} I have found some references to books on TEM history by Marton (1968) } and Hawkes (1985) but don't have these on hand.
I have a book that was published at the time of the International Congress on EM in Kyoto, Japan, in 1986, entitled "History of Electron Microscopes 1986" edited by Hiroshi Fujita. This gives a lot of information on the history of EM, mainly from a Japanese perspective. I haven't seen whether or not this has the answers to your specific questions but if there is someone near you who has a copy of this book you will find much interesting information in it. Please let me know if you cannot get hold of a copy.
Robin H Cross (Mr) Director : Electron Microscopy Unit Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 email: r.cross-at-ru.ac.za http://www.ru.ac.za/emu/index.htm
** remember ICEM-15 in Durban in September 2002 **
I am not a forensic scientist so my suggestions are general - therefore I am uncertain how the methods I give could be made robust enough for use in court. I assume that IPA is isopropyl alcohol?
Starch should be positive with the periodic acid Schiff method (PAS) and may be 'confirmed' as such by using an amylase control. Starch should also show 'Maltese Cross' birefringence under polarised light. The cellulose should also be PAS positive, resist amylase and show 'linear' birefringence.
The protein is a little more difficult to confirm absolutely but if these are skin cells they will contain cytokeratins - thus giving the possibility of using immunocytochemistry with a pancytokeratin antibody.
Good luck!
PS Why is an IR/Raman engineer doing forensic work on paper/cells?
At 16:07 2001-02-28 -0800, HAMMOND,LOMA (HP-Corvallis,ex1) wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
Congo red will stain cellulose red, and can also be viewed by fluorescence using a green (RITC-type) excitation filter. Cellulose and starch are also birefringent, which you could demonstrate by using crossed polarizers. A 1/4 wave or 1-wave plate may make this easier to see. The fluorescence of Cellulose stained with congo red is also polarized because the CR molecules line up on the oriented cellulose microfibrils. As has already been mentioned starch and cellulose stain with periodate-Schiff. If you have access to a fluorescence microsocope you might also try a fluorescence variant of PAS, by replacing schiff reagent with Lucifer Yellow CH. View with FITC-type excitation.
Iodine/potassium iodide will stain starch blue/black.
Chris
----- Original Message ----- } From: "HAMMOND,LOMA (HP-Corvallis,ex1)" {loma_hammond-at-hp.com} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 01, 2001 12:07 AM
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Fungal people, Although vapor fixation works sometimes and immersion fixation also works sometimes, often fungi samples are too delicate to handle with routine techniques. Just immersion in an aqueous solution can damage the structures. Repeated washings also can wash away delicate conidia. CPD causes shrinkage which may not be a major concern with some samples but can be a real problem with delicate ones like fungal hyphae. Really the very best way to deal with many fungi samples is cryo SEM. True you may have to coat a bit longer than normal, etc. but the structures, when instantly frozen, are most likely to be similar to the living state. Unfortunately many people do not have access to Cryo -SEM equipment so must settle for the next best approach....fixation and drying by one of the methods already mentioned.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
Microlisters,
While the simpler and less time consuming suggestions already mentioned in this thread may work in some cases, I found out a long time ago that the only way to deal with the "fungal jungle", eg. fungi cultured on agar plates, is to hit 'em with the ol' OTO method, preceded by glutaraldehye fixation. Quick outline is as follows:
Glut. fixation, your favorite buffer Buffer rinses Osmium tetroxide, 2% Water rinses Thiocarbohydrazide, sat. soln.,filtered water rinses Osmium tetroxide, 2% Water rinses dehydration series, EtOH, or acetone CPD metal coating? Maybe, maybe not. Try without first.
This eliminates the charging, and fixation is quite good.
Classic reference for this method:
"Ligand-mediated osmium binding:Its application in coating biological specimens for scanning electron microscopy". Kelley, Robert O., A.F. Dekker and John G. Bluemink. J. Ultrastrucute Research 45:254-258 (1973).
See also:
"Non-coating techniques to render biological specimens conductive/1980 update". Judith A. Murphy. Scanning Electron Microscopy/1980 Vol.I, p. 209-220.
You might try exposing your non-aquatic sample (bread molds, mushrooms, etc. ) to vapors of Osmium tetroxide for several hours ( a few drops of 4% OsO4 on the lid of a very small container with your sample inside or the lid over the sample directly), then air dry, mount, coat and view. As with any sample, the drying artifacts vary with the fungus. Some things look great.
good luck
Steve } ----------------------------------------------------------------- } Hello, } } Forgive me, I haven't tried to find this on my own yet... } } But have any of you used an electron microscope to get a } close-up of a fungus? Can this be done? } } Thanks...
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
As others have pointed out, phase halo is easily demonstrated. Shading-off is a different matter, at least for me. Both artifacts are the result of incomplete segregation of the direct and diffracted light to the conjugate and complementary areas respectively of the phase plate. Shading-off is described in "Advanced Light Microscopy" vol. 2 pg. 15-20 by Maksymilian Pluta. Shading-off appears as a brighter central area becoming darker toward the edge in positive phase contrast, and just the reverse for negative phase contrast. Pluta has illustrations of this on pages 18 and 19, but he doesn't say what the object is. It looks like a crystal of some kind.
