish, our microscopes will do well! Have a great Holidays Season! Sergey
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
POSTDOCTORAL RESEARCH FELLOWSHIP in ELECTRON DIFFRACTION
Structure Factor Determination using Convergent Beam Electron Diffraction
Applications are invited for a postdoctoral research fellowship of up to 3 years duration in the above area funded by the Australian Research Council.
The post will involve the theoretical and experimental development of a new method for the direct measurement of both the phase and amplitude of crystal structure factors from convergent beam electron diffraction patterns. The position is based at the School of Physics and Materials Engineering, Monash University, in Melbourne, but is likely to involve travel to the University of Cambridge and McMaster University for collaborative work.
Applicants should have, or have nearly completed, a doctorate in physics or a related discipline and have experience in advanced transmission electron microscopy. A background in electron scattering theory and/or convergent beam electron diffraction is highly desirable.
Monash University is a member of Australia's leading "Group of Eight" research-intensive universities and is to be the site of The Australian Synchrotron. The School of Physics and Materials Engineering has an annual research income of ~$4.5million with strong research groups in diffraction physics and its application to materials science. Its electron microscope facilities include 3 TEMs and 2 SEMs, with a UHV LEEM and advanced, energy-filtered FEG-TEM to be acquired in 2004.
The appointment will be made at Academic Level B (AUD $54,864 - $65,152 per annum) or Academic Level A (AUD $48,554 - 52,121), depending on experience.
Enquiries and formal applications, including a full CV with the contact details of at least two referees, should be addressed to Dr. Joanne Etheridge, School of Physics and Materials Engineering, Building 69, Monash University, Victoria 3800, Australia. Tel: +61 (0)3 9905 1836, Fax: +61 (0)3 9905 4940 or joanne.etheridge-at-spme.monash.edu.au. The closing date for applications
_______________________________________________
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 19 09:45:25 2003
Welcome to TEM land. I don't know if I have a passport for you but I'll try to be some help.
First, I have to second the comment of Gib Ahlstrand. You would probably be best served by starting with tungsten and working your way up to LaB6. I'm not certain that CeBix would offer much in the way of advantages for your application but that would be a topic for the FEI folks to weigh in on. At EBS we can provide 3 levels of tungsten filament as well as Denka LaB6.
In tungsten we offer standard loop, AR loop and pointed filaments. The standard loop is a good choice but the AR loop will provide greater stability, which might be good for you being that you're just starting out. Once you've mastered the AR, you could then move up to our SG (pointed) model. These filaments are much brighter but are also more delicate. The cost for tungsten in all cases is a fraction of the cost for LaB6.
Once you're comfortable with tungsten, you can then go on to LaB6, if you feel you need them. That is a choice you will make based on the requirements of the work you're doing. For TEM applications sharp LaB6 tips are generally recommended. The best choice in a sharp is a 60 degree cone angle, 10 degree tip radius. We also offer 5 degree tip radius for a brighter beam. But again, you might want to start with a standard (round) LaB6 tip that won't be as bright but will be more forgiving. Just remember that in the case of both tungsten and LaB6, the sharper the tip/the brighter the beam/the shorter the lifetime.
I hope this helps.
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Tel: 413 786-9322 Fax: 413 789-2786 {mailto:mnesta-at-ebsciences.com} {http://www.ebsciences.com} "Adding Brilliance to Your Vision"
-----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: Thursday, December 18, 2003 10:53 AM To: microscopy-at-msa.microscopy.com
In the past everyone has been very helpful so once again I am returning to the well of knowledge for advice:
I have a Philips 400 series TEM (used) currently being installed. To my surprise I found out it has a LaB6 filament. It has been suggested that I consider a CeBix filament. In either case it was suggested I use either the Mini Vogel or Denka style.
I'm light / SEM microscopist, TEM is a largely explored country to me. I anticipate using the TEM for particle sizing and polymer phase studies and I don't expect to exceed 100K magnification at 80 to 100 kv. I would like to get the best value in terms of performance, ease of use, working life for the money. Any advise, experience, recommendations on type, filament material, tip configuration, special awareness of problems or advantages would be welcome.
PS: where can I get a passport to TEM land? :-)
Thanks . . . . .
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 19 13:34:27 2003
Email: zhaoyu-at-biochem.ualberta.ca Name: Zhaoyu Li
Organization: Department of Biochemistry, University of Alberta
Title-Subject: [Microscopy] [Filtered] MListserver: immunogold staining of EM with anti-phospholipids antibody
Question: I am trying to do immunogold staining of EM with anti-phospholipids antibody for liver tissues. But some lipids staining steps(OsO4) and/or ethonal dehydration steps seemed blocked or reduced the binding sites of anti-phospholipids antibody. I cann't get specific binding. For sure, the antibody works well in the fluorescent staining. So, my question is how to skip or replace these steps? or any other methods could be used in this case?
the self absorption of X-rays in the specimen is (in most cases) the main process of all influences to generate the X-rays with electron excitation. If the fine focused electron beam is directed to an unknown surface tilt, the absorption effect will be unknown. Quantitative results vary, if there is no estimation of the changed absorption path in specimen.
Operators are thinking very common, if scanning across a larger area of an irregular surface (rough or porous), all regions with more absorption are going to adjust with these regions, which are characteristic for lower absorption. The result should be an spectrum with same absorption (and same results) like the polished specimen of identical element content.
But this isn't true!!!
Because of the not linear effects of absorption (e-function), these regions, which have higher absorption, influence the final spectrum more than the others. Because of that, higher absorption always occur for rough (and porous) surfaces, compared to a flat specimen with same element concentrations. It's possible to prove this fact with mathematics, but not trivial to understand.
The opposite is with particles on top of a surface. There it is more easy to understand, that excited X-rays have lower absorption effects, in most cases.
"Dusevich, Vladimir" schrieb am 17.12.03 08:54:39: } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Many thanks for a number of replies. } Unfortunately I formulated my question too fuzzy. I am not interested in the } measurements of porosity utilizing EDS. What I am interested in is the } effect of porosity (or nanoporosity) on EDS measurements. It seems } that an increase in porosity should lead to a decrease in the intensity of X-rays, } and that this dependence should not be linear. It's only my gut feeling, } I do not want to carry out calculations. I am sure all these calculations were } done a long time ago, and I'll appreciate any lead to a proper publication } and/or a short explanation of the problem. } } Thanks, } } Vladimir } } } ________________________________ } } } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] } Sent: Wed 12/10/2003 2:57 PM } To: Microscopy (E-mail) } Subject: [Microscopy] Microanalysis of porous material } } } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello all, } I have samples of the same material with suspected } difference in porosity, size of pores could be from } a few nanometers to a few hundred nanometers. } } Is it possible to detect porosity with microanalysis } (EDS)? How intensity could change because of porosity? } Is it possible to calculate the "detection limit" } of porosity? } } Thank you, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 22 08:33:28 2003
The real question is wether the lipids are still there. The dehydration and infiltration steps are well known to result in significant loss of lipids. You may be forced to use cryo-fixation followed by low temperature freeze-substitution and embedding or conventional chemical fixation followed by cryo-sectioning.
At 10:13 AM 12/21/2003 -0600, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Title-Subject: [Microscopy] [Filtered] Jeol 5400 SEM Image Quality Questions
Question: Hello, We have recently installed a Jeol 5400 SEM with a Voyager EDS system (have not installed the harware for the EDS yet). The image quality on the SEM begins to degrade significantly above aroun 5000x (unable to focus and the "jaggies" begin to show up significantly). The image quality up to about 2500x is good. These jaggies appear to be at 60 hz. Everything is plugged into a single outlet 215V with a stepdown transformer to 100v.
I would assume that nothing can be done to asess any other imagining issues with the scope until this power/field disturbance is corrected? The circuit for the power source runs a considerable distance (probably 100'+) from the SEM to the breaker box through metal conduit.
Related to the above, does anyone have any "preferred" procedures/sequences for aligning and adjusting this scope, with regard to the selectable apertures and tilt/shift. The manuals methods do not seem to be very effective.
Thanks in advance for your help. Charles Apex Technologies Inc.
Email: michael.didie-at-gmx.de Name: Michael DidiÈ
Organization: Pharmakology/Hamburg
Title-Subject: [Microscopy] [Filtered] Cryotome Cuts Without Foldings?
Question: Hello everyone,
We just got a nice and shiny new Leica Cryotome and i tried to make some cuts. Most of the time i'll cut whole hearts with tissue implants. I usually embed the hearts in Tissue Tec before cutting them. The thickness of the cuts ranges between 5 and 10 µm. Now i had the problem, that the cuts start to fold and neither using a new blade nor changing the angel of the blade produced better results. Has anyone any idea hoe to solve that problem?
Michael, You don't say whether you use an anti-roll plate or if you guide your sections, as they are cut, with brushes. Either method should help. I have switched to freezing my samples in a 1:1 mixture of OCT and 20% sucrose (in PBS) (based on Barthel & Raumond (1990) J Histochem Cytochem 38(9) 1383-1388). It gives wonderful histology and cuts very smoothly. You may need to cut at a slightly colder temp, since the blocks tend to be softer than plain OCT. We usually cut at about -20. 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
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 23 14:42:09 2003
We have a user doing SEM of nylon, with embedded bits. We'd like to chemically etch the nylon, which is something of an entertaining problem, since nylon is used to mask things against etchants. Does anyone have a recipe for something that will etch nylon and not silica? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 23 15:16:34 2003
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Most probable causes are:
1. Ground loop - you may have more than one line to ground, forming a loop which acts as an aerial to pick up mains frequency. Can be difficult to locate. I would first check the conduit for the power cable for being grounded at both ends. Also check any fluorescent lights in the vicinity - a not infrequent problem is neutral wires, trapped in the fittings and connecting to ground.
2. There might be a field being radiated from something in the area which cannot be fixed. In this case, there are field cancellation systems, which have one or more sensors on the SEM and X/Y/Z field coils on the walls of the SEM room to generate an opposing field.
I would suggest you contact your local JEOL customer support office for advice.
Best regards, -- Larry Stoter JEOL (UK) Ltd NOTE - any message other than plain text will be automatically deleted :-)
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 23 16:22:46 2003
It sounds as if you have a magnetic field problem, but it could be vibration, this you will be able to check this out yourself. In the US 60 cycles main supply means that ALL problems tend to be of 60 cycles.
1. Run the microscope at the highest kV and place the sample 5mm from the final lens (WD 5mm) 2. Observe the image quality a) in TV mode does a wave float up or down the screen (evidence of a field)? b) are you able to see the raged edges in the image at a slow scan(evidence of a field or vibration or an electronics problem)? 3. Move the specimen down to 25mm from the lens (WD 25mm) and repeat a and b above.
Problems at a short working distance, where the specimen is partly protected by the lens field, are to be considered gross. If the problem is vastly reduced at a short working distance then you have a field problem. If the problem is exactly the same at the two levels you have an electronics or vibration problem.
Almost any building which houses large equipment is likely to cause field problems. HOWEVER I have seen this type of problem in a laboratory where, in a newly operational adjoining office building, investigation found most plugs were incorrectly wired causing the main input cable to overheat. This cable ran alongside the input cable to the building housing the EM, we fixed one problem that fixed the other!
If you are lucky the property of a field to fall off by the inverse square law will enable you to find the best position for your new SEM within the desired room. "Simply" moving the column by a few feet may help? Another route is to take your input from a clean supply, that is one where you are the only user. I would also make sure that your earth is not shared by any other equipment.
Hope this helps? Seasons Greetings.
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "by way of MicroscopyListserver" {charles-at-3dcircuits.com} To: {microscopy-at-ns.microscopy.com} Sent: Monday, December 22, 2003 10:15 PM
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Philip Oshel wrote: ========================================================
Materials micromavens,
We have a user doing SEM of nylon, with embedded bits. We'd like to chemically etch the nylon, which is something of an entertaining problem, since nylon is used to mask things against etchants. Does anyone have a recipe for something that will etch nylon and not silica? Thanks. ======================================================= I am assuming this is a nylon that has been filled to some degree with "bits" of silica and your objective is to see the uniformity of dispersion of the silica "bits" as well as any orientation they might be exhibiting.
This is easily done in a plasma etcher such as the SPI Plasma Prep II plasma etcher, as shown and described on URL http://www.2spi.com/catalog/instruments/etchers1.shtml [Note: This can not be done with sputter etching.]
Using an oxygen plasma, the nylon will be etched away, leaving the silica "bits" protruding over the surface of the polymer, giving excellent topographical variation for good contrast.
Disclaimer: SPI Supplies is the manufacturer of the Plasma Prep II plasma etcher/asher/cleaner.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 24 11:21:38 2003
Thanks. Samples of the material have been sent to the plasma folks on campus, but they haven't as yet come back. Which is why we're looking for a chemical etchant. Oxygen plasma etching was an early thought -- seems like the best way to go, if ...
Phil
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Philip Oshel wrote: } ======================================================== } } } Materials micromavens, } } We have a user doing SEM of nylon, with embedded bits. We'd like to } chemically etch the nylon, which is something of an entertaining problem, } since nylon is used to mask things against etchants. Does anyone have a } recipe for something that will etch nylon and not silica? Thanks. } ======================================================= } I am assuming this is a nylon that has been filled to some degree with } "bits" of silica and your objective is to see the uniformity of dispersion } of the silica "bits" as well as any orientation they might be exhibiting. } } This is easily done in a plasma etcher such as the SPI Plasma Prep II } plasma etcher, as shown and described on URL } http://www.2spi.com/catalog/instruments/etchers1.shtml [Note: This can } not be done with sputter etching.] } } Using an oxygen plasma, the nylon will be etched away, leaving the silica } "bits" protruding over the surface of the polymer, giving excellent } topographical variation for good contrast. } } Disclaimer: SPI Supplies is the manufacturer of the Plasma Prep II plasma } etcher/asher/cleaner. } } 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 } ######################## } ============================================
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 27 18:19:35 2003
Email: Hbrinkies-at-groupwise.swin.edu.au Name: Hans G Brinkies
Organization: Hawthorn (Australia)-Universtity of Technology
Title-Subject: [Microscopy] [Filtered] High Vacuum/Variable Pressure SEM
Question: I am finding myself in a very difficult position to advise important people at the university (who are in charge of giving money to needy electron microscopists) and ask you for some assistance from experienced operators of SEMs, which are being used in a variable pressure mode (not manufacturers or agents please).
We are in the process of purchasing a new SEM. Having been successful to obtain a research grand, other departments interested in microscopy need to 'cough up' additional funds. They are only willing to do so, if they can use the new SEM for their research projects, whenever it is required.
The problem: one group (the majority) wants to use the SEM in the high vacuum, high resolution mode (approximately. 2 to 8 nm), the other group needs variable pressure applications for relatively moist (oil, water, tar containing samples). And the third group wants to carried out micro-lithography in a SEM
With now more than 35 years experience in SEM applications I still believe that one can only make the SEM available on short notice if one has dedicated units for above applications. I am worried that one needs a lot of 'elbow grease' to clean a SEM after having used it for 'dirty' samples (or let's say industrial specimens) before one can use it again for high resolution work.
I know what my decision would be having two operational SEMs in my laboratory, one JSM840 and an old ETEC Autoscan (believe me, it is still working).
X-mas cheers etc.
Hans Brinkies Professional Officer, Electron Microscopy and Metallography Swinburne, University of Technology School of Engineering and Science Industrial Microscopy Laboratory P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
Jim I don't see any religious content in my message. Is "Holidays Season" and "New Year" has religious content to you? If so, I am apologize and I could take back my good wishes. Have a good what? Holidays are religious, OK, DAY! Sergey.
At 08:59 AM 12/19/2003, you wrote: } Sergey - } } Is the listserv really appropriate for religious messages? } } JQuinn } } } } From MicroscopyL-request-at-ns.microscopy.com Fri Dec 19 01:28:31 2003 } } X-Authentication-Warning: ns.microscopy.com: mail set sender to } MicroscopyL-request-at-ns.microscopy.com using -f
I would say JQuinn's confusion about a classic American dilemma and the Microscopy ListServer regs have caused his snit.
Microscopists I know revere with pantheistic fervor forces that keep their EM(s) producing heavenly results.
Krepkogo zdorovia, schastia, lubvi.
Vsego Nailuchego,
Vincent & Alla
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, December 27, 2003 5:39 PM
Hans Of course your ETEC is still working! No reason why it shouldn't be.. They are also very tolerant of dirty samples, but you can't get variable pressure. I'm not terribly familiar with the variable pressure/ESEM in practice, but I think your e-beam lithography folks are not going to want to do a lot of sharing. They may want a cleaner vacuum than the hi res folks. Hopefully others can give you first hand experience on the trade-offs between low vac and clean operation.
Ken Converse owner Quality Images third party SEM service Delta, PA
by way of MicroscopyListserver wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } ------------------------------------------------------------------------- } } Email: Hbrinkies-at-groupwise.swin.edu.au } Name: Hans G Brinkies } } Organization: Hawthorn (Australia)-Universtity of Technology } } Title-Subject: [Microscopy] [Filtered] High Vacuum/Variable Pressure SEM } } Question: I am finding myself in a very difficult position to advise } important people at the university (who are in charge of } giving money to needy electron microscopists) and ask you for } some assistance from experienced operators of SEMs, } which are being used in a variable pressure mode } (not manufacturers or agents please). } } We are in the process of purchasing a new SEM. } Having been successful to obtain a research grand, } other departments interested in microscopy need to } 'cough up' additional funds. They are only willing to } do so, if they can use the new SEM for their research } projects, whenever it is required. } } The problem: one group (the majority) wants to use the SEM } in the high vacuum, high resolution mode } (approximately. 2 to 8 nm), the other group needs } variable pressure applications for relatively moist } (oil, water, tar containing samples). And the third group } wants to carried out micro-lithography in a SEM } } With now more than 35 years experience in SEM applications } I still believe that one can only make the SEM available } on short notice if one has dedicated units for above } applications. I am worried that one needs a lot of } 'elbow grease' to clean a SEM after having used it for } 'dirty' samples (or let's say industrial specimens) before } one can use it again for high resolution work. } } I know what my decision would be having two operational } SEMs in my laboratory, one JSM840 and an old } ETEC Autoscan (believe me, it is still working). } } X-mas cheers etc. } } Hans Brinkies } Professional Officer, Electron Microscopy and Metallography } Swinburne, University of Technology } School of Engineering and Science } Industrial Microscopy Laboratory } P.O.Box 218 - Hawthorn - Vic -3122 - Australia } Phone: +61 3 9214 8657 } Fax: +61 3 9214 8264 } Email: Hbrinkies-at-swin.edu.au } } } --------------------------------------------------------------------------- } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 29 21:12:47 2004
Question: Hello everyone, We are currently installing a Jeol 5400 with a Noran Voyager running Version 3.7 software. We had to obtain the Sparc Station and install the software from scratch, since the Sparc box had been removed from the system when we purchased it. I cannot say enough GOOD things about the folks at Noran regarding supporting their product and license agreement. Since we had the original documentation, hardware and work orders, they have provided the software (v 3.7), manuals, etc. for minimal cost and answered numerous harware questions on this used instrument. The technical staff also seems to have a lot of knowledge about the hardware and software (thanks Philip, are helpful, prompt and friendly.
We have installed the software on the sparc and checked out the voyager cpu box as much as possible. (we have not yet installed the detector). We intend on getting a field engineer to look at the system but were tryin to debug it as much as possible beforehand.
My question is regarding installation and setup of the Voyager software with regard to the "Image Calibration". It appears that there is NO image calibration file present when the software is installed "from scratch". We assume this prevents any sort of image acquisition at all. I was told by the technical rep that this was best left to the field engineer. However being curious and stubborn, I was wondering if this file is something we can create. We have a printout that lists all the original calibration parameters (a whole list of about 30 items). The calibration file is apparently entitled "wima.column1.cal" in the "usr/voyager/tbles" directory (there is no such file or directory when voyager v3.7 is installed from scratch). Does anyone have a "template" that could be used for this file, or suggestions on creating one.
As a related question, can the voyager software be used to acquire standard SEM images without the EDS detector being installed.
Thanks in advance for any help. Also thanyou to everyone who helped with the Jeol 5400 "jaggies" question. Charles Apex Technologies, Inc. 919-463-7585
Have a good holey day. (for those using holey carbon film)
for continuous carbon film users and other occultists: Have a good unholiday.
In Sweden you hear the mysterious "Good continuation (god fortsaettning)" (of whatever you are doing at the moment, I guess.)
So: God fortsaettning to everybody,
Philip
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em
'This winter I am giving courses to three students, of whom one is only moderately prepared, the other less than moderately, and the third lacks both preparation and ability. Such are the burdens ...' Carl Friedrich Gauss (1810) ______________________________________________
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, December 28, 2003 2:39 AM
Two Postdoctoral Fellowships are now available in the Supramolecular Structure and Function Group of the Division of Bioengineering and Physical Science at the National Institutes of Health. This program develops EM methods, including electron tomography, cryo-EM, energy-filtered imaging, STEM and EELS, and applies them to biomedical research in collaboration with NIH scientists.
Currently, our laboratory is focusing on (1) electron tomography to visualize 3-dimensional subcellular structures, and (2) electron spectroscopic imaging of cells and macromolecular assemblies.
The positions provide an excellent opportunity to work with state-of-the-art instrumentation, including a 300 kV field-emission TEM (Tecnai TF30) equipped with an energy-filter.
Applicants should send their curriculum vitae to:
Dr. Richard Leapman Division of Bioengineering & Physical Science Building 13, Room 3N17 National Institutes of Health 9000 Rockville Pike Bethesda, MD 20892 Tel: 301-496-2599 Fax: 301-435-4699 email: leapman-at-helix.nih.gov
http://www.nih.gov/od/ors/dbeps/ssfr/
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 30 12:00:21 2004
Alby On Martedì, dic 30, 2003, at 11:08 Europe/Rome, Philip Koeck wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Have a good holey day. (for those using holey carbon film) } } for continuous carbon film users and other occultists: Have a good } unholiday. } } In Sweden you hear the mysterious "Good continuation (god } fortsaettning)" } (of whatever you are doing at the moment, I guess.) } } So: God fortsaettning to everybody, } } Philip } } Philip Koeck } Svdertvrns Hvgskola and } Karolinska Institutet } Dept. of Bioscience at Novum } S-14157 Huddinge } Sweden } phone: +46-8-6089186 } fax: +46-8-6089290 } http://www.biosci.ki.se/em } } 'This winter I am giving courses to three students, of whom one is only } moderately prepared, } the other less than moderately, and the third lacks both preparation } and } ability. } Such are the burdens ...' Carl Friedrich Gauss (1810) } ______________________________________________ } } } ----- Original Message ----- } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Sunday, December 28, 2003 2:39 AM } Subject: [Microscopy] Re: Greetings } } } } } } } } ---------------------------------------------------------------------- } } ---- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } ---- } ----- } } } } Jim } } I don't see any religious content in my message. Is "Holidays Season" } } and } } "New Year" has religious content to you? If so, I am apologize and I } } could take back my good wishes. Have a good what? Holidays are } religious, } } OK, DAY! Sergey. } } } } At 08:59 AM 12/19/2003, you wrote: } } } Sergey - } } } } } } Is the listserv really appropriate for religious messages? } } } } } } JQuinn } } } } } } } } } } From MicroscopyL-request-at-ns.microscopy.com Fri Dec 19 01:28:31 2003 } } } } X-Authentication-Warning: ns.microscopy.com: mail set sender to } } } MicroscopyL-request-at-ns.microscopy.com using -f
Dear Hans, I have been running a Hitachi S-3000N for the last year and I will say that it will do most SEM jobs, both high vacuum and variable pressure, very well with no compromises. We regularly look at wet samples, high-resolution, then high-res, low kV and back to wet in a single day. However, if you want field-emmission-type resolution, you will have to get a field-emmission microscope. Mine resolves 3 nm. The variable-pressure mode converts in two mouse clicks and takes no more time than a sample change. The variable-pressure mode keeps the scope clean and can be used overnight to clean-up after any dirty samples. I would recommend a cold stage for true water-containing samples. We have been using it for every type of sample, hi-res, low kV, x-ray mapping and I couldn't imagine doing without its capabilities now. If I could only have one SEM, this would be it. If I could have two, I would also get a field-emmission for higher resolution. Good luck and congratulations on a new baby. PS I used to run an ETEC, too. MM Mary Mager Electron Microscopist Metals and Materials Eng. University of British Columbia 6350 Stores Road Vancouver, BC V6T 1Z4 CANADA Tel: 604-822-5648 Fas: 6004-822-3619 ----- Original Message ----- } From: "by way of MicroscopyListserver" {Hbrinkies-at-groupwise.swin.edu.au} To: {microscopy-at-ns.microscopy.com} Sent: Saturday, December 27, 2003 4:33 PM
While we're at it...: Bonne année 2004 à tous et à toutes!