My point in bringing this up is to solicit others ideas on how best to teach students (as well as ourselves) to interpret the phase contrast image. The phase image has these artifacts superimposed on it. The phase halo can mislead a novice but actually help an expert. Shading-off is more subtle in a complex specimen than it is in Pluta's example, and I have never seriously considered how it affects the phase image. I wonder if there is a biological sample that clearly shows both artifacts? Any thoughts and suggestions on this?
Bob
_____________________________________________________________ C. Robert Bagnell Jr., Ph.D., Res.Assoc.Prof. Microscopy Services Laboratory Department of Pathology & Laboratory Medicine CB #7525 UNC-CH, Chapel Hill, N.C. 27599 ph 919-966-2413 fx 919-966-6718 http://www.pathology.med.unc.edu/path/microscopy/welcome.htm
Email: kuznetsv-at-geo.komisc.ru Name: Chuprov Georgy School: The Institute of Geology State: Republic of Komi
Question: Please, tell me. Where I can find a computer program for plot of stereografic projections of optical axes (were tested by optical microscop)?
I have one of those very basic "thought" questions on a procedure that I've always taken for granted, but am now not so sure about. I had been trained from Day One to always focus a TEM with the beam at crossover, then to spread the beam until the exposure intensity is reached. The idea seems to be that small focusing errors at crossover will be corrected as the beam is spread and "depth of focus" is increased somehow.
The reason I'm asking is that in recent years, almost no one else I've run across focuses in this way, preferring instead to focus with the beam already spread to the point at which the photograph will be taken (at least as long as the image is bright enough to see for focusing). Out of curiosity, I ran tests using both methods and have noted no clear differences.
Does anyone have any thoughts on this? Any ideas or theories about why one method would be superior to the other?
Sincerely, Curious in Columbia
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Dr. Ahlstrand's technique is excellent for imaging hyphal structures and some spore bodies while the stick on method of Dr. Mary Mager works well for certain individual spores. However, imaging of fruiting body structures (conidiophores, etc.) are often best prepared by using Osmium vapor fixation followed simply by air-drying (AD). One can also add minimal solvent exchange before AD, but some structural collapse will be observed in any event. "One size does not fit all" for fungi, sometimes even within one species.
We have a new ESEM now and hopefully combinations of low vacuum technology and various wet modes will improve imaging for our mycologists.
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Microlisters,
While the simpler and less time consuming suggestions already mentioned in this thread may work in some cases, I found out a long time ago that the only way to deal with the "fungal jungle", eg. fungi cultured on agar plates, is to hit 'em with the ol' OTO method, preceded by glutaraldehye fixation. Quick outline is as follows:
Glut. fixation, your favorite buffer Buffer rinses Osmium tetroxide, 2% Water rinses Thiocarbohydrazide, sat. soln.,filtered water rinses Osmium tetroxide, 2% Water rinses dehydration series, EtOH, or acetone CPD metal coating? Maybe, maybe not. Try without first.
This eliminates the charging, and fixation is quite good.
Classic reference for this method:
"Ligand-mediated osmium binding:Its application in coating biological specimens for scanning electron microscopy". Kelley, Robert O., A.F. Dekker and John G. Bluemink. J. Ultrastructure Research 45:254-258 (1973).
See also:
"Non-coating techniques to render biological specimens conductive/1980 update". Judith A. Murphy. Scanning Electron Microscopy/1980 Vol.I, p. 209-220.
You might try exposing your non-aquatic sample (bread molds, mushrooms, etc. ) to vapors of Osmium tetroxide for several hours ( a few drops of 4% OsO4 on the lid of a very small container with your sample inside or the lid over the sample directly), then air dry, mount, coat and view. As with any sample, the drying artifacts vary with the fungus. Some things look great.
good luck
Steve -----------------------------------------------------------------
Dear rad0, I have looked at fungus and fungus spores on my SEM and they are very simple to prepare. Just stick them to a stub by sticky tab or glue and then gold coat. They are very robust and they don't need any fixing or dehydration.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} ----------------------------------------------------------------- } Hello, } } Forgive me, I haven't tried to find this on my own yet... } } But have any of you used an electron microscope to get a } close-up of a fungus? Can this be done? } } Thanks...
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
Bruce F. Ingber, Biologist USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
You might guess from the lack of response that quantitative stereoscopy using TEM is not that popular a pastime. I was involved some 20 years ago and can offer some comments which were relevant then.
There is a big difference between TEM and SEM stereo analysis. In SEM images you have a lower surface that can be seen easily, in TEM images you have a view through both surfaces and the specimen, the surfaces are very hard to distinguish. It is relatively easy to calculate the height from a fixed surface as in a SEM micrograph. Calculating the height from an 'invisible' surface, as in a TEM micrograph, is very difficult. It's OK to think of something like a dislocation running through the specimen and then assume that it ends at either surface but very few examples go through from surface to surface and thus are considerably more difficult.
Height measurements are made from stereo pairs by measuring the parallax between the two tilted images. Provided the tilt axis is known and the magnifications corrected, for any change in focus current, then the height difference between two points can be calculated. This is a long way from reconstructing a 3D image.
It is relatively easy to view stereo images using a stereo viewer but not everyone can do it. You need two eyes of equal strength and a certain type of brain. From memory the mapmakers said 50% of the population could see stereo properly and 10% could make accurate measurements.
There were programs that would allow 2 stereo micrographs to be placed side by side on a graphics tablet and then the same points marked on both micrographs. The height differences between these points were then calculated using the first pair of points as a reference height. I don't know if these are still available for modern computers (they ran on an Apple 11+).