Marc
On Tuesday, December 30, 2003, at 12:57 PM, diaspro wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } In Italy we have } } Auguri di Buone Feste e Felice Anno Nuovo } } Alby } On Martedì, dic 30, 2003, at 11:08 Europe/Rome, Philip Koeck wrote: } } } } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } --------- } } } } Have a good holey day. (for those using holey carbon film) } } } } for continuous carbon film users and other occultists: Have a good } } unholiday. } } } } In Sweden you hear the mysterious "Good continuation (god } } fortsaettning)" } } (of whatever you are doing at the moment, I guess.) } } } } So: God fortsaettning to everybody, } } } } Philip } } } } Philip Koeck } } Svdertvrns Hvgskola and } } Karolinska Institutet } } Dept. of Bioscience at Novum } } S-14157 Huddinge } } Sweden } } phone: +46-8-6089186 } } fax: +46-8-6089290 } } http://www.biosci.ki.se/em } } } } 'This winter I am giving courses to three students, of whom one is } } only } } moderately prepared, } } the other less than moderately, and the third lacks both preparation } } and } } ability. } } Such are the burdens ...' Carl Friedrich Gauss (1810) } } ______________________________________________ } } } } } } ----- Original Message ----- } } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Sunday, December 28, 2003 2:39 AM } } Subject: [Microscopy] Re: Greetings } } } } } } } } } } } } } --------------------------------------------------------------------- } } } ----- } } ---- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } --------------------------------------------------------------------- } } } ----- } } ----- } } } } } } Jim } } } I don't see any religious content in my message. Is "Holidays } } } Season" and } } } "New Year" has religious content to you? If so, I am apologize and } } } I } } } could take back my good wishes. Have a good what? Holidays are } } religious, } } } OK, DAY! Sergey. } } } } } } At 08:59 AM 12/19/2003, you wrote: } } } } Sergey - } } } } } } } } Is the listserv really appropriate for religious messages? } } } } } } } } JQuinn } } } } } } } } } } } } } From MicroscopyL-request-at-ns.microscopy.com Fri Dec 19 01:28:31 } } } } } 2003 } } } } } X-Authentication-Warning: ns.microscopy.com: mail set sender to } } } } MicroscopyL-request-at-ns.microscopy.com using -f
There are lots of ridiculous protests against Christmas celebrations that make that of JQuinn pale:
In Dec, 2001, a group of animal rights protestors in Cambridge announced that they intended to sue "Christianity as a whole" and anyone who celebrates Christmas. The shock announcement comes after years of protesting against Christmas which, they say, causes unnecessary cruelty to turkeys.
In November 2003, the Colorado ACLU threatened to sue to ban Christmas because it claimed it made Jews 'uneasy'.
And best of all:
Cincinnati attorney Richard Ganulin filed a lawsuit on 1998-AUG-4 in U.S. district court, 6 asking that the federal government be required to not declare future DEC-25 holidays. He feels that "Christmas is a religious holiday and the Congress of the United States is not constitutionally permitted to endorse or aid any religion, purposefully or otherwise, or [promote] entanglement between our government and religious beliefs." Judge Susan Dlott dismissed the suit. Judge Dlott decided "that Christmas can be observed as a federal holiday because non-Christians also mark the holiday by celebrating the arrival of Santa Claus. Since nonreligious people also observe the holiday, giving federal workers a day off for Christmas does not elevate one religion over another."
So, it looks like we need to be grateful to a US District Judge for saving Christmas. From a devout Raelian (only kidding) MERRY CHRISTMAS and HAPPY NEW YEAR!
John Mardinly Intel
-----Original Message----- } From: vcrvince [mailto:vcrvince-at-sbcglobal.net] Sent: Sunday, December 28, 2003 12:18 AM To: Microscopy-at-sparc5.microscopy.com; Sergey Ryazantsev
Hau'oli Makihiki Hou!
Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 31 21:10:43 2004
Has anyone seen a source of micro probes for SEM that allow electrical contact to a SEM chamber specimen? I need very precise positioning--like within 0.15u or better and 0.05u repeatability and stability.
I need two contact probes.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 1 10:38:10 2005
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Gary Gaugler wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Would someone point me to the listserver } for failure analysis discussion of metallurgical and } integrated circuit devices? } } tnx, } gary g.
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 2 10:09:20 2004
I've seen hot/cold stages from Deben that look like they could hold a standard pin stub specimen. However, they seem to be optimized for low temperature work. Does anyone know of some other supplier that makes SEM specimen holders that will heat up to about 200C and perhaps cool to -25C?
The unit would need to mate to the stage on a LEO Supra 55VP's specimen interchange stage or on a FEI Sirion 400, under similar circumstances.
Gatan makes a series of heated stages but they seem more for stress testing and bulk specimens. I will have an IC chip thermal expoxy'd to a 12mm diameter Al pin stub. I need to be able to heat this specimen in the SEM and at any tilt and WD that the SEM will support. Temperature stability could be as bad as +-5C. That is OK.
Specimen holder and specimen changeout via slide out chamber door is OK. Specimen interchange lock does not have to be used.
thanks for any ideas and leads, gary g.
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 2 11:27:25 2004
In the latest issue of Microscopy Today there is an article on pseudo-coloring of images. Since this is not something that I normally do, I read the article with interest. I was surprised to find that the author says that biologists nearly always use the "spectrum" color table for the pseudo-coloring of grayscale images. I had thought that those who study visual perception had found that among pseudo-color scales the "thermal" scale is much better than all the others at providing a good intuitive reading of the image. This is, I think, the scale that Photoshop calls "Black Body". Can someone please clear up my confusion. -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 2 11:58:48 2004
Here is the table of contents for the January/February 2004 issue of Microscopy Today.
New Subscriptions via http://www.microscopy-today.com only, please. New subscriptions will close on Thursday 8 January for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting.
Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA.
MSA members anywhere have free subscriptions.
Non-MSA, non-North American subscriptions have been reduced from $80 or $110US to $35US. Additional details in the magazine or on our web site.
THIS WILL BE THE LAST ISSUE NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED OR NOT-CURRENTLY-SUBSCRIBED INDIVIDUALS WILL RECEIVE!
Listers,
Here is the table of contents for the November/December 2003 issue of Microscopy Today. This issue is mailed with the Call for Papers for the 2004 Microscopy and Microanalysis meeting
New Subscriptions via http://www.microscopy-today.com only, please New subscriptions will close on Tuesday 11 November for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting.
Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA.
MSA members anywhere have free subscriptions.
Non-MSA, non-North American subscriptions have been reduced from $80 or $110US to $35US. Additional details in the magazine or on our web site.
PURGING OF NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED OR NOT-CURRENTLY-SUBSCRIBED INDIVIDUALS IS UNDERWAY!
January/February 2004 Carmichael: Correlating Fluorescence Microscopy with Electron Microscopy P.E. Batson: Electron Microscopy Enters a New Era Using Aberration __Correction Paula Allan-Wojtas: Microscopy and Imaging of Foods The Whys and Hows Jerry Sedgewick: Image Stitching Using Photoshop Michael Bode: A Few Thoughts About Image File Storage Paul Beauregard: Behavior of Particle Size Distributions, Means and BET __Values in Ideal and Non-Ideal Morphology Systems in a TEM Katerina Moloni: The Moving Finger Writes: Carbon Nanotubes as AFM Probe __Tips Robert M. Zucker: Confocal Microscopy System Performance: Axial Resolution Shane Roberts, Daniel Flatoff: Enhanced Sample Preparation of Cu Low-k __Semiconductors via Mechanical Polishing and Ion Beam Etching Luc Harmsen: The Year That Was! Microscopy in Southern Africa Robert P. Apkarian: Comments on Cryo High Resolution Scanning Electron __Microscopy Jose A. Mascorro: Propylene Oxide: To Use or Not to Use in Biological Tissue __Processing M. T. Postek & A. E. Vladár: Is Low Accelerating Voltage Always the Best for __Semiconductor Inspection and Metrology?
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 2 12:10:04 2004
Yes it is unfortunately true that many (most?) biologists do use the "spectrum" color scale, largely because it makes "prettier-looking" images. It the cases where they are trying to illustrate quantitative contrast this is not only grossly misleading but it is usually plain wrong and can produce horrific artifacts! The worst offenders are chiefly light microscopists who are trying to represent weak flourescence contrast and for some reason think it shows up "better" with a spectrum scale. In the STEM/X-Ray/EELS biological microanalysis field most of us use some variation of the "black Body" scale which of course more closely parallels the greyscale that is intuitively quantitative anyway (black = 0, shades of grey through white represent more positive values). Relative contrast or non-linear scaling can be achieved by manipuating the scale either continuously or by introducing discontinuities to other scales; of course color then becomes essential (a) because the human eye can perceive considerably more colors than levels of grey, and (b) one can extend the scale over a far greater dynamic range(s); most monitors only display 8-bit levels of grey (24-bit color) but data are often 32-bit or more in dynamic range.
The topic of visual perception of data is a fascinating one indeed and has been addressed in several treatises over the years. However in my experience, I have found the most (subjectively) pleasing results to come from visual artists (painters) who seem to have a natural instinct for such representations. A visit to any good art museum should convince most people of this!
Sorry if this is a rather brief and simplistic answer to your question but maybe it helps some!
Peter
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
the most common use of Pseudo color is to enhance the contrast of the images and make small details more visible.
A little background:
Computer monitors are normally set to "True Color". On most graphics cards that means 32 bit of information per pixel, or "Millions of colors" as they say. However, each pixel is represented by 3 colors (Red, Green, and Blue), and each of these colors can take on an 8 bit value. 8 bits mean, that there are 256 shades of each color available, which can be combined to give you the "millions of colors" (256 x 256 x 256). What is not so obvious, that for gray levels you need to combine the 3 colors in at the same strength, i.e. black is 0,0,0, medium gray is 128,128,128 and white is 255,255,255. This shows, that even if your monitor can display millions of colors, it can usually only show 256 levels of gray. Take into account, that the human eye can distinguish perhaps 50 or so levels of gray and modern cameras can provide anywhere from 4000 to 64,000 levels of gray, and the need for different color schemes becomes obvious.
Enter the pseudo colors.
There are as many pseudo color schemes as you can think of. Several have become "standards". Among them definitely the "black body" scheme, and the "spectrum" color scheme. The "black body" is perhaps more intuitive, as it basically goes from Red to White. This provides a linear scale, which is easy to understand to anybody who has seen a metal heated (and perhaps burned himself or herself), and it is probably easier to discern small contrasts both in the red and the bright parts of the spectrum than in b/w images. However, it does not make full use of the capabilities of a monitor. The "spectrum" pseudo color, on the other hand, makes full use of the availble color spectrum, perhaps at the price of an intuitive understanding. It may be better suited to images that have "many" gray levels, which all need to be discerned. If used wrongly, however, the spectrum pseudo color can also lead to misleading coloring.
I hope this helps.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Alwyn Eades [mailto:jae5-at-lehigh.edu] Sent: Friday, January 02, 2004 10:28 To: MSA listserver
In the latest issue of Microscopy Today there is an article on pseudo-coloring of images. Since this is not something that I normally do, I read the article with interest. I was surprised to find that the author says that biologists nearly always use the "spectrum" color table for the pseudo-coloring of grayscale images. I had thought that those who study visual perception had found that among pseudo-color scales the "thermal" scale is much better than all the others at providing a good intuitive reading of the image. This is, I think, the scale that Photoshop calls "Black Body". Can someone please clear up my confusion. -- ......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 3 07:27:59 2004
} ... } ... In the STEM/X-Ray/EELS biological microanalysis field } most of us use some variation of the "black Body" scale } which of course more closely parallels the greyscale that } is intuitively quantitative anyway (black = 0, shades of } grey through white represent more positive values).
I remember an M&M '99 session, which introduced a pseudo-color scale for quantitative images (e.g., elemental distributions, maps). That is, ranges of color for representing "orders of magnitude" ... or ranges we might refer to as "major", "minor", "trace", or "undetected". The session was intended to be its introduction, such that its color ranges would become familiar to, and used by all, as so that quantitative images could be actually compared. I thought it was interesting concept at the time, but also felt it needed some refinement. Unfortunately my inadequate notetaking didn't allow for me to ever find the color table and download it.
I have no idea if it was mentioned in the MT article, but the people who had introduced the color scheme were from NIH (or, was it NIST?). I believe it is too bad the color scheme never did rise to common use. That is, even if I did feel it still needed some refinement, it would be a good thing if quantitative images could all be compared.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 3 10:27:30 2004
} In Dec, 2001, a group of animal rights protestors in Cambridge announced that } they intended to sue "Christianity as a whole" and anyone who celebrates } Christmas. The shock announcement comes after years of protesting against } Christmas which, they say, causes unnecessary cruelty to turkeys.
And ignore the use of reindeer as beasts of burden? Or use of swine as ham? Sounds pretty discriminatory to me.
Some people think the New Year is when it is because it was the celebration of Jesus's circumcision. If we think this is bad (e.g. castration ritual), do we refuse to follow the calendar? BTW, the holiday of New Year has become almost global.
This is a time of year when we should take some time off and relax. This message is apropos to this bboard specifically because for a few days I didn't think about microscopy at all. I read novels, slept, ate ham and argued with family over the Iraq war and mostly trivial stuff. This is happy holidays.
Now, can we get back to microscopy and drop the holiday stuff? Even if some of us won't be completing our Christmas celebrations until the 6th?
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 3 11:40:17 2004
Question: Hello there, I am a starter who wishes to get her grandchildren interested in a world beyond TV & computer games. I started using computers when I purchased my first 128K Mac back in 1985 . My present Mac is a G3 192MB ram & 40 GB hard drive.
I recently aquired a second hand "MOTIC Biological Series B1 223A " but I have found that I cannot use my lovely Fuji S602Z digital camera to take photos.
Do you have any ideas which will enable me to combine the use of the hardware that I possess? I feel that the hardest part is getting software that will enable me to join up to the Macintosh even if I purchased a new camera.
I would really like to take the photos digitally but is it impossible with my present configuration? i would appreciate any comments please
} Email: faj-at-highway1.com.au } Name: Faye Taylor } } Organization: Amateur } } Education: Undergraduate College } } Location: Perth, Western Australia } } Question: Hello there, } I am a starter who wishes to get her grandchildren interested in a } world beyond TV & computer games. I started using computers when I } purchased my first 128K Mac back in 1985 . My present Mac is a G3 } 192MB ram & 40 GB hard drive. } } I recently aquired a second hand } "MOTIC Biological Series B1 223A " but I have found that I cannot } use my lovely Fuji S602Z digital camera to take photos. } } Do you have any ideas which will enable me to combine the use of the } hardware that I possess? } I feel that the hardest part is getting software that will enable me } to join up to the Macintosh even if I purchased a new camera. } } I would really like to take the photos digitally but is it } impossible with my present configuration? } i would appreciate any comments please
Faye -
You don't say WHY you can't take photos with your equipment! I suggest that you contact microscopeworld.com. They sell many Motic scopes under the U.S. brand name "National", plus camera connectors, so you should be able to get specific advice on your problem.
You'll find abundant microscopy resources for the grandkids at the MICRO website; URL below.
Caroline
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 3 15:23:11 2004
Friends, here is the table of content of the last 2003 issue of J.Microscopy (OXF) on Microscopy in the Nanobioscience.
215 Foreword A. Diaspro
217 Polysaccharide properties probed with atomic force microscopy N. I. Abu-Lail, T. A. Camesano
239 Encapsulated yeast cells inside Paramecium primaurelia: a model system for protection capability of polyelectrolyte shells S. Krol, O. Cavalleri, P. Ramoino, A. Gliozzi, A. Diaspro
244 Insights into the regulation of transcription by scanning force microscopy R. T. Dame, C. Wyman, N. Goosen
254 Monitoring enzymatic reactions in nanolitre wells I. T. Young, R. Moerman, L. R. Van Den Doel, V. Iordanov, A. Kroon, H. R. C. Dietrich, G. W. K. Van Dedem, A. Bossche, B. L. Gray, L. Sarro, P. W. Verbeek, L. J. Van Vliet
264 The molecular machines of DNA repair: scanning force microscopy analysis of their architecture A. Janiijevi, D. Ristic, C. Wyman
273 TectoRNA and 'kissing-loop' RNA: atomic force microscopy of self-assembling RNA structures H. G. Hansma, E. Oroudjev, S. Baudrey, L. Jaeger
280 The nacre protein perlucin nucleates growth of calcium carbonate crystals S. Blank, M. Arnoldi, S. Khoshnavaz, L. Treccani, M. Kuntz, K. Mann, G. Grathwohl, M. Fritz
292 Atomic force microscopy study of living diatoms in ambient conditions I. C. Gebeshuber, J. H. Kindt, J. B. Thompson, Y. Del Amo, H. Stachelberger, M. A. Brzezinski, G. D. Stucky, D. E. Morse, P. K. Hansma
300 Self-assembly and recrystallization of bacterial S-layer proteins at silicon supports imaged in real time by atomic force microscopy E. S. Györvary, O. Stein, D. Pum, U. B. Sleytr
307 Fluorescent resolution target for super-resolution microscopy P. R. H. Stark, L. J. Rinko, D. N. Larson
I hope is of interest
All my best ALby
....................................................................... ................................... .. non basta, resistere, resistere...resistere ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Alberto Diaspro Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480 fax 010314218 diaspro-at-fisica.unige.it, http://www.lambs.it http://wiley.com/cda/product/0,,0471409200,00.html
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 4 17:50:49 2004
I meant the "visual perception" of quantitation which is what Alwyn's initial observation referred to. Of course I agree totally with you that its easer to distinguish different colors from one another than shades of any color or grey. The question is rather "is bright green more or less than yellow?" Any quantitative color scale must also have factored in parameters such as Hue, Saturation abd Brightness; this is one of the main failings of the "spectrum scale" - it doesn't!
Cheers etc
Peter
} Hello Peter, } } in actuality, the situation is a bit more complex than that. I am looking at } the LUT right now that we have in our analySIS software, and you are of } course right that a pixel with no intensity (intensity 0) is displayed as } black (0,0,0). However, the intensity "1" is displayed as (R:41, G:0, B:0). } Technically, it goes from black to white, but realistically, it goes from } "dark red" to white. } } The situation becomes mor complex at the other end. To make yellow, you have } to add in a green component, and to make white you also have to add in blue. } So, it is not straightforward "black to white" or "red to white". Other } colors get mixed in at the higher intensities to make yellow to white. } } As for qantitation: I am not sure, what you are referring to. Quantitaion is } best done on the b/w images with the help of a computer, which has no } problems distinguishing intensity 2976 from 2977. For the display of small } contrasts for the human eye I agree with you, but there an even better } choice is the spectrum LUT. It's easier to distinguish yellow from green } than it is to distinguish "dark orange" from "darker orange". } } mike } } } } -----Original Message----- } From: Peter Ingram [mailto:p.ingram-at-cellbio.duke.edu] } Sent: Friday, January 02, 2004 12:03 } To: Mike Bode } Subject: [Microscopy] RE: Psuedo color } } } } Actually the "black body" scale goes from black to white, albeit } through red, orange and yellow, not simply red to white - a vital } distinction when quantitation is involved! } } Peter } } } } --------------------------------------------------------------------------- } --- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology Duke University Medical Center Box 90319 DURHAM NC 27708-0319
Gary- I have been using a dual microprobe I purchased from Ernest F. Fullam Inc.:
900 Albany Shaker Rd. Latham NY 12110-1491 800-833-4024, 518-785-5533
Part #15855 dual, o-ring sealed manipulator. They had suggested that I purchase micromanipulators that had metal bellows rather than o-ring seals since I have a FE-SEM, but since all of the other seals on my specimen chamber were o-rings, including detectors that run in and out, I opted for the less expensive o-ring sealed option, and I have had no trouble with them. I cannot say if the microprobes I have will meet your positioning accuracy requirements as I am currently using fairly blunt tips. I also do not have verniers to check reproducibility. I just watch where I am placing them using the SEM. The probes are essentially the same as one might find on an electrical probe station for testing devices. The one problem I do have with them is that since I work with the probe tips near the pole piece, the probes induce a significant aberration. I do not know if the set screws holding the tips in are magnetic, making the problem worse or not. I do know that there are magnetic knurled nuts about 2" up the shaft. You may need to specify the materials you want the probe arms made out of, as well as working at longer WD/higher kV. I haven't bothered to correct this, or to move the probes further from the pole piece, as the resolution is adequate for my experiments. I have found Fullam to be very helpful and open to customizing their products for individual needs. They have a web site at: http://www.fullam.com/. Good luck.
Sincerely, Matthew Ervin, Ph.D. (301)394-0017 phone, (301)394-1559 fax MErvin-at-ARL.Army.mil
M/S: AMSRL-SE-RL US Army Research Laboratory 2800 Powder Mill Road Adelphi, MD 20783-1197
Disclaimer: The opinions and views expressed above are those of the author and do not necessarily represent those of the U.S. Army Research Laboratory or any other government agency
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Wednesday, December 31, 2003 10:11 PM To: MSA listserver
Hi all:
Has anyone seen a source of micro probes for SEM that allow electrical contact to a SEM chamber specimen? I need very precise positioning--like within 0.15u or better and 0.05u repeatability and stability.
I need two contact probes.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 09:42:19 2004
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 10:02:05 2004
I suggest that you check out "Polymer Microscopy", 2nd edition, by Sawyer & Grubb. I believe that they addressed this issue.
Cheers,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Philip Oshel {peoshel-at-wisc.edu} To: Microscopy-at-sparc5.microscopy.com cc: Subject: [Microscopy] nylon SEM 12/23/03 02:55 PM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Materials micromavens,
We have a user doing SEM of nylon, with embedded bits. We'd like to chemically etch the nylon, which is something of an entertaining problem, since nylon is used to mask things against etchants. Does anyone have a recipe for something that will etch nylon and not silica? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 10:34:32 2004
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 10:44:59 2004
We here at the Neurotoxicology Labs at Rutgers University have an ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be dying. Coolwell went out of business years ago, and Zeiss pointed me to Lytron, Inc. as the company who bought Coolwell's stuff when it went under. I have tried to contact Lytron online about repairs or a possible replacement for this chiller, but they don't seem to be paying much attention to their email. I am going to call them directly, of course, but what I would like to know is if anyone can recommend another company or particular chiller model that would be appropriate for our EM, so that if Lytron continues to blow me off I will have other sources that I can try.
Hope you all had a happy holiday!
Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxcology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 10:52:17 2004
Many thanks to you for all your hard work over the past years. I very much appreciate the service that you provide for the microscopy/microanalysis community.
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Nestor J. Zaluzec" {zaluzec-at-microscopy.c To: microscopy-at-ns.microscopy.com om} cc: Subject: [Microscopy] Administrivia: Archives for 2003 Now On-Line 01/01/04 10:37 AM
I have a Zeiss EM900 and my Coolwell also just bit the dust about 2months ago. I went with a Haskris Model 075 chiller, which includes Option (K), a 220V interlock as specified by LEO. I have two other Haskris chillers that have been very reliable (on a JEOL SEM and on a Philips TEM).
Here is the contact info:
Doug Wagner Haskris Co. 100 Kelly Street Elk Grove Village, IL 60007 847-956-6420, x243 (tel) 847-956-6595 (fax) doug-at-haskris.com
Good luck! Ann
++++++++++++++++++++++++++++++++++++++ Ann Hein Lehman Assistant Director, Electron Microscopy Facility Mailstop: LSC-314 Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 e. ann.lehman-at-trincoll.edu f. 860-297-2538 www.trincoll.edu/~alehman
-----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] Sent: Monday, January 05, 2004 11:54 AM To: Microscopy-at-sparc5.microscopy.com
Hi, all-
We here at the Neurotoxicology Labs at Rutgers University have an ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be dying. Coolwell went out of business years ago, and Zeiss pointed me to
Lytron, Inc. as the company who bought Coolwell's stuff when it went under. I have tried to contact Lytron online about repairs or a possible replacement for this chiller, but they don't seem to be paying much attention to their email. I am going to call them directly, of course, but what I would like to know is if anyone can recommend another
company or particular chiller model that would be appropriate for our EM, so that if Lytron continues to blow me off I will have other sources that I can try.
Hope you all had a happy holiday!
Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxcology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 11:39:32 2004
Hakris is a reasonably reliable chiller. I've had several on EM diff pumps. They also make a 115V version that I used on a Hitach S4500 FESEM since my lab had no chilled water supply. Unless strapped for cash, junk the old one. If it fails it can make a nasty mess of things. I think I spent 2K$ [American] for the one I bought 6 years ago.
Happy new year. [Can I say that?]
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Lehman, Ann [mailto:Ann.Lehman-at-trincoll.edu] Sent: Monday, January 05, 2004 12:39 PM To: Kathleen Roberts; Microscopy-at-sparc5.microscopy.com
Dear Kathleen,
I have a Zeiss EM900 and my Coolwell also just bit the dust about 2months ago. I went with a Haskris Model 075 chiller, which includes Option (K), a 220V interlock as specified by LEO. I have two other Haskris chillers that have been very reliable (on a JEOL SEM and on a Philips TEM).
Here is the contact info:
Doug Wagner Haskris Co. 100 Kelly Street Elk Grove Village, IL 60007 847-956-6420, x243 (tel) 847-956-6595 (fax) doug-at-haskris.com
Good luck! Ann
++++++++++++++++++++++++++++++++++++++ Ann Hein Lehman Assistant Director, Electron Microscopy Facility Mailstop: LSC-314 Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 e. ann.lehman-at-trincoll.edu f. 860-297-2538 www.trincoll.edu/~alehman
-----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] Sent: Monday, January 05, 2004 11:54 AM To: Microscopy-at-sparc5.microscopy.com
Hi, all-
We here at the Neurotoxicology Labs at Rutgers University have an ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be dying. Coolwell went out of business years ago, and Zeiss pointed me to
Lytron, Inc. as the company who bought Coolwell's stuff when it went under. I have tried to contact Lytron online about repairs or a possible replacement for this chiller, but they don't seem to be paying much attention to their email. I am going to call them directly, of course, but what I would like to know is if anyone can recommend another
company or particular chiller model that would be appropriate for our EM, so that if Lytron continues to blow me off I will have other sources that I can try.
Hope you all had a happy holiday!
Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxcology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:07:58 2004
Oooh, thank you! You just made things a lot easier for me. :o) I guess I should talk to LEO as well to see what options I need, unless your Zeiss and mine are similar enough?
God, how I love listservs....thank you, Nestor!
Thanks again- Kathleen Neurotoxicology Labs Rutgers University
Lehman, Ann wrote:
} Dear Kathleen, } } I have a Zeiss EM900 and my Coolwell also just bit the dust about } 2months ago. I went with a Haskris Model 075 chiller, which includes } Option (K), a 220V interlock as specified by LEO. I have two other } Haskris chillers that have been very reliable (on a JEOL SEM and on a } Philips TEM). } } Here is the contact info: } } Doug Wagner } Haskris Co. } 100 Kelly Street } Elk Grove Village, IL 60007 } 847-956-6420, x243 (tel) } 847-956-6595 (fax) } doug-at-haskris.com } } Good luck! } Ann } } ++++++++++++++++++++++++++++++++++++++ } Ann Hein Lehman } Assistant Director, Electron Microscopy Facility } Mailstop: LSC-314 } Trinity College } 300 Summit Street } Hartford, CT 06106 } v. 860-297-4289 } e. ann.lehman-at-trincoll.edu } f. 860-297-2538 } www.trincoll.edu/~alehman } } } -----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] } Sent: Monday, January 05, 2004 11:54 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: [Microscopy] chiller for Zeiss EM 10CA } } } } ------------------------------------------------------------------------ } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:11:23 2004
Yes, Dr. Bonder is still at Rutgers-I just did a search for him on Rutgers' website. I don't know him personally, though, as he is on the Newark campus, where I have never been, and I am on the New Brunswick campus. Worlds apart... :o)
Kathleen
Pat Connelly wrote:
} Kathleen, } WE have been using a Haskris Co. water chiller (RO 75) since 1996 for } my Phillips 200 TEM. It works very well and the only complaint that I } have had is that we use a timer to shut down our ancient scope at } night and the one on the chiller keeps dying - it just stays on. } } This company was recommended to me when our previous chiller, also a } Haskris,died after 25 years or so, by a refrigeration specialist who } does some contract work here. } } Do you know if Dr. Ed Bonder is still at Rutgers? He was in EM the } last time I had heard from him but I do not know the exact department } - Biology? He was a grad student here at Penn a few decades ago! } } Pat Connelly } Research Specialist
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:21:08 2004
To all who wrote in reply to my question- } } Thank you for the information, you all have made my life much easier. :o) } } Now I can sit down with my boss and be able to give him some real } information about replacing this poor thing, instead of "I'm still } searching for a source..." } } Thanks again- } Kathleen } Neurotoxicology Labs } Rutgers University
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:26:57 2004
Ouch! That is an expensive repair...would LEO install a temp. sensor on a 'scope that is so old? My impression was that LEO didn't service Zeiss 'scopes anymore.
Thanks for the information- Kathleen
Ken Tiekotter wrote:
} Dear Kathleen, } } I just had a major life change as my Coolwell went down, was repaired, and } crashed again for the third and final time. The issue was exasperated by a } series of facilities failures. The outcome was about an $18k repair bill to } include a new Haskris chiller (~$5600.00) } } Unfortunately, the Zeiss EM 10CA does not a temperature sensor to determine } glycol temperature, but rather only flow rate. The repair included a } complete overhaul of the column because oil vapors were not condensing in } the diffusion pump and consequently went everywhere in the column. } } After almost 20 years with my beloved EM10, the hospital decided to donated } the scope and close my lab. You may want to check with Zeiss (LEO) about } installing a temperature sensor and automatic relay to shut the HV value if } the circulator temperature gets too high. } } Best wishes to you and your EM10! } Ken } } _______________________________________ } Kenneth L. Tiekotter, Adjunct Professor } The University of Portland } Department of Biology } 5000 N Willamette Blvd. } Portland, OR 97203 USA } } Director, MicroImaging Dx Center } Legacy Portland Hospitals } Legacy Holladay Park Medical Center } 1225 NE 2nd Avenue } Portland, OR 97232 USA } } Tel.: 503.413.5391 } } } } } } On 1/5/04 8:54 AM, "Kathleen Roberts" {kgrobert-at-rci.rutgers.edu} wrote: } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:40:57 2004
I will admit that I am not entirely sure what is wrong with it. All I know is that when I turn the 'scope on, it's fine for the first half hour or so...then a buzzer goes off-there is no indicator light to say what that buzzer is for on the 'scope, but my boss tells me that it's an overtemp alarm. If you look at the temp gauge on the chiller, it's reading way above the overtemp limit.
There was one occasion when a pump in the house distilled water system was dying, and when it was replaced, the 'scope stopped buzzing. Now, however, the distilled water system appears to be fine, and the 'scope is buzzing again, so I am assuming that it is the chiller.
I did get a local HVAC repair guy in (from the same company that resurrected our old cryostat), but he said that he couldn't do anything without the refrigeration and other specifications for that chiller. I managed to get a couple of diagrams from someone else on this list (his name escapes my memory for the moment, but thanks again anyway!), but the HVAC guy said that it wasn't enough. As Lytron isn't willing to help, I'm going to give up and get a new chiller, as this one is pretty old anyway.
Thanks, Kathleen Neurotoxicology Labs Rutgers University
McClintock, Joel wrote:
} Kathleen, } } I take care of a Zeiss 902 with a coolwell chiller. I have fixed this thing several times. What is the problem with it? Often the temp sensor goes haywire. I have found it is often the switch. I have replaced it for around $7. As others have recommended Haskris is my favorite. If money is tight I may be able to direct you to a used one. } } Joel McClintock } EM Specialist } U of Kentucky } 859-257-1242 } } -----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] } Sent: Mon 1/5/2004 11:54 AM } To: Microscopy-at-sparc5.microscopy.com } Cc: } Subject: [Microscopy] chiller for Zeiss EM 10CA } } } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi, all- } } We here at the Neurotoxicology Labs at Rutgers University have an } ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be } dying. Coolwell went out of business years ago, and Zeiss pointed me to } Lytron, Inc. as the company who bought Coolwell's stuff when it went } under. I have tried to contact Lytron online about repairs or a } possible replacement for this chiller, but they don't seem to be paying } much attention to their email. I am going to call them directly, of } course, but what I would like to know is if anyone can recommend another } company or particular chiller model that would be appropriate for our } EM, so that if Lytron continues to blow me off I will have other } sources that I can try. } } Hope you all had a happy holiday! } } Thanks in advance- } Kathleen Roberts } Principal Lab Technician } Neurotoxcology Labs } Dept of Pharmacology & Toxicology } Ernest Mario School of Pharmacy } Rutgers University } 41 B Gordon Rd } Piscataway, NJ 08854 } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 13:47:21 2004
Gary, I have seen presentation on two micro-manipulator type systems: Zyvex - a MEMS based system (www.zyvex.com), and Klendiek (http://kleindiek.com, http://www.ascendinstruments.com/product_nanoprober.asp) a micro-motor based system distributed in USA by Ascend Instruments. They are geared to manipulate specimen inside a FIB or SEM chamber and both can be used for electrical measurements. The problem is that they are expensive (~100K). Since I have not actually used either one of them I cannot recommend them. We are thinking of a versatile solution that would allow TEM sample extraction, transfer, in DB-FIB and also allow us to perform occasional electrical measurements on powered devices.
Regards and Happy New Year to all.
Jerzy
PS: If you get other replies, please post them since I would also be interested in a less expensive system but with less flexibility. I do not have any vested interest in either company, we only had sales presentations from both distributors.
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 512 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-amd.com ******************************************************
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Wednesday, December 31, 2003 9:11 PM To: MSA listserver
Hi all:
Has anyone seen a source of micro probes for SEM that allow electrical contact to a SEM chamber specimen? I need very precise positioning--like within 0.15u or better and 0.05u repeatability and stability.
I need two contact probes.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 16:05:53 2004
Another source is Omniprobe, http://www.omniprobe.com/. Click the Products nav button. They can do electrical testing, TEM lift-out, mechanical testing, etc. I have no financial interest in the company.
jerzy.gazda-at-amd.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Gary, } I have seen presentation on two micro-manipulator type systems: Zyvex - a MEMS based system (www.zyvex.com), and Klendiek (http://kleindiek.com, http://www.ascendinstruments.com/product_nanoprober.asp) a micro-motor based system distributed in USA by Ascend Instruments. They are geared to manipulate specimen inside a FIB or SEM chamber and both can be used for electrical measurements. The problem is that they are expensive (~100K). Since I have not actually used either one of them I cannot recommend them. We are thinking of a versatile solution that would allow TEM sample extraction, transfer, in DB-FIB and also allow us to perform occasional electrical measurements on powered devices. } } Regards and Happy New Year to all. } } Jerzy } } PS: If you get other replies, please post them since I would also be interested in a less expensive system but with less flexibility. I do not have any vested interest in either company, we only had sales presentations from both distributors. } } ****************************************************** } Jerzy Gazda, Ph.D. Advanced Micro Devices } Supervising Engineer 5204 E. Ben White Blvd. - MS 512 } PCAL - AIM Section Austin, TX 78741 } TEL: 1-800-538-8450, Ext. 51453 } jerzy.gazda-at-amd.com } ****************************************************** } } } -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Wednesday, December 31, 2003 9:11 PM } To: MSA listserver } Subject: [Microscopy] micro probes } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi all: } } Has anyone seen a source of micro probes for SEM that allow electrical } contact to a SEM chamber specimen? I need very precise } positioning--like within 0.15u or better and 0.05u repeatability and } stability. } } I need two contact probes. } } gary g.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 16:34:46 2004
Email: letitsnow-at-antelecom.net Name: Elizabeth Colvin
Organization: Homeschool
Education: 9-12th Grade High School
Location: Pearblossom, CA-L.A. county
Question: I have obtained several unstained blood smears. I have Wright's stain and methylene blue available. Can you please tell me how long to leave the stain on the slide before rinsing. And do I rinse with distilled water. Thank you.
Good question from Joel. The alarm (buzzer)you are hearing more than likely is the chiller fluid flow indicator on the EM10. The pressure must be maintained at 1.5 - 2liters per minute. The flow indicator is located on the right-hand side of the column inside the gray hinged 'door'. These is a small glass window to show where the float is in relationship to the flow of fluid: the higher the float, the greater the number of liters/minute.
On the front of the Coolwell chiller is also a flow indicator, which should be adjusted to meet the 1.5 - 2 liter flow on the microscope. Also check to see if the temperature gauge on the Coolwell remains the same or fluctuates. It could be the chiller is fine, but the pump is going out.
Ken
Kathleen Roberts wrote:
} } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
--------------------------------------- Kenneth L, Tiekotter, Adjunct Professor Dept. of Biology The University of Portland 5000 N Willamette Blvd, Portland, OR 97203 USA
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 17:16:32 2004
Dear List, I am posting the following for Dr. Jensen, the head of our cryo-EM group:
------------------------------------------------------------------------ ------------------------ I'm looking for an image processing scientist/computer programmer to help with our expanding biological electron tomography projects here at Caltech. We are currently imaging many specimens including cells, viruses, and purified protein complexes with a state-of-the-art 300kV, helium-cooled, energy-filtered, automated, FEG TEM. We are in the process now of purchasing a large new supercomputer for the structural biology groups. Duties would include applying existing programs as well as developing new software for image processing needs, handling large amounts of image data, managing processes on our supercomputer, working with students to help them solve image processing problems, and being a creative member of a growing scientific team. Minimum qualifications are a bachelors degree, strong programming skills, mathematical aptitude, an ability to work well with others, and enthusiasm for biology research. Graduate education or extended experience in related fields is preferred. Interested persons seeking either a post-doctoral position or a permanent staff position are encouraged to apply. Salary will be commensurate with qualifications. CalTech is located in Pasadena, California (a suburb of Los Angeles) next to the San Gabriel mountains, and offers an extraordinarily rich intellectual environment for computationally-inclined scientists, all within a sunny, affordable, diverse community that will make you want to stay. Please send CV and three letters of reference to jensen-at-caltech.edu or
Dr. Grant Jensen California Institute of Technology Biology Division, Mailcode 114-96 1200 E. California Blvd. Pasadena, CA 91125 ------------------------------------------------------------------------ --------------------------
Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 17:40:32 2004
Just to add to what Ken said just below, if the flow alarm is in fact what you are hearing, then just check the various filters in the cooling water system, probably one in the chiller tank, probably another one on the input side to the microscope. Or, the lines may be plugged up somewhere with crud such as algae or corrosion products. After checking the filters and cleaning or replacing them, if problem persists may have to have scope and/or delivery lines flushed to clear them. I had to do that once for in an SEM's interior cooling lines.
Hope this helps! -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra ---------------------------------------------------------------------------- } Kathleen, } } Good question from Joel. The alarm (buzzer)you are hearing more than } likely is the chiller fluid flow indicator on the EM10. The pressure } must be maintained at 1.5 - 2liters per minute. The flow indicator is } located on the right-hand side of the column inside the gray hinged } 'door'. These is a small glass window to show where the float is in } relationship to the flow of fluid: the higher the float, the greater } the number of liters/minute. } } On the front of the Coolwell chiller is also a flow indicator, which } should be adjusted to meet the 1.5 - 2 liter flow on the microscope. } Also check to see if the temperature gauge on the Coolwell remains the } same or fluctuates. It could be the chiller is fine, but the pump is } going out. } } Ken } } Kathleen Roberts wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------ } } Joel- } } } } I will admit that I am not entirely sure what is wrong with it. All I } } know is that when I turn the 'scope on, it's fine for the first half } } hour or so...then a buzzer goes off-there is no indicator light to say } } what that buzzer is for on the 'scope, but my boss tells me that it's } an } } overtemp alarm. If you look at the temp gauge on the chiller, it's } } reading way above the overtemp limit. } } } } There was one occasion when a pump in the house distilled water system } } was dying, and when it was replaced, the 'scope stopped buzzing. Now, } } however, the distilled water system appears to be fine, and the 'scope } } is buzzing again, so I am assuming that it is the chiller. } } } } I did get a local HVAC repair guy in (from the same company that } } resurrected our old cryostat), but he said that he couldn't do anything } } without the refrigeration and other specifications for that chiller. I } } managed to get a couple of diagrams from someone else on this list (his } } name escapes my memory for the moment, but thanks again anyway!), but } } the HVAC guy said that it wasn't enough. As Lytron isn't willing to } } help, I'm going to give up and get a new chiller, as this one is pretty } } old anyway. } } } } Thanks, } } Kathleen } } Neurotoxicology Labs } } Rutgers University } }
} --------------------------------------- } Kenneth L, Tiekotter, Adjunct Professor } Dept. of Biology } The University of Portland } 5000 N Willamette Blvd, } Portland, OR 97203 USA
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 5 17:41:59 2004
I did not want to imply that the "spectrum" scale is "perfect". There are many color scales and depending on what you want to see or show, one or the other might be better.
You bring up a good point, though: familiarity with the scale. Everybody can interpret a black and white scale, and the thermal scale is also very intuitive. Once we get to more colors, I would say that the spectrum scale is familiar to most people, red on one end, blue on the other. Other scales need more explanation or a color scale bar on the Image.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== 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: Peter Ingram [mailto:p.ingram-at-cellbio.duke.edu] Sent: Sunday, January 04, 2004 16:54 To: Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
Steven; I was kidding about being a Raelian. I am agnostic. I celebrate the spirit of Christmas, not the religious part. My wife is Turkish, and is a Moslem. She loves Christmas, as does our daughter. We are presently being visited by my wife's mother and sister. Despite the fact that they are both Moslem, and neither of them speaks English, they loved their first Christmas. When my wife celebrates her holidays, I celebrate with her. My brother's wife is Jewish (another mixed marriage-agnostic and Jewish). Their family celebrates both Hanukah and Christmas. Their two kids love having two holidays to celebrate. As for the spirit of Christmas, I am sorry to hear that it has eluded you. You can give gifts to loved ones any time, not just Christmas. You can try to bring more cheer into peoples lives any time, not just Christmas. You can get together with your family any time, not just Christmas. You can put up special decorations any time, not just Christmas, and you can listen to wonderful music any time, not just Christmas. It's just that human nature is such that we sometimes need a special stimulus to prioritize all this in a hectic world, and Christmas does just that-and more. Finally, if you have ever looked in to the eyes of a young child contemplating Santa Claus' imminent arrival, and shared that wonder, then you would know that this a wonderful thing to have. Steven, you have some homework to do. You need to find out what Christmas Spirit is. If you do it right, you will have a Happy New Year.
John Mardinly 408-765-2346 877-277-1182
-----Original Message----- } From: Steven E. Slap [mailto:siksik03-at-comcast.net] Sent: Monday, January 05, 2004 5:01 AM To: Mardinly, John; Microscopy-at-sparc5.microscopy.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 01:59:15 2004
} You bring up a good point, though: familiarity with the scale.
And one (little) cent more : I had a look a few weeks ago to the biology manual from my daughter (secondary school, french 3°, i. e. 15 years old), an all the EM and SEM pictures were in pseudo color, whith different color rules from one picture to an other and without any mention that it was "false colors" and that the true signal was a monochrome level variation ! I've than understood why a student asked my once, why we dont't have color images on the SEM ... " Not enough monney to pay it ?" asked he ! Familarity with a "false" color scale, can be an obstacle to understand the way the images was obtained (and to understand the image itself, perheps).
So pseudo color, why not of coarse, but with a color scale along a border of the picture, like the micron bar, as is often done on AFM images.
J. Faerber IPCMS Strasbourg France
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 03:36:16 2004
John I was thinking, it's only my family is "so creative". When we came to US my kids very quickly figured out what to do: ever since we do celebrate everything. Catholic Christmas, New Year, then Russian Christmas, then Russian "Old" New Year... The whole point there was to have gifts for every holiday... As far as I remember my kids also enjoyed some Jewish holidays when they had school off... I really like you description: "the spirit of Christmas". I think, good spirit should unify us, not separate by religious believe... Sergey.
At 11:48 PM 1/5/2004 -0800, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 05:30:26 2004
I'll also put in my $.02 worth (Cdn) for Haskris chillers. I've owned both Haskris and Neslab ones over the years, but what I really like about the Haskris models I've used is the fact that the water tank/reservoir has a removable lid, so you can always look inside and visually inspect the condition of the water. The Neslab one we have now, though reasonably reliable and all, has a completely sealed water tank with just a narrow little filler neck, so you can never see what's going on inside. Interestingly, both companies use the same water pumps supplied by a third company somewhere in Indiana, I believe. These pumps are rebuildable and replaceable of course, as are the electric motors that power them, so it's often possible to keep a chiller running for a very long time before it actually has to be replaced. To choose an appropriate model for your particular application you just need to know how much water (usually gallons/minute) and at what temperature your particular instrument needs it, then match that up to the model listing. There's not much point in greatly exceeding the needs of your instrument with a bigger chiller than necessary, since the motor/pump will be running constantly anyway. Some folks will let one big chiller cool several instruments. No doubt this is pretty cost-effective, but the down-side is apparent when the chiller craps out and you now have several instruments down instead of just one.......
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] Sent: Monday, January 05, 2004 12:54 PM To: Microscopy-at-sparc5.microscopy.com
Hi, all-
We here at the Neurotoxicology Labs at Rutgers University have an ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be dying. Coolwell went out of business years ago, and Zeiss pointed me to Lytron, Inc. as the company who bought Coolwell's stuff when it went under. I have tried to contact Lytron online about repairs or a possible replacement for this chiller, but they don't seem to be paying much attention to their email. I am going to call them directly, of course, but what I would like to know is if anyone can recommend another company or particular chiller model that would be appropriate for our EM, so that if Lytron continues to blow me off I will have other sources that I can try.
Hope you all had a happy holiday!
Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxcology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 05:50:44 2004
I'm searching for some decent disposable blades for our Leica Microtome to get sections of living hearts. I heard that Feather Blades should do the trick. Has anyone experience with getting sections of living cardiomyoctes and could give me an address of a supplier of blades, preferrably in Germany/Europe ?
Thanks
Michael Didié
-- +++ GMX - die erste Adresse für Mail, Message, More +++ Neu: Preissenkung für MMS und FreeMMS! http://www.gmx.net
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 07:29:15 2004
Although these are debates that should take place somewhere, this is not the forum for them. Unless one can tie a microscope to this line of remarks I would rather not see this on this listserver.
Just my opinion.
Peter Tomic Agere Systems
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, January 06, 2004 2:48 AM To: Steven E. Slap; Microscopy-at-sparc5.microscopy.com Cc: Microscopy-at-MSA.Microscopy.Com
Steven; I was kidding about being a Raelian. I am agnostic. I celebrate the spirit of Christmas, not the religious part. My wife is Turkish, and is a Moslem. She loves Christmas, as does our daughter. We are presently being visited by my wife's mother and sister. Despite the fact that they are both Moslem, and neither of them speaks English, they loved their first Christmas. When my wife celebrates her holidays, I celebrate with her. My brother's wife is Jewish (another mixed marriage-agnostic and Jewish). Their family celebrates both Hanukah and Christmas. Their two kids love having two holidays to celebrate. As for the spirit of Christmas, I am sorry to hear that it has eluded you. You can give gifts to loved ones any time, not just Christmas. You can try to bring more cheer into peoples lives any time, not just Christmas. You can get together with your family any time, not just Christmas. You can put up special decorations any time, not just Christmas, and you can listen to wonderful music any time, not just Christmas. It's just that human nature is such that we sometimes need a special stimulus to prioritize all this in a hectic world, and Christmas does just that-and more. Finally, if you have ever looked in to the eyes of a young child contemplating Santa Claus' imminent arrival, and shared that wonder, then you would know that this a wonderful thing to have. Steven, you have some homework to do. You need to find out what Christmas Spirit is. If you do it right, you will have a Happy New Year.
John Mardinly 408-765-2346 877-277-1182
-----Original Message----- } From: Steven E. Slap [mailto:siksik03-at-comcast.net] Sent: Monday, January 05, 2004 5:01 AM To: Mardinly, John; Microscopy-at-sparc5.microscopy.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:12:53 2004
Question: I am looking for procedures for various microchemical tests such as protein. Where would be a good resource for finding such procedures? Thanks you for your time,
A number of years ago I took a SEM course and I was told the following:
It seems a number of students comment to their professor that they would like a real time false color display on the SEM. At the time this was not a inexpensive request or total practical. The next day the students found a coffee mug full of Sharpie color markers next to the glass CRT screen and a note welcoming them to "Sharpie Color Technology."
Stay Safe................
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:36:44 2004
Hi All, I know we are not a theological forum ( and I'm not one, I'm just a SEM tech) I agree with the last mail about unifying us. This is only a way as we can Unify Religion with Science including Microscopy Microscopy and Science has discovered a world full of distinctive marks of intelligent design so we can not deny the chance of the existence of a creator, so if we want to give honor to him we have to make sure that we are worshipping him in a good way.
As much of us are from different cultures, we also have to be united, an example of unity is the language, in this case English. But What about religions, does exist a language in which we agree about belief? Even when some religious people have influenced wars, Yes, does exist : is the True. And I'm not being ambiguous I'm talking about the true about the date of Christ birth ( where Christmas is originated), the true about Mexican traditions, I mean all of them over the world, dates, even about the origin of the live (because religions talk about the creator). Knowing the true about our own traditions and beliefs and talk about them to others will unify us.
I agree about giving gifts, I enjoy accepting them and I deeply appreciate when people give them at any day of the year, without expectation.
Sincerely
Rafael Peña SEM Tech Process Enginering Tel 83297100 Ext 4721 Monterrey Nl Mexico
----- Forwarded by Rafael Pena/MONT3/MFG/KEMET/US on 01/06/04 08:29 AM -----
"Tomic, Peter (Peter)" {ptomic-at-agere.com} 01/06/04 07:32 AM
To: {Microscopy-at-sparc5.microscopy.com} cc: {Microscopy-at-MSA.Microscopy.Com} , (bcc: Rafael Pena/MONT3/MFG/KEMET/US) bcc: Rafael Pena/MONT3/MFG/KEMET/US Subject: [Microscopy] RE: RE: Dec. 25
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Although these are debates that should take place somewhere, this is not the forum for them. Unless one can tie a microscope to this line of remarks I would rather not see this on this listserver.
Just my opinion.
Peter Tomic Agere Systems
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, January 06, 2004 2:48 AM To: Steven E. Slap; Microscopy-at-sparc5.microscopy.com Cc: Microscopy-at-MSA.Microscopy.Com
Steven; I was kidding about being a Raelian. I am agnostic. I celebrate the spirit of Christmas, not the religious part. My wife is Turkish, and is a Moslem. She loves Christmas, as does our daughter. We are presently being visited by my wife's mother and sister. Despite the fact that they are both Moslem, and neither of them speaks English, they loved their first Christmas. When my wife celebrates her holidays, I celebrate with her. My brother's wife is Jewish (another mixed marriage-agnostic and Jewish). Their family celebrates both Hanukah and Christmas. Their two kids love having two holidays to celebrate. As for the spirit of Christmas, I am sorry to hear that it has eluded you. You can give gifts to loved ones any time, not just Christmas. You can try to bring more cheer into peoples lives any time, not just Christmas. You can get together with your family any time, not just Christmas. You can put up special decorations any time, not just Christmas, and you can listen to wonderful music any time, not just Christmas. It's just that human nature is such that we sometimes need a special stimulus to prioritize all this in a hectic world, and Christmas does just that-and more. Finally, if you have ever looked in to the eyes of a young child contemplating Santa Claus' imminent arrival, and shared that wonder, then you would know that this a wonderful thing to have. Steven, you have some homework to do. You need to find out what Christmas Spirit is. If you do it right, you will have a Happy New Year.
John Mardinly 408-765-2346 877-277-1182
-----Original Message----- } From: Steven E. Slap [mailto:siksik03-at-comcast.net] Sent: Monday, January 05, 2004 5:01 AM To: Mardinly, John; Microscopy-at-sparc5.microscopy.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:36:44 2004
Hi All, I know we are not a theological forum ( and I'm not one, I'm just a SEM tech) I agree with the last mail about unifying us. This is only a way as we can Unify Religion with Science including Microscopy Microscopy and Science has discovered a world full of distinctive marks of intelligent design so we can not deny the chance of the existence of a creator, so if we want to give honor to him we have to make sure that we are worshipping him in a good way.
As much of us are from different cultures, we also have to be united, an example of unity is the language, in this case English. But What about religions, does exist a language in which we agree about belief? Even when some religious people have influenced wars, Yes, does exist : is the True. And I'm not being ambiguous I'm talking about the true about the date of Christ birth ( where Christmas is originated), the true about Mexican traditions, I mean all of them over the world, dates, even about the origin of the live (because religions talk about the creator). Knowing the true about our own traditions and beliefs and talk about them to others will unify us.
I agree about giving gifts, I enjoy accepting them and I deeply appreciate when people give them at any day of the year, without expectation.