Stereo viewers are available that will project a 'floating' point of light in the specimen that you can position onto a feature and then read out the height from a micrometer (it will need to be scaled to get the real height). We modified one of these viewers to record and calculate these heights (and X,Y coordinates) automatically), but it it still a lot of work to produce even a simple 3D image.
I am not aware of any image recognition program that will analyse your two images and produce a 3D image from them. (Well if that doesn't get details sent in nothing will!)
If you want some references then in the mid 70s Hudson and Makin descibed the how to select the tilt angle taking into account the film thickness and the final magnification. Much of the work at that time was being done by Alan Boyde. I may be more use with references when I get my office back - contact me off list in a few days if you want 20-25 year old refences.
Good luck, Ron
On Wed, 28 Feb 2001 10:03:56 -0700 Rodney McCabe {rmccabe-at-lanl.gov} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues: } } I am interested in doing quantitative stereo microscopy using a TEM and am } looking for software that may be helpful in such an endeavor. I anticipate } the primary application will be examining the spatial arrangement of linear } defects. I have not been able to find a software designed directly for } quantitative stereo TEM. I have come across one commercial software that } can be used for SEM surface topography that may be applicable. It seems } like there must be other software packages out there used for applications } such as surface mapping, electron tomography, or even astronomy that could } be useful. Also, as I am new to stereo microscopy, any words of wisdom } would be appreciated. } } Thanks, } } Rod } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
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Pete Augustus Caswell Technology
Hi Listers,
I'm preparing a talk which is to include a timeline giving dates and names for some relevant TEM developments. I'm trying to find answers to two trivia questions with great importance to microscopy:
1) Who was the first to knowingly image a dislocation in a TEM, when was it done and where was it published? On this, I have found references as far back as an article by Heidenreich in J. Appl. Phys, 1949, but don't have this journal going back that far.
2) Who developed the solid state detector for energy analysis of x-rays (EDS), when and where published?
I have found some references to books on TEM history by Marton (1968) and Hawkes (1985) but don't have these on hand.
Thanks,
Wharton
*********************** Wharton Sinkler UOP LLC Des Plaines, IL
For fragile conidiophores and their associated conidial chains, it is still possible to use critical point drying after vapor fixation with OsO4.
A fairly simple dehydration technique that avoids the solution changes -- and turbulence -- that often lead to loss of the conidia from aerial conidiophores (and the air drying that often does the same), is described at Can. J. Microbiol. 29:653-658 (1983).
It's an adaptation of a slick idea published in Naturwissenshaften 17:402-403, for TEM, in 1962(!).
I always "heard" to focus the image with the beam in an overfocus condition (past crossover) for improved beam coherence. I find it difficult to focus at crossover anyway. Also, I don't think you can accurately assess focus or astigmatism with the beam at intensities suitable for most photography. Depends on the magnification and a host of other things, of course, such as your exposure time. I don't think there's any justification to focus at exactly the same intensity at which you will photograph. I too await further enlightenment from the list....
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } } I have one of those very basic "thought" questions on a procedure that I've } always taken for granted, but am now not so sure about. I had been trained } from Day One to always focus a TEM with the beam at crossover, then to } spread the beam until the exposure intensity is reached. The idea seems to } be that small focusing errors at crossover will be corrected as the beam is } spread and "depth of focus" is increased somehow. } } The reason I'm asking is that in recent years, almost no one else I've run } across focuses in this way, preferring instead to focus with the beam } already spread to the point at which the photograph will be taken (at least } as long as the image is bright enough to see for focusing). Out of } curiosity, I ran tests using both methods and have noted no clear } differences. } } Does anyone have any thoughts on this? Any ideas or theories about why one } method would be superior to the other? } } Sincerely, } Curious in Columbia } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } }
University of Central Florida Advanced Materials Processing and Analysis Center (AMPAC) Materials Characterization Facility (MCF)
The Advanced Materials Processing and Analysis Center (AMPAC) is seeking candidates for up to two positions as Assistant in Research to support the Materials Characterization Facility (MCF). Each individual must have at least a Bachelor degree from an accredited institution in an appropriate area of study related to surface science, materials characterization, ion beam analysis, or vacuum science, and preferably three years of experience in any of the above mentioned areas. A graduate degree in the aforementioned areas is preferred. The MCF houses an RBS, a heavy ion beam system, an Auger, an XPS, two FIBs and four SIMS instruments. Opportunities also exist with collaborative interaction with Lucent Technologies, Orlando, which houses an Auger, four FIB systems including a FIB/SIMS and two dedicated SIMS tools.
The person will be responsible for establishing, maintaining and operating ion beam and/or surface science laboratory infrastructure. Depending on the experience, the person should be able to instruct students, faculty, and staff on the use of equipment and in the interpretation of data, conduct independent research and develop surface analysis techniques. AMPAC is particularly interested in individuals desiring an academic setting to further their professional goals. Salary will be commensurate with experience. The applications will be reviewed beginning March 19, 2001 and will continue to be reviewed until the position is filled.
Applicants should send a vitae and a list of three references to Dr. Lucille Giannuzzi, Director MCF, 12443 Research Parkway, Suite 305, Orlando, FL 32826. UCF is an equal opportunity/affirmative action employer. As an agency of the State of Florida, all application materials and selection procedures are available for public review.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
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I have focused at a higher beam intensity than I use for exposing the micrograph for many years, and, in fact continue to do so on our Philips EM201. When we got a new Philips TwinLens CM12S in December of 1988, we started to have problems with slightly out of focus micrographs across the board - all experienced biological microscopists - about 10-12 of us - had the same problem. Philips, to their credit, tried repeatedly to find and solve the problem. They sent high level engineers, applications experts, attached a high mag TV camera, changed all of the high tension cabling, insulator, wehnelt, column liner, etc. over the course of nearly a year. Towards the end of 1989, in desperation and because the biological TEM applications experts were all busy, a materials science applications engineer came to the lab and made beautiful, perfectly focused micrographs. We could not duplicate his performance. Finally, he asked to watch one of our most experienced and well published microscopists work. When the Professor increased beam intensity to focus and stigmate, the applications engineer went ballistic! he maintained that the focus MUST be done at the intensity at which the micrograph is made. We did argue, but also agreed to test the hypothesis. He was, of course, correct (IN THIS PARTICULAR CASE). As we later found out, changing the C2 lens current also has a subtle effect on the objective lens current, changing focus enough to cause problems. Our service engineer was later able to demonstrate the effect (although it doesn't show up when monitoring the lens current page) and this strange phenomenon was written off to a design compromise which, apparently, physicists and materials scientists are used to and expect, but which biologists and cell biologists had never heard of. Something to do with short focal length objective lenses, low contrast which makes focusing more difficult, and compromises required to allow STEM and TEM to co-exist in the same column and be switched back and forth with a minimum of re-alignment. Anyone else have this experience?? William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
I spread the beam as much as possible in order to be able to see the changes during the focusing (usually the intensity is less than necessary to make a photograph, so, I have to condense the beam a little bit for photographing). I am using "three-chick" technique: rotating focus-control knob in both directions to find the position when you will clearly see the transition: "owerfocus-infocus-underfocus" on the background's granules. This technique works for me from x40K and up. At the lower magnification, I am using wobbler to focus (there is a "blind spot" at x30-40K). The "three-chick" technique is a good test how your instrument is aligned. The number of "clicks" depends from current setup and model of coarse. I simply could not focus correctly if the beam is condensed too much.
Sergey
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Well, I got my answer----right from one of the people who initially trained me back in the '80's (and, in fact, is responsible for me being in the field in the first place----that'll teach him).
Here is John Bozzola's reply to my post on focusing TEM's at beam crossover versus a defocused beam: "(T)his is a holdover from (our Hitachi) HU11A when illumination was a problem at the higher mags and when no focus wobbler was available for low mag work. On the other hand, it still DOES work for individuals who have diminishing eyesight and need the extra illumination and minimal depth of field. Unfortunately, it greatly increases the possibility of specimen damage and should be used only on hardy specimens.
"The bottom line is that if you have normal (or acute) vision, you won't need to do this in modern TEMs. Of course, I sometimes still "check" my focus by doing this..... old habit, I guess."
Now this is very interesting to me, because the first TEM I ever used was the Hitachi HU11AB. I was trained to focus carefully at crossover, then defocus until the proper exposure intensity was reached. Somehow over the years, I ended up with the notion that there was a property of electron "optics" that made this the preferred method of focusing because it minimized focusing errors by increasing the depth of focus. I really don't recall where this notion came from, but I trained dozens of students and others in this way of focusing. Now I wonder how many others were trained by them to do the same thing and will be just as stubborn about it as I have been.
Probably not much harm done, really, but this reminds me of one of my favorite stories about a woman cooking a ham for a family reunion picnic. Her daughter watched her mother cut off the end of the ham before putting it in a pan in the oven, and she asked why she did that. Her mother responded that that's how she was taught by HER mother, but she didn't really know why. She became curious, and at the reunion she asked her mother why she had always cut the ends off hams before cooking them. The response was that she had also been taught that way by her mother. Both curious now, they hunted up Great-grandmother at the picnic and asked her why she had cooked hams with the ends cut off. Great-grandmother answered, "I only had one pan and it was too short for the ham."
Lesson learned. Again.
Cheers, Randy
Randy Tindall Electron Microscopy Core University of Missouri
In terms of contrast transfer theory (i.e. when linear imaging allows approximating the imaging process as convolution with a point spread function), the lens transfer functon doesn't depend on convergence. So for example you shouldn't effect the defocus of the image by slight changes of the illumination convergence.
However, the convergence imposes a damping on the total contrast transfer, which gets more severe when convergence is greater. For details on this 'spatial coherence envelope' see any text on HREM. Because of this, the image quality will improve if you record with a less condensed beam (better coherence).
With a field emission gun, you won't be able to image at crossover, because the source will almost certainly be too small - these guns give the most coherent illumination available.
For a thermionic gun, the limiting factor will be: if the convergence is too great at crossover, you won't see much because everything will be blurred out by the coherence limitation of the incident irradiation. If this is the case, you should either change condenser apertures to decrease convergence (then you can focus, and/or stigmate at crossover), or spread the beam to improve coherence.
As far as I can see, there is no reason why one should or should not adjust or even record images at crossover (except that if the filament is a bit undersaturated, you will get uneven illumination). Getting the best possible coherence is important, but it doesn't really matter how one achieves this optically. Decreasing intensity is therefore good, to the extent that sample drift doesn't become too much a limiting factor.