Sincerely
Rafael Peña SEM Tech Process Enginering Tel 83297100 Ext 4721 Monterrey Nl Mexico
----- Forwarded by Rafael Pena/MONT3/MFG/KEMET/US on 01/06/04 08:29 AM -----
"Tomic, Peter (Peter)" {ptomic-at-agere.com} 01/06/04 07:32 AM
To: {Microscopy-at-sparc5.microscopy.com} cc: {Microscopy-at-MSA.Microscopy.Com} , (bcc: Rafael Pena/MONT3/MFG/KEMET/US) bcc: Rafael Pena/MONT3/MFG/KEMET/US Subject: [Microscopy] RE: RE: Dec. 25
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Although these are debates that should take place somewhere, this is not the forum for them. Unless one can tie a microscope to this line of remarks I would rather not see this on this listserver.
Just my opinion.
Peter Tomic Agere Systems
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, January 06, 2004 2:48 AM To: Steven E. Slap; Microscopy-at-sparc5.microscopy.com Cc: Microscopy-at-MSA.Microscopy.Com
Steven; I was kidding about being a Raelian. I am agnostic. I celebrate the spirit of Christmas, not the religious part. My wife is Turkish, and is a Moslem. She loves Christmas, as does our daughter. We are presently being visited by my wife's mother and sister. Despite the fact that they are both Moslem, and neither of them speaks English, they loved their first Christmas. When my wife celebrates her holidays, I celebrate with her. My brother's wife is Jewish (another mixed marriage-agnostic and Jewish). Their family celebrates both Hanukah and Christmas. Their two kids love having two holidays to celebrate. As for the spirit of Christmas, I am sorry to hear that it has eluded you. You can give gifts to loved ones any time, not just Christmas. You can try to bring more cheer into peoples lives any time, not just Christmas. You can get together with your family any time, not just Christmas. You can put up special decorations any time, not just Christmas, and you can listen to wonderful music any time, not just Christmas. It's just that human nature is such that we sometimes need a special stimulus to prioritize all this in a hectic world, and Christmas does just that-and more. Finally, if you have ever looked in to the eyes of a young child contemplating Santa Claus' imminent arrival, and shared that wonder, then you would know that this a wonderful thing to have. Steven, you have some homework to do. You need to find out what Christmas Spirit is. If you do it right, you will have a Happy New Year.
John Mardinly 408-765-2346 877-277-1182
-----Original Message----- } From: Steven E. Slap [mailto:siksik03-at-comcast.net] Sent: Monday, January 05, 2004 5:01 AM To: Mardinly, John; Microscopy-at-sparc5.microscopy.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:49:58 2004
} The next day the students found a coffee mug full of } Sharpie color markers next to the glass CRT screen and } a note welcoming them to "Sharpie Color Technology."
Ah, yes... Interactive computer graphics.
Bruce Girrell
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:54:27 2004
To the listserver, Is there an easier way to make holey carbon (small 1.5 um) films, other than steaming formvar/evaporating carbon and dissolving formvar with solvents? If not who sells good small holed pure carbon films.
thanks mike d
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 08:58:33 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I heard that Feather Blades should do the trick. Has anyone experience with } getting sections of living cardiomyoctes and could give me an address of a } supplier of blades, preferrably in Germany/Europe ? } } Thanks } } Michael Didié } } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 09:40:53 2004
Handbook of Chemical Microscopy, Vol II. (1940) E. M. Chamot and C. W. Mason
(1989 Reprints available from McCrone Research Institute, Chicago, IL.)
Karl Hagglund (513) 634-0146
} From: lookerr-at-battelle.org (by way of Ask-A-Microscopist) on 01/06/2004 02:15 PM GMT
lookerr-at-battelle.org To: microscopy-at-ns.microscopy.com (by way of Cc: (bcc: Karl Hagglund-KW/PGI) Ask-A-Microscopist) Subject: [Microscopy] AskAMicroscopist: microchemical tests
01/06/2004 09:15 AM
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Email: lookerr-at-battelle.org Name: Ron Looker
Organization: Battelle
Education: Graduate College
Location: Columbus, OH
Question: I am looking for procedures for various microchemical tests such as protein. Where would be a good resource for finding such procedures? Thanks you for your time,
Michael, Assuming your message has lost a little in translation you might like to go to the Leica's own web site www.leica-microsystems.com. They their own (feather) microtome blades but are you actually using a vibratome?
I have just seen an ad in Microscopy and analysis Jan 2004 for a new live cell cutting module for their microdissection kit for live tissue cultures. (I have no commercial interest)
Gill Brown
GlaxoSmithKline Medicines Research Centre, STEVENAGE,
""Michael Didié"" {Michael.Didie-at-gmx.de}
06-Jan-2004 11:53
To: Microscopy
cc: Subject: [Microscopy] Feather Blades
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello everyone,
I'm searching for some decent disposable blades for our Leica Microtome to get sections of living hearts. I heard that Feather Blades should do the trick. Has anyone experience with getting sections of living cardiomyoctes and could give me an address of a supplier of blades, preferrably in Germany/Europe ?
Thanks
Michael Didié
-- +++ GMX - die erste Adresse für Mail, Message, More +++ Neu: Preissenkung für MMS und FreeMMS! http://www.gmx.net
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 10:04:41 2004
There is a thoughtful, well-presented magazine called Science & Spirit that addresses the intersection under discussion, which may be an appropriate venue for this dialogue. (It might make a good article for them if anyone wants to pursue it!)
-- *************************************************************** Do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro Journeys in Microspace (Columbia University Press, 1995) http://www.lsc.org/antarctica/front.html
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 10:10:32 2004
Well, knowing my boss, he will want to start with the smaller stuff -all the suggestions of what to check on the Coolwell that you and everyone else has been sending me (thank you oh so VERY much, everyone!)-and then when all those have been exhausted, go buy a Haskris. So, if it is not too much trouble, could you please dig up & send me that Grainger part number, just in case?
I will check everyone's suggestions and see if they work. At the very least, it would be nice to be at least somewhat functional until we get the new chiller in, assuming that the Facilities people here can figure the Coolwell out without diagrams. :o)
Muchas gracias, Kathleen Neurotoxicology Labs Rutgers University
McClintock, Joel wrote:
} Kathleen, } } This sounds real familiar. I suspect the switch is your problem with this chiller. No matter what chiller you have the compressor comes on then off, back and forth. On the front of this chiller right in the middle should be a temp adjustment, don't recall exact wording, but if you take a flat blade screw driver and move it around you should hear the cmpressor come on. If you do watch the temperature and see if it goes down. If so the compressor is working. Your problem then is something is not telling it ot come on properly. In my experience it is the switch from the thermocouple. Coolwells are really sensitive and when operating correctly keep the water temp pretty steady. THe compressor comes off and on, on and off quite often. The thermocouple initiates this. The thermocouple is stuck in the water and a copper line with a flat copper plate at it's end. This plate buts up to a switch. The switch has a little button on it and is pretty sensitive. As the water temp changes the copper swells and shrinks turning the switch off and on respectively. If the switch is sticky or fails things don't work. Doesn't take much as you might imagine the copper moves very little. I can dig up the number for this switch if you need it as I got it through a company called Grainger. Sound like you are going with a new one though. Good luck. } } One thing to keep in mind is whether you have a Haskris chiller or coolwell over time maintenance and/or repair will be necessary. Over the years I have called Haskris many times and their phone support is very very good. The only way to get any help on a Coolwell is if you had a time machine and even then I would not be hopeful. The design of Haskris is service friendly. } } Joel } } -----Original Message----- } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] } Sent: Mon 1/5/2004 2:50 PM } To: McClintock, Joel; Microscopy-at-sparc5.microscopy.com } Cc: } Subject: [Microscopy] Re: chiller for Zeiss EM 10CA } } } } Joel- } } I will admit that I am not entirely sure what is wrong with it. All I } know is that when I turn the 'scope on, it's fine for the first half } hour or so...then a buzzer goes off-there is no indicator light to say } what that buzzer is for on the 'scope, but my boss tells me that it's an } overtemp alarm. If you look at the temp gauge on the chiller, it's } reading way above the overtemp limit. } } There was one occasion when a pump in the house distilled water system } was dying, and when it was replaced, the 'scope stopped buzzing. Now, } however, the distilled water system appears to be fine, and the 'scope } is buzzing again, so I am assuming that it is the chiller. } } I did get a local HVAC repair guy in (from the same company that } resurrected our old cryostat), but he said that he couldn't do anything } without the refrigeration and other specifications for that chiller. I } managed to get a couple of diagrams from someone else on this list (his } name escapes my memory for the moment, but thanks again anyway!), but } the HVAC guy said that it wasn't enough. As Lytron isn't willing to } help, I'm going to give up and get a new chiller, as this one is pretty } old anyway. } } Thanks, } Kathleen } Neurotoxicology Labs } Rutgers University
} } McClintock, Joel wrote: } } } Kathleen, } } } } I take care of a Zeiss 902 with a coolwell chiller. I have fixed this thing several times. What is the problem with it? Often the temp sensor goes haywire. I have found it is often the switch. I have replaced it for around $7. As others have recommended Haskris is my favorite. If money is tight I may be able to direct you to a used one. } } } } Joel McClintock } } EM Specialist } } U of Kentucky } } 859-257-1242 } } } } -----Original Message----- } } From: Kathleen Roberts [mailto:kgrobert-at-rci.rutgers.edu] } } Sent: Mon 1/5/2004 11:54 AM } } To: Microscopy-at-sparc5.microscopy.com } } Cc: } } Subject: [Microscopy] chiller for Zeiss EM 10CA } } } } } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } Hi, all- } } } } We here at the Neurotoxicology Labs at Rutgers University have an } } ancient Coolwell chiller for our Zeiss EM 10CA, and it appears to be } } dying. Coolwell went out of business years ago, and Zeiss pointed me to } } Lytron, Inc. as the company who bought Coolwell's stuff when it went } } under. I have tried to contact Lytron online about repairs or a } } possible replacement for this chiller, but they don't seem to be paying } } much attention to their email. I am going to call them directly, of } } course, but what I would like to know is if anyone can recommend another } } company or particular chiller model that would be appropriate for our } } EM, so that if Lytron continues to blow me off I will have other } } sources that I can try. } } } } Hope you all had a happy holiday! } } } } Thanks in advance- } } Kathleen Roberts } } Principal Lab Technician } } Neurotoxcology Labs } } Dept of Pharmacology & Toxicology } } Ernest Mario School of Pharmacy } } Rutgers University } } 41 B Gordon Rd } } Piscataway, NJ 08854 } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 11:05:49 2004
Group, FYI, I have posted the availability of a Kevex Sigma Gold EDS processor and a Kevex 4855 Digital Beam Control Interface to the surplus equipment list. Cheers, Tom
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 11:07:24 2004
Morning Elizabeth, If you have Wright's Stain you should do the following. 1. find two glass rods that will reach across a dish/bowl on which you can support a slide with smear up. 2. Wright's Stain is dissolved in methanol (or should be! - 0.5g Wright's in 100ml (~ { 3oz) - mixed by 'trituration' (grinding with the end of a round-bottom test tube) in small amounts of the alcohol until all is dissolved!). It is delivered on the horizontal dry smear slowly, with a dropper, so that a puddle of stain covers the smear (& perhaps the entire slide). Leave for 2.5 min. 3. Add water dropwise to the surface of the stain until IT forms a puddle that covers about 1/2 of the slide. Then blow gently, back-and-forth, on the surface of the water-stain until the two are mixed well. Total time should be about 4.5 min. 4. Rinse the water-stain off the slide in gently running water and stand the slide against an inverted glass to dry - with smear down. [If at home, DO ALL staining, rinsing and drying on aluminum foil. The dye will stain Formica surfaces. Removal requires another email.] When completely dry, a coverglass can be applied using appropriate care with a permanent mountant. 5. You can also view the smear directly with an oil immersion lens (that's the way it is done in pathology labs). Oil is placed directly on the smear and then differential counting is performed. Count 100 white blood cells, identifying each, and record the distributions. A normal smear will show something like the following: 1 basophil, 2 eosinophils, 38 neutrophils (each of the previous identified by cytoplasmic granules that are dark blue, bright red and the latter pink with a segmented nucleus respectively), and 59 lymphocytes (small cells with a round nucleus and a thin rim of cytoplasm). All red blood cells should be orange and without nuclei.
The theory is this. Methylene blue (base) and eosin (acid) are mixed in water (1:1) and combine to form a precipitate. The precipitate is dried and then dissolved(?) in methanol. After the dye thoroughly penetrates the cells in the smear, the water causes the precipitate to dissociate (based on mass action). The methylene blue and eosin are then simultaneously accessible to cellular constituents and are attracted according to their individual affinities. The rinse in excess water then removes all unbound dye. Applying the dyes separately requires much more work and gives much less satisfactory results. The above dyes belong to a group of blood dyes called "Romanovsky Stains".
Coverslipping. If you do not have an oil immersion lens, you can do the following so that you can view cells with a 40X dry objective. This will work though you will have to remove the coverslip and the oil to store the smear. To keep an oiled smear, absorb the excess oil with the tip of a piece of paper towel.
DO NOT use alcohol (will leach dyes!) to remove the rest. Wrap the slide once with good quality paper and NO Scotch tape!
DO NOT try to look at the cells when dry. The image will be saturated with diffraction rings that arise through the interaction of the microscope light and the curved surfaces of the cells - which are whole in a smear (remember?).
Hope this helps,
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson-at-wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:letitsnow-at-antelecom.net] Sent: Monday, January 05, 2004 5:38 PM To: microscopy-at-ns.microscopy.com
Email: letitsnow-at-antelecom.net Name: Elizabeth Colvin
Organization: Homeschool
Education: 9-12th Grade High School
Location: Pearblossom, CA-L.A. county
Question: I have obtained several unstained blood smears. I have Wright's stain and methylene blue available. Can you please tell me how long to leave the stain on the slide before rinsing. And do I rinse with distilled water. Thank you.
Please, PLEASE, can the next person to add to this thread spell PSEUDO correctly! ..please... (if it's not too much to ask...) (whimper) :-)
Mike O'Keefe
Bruce Girrell wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } The next day the students found a coffee mug full of } } Sharpie color markers next to the glass CRT screen and } } a note welcoming them to "Sharpie Color Technology." } } Ah, yes... Interactive computer graphics. } } Bruce Girrell
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 12:12:03 2004
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003 .
We use a manual processing method (no nitrogen-burst agitation). Although we have now managed to essentially eliminate the background fog and "mottled" appearance, the negatives appear darker (denser) than with the previous formulation.
These darker negatives can be printed satisfactorily in the darkroom. However, we digitize our images and archive them on-line. With the "New Formulation" film and denser negative we have noticed scan lines in our
digitized images. The darker the negative, the more apparent the scan lines. We are currently investigating the plate camera emulsion selector positions on our TEM with the hope that concurrent adjustments of exposure time/emulsion setting/darkroom technique will result in a 'less dense' negative.
Has anyone else run in to this problem when digitizing their "denser" negatives? Does anyone have any thoughts beyond what we are trying?
Thank you in advance for your assistance.
Catherine Powell
Catherine Powell, Ph.D. Division of Surgical Pathology Department of Laboratory Medicine, Saint John Regional Hospital P.O. Box 2100, Saint John, NB Canada E2K 4G9 phone (506) 648-7183 FAX (506) 648-6514 email powca-at-reg2.health.nb.ca
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 12:55:28 2004
We've been through the ringer with this film and have finally come to terms with it. However, during the last three changes of D-19 developer we have gotten significantly darker negatives (including the data bar, which is added by the scope independently of negative exposure in our JEOL 1200EX), for some reason we don't yet understand. We didn't change anything in the way we mixed our developer or exposed our films, and we have no reason to think the developer has been changed. We solved this in the meantime by diluting the developer, since the first batch of negatives had already been exposed and it's not a good idea to shorten already short developing times---four minutes in our case. We added about 25% more water (i.e., another quart per gallon of developer), ran a couple test negatives, then proceeded normally with regard to time/temperature/agitation. This worked well for us. Of course, changing developer dilutions can affect the tonal response of films, but in this case the results were fine. If you try this, always run your own tests, of course.
We've had no problems with "scan lines". That's puzzling. They must be coming from the scanner, itself, which maybe indicates that some of the scanning elements (sorry, I forget the terminology---are they called pixels?) aren't doing their thing and are causing lines in the digitized image.
Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Dr. Catherine Powell [mailto:POWCA-at-reg2.health.nb.ca] Sent: Tuesday, January 06, 2004 12:15 PM To: Microscopy-at-MSA.Microscopy.Com
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 13:19:33 2004
Regarding the denser negs, anytime you change film in any camera system you usually need to re-calibrate camera exposure and film development. Sounds like you have done that and are getting a less dense neg that can be nicely digitized.
Regarding the scan lines flaw, you didn't mention if you are using an actual negative scanner, or a flatbed scanner to digitize the negs. But in either case, you may just need to lubricate the moving parts. In my case, I use a UMAX Vista-S8 and about once a year I need to pull the glass surfaces off to get inside and lubricate the metal bar that the scanner heads move on. They just get dry, so then they don't slide smoothly and "skip" a line here and there. Use a clean lint-free cloth with a dab of a fine oil on it, or 3-in-1 oil is what I use. In fact, I just did this lube job yesterday, and it worked!
Good luck!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra
} Our lab recently began using the "New Formulation" Kodak Electron } Microscope Film 4489 and employed a number of tips for developing this } film which we retrieved from the Listserver archive commencing Jan 2003 . } } We use a manual processing method (no nitrogen-burst agitation). } Although } we have now managed to essentially eliminate the background fog and } "mottled" appearance, the negatives appear darker (denser) than with } the previous formulation. } } These darker negatives can be printed satisfactorily in the darkroom. } However, we digitize our images and archive them on-line. With the } "New Formulation" film and denser negative we have noticed scan lines in our } digitized images. The darker the negative, the more apparent the scan } lines. } We are currently investigating the plate camera emulsion selector } positions on } our TEM with the hope that concurrent adjustments of exposure } time/emulsion setting/darkroom technique will result in a 'less dense' } negative. } } Has anyone else run in to this problem when digitizing their "denser" } negatives? Does anyone have any thoughts beyond what we are trying? } } Thank you in advance for your assistance. } } Catherine Powell } Catherine Powell, Ph.D. } Division of Surgical Pathology } Department of Laboratory Medicine, Saint John Regional Hospital } P.O. Box 2100, Saint John, NB Canada E2K 4G9 } phone (506) 648-7183 FAX (506) 648-6514 } email powca-at-reg2.health.nb.ca
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 13:32:52 2004
I haven't had any issues scanning in the darker negatives. We have an Agfa Duoscan T2500 that works well enough with these newer negatives at 1000 and 2500 ppi (the two resolutions I've tried). Before the holiday break I put together a group of images (digitally scanned ones) that had both new and old negatives included with no discernable differences between them. Other than having the scanner support discontinued its been a great and reliable scanner for lecture slides and TEM negatives (if anyone has a driver for it that works with the latest Mac OS 10.3 I would be very interested in hearing about it). As you have mentioned, it would probably be worth trying a emulsion setting series to see if that improves your results. But alas I cannot say that I've tried this yet to attempt to get the old and new density similar on the negatives.
HTH,
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: Dr. Catherine Powell [mailto:POWCA-at-reg2.health.nb.ca] } Sent: Tuesday, January 06, 2004 1:15 PM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] TEM -Need help with scanning EM film 4489 } } } } ------------------------------------------------------------------------ -- } ---- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ -- } ----- } } Our lab recently began using the "New Formulation" Kodak Electron } Microscope Film 4489 and employed a number of tips for developing this } film } which we retrieved from the Listserver archive commencing Jan 2003 . } } We use a manual processing method (no nitrogen-burst agitation). } Although } we have now managed to essentially eliminate the background fog and } "mottled" appearance, the negatives appear darker (denser) than with } the previous formulation. } } These darker negatives can be printed satisfactorily in the darkroom. } However, we digitize our images and archive them on-line. With the } "New } Formulation" film and denser negative we have noticed scan lines in our } } digitized images. The darker the negative, the more apparent the scan } lines. } We are currently investigating the plate camera emulsion selector } positions on } our TEM with the hope that concurrent adjustments of exposure } time/emulsion setting/darkroom technique will result in a 'less dense' } negative. } } Has anyone else run in to this problem when digitizing their "denser" } negatives? Does anyone have any thoughts beyond what we are trying? } } Thank you in advance for your assistance. } } Catherine Powell } } } } Catherine Powell, Ph.D. } Division of Surgical Pathology } Department of Laboratory Medicine, Saint John Regional Hospital } P.O. Box 2100, Saint John, NB Canada E2K 4G9 } phone (506) 648-7183 FAX (506) 648-6514 } email powca-at-reg2.health.nb.ca
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 13:34:03 2004
Thanks, Mike, for pointing out the proper spelling. The correct pronunciation of âpsuedoâ is as in âblue psuedo shoes.â And thereâs nothing all that astonishing about using colored markers on the screen. When we first introduced a computer (one of the original upright Macs) in the department office a couple of decades ago, to a secretary firmly set in the old ways, we had to take turns each night cleaning the whiteout correction fluid off the screen. {;-)
Anyway, to the use and abuse of pseudo color;...
It is certainly true that human vision can only distinguish a few dozen shades of grey brightness on a display screen, as compared to a few hundred colors. Note that both of these values are far less than the 256 shades of brightness or âmillionsâ of colors that the hardware typically controls. It is also true that trying to direct someoneâs attention to the âkind of darkish grey spotâ is a lot less helpful than âthe yellow-orange spotâ in an image (but of course, human words for colors arenât terribly consistent or widely agreed, either - look at any set of paint chips in the turquoise-teal-seagreen-etc. family). Pseudo- or false-color certainly has some valid uses. But it is also easily misunderstood, widely abused, and often hides more in the image than it reveals. And if the viewer is one of the 5-10% of men (or the very tiny percentage of women) who have defective color vision, it is inappropriate in any case.
Firstly, of course, the table must be shown along with a numerical scale that translates it. But even then, simple spectrum lookup table is rarely if ever a good choice. The problem is that the table is typically constructed with uniform steps in hue, going around the color wheel. But human vision is notably insensitive to changes in hue in the green part of the spectrum, and much more so at the red and blue ends and through the red-to-blue purples. A perceptually uniform hue scale (which I have never seen used) would stretch these out and compress the greens and could probably produce more than a hundred discernible colors.
More colors could be seen if they were NOT fully saturated. Changing saturation and hue in a spiral pattern, or also altering brightness along with hue and saturation, can produce color tables that varied in a gradual way and produced greater ability to distinguish changes. The gradual part is important - if the colors jump around too much in discontinuous ways, the image is badly broken up (camouflaged, in effect) and the overall sense of structure, the gestalt of the image, is hidden. To some extent this happens even with a good, gradual table. The use of the âheatâ or âthermoâ scale is an example of a gradual and visually attractive scale, which does not break up the content of the image. But it does not actually add very much to the ability to visually distinguish small changes - perhaps a 20-40% improvement over straight grey scale (which is why color tints are also used in photographic printing, to gain the same increase). Note that the brightness increases monotonically in this scale, and that it is by contributing more steps at the dark end that the increase is obtained.
For selected purposes, carefully constructed color scales can be useful to help the viewer perceive subtle differences or make comparisons from one part of an image to another. But they need to be documented, and in most cases it is also important to show the original data as well in case the color scale can produce misinterpretation or hide other information.
It has been my experience that people are not generally assisted very much by pseudo color scales, as compared to other ways to reveal subtle detail. One of the best of these is to render the surface with elevation representing the original grey scale value. We have millions of years of evolution in our brain wiring that knows how to interpret surface images, in terms of shape and roughness. Using computer graphics to generate properly rendered images with correct perspective, and adjustable viewpoints, surface characteristics, and illumination is easy with current technology and communicates very effectively. The AFM folks use this trick too, along with color scales, although in most cases in only limited ways.
John Russ (see www.DrJohnRuss.com for a schedule of upcoming workshops on image analysis)
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 13:50:49 2004
The recent thread about the use and abuse of pseudo color is only one of many issues that have to do with an important topic that underlies just about everything that we as microscopists do - namely, we LOOK AT images. But while we are typically very concerned with the performance and specifications of our microscopes, we take for granted the performance of our visual systems, to our peril.
Over the past 5 years or so I have been invited several times to give a talk on human vision and how it impacts what microscopists see (and fail to see) in images. At the repeated urging of many people, Iâve prepared the lecture in written form. Anyone who wants to read it can download the "Seeing the Scientific Image" pdf file from my website (www.DrJohnRuss.com). If someone thinks there is a logical place to publish it (too long for Microscopy Today, and not a research paper for Microscopy and Microanalysis) please let me know.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 14:12:58 2004
Silly me, in all my crabbing about this chiller, I didn't give the model? The model # is SE-075W CZ. I got some diagrams from Eckhart Dorneich, and a manual is on the way from Joel McClintock (thank you both!), but I'll take yours too, if they match my chiller. I figure you can never have too much information, and it's always good, in this lab, to have extra copies. :o)
Thank you very much- Kathleen Neurotoxicology Labs Rutgers University
Tina Carvalho wrote:
} Hi- } } What model chiller do you have? I might have the wiring and plumbing } diagrams. } } I have a 10/A and have struggled to keep the chiller going. The scope was } down for a couple of years - oil and then water in the column! - while we } used our new LEO 912 EFTEM. The expensive new instrument can be very } frustrating, and was down once for nearly 5 months! So we got the 10/A } going again, including the chiller. We all had forgotten what a gem it is, } and right now I'd sell the new one and keep the old if I had a } choice. } } We have Hakris chillers on our other two instruments, and they are } fine. We got the very first one that Haskris built for Zeiss/LEO, and they } had a fit trying to include both a pressure and flow gauge plus that } interlock that keeps it going to cool down the diff pump after you turn } off the scope, but they are quite used to it, now. They're pretty reliable } and, fortunately, not prone to that temperature sensor screwing up like } the Coolwell. } } Over the years I've found out there are a whole lot of closet Zeiss 10s } out there - the most reliable backup even if you have a fancy new } instrument! Keep it going. My next trick is to outfit it with a digital } camera. } } Aloha, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 15:03:50 2004
Dear Catherine, I found the new formulation film was more sensitive and adjusted my auto settings in the TEM to compensate. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "Dr. Catherine Powell" {POWCA-at-reg2.health.nb.ca} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Tuesday, January 06, 2004 10:14 AM
Scanning very dense negatives can cause problems related to the scanner if your scanner is using a CCD rather than a PMT. With a CCD you can not crank up the current to boost the signal like you can with a PMT. The result is that there is not enough light i.e. signal getting to the camera array. If that is the case then you must either slow down the scan speed so that the camera can collect enough signal or if this is impossible make your negatives less dense. Our ZI Photoscan 2000 has problems with negatives above an OD of 2.3 or so. Above this lines like what you described appear. I hope this helps. Good luck.