Overfocusing the illumination rather than underfocusing is better, I would say (based on experience). I'm not sure why - perhaps some of the energy spread gets filtered out more effectively - at any rate for the same size beam one has more intensity underfocused than overfocused, so something is being cut out by overfocusing.
Wharton
} -----Original Message----- } From: Matthew Lynn [SMTP:mlynn-at-miami.edu] } Sent: Thursday, March 01, 2001 2:30 PM } To: 'Microscopy-at-MSA.Microscopy.com' } Subject: RE: Focusing } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I always "heard" to focus the image with the beam in an overfocus } condition } (past crossover) for improved beam coherence. I find it difficult to } focus at } crossover anyway. Also, I don't think you can accurately assess focus or } astigmatism with the beam at intensities suitable for most photography. } Depends on the magnification and a host of other things, of course, such } as } your exposure time. I don't think there's any justification to focus at } exactly the same intensity at which you will photograph. I too await } further } enlightenment from the list.... } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } (305)284-4736 } mlynn-at-miami.edu } } } On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. } [SMTP:TindallR-at-missouri.edu] wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi all, } } } } I have one of those very basic "thought" questions on a procedure that } I've } } always taken for granted, but am now not so sure about. I had been } trained } } from Day One to always focus a TEM with the beam at crossover, then to } } spread the beam until the exposure intensity is reached. The idea seems } to } } be that small focusing errors at crossover will be corrected as the beam } is } } spread and "depth of focus" is increased somehow. } } } } The reason I'm asking is that in recent years, almost no one else I've } run } } across focuses in this way, preferring instead to focus with the beam } } already spread to the point at which the photograph will be taken (at } least } } as long as the image is bright enough to see for focusing). Out of } } curiosity, I ran tests using both methods and have noted no clear } } differences. } } } } Does anyone have any thoughts on this? Any ideas or theories about why } one } } method would be superior to the other? } } } } Sincerely, } } Curious in Columbia } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } } }
We're analysing Cu wire here for purposes better known to ourselves. We are noticing, but not able to image interesting variations in the concentration of impurities, which suggests to us, either that the original material was inhomogeneous, or that drawing the wire has caused some kind of exsolution process. Have any of you encountered smilar things in metals? Refs? Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
To add to Wharton's explanation the reason overfocus is better than underfocus is that he has a strong objective lens (as most are these days). Many years ago underfocus would have been better.
What we are trying to achieve is parallel illumination not convergent (or divergent). Modern TEMs have strong objective lenses and use the prefield (field above the specimen) as part of their probe forming optics thus if there is a crossover above the specimen (overfocus)the prefield converges the diverging rays from this to form a parallel probe. In underfocus conditions the crossover is below the specimen and the probe on the specimen is made more convergent by the objective lens prefield.
If you have a Lorentz (low field for magnetic work) pole piece or an old instrument (pre 1970ish) the objective lens is weak and does not have this prefield effect so underfocus is more parallel.
Regards, Ron
On Thu, 1 Mar 2001 15:30:22 -0600 "Sinkler, Wharton" {WSinkler-at-uop.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } In terms of contrast transfer theory (i.e. when linear imaging allows } approximating the imaging process as convolution with a point spread } function), the lens transfer functon doesn't depend on convergence. So for } example you shouldn't effect the defocus of the image by slight changes of } the illumination convergence. } } However, the convergence imposes a damping on the total contrast transfer, } which gets more severe when convergence is greater. For details on this } 'spatial coherence envelope' see any text on HREM. Because of this, the } image quality will improve if you record with a less condensed beam (better } coherence). } } With a field emission gun, you won't be able to image at crossover, because } the source will almost certainly be too small - these guns give the most } coherent illumination available. } } For a thermionic gun, the limiting factor will be: if the convergence is too } great at crossover, you won't see much because everything will be blurred } out by the coherence limitation of the incident irradiation. If this is the } case, you should either change condenser apertures to decrease convergence } (then you can focus, and/or stigmate at crossover), or spread the beam to } improve coherence. } } As far as I can see, there is no reason why one should or should not adjust } or even record images at crossover (except that if the filament is a bit } undersaturated, you will get uneven illumination). Getting the best } possible coherence is important, but it doesn't really matter how one } achieves this optically. Decreasing intensity is therefore good, to the } extent that sample drift doesn't become too much a limiting factor. } } Overfocusing the illumination rather than underfocusing is better, I would } say (based on experience). I'm not sure why - perhaps some of the energy } spread gets filtered out more effectively - at any rate for the same size } beam one has more intensity underfocused than overfocused, so something is } being cut out by overfocusing. } } Wharton } } } } -----Original Message----- } } From: Matthew Lynn [SMTP:mlynn-at-miami.edu] } } Sent: Thursday, March 01, 2001 2:30 PM } } To: 'Microscopy-at-MSA.Microscopy.com' } } Subject: RE: Focusing } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I always "heard" to focus the image with the beam in an overfocus } } condition } } (past crossover) for improved beam coherence. I find it difficult to } } focus at } } crossover anyway. Also, I don't think you can accurately assess focus or } } astigmatism with the beam at intensities suitable for most photography. } } Depends on the magnification and a host of other things, of course, such } } as } } your exposure time. I don't think there's any justification to focus at } } exactly the same intensity at which you will photograph. I too await } } further } } enlightenment from the list.... } } } } Matt } } } } Matthew J. Lynn } } Center for Advanced Microscopy } } University of Miami } } (305)284-4736 } } mlynn-at-miami.edu } } } } } } On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. } } [SMTP:TindallR-at-missouri.edu] wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hi all, } } } } } } I have one of those very basic "thought" questions on a procedure that } } I've } } } always taken for granted, but am now not so sure about. I had been } } trained } } } from Day One to always focus a TEM with the beam at crossover, then to } } } spread the beam until the exposure intensity is reached. The idea seems } } to } } } be that small focusing errors at crossover will be corrected as the beam } } is } } } spread and "depth of focus" is increased somehow. } } } } } } The reason I'm asking is that in recent years, almost no one else I've } } run } } } across focuses in this way, preferring instead to focus with the beam } } } already spread to the point at which the photograph will be taken (at } } least } } } as long as the image is bright enough to see for focusing). Out of } } } curiosity, I ran tests using both methods and have noted no clear } } } differences. } } } } } } Does anyone have any thoughts on this? Any ideas or theories about why } } one } } } method would be superior to the other? } } } } } } Sincerely, } } } Curious in Columbia } } } } } } Randy Tindall } } } EM Specialist } } } Electron Microscopy Core Facility } } } W122 Veterinary Medicine } } } University of Missouri } } } Columbia, MO 65211 } } } Tel: (573) 882-8304 } } } Fax: (573) 884-5414 } } } Email: tindallr-at-missouri.edu } } } Web: http://www.biotech.missouri.edu/emc/ } } } } } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
we are looking for a working Nanoscope III (III or IIIa) controller together with the PC boards (Digital Instruments, Santa Barbara, USA). If somebody is going to clean out its lab please contact us directly by phone, fax or e.mail
With best regards,
Dr. Dmitry Cherny, PhD, Dr.Sc.
MPI for Biophysical Chemistry, Dept. of Molecular Biology am Fassberg 11, D-37077 Gottingen, Germany tel: +49(0) 551 201 1383; fax: +49(0) 551 201 1467 e.mail: dtcherny-at-mpc186.mpibpc.gwdg.de
does anyone have any suggestions on how to assess hydronephrosis with paraffin sections of kidney using a quantitative technique. equipment we currently use for other tissues include microscope, spot camera, digitizing tablet, camera lucida, SigmaScan, and image processing software plugins for adobe photoshop,
Nonuniform distribution of impurities in metal wires is not uncommon. There may be a core-to-surface variation, which may be caused by contamination (or de-contamination) of the surface layer during drawing or annealing . Variations in impurity concentration may also relate to original inhomogeneity in the billet prior to drawing. Segregation of impurities to dislocation-rich deformed areas is also a possibility.
What are the impurity elements that you are seeing? How are the concentration variations distributed spatially? (Radial gradients, irregular patches smaller than wire diameter or larger than wire diameter, wire bends vs. straight segments, or what?)
-------Roy ==================================== Roy Arrowood, Associate Professor Metallurgical and Materials Engineering UTEP, El Paso, TX 79968-0520 (915)747-6934 arrowood-at-utep.edu
} From a practical standpoint, wouldn't it be best to focus using the same conditions that are later used for taking the images, unless there are intensity or other issues that prevent that?
A few years back, when I was doing hi-res TEM, we would carefully set the conditions on the TV screen, then turn off the room fan, move away from the column, hold our breath and take a picture.
However, this may have been peculiar to hi-res materials work, where even tiny changes in focus can have dramatic effects.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Thursday, March 01, 2001 2:30 PM To: 'mlynn-at-miami.edu'; 'Microscopy-at-MSA.Microscopy.com'
In terms of contrast transfer theory (i.e. when linear imaging allows approximating the imaging process as convolution with a point spread function), the lens transfer functon doesn't depend on convergence. So for example you shouldn't effect the defocus of the image by slight changes of the illumination convergence.
However, the convergence imposes a damping on the total contrast transfer, which gets more severe when convergence is greater. For details on this 'spatial coherence envelope' see any text on HREM. Because of this, the image quality will improve if you record with a less condensed beam (better coherence).
With a field emission gun, you won't be able to image at crossover, because the source will almost certainly be too small - these guns give the most coherent illumination available.
For a thermionic gun, the limiting factor will be: if the convergence is too great at crossover, you won't see much because everything will be blurred out by the coherence limitation of the incident irradiation. If this is the case, you should either change condenser apertures to decrease convergence (then you can focus, and/or stigmate at crossover), or spread the beam to improve coherence.
As far as I can see, there is no reason why one should or should not adjust or even record images at crossover (except that if the filament is a bit undersaturated, you will get uneven illumination). Getting the best possible coherence is important, but it doesn't really matter how one achieves this optically. Decreasing intensity is therefore good, to the extent that sample drift doesn't become too much a limiting factor.
Overfocusing the illumination rather than underfocusing is better, I would say (based on experience). I'm not sure why - perhaps some of the energy spread gets filtered out more effectively - at any rate for the same size beam one has more intensity underfocused than overfocused, so something is being cut out by overfocusing.