Bob Grassucci
At 02:14 PM 1/6/04 -0400, Dr. Catherine Powell wrote:
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Robert Grassucci Howard Hughes Medical Institute at The Wadsworth Center Empire State Plaza Albany, NY 12201-0509 Phone: (518)474-5821 Fax: (518)486-2191 E-Mail: bobg-at-wadsworth.org
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 6 17:31:44 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alchung-at-ucla.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, January 6, 2004 at 16:51:57 ---------------------------------------------------------------------------
Email: alchung-at-ucla.edu Name: Albert Chung
Organization: UCLA
Education: Graduate College
Location: Los Angeles, CA USA
Question: I am having issues with the silicon monoxide film tearing in the TEM. I have called Ted Pella about this problem and they are making a new batch as we speak, but I have about 50 silicon monoxide films on copper grids for which I cannot use. Any suggestions on how to approach this problem is greatly appreciated. I am operating the JEOL 100CX and the typical current saturation is about 80 mA.
If you need additional information, please let me know.
There is another one: you may bombard thin nylon film by some heavy ions (argon may be good) in the accelerator. It will produce very uniform holes. The size of holes strictly dependent from the energy and type of particle used. But, still: you need to eveporate carbon over and dissolve nylon (have no idea how to do so). As a matter of fact this technology widely used to produce filters for ultrafiltration.
Sergey
At 07:03 AM 1/6/2004, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Albert Chung wrote: ======================================================================== Question: I am having issues with the silicon monoxide film tearing in the TEM. I have called Ted Pella about this problem and they are making a new batch as we speak, but I have about 50 silicon monoxide films on copper grids for which I cannot use. Any suggestions on how to approach this problem is greatly appreciated. I am operating the JEOL 100CX and the typical current saturation is about 80 mA. ======================================================================== Support films of SiOx can be too thick or too thin or could have other features about them that render them unstable in the electron beam. This is of course exactly the same for carbon coated grids. That is why it is imperative to have in-house, as part of the grid coating process, a TEM for batch inspection purposes so that the customer does not end up being responsible for their own QC. And does not end up preparing a large number of grids only to find them unusable.
In the case of carbon replica films that were prepared but which are too thin, we have in the past, been able to resurrect them by applying another coating of carbon to strengthen them and to make grids that were once unstable, now stable. We have never done this with SiOx filmed grids but in theory, it might work the same way.
Disclaimer: SPI Supplies produces SiOx firmed grids as a part of our business and to our knowledge, we have not had any problems with film stability in the electron beam. On occasion, we do make a batch that flunks our own in-house TEM inspection process but those grids never make it into the hands of customers.
Chuck
PS: Remember that we are 100% paperless and the only way we can keep track of this kind of correspondence is if you reply using the reply feature of your e-mail software. That way the entire string of correspondence will be in one place.
============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 00:09:32 2004
by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id i0769WI9018789 for {LL1-at-ns.microscopy.com} ; Wed, 7 Jan 2004 00:09:32 -0600 Received: (from mail-at-localhost) by ns.microscopy.com (8.12.8/8.12.8/Submit) id i0769WFG018785 for LL1; Wed, 7 Jan 2004 00:09:32 -0600 X-Authentication-Warning: ns.microscopy.com: mail set sender to MicroscopyL-request-at-ns.microscopy.com using -f Received: from NJZ-MicroscopyListServer_filter (ns.microscopy.com [206.69.208.10]) by ns.microscopy.com (8.12.8/8.12.8) with SMTP id i0768pI9018755 for "MListServerFilteredEmail_6-at-microscopy.com"; Wed, 7 Jan 2004 00:09:11 -0600 Received: from caduceus.jf.intel.com (fmr06.intel.com [134.134.136.7]) by ns.microscopy.com (8.12.8/8.12.8) with ESMTP id i0768oI9018752 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 7 Jan 2004 00:08:50 -0600 Received: from talaria.jf.intel.com (talaria.jf.intel.com [10.7.209.7]) by caduceus.jf.intel.com (8.12.9-20030918-01/8.12.9/d: major-outer.mc,v 1.12 2003/12/18 18:58:11 root Exp $) with ESMTP id i076DUa5019496; Wed, 7 Jan 2004 06:13:30 GMT Received: from orsmsxvs040.jf.intel.com (orsmsxvs040.jf.intel.com [192.168.65.206]) by talaria.jf.intel.com (8.12.9-20030918-01/8.12.9/d: major-inner.mc,v 1.7 2003/12/18 18:58:10 root Exp $) with SMTP id i076A5Zv017386; Wed, 7 Jan 2004 06:10:20 GMT Received: from orsmsx332.amr.corp.intel.com ([192.168.65.60]) by orsmsxvs040.jf.intel.com (SAVSMTP 3.1.2.35) with SMTP id M2004010622122332399 ; Tue, 06 Jan 2004 22:12:23 -0800 Received: from orsmsx312.amr.corp.intel.com ([192.168.65.62]) by orsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(5.0.2195.5329); Tue, 6 Jan 2004 22:12:23 -0800 Received: from scsmsx402.amr.corp.intel.com ([10.3.90.16]) by orsmsx312.amr.corp.intel.com with Microsoft SMTPSVC(5.0.2195.5329); Tue, 6 Jan 2004 22:12:23 -0800 Received: from scsmsx403.amr.corp.intel.com ([10.3.90.18]) by scsmsx402.amr.corp.intel.com with Microsoft SMTPSVC(5.0.2195.5329); Tue, 6 Jan 2004 22:12:22 -0800 content-class: urn:content-classes:message MIME-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" X-MimeOLE: Produced By Microsoft Exchange V6.0.6487.1
Catherine; One of my favorite web sites, Imaging Resources http://www.imaging-resource.com has reviewed numerous film scanners. Scanning extremely dense film has always been an acid test for a scanner. The following link is to a negative scanned with a Minolta Dimage Scan Elite II Film & Slide Scanner that failed the acid test in some dark regions, and the effect there seems similar to what you describe. The site tests many other scanners with the same negatives, and many do not exhibit this artifact. http://www.imaging-resource.com/SCAN/DSEII/DSETRAA602PS.HTM
John Mardinly Intel
-----Original Message----- } From: Dr. Catherine Powell [mailto:POWCA-at-reg2.health.nb.ca] Sent: Tuesday, January 06, 2004 10:15 AM To: Microscopy-at-MSA.Microscopy.Com
Our lab recently began using the "New Formulation" Kodak Electron Microscope Film 4489 and employed a number of tips for developing this film which we retrieved from the Listserver archive commencing Jan 2003 .
We use a manual processing method (no nitrogen-burst agitation). Although we have now managed to essentially eliminate the background fog and "mottled" appearance, the negatives appear darker (denser) than with the previous formulation.
These darker negatives can be printed satisfactorily in the darkroom. However, we digitize our images and archive them on-line. With the "New Formulation" film and denser negative we have noticed scan lines in our
digitized images. The darker the negative, the more apparent the scan lines. We are currently investigating the plate camera emulsion selector positions on our TEM with the hope that concurrent adjustments of exposure time/emulsion setting/darkroom technique will result in a 'less dense' negative.
Has anyone else run in to this problem when digitizing their "denser" negatives? Does anyone have any thoughts beyond what we are trying?
Thank you in advance for your assistance.
Catherine Powell
Catherine Powell, Ph.D. Division of Surgical Pathology Department of Laboratory Medicine, Saint John Regional Hospital P.O. Box 2100, Saint John, NB Canada E2K 4G9 phone (506) 648-7183 FAX (506) 648-6514 email powca-at-reg2.health.nb.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 06:34:24 2004
We have too 2 Kevex Delta avavaible, one complete, with detector (10mm2, 138 eV, warm, but was working in July)), the other only the electronic, for spares, with 8" and 5"1/4 Bernouilli.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
If anyone has a mechanical lab balance they would like to dispose of, I could use one. I collect/repair antique phonographs as a hobby and would like to be able to true up governor weights from time to time. I've looked at E-bay but I am leery since, if a balance moves up and down, the general public thinks it works. Some units offered are missing parts. etc.
Thanks,
Ron
-----Original Message----- } From: qualityimages [mailto:qualityimages-at-netrax.net] Sent: Wednesday, January 07, 2004 7:37 AM To: Microscopy
Posting this for a friend.
Ken Converse owner Quality Images third party SEM service Delta, PA
Hi All,
I am doing some housecleaning & have the following EDS Systems free to anyone:
As many of you may know I run a training organisation that travels world wide spreading the word on SEM, TEM and EDS operation. I also have a deep interest in "Quality in Electron Microscopy"as those who picked up our paper last year would know?
Now to the point. Once again I am picking up respected journals and finding examples of what I would call poor microscopy, but in truth it also demonstrates poor quality control!
To bring one image to mind. The micrograph is of a structure which is described as being an example of a smooth surface on a biological material. But, the micrograph was taken at 20kV, where the vast majority of the information will have come from beneath the surface softening the true surface detail.
First to remove the training aspect . Operators of SEM should be taught that by manipulation of kV and working distance one may subdue or enhance surface features. To use more than 10kV on most biological samples is asking for sub surface detail, ignore this and comments on surface irregularities are null and void in my mind. (I have to say I would probably try to use 2 to 5kV if the microscope used was produced in the last 15 years!)
Now the quality aspect. By the time a paper is published a number of steps should have been taken. Working backwards, the publisher should have the paper vetted by knowledgeable scientists who would be able to pick out the problems that I see and have them corrected prior to going to print. Next back in the chain is the laboratory that was involved with the scientist; did they check the quality of the work leaving their EM unit? Stepping back again did the scientist take the micrographs or did they receive help? Either way the training of staff and operators should overcome this type of problem! But if the results the staff and visiting operators produce are not assessed how do you know that their training is inadequate?
As the pressure to perform increases and funding decreases only the cream of our laboratories will remain. In industry there is no question about following rigid "Quality" procedures and it is not too far off that this will hit the world's EM units too! I know that this is my baby but is it not about time that we woke up to these facts?
There is an area where I believe we have room for discussion; what do you think?
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob +44 (0) 7711 606967 www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 10:49:04 2004
Steve, Your post (whose spirit I agree with totally) prompted a question. If I have a biological sample sputter coated with metal, isn't all the contrast coming from the metal? Does the underlying carbon based material scatter anything much? If the carbon isn't scattering much then wouldn't the problem you used for illustration matter only at high mags where distances on the order of the coat thickness are being resolved?
As ever, Tobias
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Steve, I agree entirely with you in that training (perhaps educating is a better word) is key to good and reliable results. The example you sited happens constantly. I take great pains to not only lecture about, but prove through lab exercises, the effects that varying microscope parameters have on the final image.
Unfortunately many "trained" people ask to use our facility and are denied because their training was inadequate. They are either retrained so they have the theory to go along with twiddling the knobs or rely on our "service" option (trained staff does the actual imaging). The reputation of our facility is very important.
We cannot guarantee to get perfect results with every research system on the first try but we do our best and learn from our mistakes. At least if we understand our instruments we can concentrate on sample prep to get the best possible final results...not what the investigator thinks is there but what is ACTUALLY there.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 1/7/04 9:42 AM, "Steve Chapman" {protrain-at-emcourses.com} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Hi Listers } } As many of you may know I run a training organisation that travels world } wide spreading the word on SEM, TEM and EDS operation. I also have a deep } interest in "Quality in Electron Microscopy"as those who picked up our paper } last year would know? } } Now to the point. Once again I am picking up respected journals and finding } examples of what I would call poor microscopy, but in truth it also } demonstrates poor quality control! } } To bring one image to mind. The micrograph is of a structure which is } described as being an example of a smooth surface on a biological material. } But, the micrograph was taken at 20kV, where the vast majority of the } information will have come from beneath the surface softening the true } surface detail. } } First to remove the training aspect . Operators of SEM should be taught that } by manipulation of kV and working distance one may subdue or enhance surface } features. To use more than 10kV on most biological samples is asking for } sub surface detail, ignore this and comments on surface irregularities are } null and void in my mind. (I have to say I would probably try to use 2 to } 5kV if the microscope used was produced in the last 15 years!) } } Now the quality aspect. By the time a paper is published a number of steps } should have been taken. Working backwards, the publisher should have the } paper vetted by knowledgeable scientists who would be able to pick out the } problems that I see and have them corrected prior to going to print. Next } back in the chain is the laboratory that was involved with the scientist; } did they check the quality of the work leaving their EM unit? Stepping back } again did the scientist take the micrographs or did they receive help? } Either way the training of staff and operators should overcome this type of } problem! But if the results the staff and visiting operators produce are } not assessed how do you know that their training is inadequate? } } As the pressure to perform increases and funding decreases only the cream of } our laboratories will remain. In industry there is no question about } following rigid "Quality" procedures and it is not too far off that this } will hit the world's EM units too! I know that this is my baby but is it } not about time that we woke up to these facts? } } There is an area where I believe we have room for discussion; what do you } think? } } Steve Chapman } Senior Consultant Protrain } Electron Microscopy Training and Consultancy World Wide } Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob +44 (0) 7711 606967 } www.emcourses.com } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 14:28:28 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Put into TEM at low mag but still with an objective aperture in place, and 100 kV. Really spread the beam and turn up slowly. You may find that by gradually increasing the electron dose, you'll "harden" the film. This approach certainly works with formvar films and biological resin sections but I've never tried it with silicon monoxide, so I can't guarantee it'll work. -- Larry Stoter NOTE - any message other than plain text will be automatically deleted :-)
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 15:12:05 2004
Does anyone have a circuit diagram for an oldish Leitz microscope lamp power supply Type 301-314.001?
It's a beige box with an analog voltmeter, a mains switch, and a brighness adjust knob on the front panel.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 15:24:34 2004
Question: we used a spurr's resin prep on a sample of nematodes, after staining and heat-fixing the sections, we see dark ribbon-like structures on several specimens. Are these artifacts caused by air bubbles and can they be avoided? thank you, jamie
We believe the problem may have been caused by weak film. Silicon monoxide film should be as fresh as possible. Some of our customers have requested thicker coatings but this doesn't help. We make our film up the day it is requested by the customer to insure this problem doesn't occur. We suggest you purchase the minimum amount required at one time because new batches can be made up in a day or so.
John Arnott Disclaimer: Ladd Research produces a wide variety of EM supplies including substrates, such as silicon monoxide.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Larry Stoter" {larry-at-cymru.freewire.co.uk} To: "by way of Ask-A-Microscopist" {alchung-at-ucla.edu} ; {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, January 07, 2004 3:27 PM
Steve, et al.:
I agree there is a definte problem in terms of (some of) the microscopy work being used and published. Steve hits a very good case where failure to understand the technique can easy lead to poor data. (by microscopy I mean in the broadest sense: SEM, TEM, LM, SPM, etc. . . Anyone dealing with molecular biolgists labling molecules and wanting to use a confocal microscope already knowing exactly what kind of "picture" they want to get knows what I mean)
One of the really frightening aspects is that very very few people wish to learn microscopy - become microscopists - today. I have been watching this trend for several years now where users (students and faculty in my case) simple want images. They want to push a button out comes an image - that's all. If they could have the microscope automatically load the sample that would be even better. Now granted there are times when a simple click image will sufice - but more and more often researchers are failing to realize how far they are trying to push the capabilities of a microscopy technique. I have been told specifically they do NOT want to learn the microscope they want an image. And then you try and turn around and tell them the data they are collecting is bad science?
} Next back in the chain is the laboratory that was involved with the } scientist; did they check the quality of the work leaving their EM unit? } Stepping back again did the scientist take the micrographs or did they } receive help? Either way the training of staff and operators should overcome } this type of problem! But if the results the staff and visiting operators } produce are not assessed how do you know that their training is inadequate? }
There is a time you can attempt to argue with the "users" over scienfic quality, but running and EM Lab you can not dictate it - certainly not in academia, and not even in industry - yes, you can be asked for an evaluation of the data (and that is what peer review also does) but no one can really control what the data is used for after it leaves the lab.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Wed Jan 7 16:48:19 2004
We are having one of those debates that we microscopists seem to obsess about. The question is whether to store our saturated uranyl acetate solution (in dH2O) at room temp or at 4 C. Opinions, especially those backed by data, would be welcome. Happy New Year. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
With huge headache I've established procedure in my Lab that everyone, who uses equipment in the Lab should go through mandatory training. Without this training, person just could not enter the facility (digital lock). My training course includes intense training on data collection, interpretation and sample preparation. So, it does not insure from the "bad science", but at least I felt people knows what they doing. It means, if they will present "bad data" I know, they did it on purpose... My course is about week long (2h/day) and people's reaction are very different. Nevertheless, I noticed that majority of the users finally enjoyed "good electron microscopy", because it save them time and the quality of their images is good (and confidence is great). In general, I do agree: people becomes more and more "lazy" - they want to have results doing nothing (best scenario - machine will do). It's very pity and made me very skeptical on quality of many data published. Sergey
P.S. Knowledge is power.
At 02:20 PM 1/7/2004, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
More important - to store in the dark! At +4oC you will have lower concentration because of solubility. I prefer to store most of the chemical solutions at cold temperature. Sergey
At 02:49 PM 1/7/2004, 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 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
If it makes all of you feel any better, the phenomenon of users not wanting to learn how to use something, just wanting the results, is not limited to microscopy, and is a dangerous trend. This trend is ironically aided by the very advancing technologies that make truly understanding the theories and principles of what is happening more important than ever.
Too many devices, instruments and other systems have become "too easy to use". The advances in human interfaces, automation, computer controls, etc. have made it very easy for just about anyone to get results. The danger is that there is no way for someone who doesn't truly understand what's going on to know if the results are meaningful or not. "I got the answer, and it's what I was expecting, so it must be right!"
The problem is also not limited to "sophisticated" technology. We have users who no longer know that there is a darkness adjustment on a copy machine, much less how to use it. Not that long ago, you couldn't get two copies in a row to turn out without tweaking the adjustment. If someone does take the thing off "auto" and sets it dark, everyone else thinks the thing is broken...
If someone finds a "real" cure for this phenomenon, please share it with the world, because the problem is pretty universal.
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 03:26:44 2004
I fully agree with this fact that the problem is universal, and it is not only limited to microscopy. However, before one finds the "cure for this phenomenon", one should look for the cause of this problem.
If the new generation, i.e. the younger students and others, "are just wanting results", is it not the consequence of the ATTITUDE OF THE LARGE MAJORITY of those who trained them?
Those who trained the last two generations are now collecting the results of what they have done. The biggest reference for this generation is having rapid success, which means gaining large amounts of money and if possible a quick fame.
No university professor considers the students for their interest for science and their will to do something for the sake of humanity. They rank them on their good marks, without worrying about how they obtained it. And even if they do so, it is to REPRIMAND the student and not for TEACHING them the importance of honesty. A large percentage of students learn just to obtain a good mark, without thinking about why's and how's. And those few who do, are not appreciated by the professors, may be since they need more time that is not available.
A professorship is the art of teaching many things including highest human values to the students, I can not name many who have done that, or even have thought to do it. If they do not reflect such thoughts, at least they should avoid to discourage the very few who do have such values (and have truly come to learn something that can be helpful for the society) by treating them as naives and dealing with irony with them.
Some students tell me that the science professors are so arrogant that the students have no more appeal for scientific branches. } From my discussions with some undergraduate science students, their feeling is that if they go for science in their graduate school they could ruin their future and they will loose the chances of success!
I agree that the danger is real and from my understanding the solution to the problem is to reverse the trend from now on by changing the teaching philosophy. However, one should have the certitude of the importance of such a training to be able to have an impact on one's student.
Sousan Abolhassani, Ph.D. Laboratory for Materials Behaviour Paul Scherrer Institut 5232 Villigen PSI Switzerland
"John W. Raffensperger, Jr." wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } If it makes all of you feel any better, the phenomenon of users not } wanting to learn how to use something, just wanting the results, is not } limited to microscopy, and is a dangerous trend. This trend is } ironically aided by the very advancing technologies that make truly } understanding the theories and principles of what is happening more } important than ever. } } Too many devices, instruments and other systems have become "too easy to } use". The advances in human interfaces, automation, computer controls, } etc. have made it very easy for just about anyone to get results. The } danger is that there is no way for someone who doesn't truly understand } what's going on to know if the results are meaningful or not. "I got } the answer, and it's what I was expecting, so it must be right!" } } The problem is also not limited to "sophisticated" technology. We have } users who no longer know that there is a darkness adjustment on a copy } machine, much less how to use it. Not that long ago, you couldn't get } two copies in a row to turn out without tweaking the adjustment. If } someone does take the thing off "auto" and sets it dark, everyone else } thinks the thing is broken... } } If someone finds a "real" cure for this phenomenon, please share it with } the world, because the problem is pretty universal. } } John W. Raffensperger, Jr. } IS Manager } Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 04:32:18 2004
Our old camera has to be replaced and I am offered a new one for 1200 euros. This may not be too expensive but I wonder if I can get the same results with a cheaper webcam attached to the microscope. I understand it is just a question of removing the camera objective lense or to make it focus infinity.
My question is: can I expect a really better performance from a more expensive, purpose-built camera? Quite likely the camera will not be the limiting factor in the quality of micrographs, but other factors in the microscope and the preparation...
And, which factors should I cosider in the camera? I am thinking of sensitivity to poor light, gain, and so.
Thanks for all your comments,
Antonio D. Molina-García Inst. del Frio (CSIC) Madrid, Spain
PD. My main purpose for video recording from a microscope is to study ice crystal evolution during growth and recrystallization. Image is so, not too sharp ever, as the contrast between ice crystals is small. Also the sample thickness is larger than when using other sample types and, when the size of crystals is small, it is even difficult to get any light through...
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 04:54:16 2004
Second Announcement From Microscopy to Nanoscopy, Genoa, 9-11 February 2004. The 5th Practical and Intensive Workshop on 3D Confocal Microscopy will be held in Genoa, at LAMBS, Department of Physics, Via Dodecaneso 33, 16146 Genoa. The aim of the workshop, as usual, is to introduce researchers to Confocal Microscopy and related methods. This year we would like to show how it is possible to move from Microscopy to Nanoscopy, according to a Stefan Hells recent paper (Natur.Biotech., 21 (11), 2003, p.1347). 2004 Program includes: lessons on basic aspects of fluorescence and confocal microscopy (February 9th, 10 a.m.- 1 p.m.), lectures on applications of confocal microscopy and related methods (February 9th, 2 p.m.-7 p.m. with coffe break), experimental sessions on confocal and multiphoton architectures, experimental sessions with image analysis, processing, visualization and deconvolution software, roundtables with speakers (February 10th-11th, 8,30 a.m. - 7 p.m., including lunch break). Lecturers: Tony Wilson, Erik Manders, Alberto Diaspro, Mario Faretta, Cesare Usai. The workshop is limited to 20 students served on first-in-first-out basis. Only Monday afternoon lectures are open. The workshop fee is 250 Euros. Some grants will be available on the basis of real need. Please register or request information sending an e-mail to diaspro-at-fisica.unige.it using "confocal 5 - genoa" as subject. Please, specify if you want to attend the Workshop or only Monday afternoon lectures. Logistic help can be provided upon request. Poster can be sent upon request in pdf format. Main sponsor of the Workshop is Nikon Italia, SpA, Firenze. News on the workshop can be found at www.lambs.it, from January 13th 2004. Further sponsorship can be proposed to diaspro-at-fisica.unige.it using as subject "confocal 5 - sponsor". All my best Alberto Diaspro ....................................................................... ................................... .. non basta, resistere, resistere...resistere ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Alberto Diaspro Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480 fax 010314218 diaspro-at-fisica.unige.it, http://www.lambs.it http://wiley.com/cda/product/0,,0471409200,00.html
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 06:43:27 2004
Electron Microscopist/Laboratory Manager Position Department of Materials Science and Engineering Drexel University, Philadelphia, PA 19010
We are seeking a candidate to supervise a microscopy laboratory, which includes an FEI/Phillips XL30 field emission environmental scanning electron microscope with an EDAX-EDS and TSL-OIM, an Amray 1830/D4, and a basic TEM (we outsource most TEM work), optical microscopes, sample preparation facilities, two X-ray diffractometers, etc. We are moving towards a centralized facilities model for the entire university and anticipate the move of many of our laboratories to a new research building by 2005. In addition to instrument maintenance, user scheduling, supervising and training users, the person in this position is expected to participate in the planning and execution of tasks related to the centralized materials testing and characterization facilities and the relocation of the labs to the new building. Candidates should have demonstrated experience with electron microscopy and preferably a degree in Materials, Physics or Biology. Salary will be commensurate with qualifications and experience.
The successful candidate will join a technical staff of three within the department. Detailed information about the department can be found at http://www.materials.drexel.edu, a copy of our recent newsletter can be found at http://www.materials.drexel.edu/Newsletter/fall_03.pdf and our last annual report at http://www.materials.drexel.edu/annualreports/Annual%20Report%202002.pdf. Candidates interested in this position should please submit a CV and a short career plan to: Microscopy Hiring Committee, Drexel University, Department of Materials Science and Engineering, 3141 Chestnut Street, Philadelphia, PA 19104.
Dr. Richard Knight -- "And the men who hold high places, must be the ones to start..."
Richard Knight, Ph.D., FASM, Auxiliary Professor, Center for the Plasma Processing of Materials [CPPM], Drexel University, Dept. of Materials Science & Engineering, 3141 Chestnut Streets, Philadelphia, PA 19104
A most interesting thread, and one that is near and dear to my heart.......