Wharton
} -----Original Message----- } From: Matthew Lynn [SMTP:mlynn-at-miami.edu] } Sent: Thursday, March 01, 2001 2:30 PM } To: 'Microscopy-at-MSA.Microscopy.com' } Subject: RE: Focusing } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I always "heard" to focus the image with the beam in an overfocus } condition } (past crossover) for improved beam coherence. I find it difficult to } focus at } crossover anyway. Also, I don't think you can accurately assess focus or } astigmatism with the beam at intensities suitable for most photography. } Depends on the magnification and a host of other things, of course, such } as } your exposure time. I don't think there's any justification to focus at } exactly the same intensity at which you will photograph. I too await } further } enlightenment from the list.... } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } (305)284-4736 } mlynn-at-miami.edu } } } On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. } [SMTP:TindallR-at-missouri.edu] wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi all, } } } } I have one of those very basic "thought" questions on a procedure that } I've } } always taken for granted, but am now not so sure about. I had been } trained } } from Day One to always focus a TEM with the beam at crossover, then to } } spread the beam until the exposure intensity is reached. The idea seems } to } } be that small focusing errors at crossover will be corrected as the beam } is } } spread and "depth of focus" is increased somehow. } } } } The reason I'm asking is that in recent years, almost no one else I've } run } } across focuses in this way, preferring instead to focus with the beam } } already spread to the point at which the photograph will be taken (at } least } } as long as the image is bright enough to see for focusing). Out of } } curiosity, I ran tests using both methods and have noted no clear } } differences. } } } } Does anyone have any thoughts on this? Any ideas or theories about why } one } } method would be superior to the other? } } } } Sincerely, } } Curious in Columbia } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } } }
*************************************************** PROGAM: FOCUS ON MICROSCOPY 2001 ***************************************************
Dear colleagues,
The program of the conference "Focus on Microscopy 2001", April 1 - 4, 2001, Amsterdam, has been finalised and can be found on our Web-site
www.focusonmicroscopy.org
This conference is the 14th in a successful series of interdisciplinary meetings on 3D acquisition and 3D image processing and forms an effective meeting point for both developers and users of 3D-microscopy. The more than 120 contributions this year range from new developments in Coherent Anti-Stokes Raman Microscopy (CARS) and multi-photon excitation to the application of such techniques in biology, medicine and material sciences. This year "3D Microscopy of living cells and tissues" will receive special attention, which reflects the growing number of researchers that use GFP-techniques and life-cell imaging for their experiments.
A substantial instrument exposition including the main manufacturers in the field will accompany the conference.
Please, visit our web site http://www.focusonmicroscopy.org for further details concerning the program and the conference.
Looking forward to seeing you in 1st of April in Amsterdam, on behalf of the program committee,
We are working with embbeded cells in LR White with and without osmium. (Polimerization was acomplished at 60ºC). Our purpose is to detect gangliosides (GD3). Do you have any suggestion?
I also was told early on in my career that one should focus at the magnification the picture was to be taken at, though I find it difficult to do this in practice because the illumination is often too low. I usually compromise and focus with as little illumination as I can get away with.
I wonder whether the reason for the change in focus is not due to changes in the illumination, but changes in the specimen position. At different beam densities the degree of specimen charging may vary, thereby changing the position of the specimen in the lens field. This might explain why some microscopes are less sensitive to the specimen current density than others. Certainly the amount of specimen charging seems to be affected by the objective aperture, for example.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
Peggy: I don't know if you received any responses off-line, and I hope that Ronald Austin was able to provide you with a protocol for his microwave procedure (actually, if you have one available Ronald, I'd like a copy, please). Your question is most appropriate because that is what I am (frantically) in the midst of doing. Altho' "frantic" applies only to the deadline.... Sans a microwave procedure, I am doing the old time-tested methods. The only thing I make sure of is the buffer used to transfer the tissue from formalin. I add 6% sucrose to the buffers and carry that through until I get into osmium. Loss of membranes occurs in the formalin, and if you add the 6% sucrose it seems to help retain membranes in the final EM sample.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On 26 Feb 2001 15:06:20 -0500 (EST), "HARGER-ALLEN.MARGARET_J-at-INDIANAPOLIS.VA.GOV"-at-sparc5.microscopy.com wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Pathology lab. listers: are there any new/and/or superior techniques | for processing previously formalin fixed tissue for TEM? | Thanks, Peggy Harger-Allen | |TAB|email:harger-allen-at-indianapolis.va.gov |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
The pellets are from a hydrobiid snail found in highly mineralized warm desert springs and Posos in Mexico. I am interested primarily in the bacteria found on and in them but would also like to be able to inventory the entire contents of the pellets to help get a better picture of what part they play in their isolated ecosystem. I've done some light and SEM on samples but need to get inside to determine if the bacteria that break them down are carried internally or are picked up environmentally from the water. For sample preparation and sectioning I need to consider other contents like diatoms and stromatolite pieces and the high concentration of mineral solids in the samples. To start I plan to fix with glut/OsO4, dehydrate with acetone, infiltrate with propylene oxide and go into embed 812. After sections are imaged for non organic I planned on staining with lead citrate and comparing. Being new to TEM and not having a good idea of the precise osmolarity of the sample has left me puzzling over buffer selection (sodium cacodylate vs. phosphate), concentration, and times. We have an excellent EM technician here at NAU but it seems more my responsibility to make these determinations so any suggestions or references you could offer would be greatly appreciated. Thanks Pete Polsgrove Northern Arizona University pjp6-at-dana.nau.edu
we have just bought a Nikon CoolPix 990. It is an outstanding camera. One thing that we are disapointed it is a focusing problem when taking picture in Microscope. For any zoom setting and even in a manual mode, the micrograph borders always get out of focus, and in some situation with a Chromatic aberration. Does anyone have any idea how to eliminate this problem? The camera came with all necessary accessories for mounting in a microscope, e.g., c-mount, and MDC relay lens and remote cable.