I frequently get people new to SEMs, either through personnel changes at a customer's site or through someone's acquisition of a "new" (used) SEM. Half of my business is still servicing ETEC Autoscans and as the new systems get more and more automated, I get more and more comments on how user unfriendly the Autoscan is. My standard reply is that it is not at all unfriendly if you understand how an SEM works.
I always start a new user out by building an SEM on paper and guess what? A scanning electron microscope is not a microscope at all! The only image formed in its optics is a DEmagnified image of the tip of the filament! So, what is it? It's a signal generator with one or more signal processors attached. What I really love about SEMs, especially with an EDS attached, is that you can tell me what you want to see and I can probably show it to you. Then we have to sit down and have a long talk about what is real. Don't tell me, "the computer said...". The computer doesn't know squat. And I haven't even gotten to specimen preparation!
The problem goes beyond the need for instant gratification, but may be related to the definition of success. People aren't interested in becoming microscopists because organizations no longer hire microscopists. One must pay too much for the knowledge and experience. Most are looking for "machine operators" that can be transferred between different pieces of equipment at will or they simply equate a microscope with a copier or personal computer. It's merely another tool to be used in getting the job done. We no longer have someone designated as the "copier specialist" in the office and having a degree in computer science is not a prerequisite for operating a computer on your desktop.
In part, there is simply so much more to know about any piece of equipment or area of interest, and the areas of interest and expertise are so intertwined, that one cannot be an expert in all areas, and in part it is the "Walmartization" of the world. The dive to the bottom, the least common denominator, the lowest cost, the lowest price. Of course, if everyone worked for Walmart, no one could afford to do much shopping............
I don't have any answers. I've always liked to understand any equipment that I use, whether it's a microscope or an automobile or a dishwasher, but then, I repair things for a living. I make a living this way because other people have different interests and drives (and that often does NOT include an interest or ability to repair things). A hundred years ago, most people who owned an automobile also had a mechanic in their employ, and the mechanic often traveled with, or even drove, the car.
I believe our problem in science will also get worked out. It's probably going to take some disaster related to bad science to get people's attention, but in the end, some kind of accommodation will be worked out. It will be interesting to watch it unfold.
In the mean time, we can all keep trying to spread our knowledge and our concerns. We subscribe to this listserver because we care. Keep caring!
Ken Converse owner Quality Images third party SEM service Delta, PA
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 07:15:40 2004
I would be careful with using a webcam for professional microscopy. Indeed, you can adjust a webcam this way that it can be attached to a microscope (remove the lens and put the ccd in the focuspoint of the lightbeam), but most webcams are very limited. Limited in shutterspeed, colorrange and fo sure in pixelresolution. Webcams are often used for amateur-microscopy and astronomy and give nice results, but if you really want to start analysing the (live) images or use them for publication, I'm afraid you might get dissapointed. Also: more expensive, professional camera's will last longer ,give you nicer resultes and a better support after sale will be offered. Best regards,
Sven Terclavers
On 08-01-2004 23:33, "Antonio Molina" {ifrm111-at-if.csic.es} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Our old camera has to be replaced and I am offered a new one for 1200 euros. } This may not be too expensive but I wonder if I can get the same results } with a cheaper webcam attached to the microscope. I understand it is just a } question of removing the camera objective lense or to make it focus } infinity. } } My question is: can I expect a really better performance from a more } expensive, purpose-built camera? Quite likely the camera will not be the } limiting factor in the quality of micrographs, but other factors in the } microscope and the preparation... } } And, which factors should I cosider in the camera? I am thinking of } sensitivity to poor light, gain, and so. } } Thanks for all your comments, } } Antonio D. Molina-García } Inst. del Frio (CSIC) Madrid, Spain } } PD. My main purpose for video recording from a microscope is to study ice } crystal evolution during growth and recrystallization. Image is so, not too } sharp ever, as the contrast between ice crystals is small. Also the sample } thickness is larger than when using other sample types and, when the size } of } crystals is small, it is even difficult to get any light through... } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 07:31:01 2004
I very much agree with Sousan about what drives students and their reward systems. However, consider the fact that universities often reflect what is going on in private industry. I would bet that many researchers, scientists and engineers report to people that have scant knowledge of the science they ostensibly oversee. I call this the "MBA Phenomenon." I've observed this trend not only in microscopy. How often must someone in the lab. explain to their "management" exactly what an image is telling them? Quality should start with quality management and that means the management ought to know what they are managing.
Peter Tomic Agere Systems
Disclaimer: I am happy to report that the above conditions do not apply to me at the moment.
-----Original Message----- } From: Sousan Abolhassani [mailto:sousan.abolhassani-at-psi.ch] Sent: Thursday, January 08, 2004 4:29 AM To: John W. Raffensperger, Jr. Cc: microscopy-at-MSA.Microscopy.com
I fully agree with this fact that the problem is universal, and it is not only limited to microscopy. However, before one finds the "cure for this phenomenon", one should look for the cause of this problem.
If the new generation, i.e. the younger students and others, "are just wanting results", is it not the consequence of the ATTITUDE OF THE LARGE MAJORITY of those who trained them?
Those who trained the last two generations are now collecting the results of what they have done. The biggest reference for this generation is having rapid success, which means gaining large amounts of money and if possible a quick fame.
No university professor considers the students for their interest for science and their will to do something for the sake of humanity. They rank them on their good marks, without worrying about how they obtained it. And even if they do so, it is to REPRIMAND the student and not for TEACHING them the importance of honesty. A large percentage of students learn just to obtain a good mark, without thinking about why's and how's. And those few who do, are not appreciated by the professors, may be since they need more time that is not available.
A professorship is the art of teaching many things including highest human values to the students, I can not name many who have done that, or even have thought to do it. If they do not reflect such thoughts, at least they should avoid to discourage the very few who do have such values (and have truly come to learn something that can be helpful for the society) by treating them as naives and dealing with irony with them.
Some students tell me that the science professors are so arrogant that the students have no more appeal for scientific branches. } From my discussions with some undergraduate science students, their feeling is that if they go for science in their graduate school they could ruin their future and they will loose the chances of success!
I agree that the danger is real and from my understanding the solution to the problem is to reverse the trend from now on by changing the teaching philosophy. However, one should have the certitude of the importance of such a training to be able to have an impact on one's student.
Sousan Abolhassani, Ph.D. Laboratory for Materials Behaviour Paul Scherrer Institut 5232 Villigen PSI Switzerland
"John W. Raffensperger, Jr." wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } If it makes all of you feel any better, the phenomenon of users not } wanting to learn how to use something, just wanting the results, is not } limited to microscopy, and is a dangerous trend. This trend is } ironically aided by the very advancing technologies that make truly } understanding the theories and principles of what is happening more } important than ever. } } Too many devices, instruments and other systems have become "too easy to } use". The advances in human interfaces, automation, computer controls, } etc. have made it very easy for just about anyone to get results. The } danger is that there is no way for someone who doesn't truly understand } what's going on to know if the results are meaningful or not. "I got } the answer, and it's what I was expecting, so it must be right!" } } The problem is also not limited to "sophisticated" technology. We have } users who no longer know that there is a darkness adjustment on a copy } machine, much less how to use it. Not that long ago, you couldn't get } two copies in a row to turn out without tweaking the adjustment. If } someone does take the thing off "auto" and sets it dark, everyone else } thinks the thing is broken... } } If someone finds a "real" cure for this phenomenon, please share it with } the world, because the problem is pretty universal. } } John W. Raffensperger, Jr. } IS Manager } Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 08:13:05 2004
----- Original Message ----- } From: jamielange-at-wi.rr.com (by way of MicroscopyListServer)
The power of semantics
I am convinced that one of the reasons for a decline in the quality of microscopy, especially in industry, is semantic. Over the last few years it has become fashionable for managers to refer to instruments as "tools". They no longer distinguish between different kinds of equipment - they are all tools. Tools include: lathes, electron microscopes, fork lift trucks, x-ray diffraction units,.....
When a person using a tool leaves or is promoted, you need another person to operate the tool. So you take a person from this tool and put them on to that tool. Give them a couple of hours to read the manual and away you go.
I can only assume that machinists are complaining about the decline in the quality of work done in machine shops. All the ex-SEM operators must be doing a lousy job of operating the milling machines.
-- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 08:39:34 2004
In a message dated 1/8/04 9:28:40 AM, jae5-at-lehigh.edu writes:
} I can only assume that machinists are complaining about the decline in } } the quality of work done in machine shops. All the ex-SEM operators } must be doing a lousy job of operating the milling machines.
Alwyn, I don't think the analogy holds. Machine tools have gotten "smarter" in recent years. Numerical control and feedback devices have made it possible for information to flow directly from the design computers and CAD programs (which in turn have replaced the traditional draftsman) to control the machining process. In some cases the "operators" aren't even present, or just keep watch over a large array of machines in case of malfunctions or to load raw material. As the microscopes have evolved, they have for the most part not become easier to operate. Oh sure, some "little" things like adjusting astigmatism, saturating filaments, even focusing, have been automated. But not the "big" things like taking a meaningful picture. In fact, it has arguably become more complicated to use the modern microscopes because they offer a much broader range of possibilities than the old ones did. More imaging modes, more types of detectors, etc., create a greater demand for insight and knowledge on the part of the operator.
John Russ (visit www.DrJohnRuss.com for a schedule of upcoming image analysis workshops and other information)
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 08:50:32 2004
Mike Delannoy asked whether there is an easier way to make holey carbon films than Formvar, steam, create the holes in the damaged Formvar film, carbon coat the holey Formvar and dissolve the Formvar away to leave the holey carbon film. If there is, we'd like to know about it! This is the easiest method about which we know.
I must stress, however, that making holey and lacey carbon coated grids is pure art. People who have decades of experience making support films still have difficulty at times with the reproducibility of the method, and the details are entirely a matter of getting the "feel" of the method. Details of our methods are described on
http://www.2spi.com/catalog/grids/cusctgrd.html
and the pages linked to it.
An easier way to obtain holey carbon support films with a known and precise distribution of holes is the Quantifoil Micromachined Holey Carbon Grids, described on
Disclaimer: SPI Supplies has a very active business making coated grids for customers throughout the world. We also sell grids and other supplies for customers who prefer to coat their own grids, and we sell the Quantifoil Holey Carbon Grids.
Andy
Andrew W. Blackwood, Ph.D. Vice President, Technical SPI Supplies P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 X108 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:25:16 2004
While I see your point, it is much easier to at produce something with today's "tools", even your "tools". We can now use PhotoShop or some other application, load up some Image Tool filters and away you go with image analysis. My first IA package was very cumbersone and it took some major training just to load the images and navigate through the software. Presumably, you would pick some understanding of the subject matter through osmosis, solid state diffusion or entropy while learning how to operate the software. Now the software is so easy, anyone can sit down and after a few minutes start cranking out numbers.... for as meaningful or meaningless as they may be.
With SEMs, my first one filled a room and producing a photo was a real chore. You had to have an intimate knowledge of the controls and operating conditions to get even a poor quality photo. Now you can train a chipmunk (borrowed from one of my associates) to run an SEM. We have seen PhD's operate the SEM as a machine and have absolutely zero fundamental knowledge of the images or EDS data. I am thankful that I can sit down at my SEM and get a great photo in 5 minutes, but that means anyone else can too.
Al Stone ASTON
ps. no offense to anyone with a PhD, the point is that just because you can drive a car doesn't mean you understand the rules of the road or know how to travel cross country
At 08:42 AM 1/8/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:25:52 2004
My favorite quote fits well in this discussion: "It is not enough to believe what you see. You must also understand what you see. - Leonardo da Vinci"
I think it has to do a bit with what John just brought up in regards to the evolution of the microscopes. They have become "easier" to operate, and in doing so (computer control, mouse operated saturation...) there is a great disconnect with how the 'system' works. Users learning on an old EM300 for example have more interactions with the parameters on the microscope and thus are in a better position to 'understand' what they are doing in the process of collecting an image.
It is difficult to stress integrity, understanding when teaching. Even harder is the problem educators have in providing evaluation of the process, esp when there is a definitive end product (a lab report with images for example). I would much prefer to be able to grade students on their process than on the end results, and to provide an objective progress evaluation that translates onto a final grade. The biggest problem is that each student is different, has different learning speeds, different ways of coming to the same end point. So many variables to deal with. I find that the most effective and simple question I can present to students and faculty using the facility is "How do you know [ That ] is what you are looking at?" And sometimes it becomes a significant frustration level when the individual cannot convince me. But when they CAN convince me they wind up becoming much more confident and it forces them to find supporting evidence that they were too lazy or busy to look for earlier, and then they (more often than not) are much better at looking (literature sourcing) BEFORE starting their next project.
Well it is a new year and for some of us a new Semester. Time to start doing what ever we can do to reverse the trend!
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] } Sent: Thursday, January 08, 2004 9:42 AM } To: jae5-at-lehigh.edu; microscopy-at-msa.microscopy.com } Subject: [Microscopy] Re: Re: Knowledge and Quality } } } } ------------------------------------------------------------------------ -- } ---- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ -- } ----- } } } In a message dated 1/8/04 9:28:40 AM, jae5-at-lehigh.edu writes: } } } I can only assume that machinists are complaining about the decline in } } } } the quality of work done in machine shops. All the ex-SEM operators } } must be doing a lousy job of operating the milling machines. } } Alwyn, I don't think the analogy holds. Machine tools have gotten } "smarter" } in recent years. Numerical control and feedback devices have made it } possible } for information to flow directly from the design computers and CAD } programs } (which in turn have replaced the traditional draftsman) to control the } machining } process. In some cases the "operators" aren't even present, or just keep } watch } over a large array of machines in case of malfunctions or to load raw } material. As the microscopes have evolved, they have for the most part not } become } easier to operate. Oh sure, some "little" things like adjusting } astigmatism, } saturating filaments, even focusing, have been automated. But not the } "big" things } like taking a meaningful picture. In fact, it has arguably become more } complicated to use the modern microscopes because they offer a much } broader range of } possibilities than the old ones did. More imaging modes, more types of } detectors, etc., create a greater demand for insight and knowledge on the } part of } the operator. } } John Russ } (visit www.DrJohnRuss.com for a schedule of upcoming image analysis } workshops } and other information)
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:28:16 2004
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Thursday, January 08, 2004 8:42 AM
In a message dated 1/8/04 9:28:40 AM, jae5-at-lehigh.edu writes:
} I can only assume that machinists are complaining about the decline in } } the quality of work done in machine shops. All the ex-SEM operators } must be doing a lousy job of operating the milling machines.
Alwyn, I don't think the analogy holds. Machine tools have gotten "smarter" in recent years.
-Reply:
I agree with Alwyn. This is the exact phenomenon I was referring to, and Machine Tools are one of the specific examples I had in mind with my original comments to this thread.
As you stated, there are shops where those involved truly understand what's going on, and things work very well.
In other shops, while these "tools" have indeed become "smarter", the quality of the output has decreased. This is because, as another post stated, these newer machines are viewed as a "tool", and an "operator" is assigned. Because of the intelligence of the machine, the "operator" who is no longer a "machinist" can get a result. It is often not an optimal result, either in terms of quality, and/or in terms of production rate. But a result was obtained. Compounding this, management has seen this situation "evolve" gradually, so they don't realize how much the "lower priced" help is actually costing them vs. a "real" machinist.
Add CAM instructions coming from an engineer who has never even operated a machine, and things get even more fun. Ask me about the $100,000+ damage one of these "wonder kids" did when they crashed a brand new machining center. The simulation ran perfectly. Too bad the engineer forgot that something had to hold the work piece, and this something was 2" think...
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:36:59 2004
We have a Zeiss 109 TEM available. Scope includes the unique SEM attachment (gives SEM capability with two SEM stages that fit standard Zeiss pin mounts). Turbo pumped; extra gun assembly, filaments, water chiller, manuals, and original orange paint on column included. Has been under full Zeiss/Leo service contract and in use until last month.
Needs to be removed as soon as possible to make room for a new scope.
Preference given to academic institutions. If you want it, you need to make arrangements for shipping, or pick it up in New London, CT. Contact me offline for further information.
Page Owen
************************ T. Page Owen, Jr., Chair Department of Botany Connecticut College New London, CT 06320 860-439-2147 tpowe-at-conncoll.edu ************************
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:46:30 2004
Without a picture of what you are seeing advice is difficult. My guess is that the section is not flat on the slide and water/stain is getting underneath a wrinkle in the section and the result is the "ribbon" you are seeing. Suggestions to solve the problem:
1. A sharp knife. If you are using glass knives use only knives made "fresh". Glass, being a super-cooled liquid, flows and older knives are less sharp. Try it! 2. De-wrinkle the sections with 1,2 dicholorethane or chloroform. 3. Float the sections on a larger drop of water on a hot plate. The larger drop will take longer to evaporate and give the section more time to be expanded/dewrinkled by heat. 4 Thinner sections? I don't know how thick your sections are but 1-2 microns is the range to shot for.
Geoff
by way of MicroscopyListServer wrote:
} Email: jamielange-at-wi.rr.com } Name: jamie lange } } Organization: university of wisconsin } } Education: Graduate College } } Location: milwaukee, wi. usa } } Question: we used a spurr's resin prep on a sample of nematodes, after } staining and heat-fixing the sections, we see dark ribbon-like } structures on several specimens. Are these artifacts caused by air } bubbles and can they be avoided? thank you, } jamie } } --------------------------------------------------------------------------- } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 09:52:27 2004
First of all EVERYONE must realise that the image that they see (even though they pressed the SE or SEI button) is a mixed SE+BSE image. This means surface detail from the SE and sub surface detail from the BSE. As you put the kV up the BSE dominates more and of course as you take the kV down the BSE are reduced allowing the SE to dominate. I should point out that you will often see BSE information even at 2kV or less; this being the result of the SE3 contribution.
Place a thin coating on the surface of a specimen and you increase the coefficient of emission, the metal being the SE emitter rather than the original biological material. You do image the specimen surface as that topography has been followed by the coating procedure. However the BSE come from a far greater depth below the surface and at any kV, under the wrong circumstances of WD and spot size, these electrons will contribute to the image. Should there be structure of differing density, by way of the BSE, this will show as "shadows" in the background or may even dominate the image.
In a field emission instrument, of the type where the above lens detector is available, it is possible to screen out the BSE contribution and display a pretty pure SE image. I have to say in my 40 years in the business we have more often gone for "information" rather than "resolution" therefore I do try to include a degree of BSE to add contrast to the image.
Hope this helps?
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "Tobias Baskin" {baskin-at-bio.umass.edu} To: "Steve Chapman" {protrain-at-emcourses.com} ; {microscopy-at-MSA.microscopy.com} Sent: Wednesday, January 07, 2004 4:51 PM
It appears we have a change in how quality is monitored over the years of SEM evolution. As may be the case where older SEMs require understanding of how a SEM works to get an image, to the extent that newer SEMs do not require this, those operating newer SEMs may not require the prior extent of knowledge to produce images. Where once the decision to take images were made by those who knew enough to understand and self monitor quality, that appears not to be the case anymore. Advances in technology have allowed images to be taken by operators who do not understand what they are looking at; one can not expect an unknowing individual to monitor quality.
We can address the quality issue with education or attitude (as proposed by others on this board). This is a head on approach; a less direct method would be to change the decision rights or incentives. For example, operators should not be given the decision right of what to take an image of. For example, operators should not be rewarded simply on the number of images taken.
The point is that monitoring of quality has changed, so we should consider changing decision rights and incentives/rewards. If these three components of organizational architecture are not in balance, results will be unsatisfactory.
Sincerely, John Moore Montara Industries 919-434-8457
Disclaimers: 1) This framework is described in a book titled "Managerial Economics and Organizational Architecture", a text that I studied while obtaining my MBA at the University of Rochester, Class of 2000.
2) In defense of my education and this technique, I cite first that the poster from Agere on this topic reported not suffering certain difficulties, and second that in my recent job search I found Agere to be hiring MBA's (see monster.com directemployers.com, hotjobs.com or flipdog.com).
3) I am not a SEM expert, nor do I have direct experience in this field. I began following this message board in the interest of selling a defect review tool that I made a speculative investment in: a SEMVision by Applied Materials.
----- Original Message ----- } From: "Sousan Abolhassani" {sousan.abolhassani-at-psi.ch} To: "John W. Raffensperger, Jr." {johnr-at-helwigcp.com} Cc: {microscopy-at-MSA.Microscopy.com} Sent: Thursday, January 08, 2004 4:28 AM
I fully agree with this fact that the problem is universal, and it is not only limited to microscopy. However, before one finds the "cure for this phenomenon", one should look for the cause of this problem.
If the new generation, i.e. the younger students and others, "are just wanting results", is it not the consequence of the ATTITUDE OF THE LARGE MAJORITY of those who trained them?
Those who trained the last two generations are now collecting the results of what they have done. The biggest reference for this generation is having rapid success, which means gaining large amounts of money and if possible a quick fame.
No university professor considers the students for their interest for science and their will to do something for the sake of humanity. They rank them on their good marks, without worrying about how they obtained it. And even if they do so, it is to REPRIMAND the student and not for TEACHING them the importance of honesty. A large percentage of students learn just to obtain a good mark, without thinking about why's and how's. And those few who do, are not appreciated by the professors, may be since they need more time that is not available.
A professorship is the art of teaching many things including highest human values to the students, I can not name many who have done that, or even have thought to do it. If they do not reflect such thoughts, at least they should avoid to discourage the very few who do have such values (and have truly come to learn something that can be helpful for the society) by treating them as naives and dealing with irony with them.
Some students tell me that the science professors are so arrogant that the students have no more appeal for scientific branches. } From my discussions with some undergraduate science students, their feeling is that if they go for science in their graduate school they could ruin their future and they will loose the chances of success!
I agree that the danger is real and from my understanding the solution to the problem is to reverse the trend from now on by changing the teaching philosophy. However, one should have the certitude of the importance of such a training to be able to have an impact on one's student.
Sousan Abolhassani, Ph.D. Laboratory for Materials Behaviour Paul Scherrer Institut 5232 Villigen PSI Switzerland
"John W. Raffensperger, Jr." wrote: } } -------------------------------------------------------------------------- ---- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------------- ----- } } If it makes all of you feel any better, the phenomenon of users not } wanting to learn how to use something, just wanting the results, is not } limited to microscopy, and is a dangerous trend. This trend is } ironically aided by the very advancing technologies that make truly } understanding the theories and principles of what is happening more } important than ever. } } Too many devices, instruments and other systems have become "too easy to } use". The advances in human interfaces, automation, computer controls, } etc. have made it very easy for just about anyone to get results. The } danger is that there is no way for someone who doesn't truly understand } what's going on to know if the results are meaningful or not. "I got } the answer, and it's what I was expecting, so it must be right!" } } The problem is also not limited to "sophisticated" technology. We have } users who no longer know that there is a darkness adjustment on a copy } machine, much less how to use it. Not that long ago, you couldn't get } two copies in a row to turn out without tweaking the adjustment. If } someone does take the thing off "auto" and sets it dark, everyone else } thinks the thing is broken... } } If someone finds a "real" cure for this phenomenon, please share it with } the world, because the problem is pretty universal. } } John W. Raffensperger, Jr. } IS Manager } Helwig Carbon Products, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 10:39:09 2004
To Joel, Eckhart, Kenneth, Peter, and anyone else who gave me advice-
OK, I finally had a chance to sit down and fiddle with the chiller, and here is what I have.
I turned on my Zeiss EM 10CA and checked the flowmeter in the column: about 2.2 L/min, if I am reading it right. There was no gauge for the chiller flow rate, but there is a pressure gauge on the front of the chiller, and it read about 39 psi. This reading did not change through the entire test, and neither did the EM flowmeter.
Chiller temperature reading was about 75 degrees F at start. It needs to be 68 degrees F, according to the EM's manual. Only the "Cool" light was on...all others, including the "Compressor" light, were off.
I turned the temperature screw to the left to lower the temp setting. No response from the compressor. Turned it down a second time....still no response. After about 5 minutes the Compressor light turned on, and the temperature started to drop, finally. The temperature went all the way down to 53 degrees F, at which point the "Lo Temp" light went on, the "Compressor" light went off, and the "Heat" light went on. I turned up the temp screw a little bit.
At about 60 degrees F, the "Heat" and "Lo Temp" lights went off and the "Cool" and "Pump" lights are on (actually the "Pump" light never turned off). It rose to 68 degrees F....and continued to rise. I let it get to about 79 degrees F, and the compressor never turned back on. I tried turning the screw back to the left to drop the temp setting...it never went back on.
At this point, I decided to turn the EM 'scope off....though the flow rate never dropped, and the buzzer never went off. The temperature gauge on the chiller continued to rise, even after turning the 'scope off. It was reading about 90 degrees F when I left to come write all of you.
Just now went back in to check the thing...the compressor finally came on...it's reading about 53 degrees F and the "Lo Temp" light is on, and the "Compressor" light is off.
So I think it's the switch....or perhaps the "Hi Temp" sensor? Or both? I have no idea how all the sensors and switches are connected...even with the diagrams and manual that were sent, as I have no idea how to read such things (been eons since I did circuits in Physics class...). What do you all think?
Thank you all so much for your help- Kathleen Neurotoxicology Labs Rutgers University
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 12:23:55 2004
I've been reading the iterations on this thread and it seems to me that it would be useful to separate the chiller from the TEM to test the chiller. Kathleen, if you have a carbon evaporator that has a diffusion pump you can cool it with the Coolwell long enough to tell if the Coolwell has a problem. If the Coolwell tests okay on another heat load (the carbon evaporator) then your problem is in the TEM.
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
At 11:48 AM 1/8/2004 -0500, Kathleen Roberts wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 13:00:36 2004
Well some interesting stuff, I was particularly taken by Peter Tomic's comments
"Quality should start with quality management and that means the management ought to know what they are managing."
Via MSA and mostly privately I have had mail from, judging by the mailers enthusiasm, pretty good EM units. However I have probably visited more EM units in the world than most of you and I have to say in very many cases they are just not up to the standards that I would set. Here Peter's comment hits home to me! You see, as an example, in most units people were trained when the SEM, TEM or EDS system first arrived in the lab. Now they have new equipment and I bet they have rarely been trained to operate using the techniques opened up by the new SEM or TEM? Once trained you know how to use them is what I see. Guess what, a customer born on a Cambridge 100 SEM still used their new sexy Field Emission SEM in just the same way. A thick gold coating and 20kV plus was the way to do it, quote "we have always done it this way". We produced amazing results uncoated at 150,000X at 1.5kV! Do I make my point? This was a very highly rated government research laboratory that by perception would be rated as in the top 5 in a country very well respected for its EM work!
We all believe we run a good unit, my point would be "how do you know?" Do you have management procedures that are up to date, do you routinely check the performance of your staff, do you look at the output your users produce, etc etc? Take a look at www.iaaem.com/mppa1 here you will find a quality procedure that has been proposed by a group of scientists and Quality experts from across the world. Answer the questions and total up the points for your laboratory and lets see how far you area away from Total Quality Management. What would you suggest is included and maybe together we will be able to develop procedures that would help sort out what you all, almost, seem to agree is a bit of a mess!
I am working overseas (Egypt) for the next week so have fun on your own for a little while.
Regards
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 13:44:05 2004
I want to give away the following to a university or non-profit lab or group:
Plexiglass racks for holding 3.25" x 4" (TEM) and 4" x 5" (Polaroid) negative film; and hard rubber tanks for processing negatives.
Interested parties should contact me off-line. Commercial/industrial users need not respond.
Thanks,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 14:04:04 2004
Good idea, except that I don't have a carbon evaporator. :o)
Thanks anyway, Kathleen Neurotox Labs Rutgers University
Owen P. Mills wrote:
} I've been reading the iterations on this thread and it seems to me } that it would be useful to separate the chiller from the TEM to test } the chiller. Kathleen, if you have a carbon evaporator that has a } diffusion pump you can cool it with the Coolwell long enough to tell } if the Coolwell has a problem. If the Coolwell tests okay on another } heat load (the carbon evaporator) then your problem is in the TEM. } } Owen } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills } } At 11:48 AM 1/8/2004 -0500, Kathleen Roberts wrote: } } } } ------------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } } } To Joel, Eckhart, Kenneth, Peter, and anyone else who gave me advice- } } } } OK, I finally had a chance to sit down and fiddle with the chiller, } } and here is what I have. } } } } I turned on my Zeiss EM 10CA and checked the flowmeter in the column: } } about 2.2 L/min, if I am reading it right. There was no gauge for } } the chiller flow rate, but there is a pressure gauge on the front of } } the chiller, and it read about 39 psi. This reading did not change } } through the entire test, and neither did the EM flowmeter. } } Chiller temperature reading was about 75 degrees F at start. It } } needs to be 68 degrees F, according to the EM's manual. Only the } } "Cool" light was on...all others, including the "Compressor" light, } } were off. } } } } I turned the temperature screw to the left to lower the temp setting. } } No response from the compressor. Turned it down a second } } time....still no response. After about 5 minutes the Compressor } } light turned on, and the temperature started to drop, finally. The } } temperature went all the way down to 53 degrees F, at which point the } } "Lo Temp" light went on, the "Compressor" light went off, and the } } "Heat" light went on. I turned up the temp screw a little bit. } } } } At about 60 degrees F, the "Heat" and "Lo Temp" lights went off and } } the "Cool" and "Pump" lights are on (actually the "Pump" light never } } turned off). It rose to 68 degrees F....and continued to rise. I } } let it get to about 79 degrees F, and the compressor never turned } } back on. I tried turning the screw back to the left to drop the temp } } setting...it never went back on. } } } } At this point, I decided to turn the EM 'scope off....though the flow } } rate never dropped, and the buzzer never went off. The temperature } } gauge on the chiller continued to rise, even after turning the 'scope } } off. It was reading about 90 degrees F when I left to come write all } } of you. } } } } Just now went back in to check the thing...the compressor finally } } came on...it's reading about 53 degrees F and the "Lo Temp" light is } } on, and the "Compressor" light is off. } } So I think it's the switch....or perhaps the "Hi Temp" sensor? Or } } both? I have no idea how all the sensors and switches are } } connected...even with the diagrams and manual that were sent, as I } } have no idea how to read such things (been eons since I did circuits } } in Physics class...). What do you all think? } } } } Thank you all so much for your help- } } Kathleen } } Neurotoxicology Labs } } Rutgers University } } } } } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 14:12:46 2004
In my experience, CCTV cameras as used in security applications work better than webcams, but the resolution of both is not that great and you may spend considerable time on the mechanical aspects of the interfacing and be disappointed with the result.
1200 euros may be well worth it for something that you can just unpack from the box and be using in a few minutes.
cheers
rtch
Send reply to: "Antonio Molina" {ifrm111-at-if.csic.es} } From: "Antonio Molina" {ifrm111-at-if.csic.es} To: {Microscopy-at-MSA.Microscopy.Com}
Most people store UA in the refrigerator perhaps without understanding why. UA is photosensitive and degraded by light (especially fluorescent lighting that contains UV). If you store aqueous solutions in the light they will eventually precipitate --first long the sides of the container and then on the bottom. I have no data, just based on observation.
} We are having one of those debates that we microscopists seem to } obsess about. The question is whether to store our saturated uranyl } acetate solution (in dH2O) at room temp or at 4 C. Opinions, } especially those backed by data, would be welcome. Happy New Year. } Tom } } } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 14:31:49 2004
Thank you for the honorable mention. If my boss is on this listserver I'll have to polish up the old resume. It's so much fun to be quoted. I feel like Hemingway.
Once again, I don't have ANY of these issues HERE.
I needed a good laugh today.
Peter Tomic Unknown Corporation, Inc. Anytown, USA
-----Original Message----- } From: Steve Chapman [mailto:protrain-at-emcourses.com] Sent: Thursday, January 08, 2004 1:59 PM To: MSA
Hi Listers
Well some interesting stuff, I was particularly taken by Peter Tomic's comments
"Quality should start with quality management and that means the management ought to know what they are managing."
Via MSA and mostly privately I have had mail from, judging by the mailers enthusiasm, pretty good EM units. However I have probably visited more EM units in the world than most of you and I have to say in very many cases they are just not up to the standards that I would set. Here Peter's comment hits home to me! You see, as an example, in most units people were trained when the SEM, TEM or EDS system first arrived in the lab. Now they have new equipment and I bet they have rarely been trained to operate using the techniques opened up by the new SEM or TEM? Once trained you know how to use them is what I see. Guess what, a customer born on a Cambridge 100 SEM still used their new sexy Field Emission SEM in just the same way. A thick gold coating and 20kV plus was the way to do it, quote "we have always done it this way". We produced amazing results uncoated at 150,000X at 1.5kV! Do I make my point? This was a very highly rated government research laboratory that by perception would be rated as in the top 5 in a country very well respected for its EM work!
We all believe we run a good unit, my point would be "how do you know?" Do you have management procedures that are up to date, do you routinely check the performance of your staff, do you look at the output your users produce, etc etc? Take a look at www.iaaem.com/mppa1 here you will find a quality procedure that has been proposed by a group of scientists and Quality experts from across the world. Answer the questions and total up the points for your laboratory and lets see how far you area away from Total Quality Management. What would you suggest is included and maybe together we will be able to develop procedures that would help sort out what you all, almost, seem to agree is a bit of a mess!
I am working overseas (Egypt) for the next week so have fun on your own for a little while.
Regards
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 14:48:50 2004
Yes, alas, we have also seen the "take the picture and run" mentality here as well.
Like most microscopists who came up through the system, most of us eagerly learned everything related to microscopy (specimen prep, knife making, ultramicrotomy, alignment, optimization of the scope, darkroom, image interpretation, etc). It was fun and we enjoyed learning and making discoveries. Now, it seems the thrill is gone and the object is to get into the job market and make big bucks....
MONEY is the motivator and it can be used to influence people. For example, I point out that microscopy is a marketable skill and I prove it by giving the names of our former students (in biological and physical sciences) who got jobs in their discipline since they could do microscopy. An employer will generally hire the individual with the better set of skills. Unless they are incredibly naive or just plain stupid (in which case you wouldn't want them using your equipment anyway) they will realize the value in learning the discipline.
More immediately, I point out that it is CHEAPER for them to do their own microscopy than to hire it out.
-- ############################################################## 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 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 15:10:24 2004
I was wondering if someone knows where I can find an monochromator for spectral scanning 400-700 nm that can be attached to a Zeiss microscope (Axioskop) for transmitted light imaging ( scanning microphotometry).
On Jan 8, 2004, at 6:42 AM, DrJohnRuss-at-aol.com wrote:
} Alwyn, I don't think the analogy holds. Machine tools have gotten } "smarter" } in recent years. Numerical control and feedback devices have made it } possible } for information to flow directly from the design computers and CAD } programs } (which in turn have replaced the traditional draftsman) to control the } machining } process. In some cases the "operators" aren't even present, or just } keep watch } over a large array of machines in case of malfunctions or to load raw } material. Dear John, So now the machine tools are capable of performing as well as the automated tomography packages we use? In our case, we have to watch the progress of the program carefully to see that the auto-tracking, auto-focussing, etc. is working properly, and lately we have also found that the file made from the tilt series has values that do not match the exposure we set (to the extent that some of the images were blank). The take-home lesson is that there still must be knowledgeable oversight, especially with regard to automated processes, to assure the quality of the results. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 15:14:33 2004
A lab user has asked me to find some place he can get freeze fracture work done. Close to San Francisco bay area would be best. Reply to me and I will pass on the contact info.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 15:34:44 2004
Dear Kathleen, I have had good luck getting a local HVAC (Heating, Ventilation and Air conditioning) or Air Conditioning repair shop to fix my Haskris water chillers. Haskris are good but they do occasionally break down. Sounds like one of your sensors or accuators is stuck. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Kathleen Roberts" {kgrobert-at-rci.rutgers.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 08, 2004 8:48 AM
Thank you for the advice...I figure $10 isn't too much to spend to try and fix this thing....if the repair cost starts to escalate, we'll junk it and call Haskris. :o)
Kathleen Neurotoxicology Labs Rutgers University
Webster, Paul wrote:
} Hi Kathleen, } } A word of warning if you plan to try to fix the chiller. We spent nearly $7,000 in a year of repairing our Coolwell chiller before it finally failed for good and we had to buy a new machine. When the second one started giving problems, we used the repair money to buy a new one immediately. } } The first one started by developing a leak in the compressor, then the pump failed and finally the thermostat switch. Getting a similar thermostat switch to the one that failed was impossible. We searched through Grainger, talked with refrigeration engineers and could only come up with a close approximation of what we needed. } } We had to suffer as the less sensitive thermostat switch attempted to hold the water temperature steady but with much less control. In the end it was when some other part of the machine failed that we decided to replace it. Looking back, the time and expense needed to repair the machine was not worth it. } } If you do find your chiller has only a small problem then try fixing it and hope the costs don't escalate. } } Regards, } } Paul Webster. } } } } } Paul Webster, Ph.D. } House Ear Institute } 2100 West Third Street } Los Angeles } CA 90057 } phone (213) 273 8026 } fax (213) 413 6739 } email: pwebster-at-hei.org } } } } } } ---------- } } From: Kathleen Roberts } } Sent: Thursday, January 8, 2004 12:13 PM } } To: Owen P. Mills } } Cc: Microscopy-at-sparc5.microscopy.com } } Subject: [Microscopy] Re: Coolwell chiller testing } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 16:00:34 2004
Nope, no SEM and no BIG waterbath. I'm going to call Facilities here at RU-they may be able to devise something to test it.
Thanks- Kathleen Neurotoxicology Labs Rutgers University
Owen P. Mills wrote:
} Kathleen, } } Any heat source that can be water cooled will work, anything with a a } diffusion pump. SEM? Got a BIG hot water bath? Coil copper tubing } in the bath, turn the bath heat way up, connect the chiller lines to } the ends of the coil of copper tubing. Should be a low tech test of } the chiller. } } Owen } } At 03:13 PM 1/8/2004 -0500, you wrote: } } } Owen- } } } } Good idea, except that I don't have a carbon evaporator. :o) } } Thanks anyway, } } Kathleen } } Neurotox Labs } } Rutgers University } } } } Owen P. Mills wrote: } } } } } I've been reading the iterations on this thread and it seems to me } } } that it would be useful to separate the chiller from the TEM to test } } } the chiller. Kathleen, if you have a carbon evaporator that has a } } } diffusion pump you can cool it with the Coolwell long enough to tell } } } if the Coolwell has a problem. If the Coolwell tests okay on } } } another heat load (the carbon evaporator) then your problem is in } } } the TEM. } } } } } } Owen } } } } } } Owen P. Mills } } } Electron Optics Engineer } } } Materials Science & Engineering } } } Michigan Technological University } } } Rm 512 M&M Bldg. } } } Houghton, MI 49931 } } } PH 906-369-1875 } } } FAX 906-487-2934 } } } mailto:opmills-at-mtu.edu } } } http://www.mm.mtu.edu/~opmills } } } } } } At 11:48 AM 1/8/2004 -0500, Kathleen Roberts wrote: } } } } } } } } } } ------------------------------------------------------------------------------ } } } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } } America } } } } To Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } On-Line Help } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } ------------------------------------------------------------------------------- } } } } } } } } } } } } To Joel, Eckhart, Kenneth, Peter, and anyone else who gave me advice- } } } } } } } } OK, I finally had a chance to sit down and fiddle with the chiller, } } } } and here is what I have. } } } } } } } } I turned on my Zeiss EM 10CA and checked the flowmeter in the } } } } column: about 2.2 L/min, if I am reading it right. There was no } } } } gauge for the chiller flow rate, but there is a pressure gauge on } } } } the front of the chiller, and it read about 39 psi. This reading } } } } did not change through the entire test, and neither did the EM } } } } flowmeter. } } } } Chiller temperature reading was about 75 degrees F at start. It } } } } needs to be 68 degrees F, according to the EM's manual. Only the } } } } "Cool" light was on...all others, including the "Compressor" light, } } } } were off. } } } } } } } } I turned the temperature screw to the left to lower the temp setting. } } } } No response from the compressor. Turned it down a second } } } } time....still no response. After about 5 minutes the Compressor } } } } light turned on, and the temperature started to drop, finally. The } } } } temperature went all the way down to 53 degrees F, at which point } } } } the "Lo Temp" light went on, the "Compressor" light went off, and } } } } the "Heat" light went on. I turned up the temp screw a little bit. } } } } } } } } At about 60 degrees F, the "Heat" and "Lo Temp" lights went off and } } } } the "Cool" and "Pump" lights are on (actually the "Pump" light } } } } never turned off). It rose to 68 degrees F....and continued to } } } } rise. I let it get to about 79 degrees F, and the compressor never } } } } turned back on. I tried turning the screw back to the left to drop } } } } the temp setting...it never went back on. } } } } } } } } At this point, I decided to turn the EM 'scope off....though the } } } } flow rate never dropped, and the buzzer never went off. The } } } } temperature gauge on the chiller continued to rise, even after } } } } turning the 'scope off. It was reading about 90 degrees F when I } } } } left to come write all of you. } } } } } } } } Just now went back in to check the thing...the compressor finally } } } } came on...it's reading about 53 degrees F and the "Lo Temp" light } } } } is on, and the "Compressor" light is off. } } } } So I think it's the switch....or perhaps the "Hi Temp" sensor? Or } } } } both? I have no idea how all the sensors and switches are } } } } connected...even with the diagrams and manual that were sent, as I } } } } have no idea how to read such things (been eons since I did } } } } circuits in Physics class...). What do you all think? } } } } } } } } Thank you all so much for your help- } } } } Kathleen } } } } Neurotoxicology Labs } } } } Rutgers University } } } } } } } } } } Owen P. Mills } } } Electron Optics Engineer } } } Materials Science & Engineering } } } Michigan Technological University } } } Rm 512 M&M Bldg. } } } Houghton, MI 49931 } } } PH 906-369-1875 } } } FAX 906-487-2934 } } } mailto:opmills-at-mtu.edu } } } http://www.mm.mtu.edu/~opmills } } } } } } Owen P. Mills } Electron Optics Engineer } Materials Science & Engineering } Michigan Technological University } Rm 512 M&M Bldg. } Houghton, MI 49931 } PH 906-369-1875 } FAX 906-487-2934 } mailto:opmills-at-mtu.edu } http://www.mm.mtu.edu/~opmills }
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 16:02:07 2004
In a message dated 1/8/04 4:44:05 PM, tivol-at-caltech.edu writes:
} The take-home lesson is that there still must be knowledgeable } oversight, especially with regard to automated processes, to assure the } quality of the results
You'll get no argument from me about that! The point I was trying to make is that as the tools have become more complex, many of the tasks that we once dealt with manually (I learned electron microscopy on a Siemens 1A, right on the Caltech campus, back in the '50's - and as a result I really know what alignment means, not just how to push a button) are now so automated that they are hard to control. We may (but may not) be able to spot problems, but the casual user (shudder) will not know how to correct them.
I think this thread has been interesting primarily in that everyone who has commented has been in basic agreement that far too many people who use microscopes understand them, or the ancillary techniques of specimen preparation, image analysis, etc., well enough to keep out of trouble or get really optimum results. Clearly they have other priorities than learning all that stuff. I've taught image analysis courses now to something approaching 3500 people. Even assuming they all learned everything I wanted them to, that is a drop in the bucket. And the people who really need it most don't come - at best they send a technician whom they can subsequently tell to do the work. But that's another rant.
John Russ
John Russ
From MicroscopyL-request-at-ns.microscopy.com Thu Jan 8 17:36:30 2004
} A lab user has asked me to find some place he can get freeze fracture } work } done. Close to San Francisco bay area would be best. Reply to me and I } will } pass on the contact info. } Dear Jon, If Kent McDonald does not have freeze-fracture, he probably knows someone in or near Berkeley who does. I don't have his email address immediately at hand. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 9 03:06:04 2004
The same is with EPMA (Electron Probe Microanalysis) with EDS. Unfortunately m o s t manufacturers moved their goals and tried to develop calculation machines as 'black box'. Because of the giant improvements in calculation speed of our computers (in last decades), only one button hit is necessary to display the complete analysis result (in most cases computer needs a tiny part of a second). The unskilled or less skilled operators believe in these results, without any concern. Even element concentration result errors of 0.1% and less are taken from the computer display as the truth. But such a very high 'accuracy' must be only the statistical part! The computer speed and modern easy to use software interfaces cover the very complex and not linear relations between measured signal and element concentration in specimen. The iteration process to get result convergence and the systematic and statistic errors with their error propagation during computing process are not visible.
I think for future, a more open software is going to be a trend. There must be a possibility to interact between the knowledge of the microanalyst and the computer program. A visible and easy understandable feedback for all computing steps is necessary. Of course, a higher skilled level of the operator is then necessary. This makes sense only, if the software give the possibility to share the knowledge with the operator, which is then really become a microanalyst.
A couple of years ago, I found in a very old German book of C. Remigius Fresenius "Anleitung zur Qualitativen Chemischen Analyse" ("Guidance to the Qualitative Chemical Analysis") - Braunschweig 1874:
"Es muss daher ein Halbwissen, wie überall so ganz besonders hier, für schlimmer als ein Nichtwissen erachtet und vor oberflächlicher Beschäftigung mit der chemischen Analyse ganz vorzüglich gewarnt werden."
Translation (I hope the feeling is transfered): Therefore a partial knowledge, like everywhere so particularly here, must be worse than even ignorance. It should be warned before superficial concern with the chemical analysis completely and excellently here.
These words are still valid and can be used nowadays for EPMA and Electron Microscopy including image interpretation, as well.
Frank Eggert
Steve Chapman wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Jan 9 05:35:11 2004
My two cents worth on a subject that has quite a few here tossing their own around.
Having spent 25 years in this area, I've had the opportunity to see the initial investigations of this practical application of sub-atomic particle physics in basic research labs as well as its resultant spread to a wide variety of industries. The use of SEM analysis has become a commodity because it is so successful in so many areas. Decades ago, many were researching the potential applications, and in virtually each case, they found the usefulness.
I can't begin to tell you the variety of areas where my customers have found the SEM useful. From missle guidance systems to electron beam lithography. Particulate analysis of howitzer oils to particulate contaminant analysis of sausage casings. The taxonomical classification of emerging bacterial species to process control for the laser produced hologram labels used for software copyright and federal IDs.
The fact is that the SEM has perhaps been too successful. We still hold in reverence the atomic physicists who can design an atomic bomb or understand the results from particle accelerators. But the SEM is a child of these processes that has found wide spread use and the demands for its application greatly exceed the number of those who really understand the underlying processes, much less the proper application of the various subtleties of its application.
I've had to deal with this over the years as the operators I deal with have changed from those individuals who first brought an SEM into a lab to the 'checklist' operators who know nothing about the instruments other than the written sequence of actions to turn it on and get an image. One of the challenges in my work is to try to encourage the SEM operators to learn and think more about the consequences of each turn of a knob or click of a mouse. As a maintenance provider, it certainly helps me if my customers have some understanding of their machines - but more importantly, it helps them do better work. SEMs aren't the only problems here - x-ray spectroscopy in fluorescence or the SEM based microanalysis is another area where operators are often fooled into thinking that results are a simple matter of a set routine.
The computerization of the instruments is furthering the problem. While the manufacturers are seeking only to play to the market, how many operators really understand what's happening when they tell their computer to bring up an FE gun? It really started with simple improvements such as electronic gun adjustments. When an operator had to physically move the gun assembly around, it made some sense that the position of the gun was important to aim the beam down the column. How many really understand the use of magnetic fields in the gun to tilt and shift the beam to alignment? Most that I see at first only know that they have to tweak these knobs and watch for an improvement in the signal.
Like it or not, this trend will continue. But it is not selective to SEMs - I see similar trends in every analytical instrument. As these instruments become more 'user friendly', they are actually lulling users into thinking that all that is needed is a brief glimpse at the user manual, which usually only describes how to push the buttons. IR, GC, LC, MS, ICP and many other techniques that involve complex physics have been reduced to a simplicity that masks their proper use primarily because those using them and buying them want simple answers. A material scientist investigating ceramics doesn't want to have to learn the sub-atomic particle interactions involved, he just wants pretty pictures that explain a manufacturing fault and justify the expense of the SEM, not to mention his job.
'Ease of use' is a marketing tool, and as such, it is a primary goal of manufacturers. I don't mean to focus on them, because it is a vicious circle - the customers are demanding it, the manufacturers simply provide it. In this process, though, what gets lost is that the proper use and interpretation of these instruments requires more than the customers are wanting to afford and more than the manufacturers are wanting to admit to.
Now Steve's attention is a little more esoteric - the quality control of a reviewed paper. But doesn't that just follow from the above? The results, rather than the process, are what matter most now. More and more we see examples in studies that are published, only to be later refuted. NASA's claim a couple of years ago about evidence of ancient bacteria in Mars based rock found in the Antarctic has, last I knew, been lost in dispute. Pons and Fleischman, and Gallo, are of course extremes, but how much of what is accepted as reputable science has later fallen as poor science. A brief look at medical headlines over the past decade or two can give a good glimpse. Science itself is supposed to ensure honest and accurate results, and the assumption of most people, scientists included, is that anything purporting to be science, promoted by supposed scientists, has some truth. Innocent until proven guilty, so to say.
Whether authored by lack of knowledge of instrumental techniques, lack of personal integrity or poor selection of measured variables, many papers get published that should have been caught by reviewers. That's assuming that those reviewers are well versed in all aspects of a particular paper. But given the wide variety of instrumental techniques available today, it can be a daunting task to find a single person expert in all of the instrumental techniques presented in a paper, not to mention the basic field of the paper and mathmatical aspects. If a reviewer isn't well versed in all aspects and techniques of a particular paper, can he be expected to catch the kind of cross-discipline problem in your example? Since much goes unsaid in virtually any paper, should reviewers be required to request all details of sample provenance - the collection, preparation and analysis?
By the way, Steve, was there any mention, in the example you cited, whether the sample was coated or not, or is it an assumption of yours that is wasn't?
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 ph. (630) 513-7093 fax (630) 513-7092 web http://www.sem.com
-----Original Message----- } From: Steve Chapman [SMTP:protrain-at-emcourses.com] Sent: Wednesday, January 07, 2004 6:42 AM To: MSA
Hi Listers
As many of you may know I run a training organisation that travels world wide spreading the word on SEM, TEM and EDS operation. I also have a deep interest in "Quality in Electron Microscopy"as those who picked up our paper last year would know?
Now to the point. Once again I am picking up respected journals and finding examples of what I would call poor microscopy, but in truth it also demonstrates poor quality control!
To bring one image to mind. The micrograph is of a structure which is described as being an example of a smooth surface on a biological material. But, the micrograph was taken at 20kV, where the vast majority of the information will have come from beneath the surface softening the true surface detail.
First to remove the training aspect . Operators of SEM should be taught that by manipulation of kV and working distance one may subdue or enhance surface features. To use more than 10kV on most biological samples is asking for sub surface detail, ignore this and comments on surface irregularities are null and void in my mind. (I have to say I would probably try to use 2 to 5kV if the microscope used was produced in the last 15 years!)
Now the quality aspect. By the time a paper is published a number of steps should have been taken. Working backwards, the publisher should have the paper vetted by knowledgeable scientists who would be able to pick out the problems that I see and have them corrected prior to going to print. Next back in the chain is the laboratory that was involved with the scientist; did they check the quality of the work leaving their EM unit? Stepping back again did the scientist take the micrographs or did they receive help? Either way the training of staff and operators should overcome this type of problem! But if the results the staff and visiting operators produce are not assessed how do you know that their training is inadequate?
As the pressure to perform increases and funding decreases only the cream of our laboratories will remain. In industry there is no question about following rigid "Quality" procedures and it is not too far off that this will hit the world's EM units too! I know that this is my baby but is it not about time that we woke up to these facts?
There is an area where I believe we have room for discussion; what do you think?
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob +44 (0) 7711 606967 www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 9 08:38:55 2004
I cannot help you with your MAC and the software but if you need an adapter for your Fuji S602Z and a microscope I can help you. We have adapters for all kind of digital cameras either for C-Mount or eye-pieces. Let me know if you want to know more about it.
Unfortunately our web site is not in english yet. However you can find the list of digital cameras we support here:
www.klughammer.de - enter the german pages, then open "Kameraadapter" - "für dig. Kameras" - go to the bottom of this page there you find "Kameraübersicht (PDF)", open it and then you get an overview of cameras.
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bwoAAM} I would really like to take the photos digitally but is it impossible bwoAAM} with my present configuration? bwoAAM} i would appreciate any comments please
Dear Jon, Caroline sent this to me; please pass it along to your colleague.
Begin forwarded message:
} Bill - } } Kent gave my old machine to a woman in SF who is running it for hire; } I don't know anything about her or the quality of her work. Look at } www.nanoanalytical.netfirms.com . } } 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/ } Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Jan 9 14:49:55 2004
Question: The lab where I currently work uses Scion Image, a freeing distributed graphical editing program. I have been having a problem with the "Set Scale" option under the "Analyze" tab. After taking a snapshot of a known scale under the microscope, I set the scale accordingly by typing in the known distance and setting the units to micrometers. When I switch to a different image and wish to use the same scale, the scale I have just calibrated has been reset to the default. Also, I have gone under the "file" tab and clicked on "record preferences," which seems to do nothing. No save box opens, and I am left with my cursor. In addition, the "revert to save option," also under the "file tab" is never illuminated.
How can I set the scale so it will be calibrated for all images open in the editing session?
Thanks and I hope someone out there has some answers.
I am trying to help a plant biology student with some plant histology on mango saplings. She is interested in looking at paraffin sections of the woody stems (LM). Does anyone have suggestions for a processing schedule? I have my processor set up for human tissues but I could easily extend the programmes to accomodate the cellular nature of the material. Having some suggestions would greatly cut down my trial and error!
Many thanks,
Evelyn Kaplan, Dept of Pathology, College of Medicine and Health Sciences, Sultan Qaboos University, Oman
From MicroscopyL-request-at-ns.microscopy.com Sat Jan 10 00:57:39 2004
Dear friends, the Italian Master on Microscopy at University I Level is starting on February 2nd, 2004. Only few positions are still available due to a delayed registration on January 19th, 2004. Details can be found at "Master Universitario di I livello in Microscopie ed Analisi Microscopiche in Biologia" http://www.studenti.unige.it/corsi/master.html and at www.lambs.it. All my best Alby
p.s. for further information, please e-mail to diaspro-at-fisica.unige.it using "microscopy master 2004" as subject.
....................................................................... ................................... .. non basta, resistere, resistere...resistere ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Alberto Diaspro Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa, Italy voice: +39-0103536426/480 fax 010314218 diaspro-at-fisica.unige.it, http://www.lambs.it http://wiley.com/cda/product/0,,0471409200,00.html
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 11 10:11:10 2004
} } } } Does anyone have a copy of the book "The Cell", 2nd Edition, by Don } } Wayne Fawcett, MD that they are willing to cell. This is a classic book } } full of electron micrographs. I am a graduate student and I will be } } taking an electron microscopy lab this semester and I am looking for 1 } } or more copies of this book. } } } } Please reply to hiswayt-at-earthlink.net } } } } Thank you.
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 11 10:33:04 2004
We occasionally embed plant samples for LM and make a few changes over what is customary for animal tissue. First of all plant samples do not get as brittle if dehydrated in a TBA (tertiary butyl alcohol)/ETOH series ending with 100% TBA.
Start with 15 min in each of 25 and 50% ETOH. Dehydrate for ~ 2-4hrs in each of the following percents: 90 ETOH - 10 TBA 80 ETOH - 20 TBA 65 ETOH - 35 TBA 45 ETOH - 55 TBA 25 ETOH -75 TBA 100% TBA - 3 changes for at least 4hrs total time
Infiltration is helped along by the following: Put samples into an oven set at a sufficiently high temperature to melt your paraffin. (I put all the cassettes into a beaker large enough to hold them so they are completely covered by TBA and then add room for the paraffin. Add solid paraffin (paraplast) to container and allowing it to gradually melt and mix with TBA. The TBA gradually evaporated. The paraplast is then changed a total of 3x over a period of a couple of days prior to embedding tissue in molds.
It is also advisable to use subbed slides or slides coated with poly L-lysine (100µg/ml in 10mM tris at pH 8.0). Plant cell walls tend to lift from the slide surface during staining.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 1/10/04 12:55 AM, "Evelyn Kaplan" {ekaplan-at-squ.edu.om} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } } } Good morning, } } I am trying to help a plant biology student with some plant histology on } mango saplings. She is interested in looking at paraffin sections of the } woody stems (LM). Does anyone have suggestions for a processing schedule? I } have my processor set up for human tissues but I could easily extend the } programmes to accomodate the cellular nature of the material. Having some } suggestions would greatly cut down my trial and error! } } Many thanks, } } Evelyn Kaplan, } Dept of Pathology, } College of Medicine and Health Sciences, } Sultan Qaboos University, } Oman } } }
From MicroscopyL-request-at-ns.microscopy.com Sun Jan 11 12:40:08 2004
} } } } } Does anyone have a copy of the book "The Cell", 2nd Edition, by Don } } } Wayne Fawcett, MD that they are willing to cell. This is a classic book } } } full of electron micrographs. I am a graduate student and I will be } } } taking an electron microscopy lab this semester and I am looking for 1 } } } or more copies of this book. } } } } } } Please reply to hiswayt-at-earthlink.net } } } } } } Thank you.
If you look at the used book search site www.abebooks.com you'll find 10 copies at prices from $9-$75. -- 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/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 09:54:06 2004
I have used these old protocols with success on woody materials. Put plant material always requires greatly extended times compared to animal tissue. Also, if you have problems with tearing of the embedded tissue during sectioning you can soften the embedded material in Gifford's solution (below). The difficulty is trying to get the harder tissue soft enough so it doesn't tear the softer tissue during sectioning.
The details we used are as follows: Fixation for 12 to 48 hours in FAA (you might also try Navashin's fixative which contains chromic acid) Infiltrate through TBA series (Johansen series) for 2 hours each step 3 changes of pure TBA over 24 hours 3 changes of Paraplast over 24 hours Embed
Soften the blocks by soaking overnight to 7 days in water at up to 38°C If this doesn't work try 2 days to 2 weeks in Gifford's (room temp in fume hood): 20 ml glacial acetic acid 80 ml 60% ethanol 5 ml glycerine You will have to try the softening times with your own tissue. If you leave it too long the soft tissue will become macerated. Let me know if you need more detail.
Good luck,
Kim
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 10:00:07 2004
We have recently acquired a Pelco Biowave and are in the process of acquainting ourselves with it. Currently, I am trying to fix two species of insects with it for SEM (Colorado potato beetle larvae and Diamond back moth larvae). A literature search has not turned up much information on microwave processing for insects. I have tried adaptations of the standard protocol (2% glut, rinse, 1% osmium, rinse, ethanol dehydration, critical point dry) at various microwave wattages, times, and with vacuum applied. So far, I have not achieved reproducible results for either insect. While I plan on more trial and error, I was wondering if anyone has a microwave protocol for insects or any advice on this.
Shannan Little Research Technician/Technicien de recherche Electron Microscopy and Image Analysis / Microscopie électronique et Analyse d'images Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 403-317-3446 Facsimile/Télécopieur: 403-382-3156 P.O. Box 3000 / CP 3000 Lethbridge, Alberta T1J 4B1 littlesm-at-em.agr.ca http://res2.agr.ca/lethbridge/emia/index_e.htm
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 10:00:47 2004
Dear all, We would be interested in users comments about EBSD systems. In particular, we are looking at the systems offered by HKL and by TSL/EDAX. Please comment on some of the following points:
* Ease of use
* Robustness/reliability of indexing when dealing with low symmetry structures
* Calculation of GB misorientations and display of crystallographically equivalent misorientations
* Possibilities for generating different types of map (e.g. orientation, GB misorientation, phase)
* Correlation with EDXS data or maps / generation of combined maps
I look forward to hearing from you. Please copy all responses to myself and to Martin Lee {m.lee-at-earthsci.gla.ac.uk}
Thanks and best wishes
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 10:32:46 2004
I have been calibrating my recently installed Philips TEM with a grating replica and I need some suggestions. At a print magnification of about 80KX I see about a 1% variation in my calculated magnification depending where I select my stop and start marks.
How much variation should I expect in magnification due to changes in lens voltage and current?
Thanks!!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 10:46:04 2004
Frank, I suspect that these variations are in the grating itself, not your TEM. They can be a bit variable, depending on stretching and buckling from the preparation process. We have a record of measurements of semiconductor standard samples going back 8 years on our Jeol 120CX TEM, and find a reproducibility better than 1% over this time. There is a change in magnification from the centre to the edge of the micrographs on our machine of about 1%, but our microscope is now pretty ancient and I would hope that newer machines are much better than this. We are about to embark on a full gauge capability test on the machine, which should be interesting. I can let you know the results of the study if you like.
-----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: 12 January 2004 16:18 To: microscopy-at-msa.microscopy.com
I have been calibrating my recently installed Philips TEM with a grating replica and I need some suggestions. At a print magnification of about 80KX I see about a 1% variation in my calculated magnification depending where I select my stop and start marks.
How much variation should I expect in magnification due to changes in lens voltage and current?
Thanks!!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 11:13:40 2004
In my experience, successful microwave fixation depends mainly upon the size of the specimen. How big are these insects? Second, if the larvae are difficult to penetrate (I have no idea), longer times may be required. You might also consider Karnofsky's fixative instead of Glutaraldehyde (I found that it improved results in many cases over a wide variety of specimen types, although I didn't try insects). I would try lengthening the primary fixation time before adjusting any of the other processing variables. Fixation temperatures should never exceed 50°C (I don't know what this translates to in watts on your Biowave), or you will get "crispy critters".
best regards, Steven Slap Microwave Consultant
At 11:04 AM -0500 1/12/04, Shannan Little wrote: } We have recently acquired a Pelco Biowave and are in the process of } acquainting ourselves with it. Currently, I am trying to fix two species } of insects with it for SEM (Colorado potato beetle larvae and Diamond } back moth larvae). A literature search has not turned up much } information on microwave processing for insects. I have tried } adaptations of the standard protocol (2% glut, rinse, 1% osmium, rinse, } ethanol dehydration, critical point dry) at various microwave wattages, } times, and with vacuum applied. So far, I have not achieved reproducible } results for either insect. While I plan on more trial and error, I was } wondering if anyone has a microwave protocol for insects or any advice } on this.
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 11:46:35 2004
FYI I just opened a new supply of Kodak Professional Rapid Fixer and found larger boxes. After the film switch not so very long ago, I decided to check the ingredients. "Solution A" now has Ammonium Sulfite, Sodium bisulfite and Sodium acetate added to what was printed on the old box. The mixing directions are the same 1999 version. "Solution B" and the CAT # 146 4106 appear to be the same.
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 11:53:21 2004
We are trying to install two cameras on a Windows XP computer.
We cannot get XP to recognize both the Sensicam PCI card and the Roper PCI card. It's completely an exclsuive or installation. We've poked around the Device driver menu and downloaded the most recent drivers.
Has anybody figured out how to install both these camera simultaneously?
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 12:01:24 2004
I would recommend the Mag-I-Cal sample for calibrating your TEM. You can do a much larger range of Magnification settings than using a replica. I think that the error is 1 to 2%. In addition, you can do rotation and camera constant calibrations with the same sample.
One thing to minimize the variation in the microscope is to always changed the lenses in the same direction to avoid hysterisis and to make sure that you are at the eucentric point so that the obj lens is the same. Again, you should bring the focus from the same direction.
If the goniometer has not bee removed or the column split, the values that you get from time to time should be very close. We had to run calibration twice a year on the TEM. I can't remember the range specifically, but I think that the variation in mag was less than 0.5% and may have been better than 0.1% when the above criteria was met.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472 Walck-at-PPG.com (412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: Monday, January 12, 2004 11:18 AM To: microscopy-at-msa.microscopy.com
I have been calibrating my recently installed Philips TEM with a grating replica and I need some suggestions. At a print magnification of about 80KX I see about a 1% variation in my calculated magnification depending where I select my stop and start marks.
How much variation should I expect in magnification due to changes in lens voltage and current?
Thanks!!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 12:53:50 2004
When we manufacture TEM or SEM gratings we make them from a replica of a master grating. When you dissolve away the replication material there is some shrinkage, but it should be very limited. The shrinkage is inherent in the manufacturing, but we cull any which show problematic shrinkage. It is very similar to our carbon substrate manufacturing.
John Arnott
Disclaimer: Ladd Research manufactures and sells the gratings, replicating materials and substrates mentioned in this email.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
----- Original Message ----- } From: {Frank.Karl-at-degussa.com} To: {microscopy-at-msa.microscopy.com} Sent: Monday, January 12, 2004 11:17 AM
Frank, If you are using film, a second variation in measurement may come from the enlarger when you print the picture. If the negative is not supported on glass, it can bow in the center and distort the measurement a bit.
It was a surprise to me to find that if I had set my "MAG. ZERO" early in the morning and then checked it later in the day, there was frequently a slight change. It was explained to me that in an old building, when there was a greater draw of electricity, a change could be expected and for really important work, I should re-calibrate. Am I just gullable?
My favorite goof has been the result of not adjusting my tilting specimen holder to read 0 degrees!
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } When we manufacture TEM or SEM gratings we make them from a replica of a } master grating. When you dissolve away the replication material there is } some shrinkage, but it should be very limited. The shrinkage is inherent in } the manufacturing, but we cull any which show problematic shrinkage. } It is very similar to our carbon substrate manufacturing. } } John Arnott } } Disclaimer: Ladd Research manufactures and sells the gratings, replicating } materials and substrates mentioned in this email. } } Ladd Research } 83 Holly Court } Williston, VT 05495 } On-line Catalog: http://www.laddresearch.com } tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) } fax: 1-802-660-8859 } e-mail: sales-at-laddresearch.com } } ----- Original Message ----- } } From: {Frank.Karl-at-degussa.com} } To: {microscopy-at-msa.microscopy.com} } Sent: Monday, January 12, 2004 11:17 AM } Subject: [Microscopy] TEM mag question } ----------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } I have been calibrating my recently installed Philips TEM with a grating } } replica and I need some suggestions. At a print magnification of about } } 80KX I see about a 1% variation in my calculated magnification depending } } where I select my stop and start marks. } } } } How much variation should I expect in magnification due to changes in lens } } voltage and current? } } } Frank Karl
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 15:39:11 2004
I wanted to know the vapor pressure of alumina (Al2O3) at 1200 C. I know that it will be very low, but an estimate would also be helpful. I looked at various places (such as CRC handbook of Physics and Chemistry, Handbook of thermophysical properties of solid materials, ASM handbook, Vol. 5), but got values above 1950C (e.g. 10^-3 torr at 1950 C).
I want to know the answer to this question since I work at vacuum levels of 10^-6 to 10^-8 torr and at 1200C. I am interested in knowing if the aluminum oxide layer on my samples evaporates under these conditions. One company representative said that it wont, but he did not have vapor pressure values to support the assertion.
thanks in advance
Rahul Panat Univ of Illinois Urbana, IL
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 17:23:47 2004
Title-Subject: [Microscopy] [Filtered] Kodak 4489 EM film
Question: Can anyone use some left-over Kodak 4489 electron microscopy film? I was about to throw out 5 boxes [2 x 10 in. 100 strips per box] during "housecleaning" the other day, but thought there might be someone who could use it. It has been refrigerated and unopened for about 12 years. [I used to use it in our old RCA EMU 3G TEM.] If someone wants it send me your mailing address and I'll ship it out to you. Otherwise, it will be recycled or trashed.
Lee Hadden Department of Biology Wingate University Wingate, NC 28174
Question: I am interested in getting quantitative analysis training for analyzing aluminum alloys and oxides. In our lab, we have an Hitachi S-3500N equipped with an Oxford EDS Spectrometer (model# 7021)that has the INCA and ISIS software packages installed on the computer. I would like to visit an independent laboratory or university for a week or two for training. If someone could assist me I would appreciate it.
} } Hello Shannan. } } I have some experience with microwave fixation of Drosophila larval } salivary glands for TEM. First, I run my cold spot -at- 15-18 degrees (this } removes the heating effect) I fix them in Karnovsky's fix power level } 1(100 watts on my MW) for 1 minute on-1minute off-1 minute on). Then I } turn the power up to 450 watts (power level 4 on my machine) and pulse for } 30 seconds on-30 seconds off-30 seconds on 3 times.They should not get } hot! I let sit on the bench in fixative for 5 minutes. I have also tried } this protocol on zebrafish larvae (with vacuum) and they are well fixed. } The insect probably has a cuticle which may hinder the penetration of the } fixative. If you can find a way to partially remove this, or inject the } fix then MW, you might have better results. } For osmium fix, I repeat the above timetable. I dehydrate under vacuum 45 } seconds -at- power level 3(350 watts) 50, 70,95% once each. I do 100% } ethanol 3 times followed by 100% acetone before infiltration in resin. } Hope this helps, JoAnn Buchanan } } } } We have recently acquired a Pelco Biowave and are in the process of } } acquainting ourselves with it. Currently, I am trying to fix two species } } of insects with it for SEM (Colorado potato beetle larvae and Diamond } } back moth larvae). A literature search has not turned up much } } information on microwave processing for insects. I have tried } } adaptations of the standard protocol (2% glut, rinse, 1% osmium, rinse, } } ethanol dehydration, critical point dry) at various microwave wattages, } } times, and with vacuum applied. So far, I have not achieved reproducible } } results for either insect. While I plan on more trial and error, I was } } wondering if anyone has a microwave protocol for insects or any advice } } on this.
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 19:07:09 2004
Michael, Have you tried rearranging the cards on the Mobo? I had the same adventure when installing a Pixera camera and Scion system on an XP system. I did not get the New Hardware Found announcement for the Pixera card until I did some card swapping.
Let me know how you get on.
Greg
Dr. G. F. Barclay Plant Science Unit, Dept of Life Sciences University of the West Indies St. Augustine, Trinidad and Tobago West Indies Phone: 868 645 3232 ext 3112/2045 Fax: 868 645 7132
} From: Michael Cammer {cammer-at-aecom.yu.edu} } To: microscopy-at-MSA.microscopy.com } Subject: [Microscopy] Sensicam QE & Roper HQ } Date: Mon, 12 Jan 2004 12:58:55 -0500 } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_________________________________________________________________ Help STOP SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 20:02:39 2004
Dear Shannan, Below is a method we have used for a few different insect types, for TEM not SEM, but the fixing steps we use might provide a useful comparison at least.
With SEM I assume you won't want to dissect your samples prior to processing (as we did) so penetration of the solutions may be more of a problem, but then the requirements for fixation for SEM are also less stringent. The microwave conditions may be useful guidelines but of course you'll have to determine the conditions for your own microwave and samples. If you haven't already got them, I strongly recommend Gary Login's text (Login, G. R., & Dvorak, A. M. (1994) " The Microwave Toolbook A Practical Guide for Microscopist" Boston: Beth Israel Hospital) and Kok and Boon's book "The Microwave Cookbook for Microscopists", Boon and Kok, Coulomb Press Leiden, 1992.
Method: Primary fix: 3% glutaraldehyde in 0.1M Na cacodylate, pH 7.4. Samples dissected out and placed into the primary fix solution (in a small plastic petri dish).
They were then put into fresh fixative (specimen containers for a Leica AFS were used inside the petri dish) and microwave irradiated as follows:
EMS lab microwave oven setup: -a dummy load of 100ml dw at 20degC in a 250 glass beaker was used (always in the same place - right rear corner for us) -temperature limiting off -100% 'power' (ie magnetron on continuously) -sample volume for all the fixing steps was 4.0ml -magnetron was pre-warmed for 2 minutes with a load of 500-600ml water before each step (unless the oven was used less than 2 minutes earlier).
1) Microwave Primary Fixation:
The sample in 4 ml of primary fix is irradiated for (7s) to give a final temperature of about 50degC - check the temperature after irradiation (and obviously before you use your actual samples if you're doing this for the first time) and alter subsequent run times if necessary. (We use the spot in the oven we deem to be receiving a steady, high level of radiation).
Allow sample to sit in fume cupboard in fix container for 3 minutes to cool it to room temp before removing fix. Replace fix with fresh and repeat the irradiation process twice. A cool dummy load must used with each run.
Leave samples in fume cupboard for 30 minutes at room temperature.
2) Rinse the samples:
Three X 10 minutes in 0.1M cacodylate buffer.
3) Secondary fixation:
Place 2% OsO4 in 0.1M cacodylate buffer into fix dish with specimens. Irradiate for 7s Leave for 40 minutes in the OsO4.
4) Rinse with 0.1M cacodylate buffer
Three X 10 minutes
The remainder of my method is for TEM preparation so you could do your usual pre-drying and drying steps then.
Regards,
Richard
} } We have recently acquired a Pelco Biowave and are in the process of } acquainting ourselves with it. Currently, I am trying to fix two species } of insects with it for SEM (Colorado potato beetle larvae and Diamond } back moth larvae). A literature search has not turned up much } information on microwave processing for insects. I have tried } adaptations of the standard protocol (2% glut, rinse, 1% osmium, rinse, } ethanol dehydration, critical point dry) at various microwave wattages, } times, and with vacuum applied. So far, I have not achieved reproducible } results for either insect. While I plan on more trial and error, I was } wondering if anyone has a microwave protocol for insects or any advice } on this. } } Shannan Little
Richard Easingwood Otago Centre for Electron Microscopy Department of Anatomy and Structural Biology School of Medical Sciences, University of Otago PO Box 913, Dunedin, NEW ZEALAND Telephone: 0064 3 479 7301 Facsimile: 0064 3 479 7254 GSM: 0064 21 222 4759 mailto:richard.easingwood-at-stonebow.otago.ac.nz Web site: http://ocem.otago.ac.nz/
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 21:28:45 2004
We swapped cards all over the place. The one in the lowest slot gets recognized, but not the other. Also, the Roper card doesn't seem to work in the highest slot, even alone. We could get either card to work with a firewire camera in another slot, but the Retiga we have isn't low noise enough for this application. I think we'll have to temporarily get a second computer in the room. Thanks.
On Tue, 13 Jan 2004, gregor barclay wrote:
} Michael, } Have you tried rearranging the cards on the Mobo? I had the same adventure } when installing a Pixera camera and Scion system on an XP system. I did not } get the New Hardware Found announcement for the Pixera card until I did some } card swapping. } } Let me know how you get on. } } Greg } } } } Dr. G. F. Barclay } Plant Science Unit, Dept of Life Sciences } University of the West Indies } St. Augustine, Trinidad and Tobago } West Indies } Phone: 868 645 3232 ext 3112/2045 } Fax: 868 645 7132 } } } } } } } From: Michael Cammer {cammer-at-aecom.yu.edu} } } To: microscopy-at-MSA.microscopy.com } } Subject: [Microscopy] Sensicam QE & Roper HQ } } Date: Mon, 12 Jan 2004 12:58:55 -0500 } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } We are trying to install two cameras on a Windows XP computer. } } } } We cannot get XP to recognize both the Sensicam PCI card and the Roper PCI } } card. It's completely an exclsuive or installation. We've poked around } } the Device driver menu and downloaded the most recent drivers. } } } } Has anybody figured out how to install both these camera simultaneously? } } } } } } } } } } ____________________________________________________________________________ } } Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of } } Med. } } Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 } } (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ } } } } } } _________________________________________________________________ } Help STOP SPAM with the new MSN 8 and get 2 months FREE* } http://join.msn.com/?page=features/junkmail } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jan 12 22:57:10 2004
We have an old Wild M8 dissecting microscope with a 1.0x objective that can be converted to 0.4x or 1.6x by clipping adapter lenses onto the bottom of the 1.0x objective. We'd like to get a second set for another M8 because they get used so much now that they are sometimes needed by both microscopes. Unfortunately, these are no longer available from Leica, instead you have to get an adapter and rather more expensive 0.4x and 1.6x objectives. These new objectives are better quality than the old adapters, but for our work, the ease of switching with the adapters and low cost outweighs the marginal increase in quality at the magnifications we're using.
Anyone willing to part with their adapters, or know of a source? I've tried ebay and several other used equipment sites with no luck so far.
0.4x adapter lens part no. 367898 1.6x adapter lens part no. 367916
Thanks much, Rosemary White
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
From MicroscopyL-request-at-ns.microscopy.com Tue Jan 13 07:23:25 2004
Richard makes some good points here. I second his reading recommendations, and want to let everyone know that there is a brand new edition of Kok and Boon, Microwaves for the Art of Microscopy, Coulomb Press, Leiden, 2003, which has some very useful new information.
I believe that the use of a dummy load is not needed in the Biowave, which has a recirculating water cooler. In any event, the end temperature of 50°C is the key, and Richard's 3 x 7 minutes should give Shannon a good starting point for time in the fixative. I would be a little concerned about using such a small volume of fixative as Richard did (4.0 ml), both because of a fear of exceeding the temperature set point and because a greater volume of fixative to specimen ratio seems prudent based upon my experience with light microscopy specimens. I generally work with about 11.0 ml of fixative.
best regards, Steven Slap Microwave Consultant
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From MicroscopyL-request-at-ns.microscopy.com Tue Jan 13 07:44:35 2004
O.k., folks following the various recommendations from the list for flatbed scanners we got an Microtek AtrixScan 2500f.
So now, anyone out there with a Microtek 2500f what is the part number and where do you get the bulbs from?
Microtek offical position is "There are no user servicable parts. You have to ship it back - at your cost - to Microtek in California for repair". Now, I am not shipping a 100lb scanner back to California for a $20 bulb replacment - let alone doing it every 6-8 months. Surely someone else out there has already come across this (especially since the bulbs never power down).
Thank you in advance for any help.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
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