Best regards,
Leonardo -- --- Leonardo Lagoeiro Departamento de Geologia Universidade Federal de Ouro Preto Ouro Preto, MG, 35400-000 Brazil E-mai: lagoeiro-at-degeo.ufop.br
The CoolPix 990 is indeed a nice camera. Unfortunately, it is not ideal for microscopy. It will work in this type of application but the results are mixed and the effort can be great.
If you zoom out to infinity, you should get best results. The key problem I have found is obtaining consistent exposure values. Preview with the puck looks good but the captured image is over exposed. Trying the best of five sometimes helps. But i have relegated the 990 to travel and snapshots. I don't think that it is ready from prime time microscopy. YMMV.
gary g.
http://photoweb.net
At 01:35 PM 3/2/01, you wrote:
} Dear All, } } we have just bought a Nikon CoolPix 990. It is an outstanding camera. One } thing that we are disapointed it is a focusing problem when taking picture } in Microscope. For any zoom setting and even in a manual mode, the } micrograph borders always get out of focus, and in some situation with a } Chromatic aberration. } Does anyone have any idea how to eliminate this problem? The camera came } with all necessary accessories for mounting in a microscope, e.g., } c-mount, and MDC relay lens and remote cable. } } } Best regards, } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br
I am trying to distinguish between neurons and glia in cortical tissue that has been embedded in epoxy resin (LX112/NMA/DDSA/DMP-30). Does anyone know how to do this on semithin sections? Toluidine-Blue staining imparts some subtle differences in the appearance of chromatin in the nuclei of neurons vs. glia, but i am looking for a staining procedure that provides a more obvious color difference (without going all the way to immunohistochemistry).
It is my understanding also that KMnO4 will react with NMA (even if i disolve the resin out??), so those procedures are out, unless I can find a substitute for KMnO4.
let me make two observations and see if you agree:
1) The Coolpix (and any other camera like it) is originally designed for snapshots. That means, that the emphasis is on "nice" pictures under normal (outside or flash) conditions. This does not necessary mean, that one can take good scientific images. This may be the reason that you had problems with exposure time. The camera may take a weighted measurement to calculate exposure time, for example only from the middle. If you take a snapshot outside, the most important feature is likely to be found somewhere around the center of the image. So it's important to get that part right. That might not apply to photography on a microscope.
2) The quality of the picture is affected by the weakest link in the chain. My suspicion is, that the lens on the camera is far inferior to anything you put on a microscope. For example, we spend a lot of time (and money) to develop and make our own lenses for our TEM cameras. I don't see the point of spending thousands of Dollars on good microscope lenses and then have all that negated by an inferior lens on the camera.
One word to Nikon: I am not trying to knock this camera, it is probably a very good camera (I don't know it myself). My remarks are general and apply to other camera manufacturers (Kodak, Canon, Fuji, etc.). This was just a thread which mentioned the Nikon camera. Plus, all these ramblings are my personal opinion.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Friday, March 02, 2001 10:20 PM To: Leonardo Lagoeiro Cc: MSA listserver
The CoolPix 990 is indeed a nice camera. Unfortunately, it is not ideal for microscopy. It will work in this type of application but the results are mixed and the effort can be great.
If you zoom out to infinity, you should get best results. The key problem I have found is obtaining consistent exposure values. Preview with the puck looks good but the captured image is over exposed. Trying the best of five sometimes helps. But i have relegated the 990 to travel and snapshots. I don't think that it is ready from prime time microscopy. YMMV.
gary g.
http://photoweb.net
At 01:35 PM 3/2/01, you wrote:
} Dear All, } } we have just bought a Nikon CoolPix 990. It is an outstanding camera. One } thing that we are disapointed it is a focusing problem when taking picture } in Microscope. For any zoom setting and even in a manual mode, the } micrograph borders always get out of focus, and in some situation with a } Chromatic aberration. } Does anyone have any idea how to eliminate this problem? The camera came } with all necessary accessories for mounting in a microscope, e.g., } c-mount, and MDC relay lens and remote cable. } } } Best regards, } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br
Does anyone have a GaAs cathodoluminescence attachment in a SEM? This means a photomultiplier optimized to 8500A, an S1 photomultiplier. I would like to look at light emitted from a GaAs diode under power. Or does anyone know of a lab that does have GaAs cathodoluminescence as a SEM attachment.
I would like to thank those colleagues who kindly provided their advice and shared their experience with us regarding the problems of vibratome brain sectioning that I posted on the list server a while ago.
We now use double-edge feather blades that we purchased from Ted Pella, the quality of sections are much improved, also sometimes it seems helpful at low amplitude rather than high one.
Xinran Liu, M.D., Ph.D.
Center for Basic Neuroscience UT Southwestern Medical Center 6000 Harry Hines Blvd., NA4. 214A Dallas, TX 75390-9111
Arron: Try using Araldrite 502 for your resin. It does not contain NMA. This should eliminate your concern with using KMnO4. The mixture I use is a 1-1 Araldrite 502 and DDSA, mix well then add your DMP-30 and mix again. Don't worry about the air bubbles.
Ron Austin (Research Associate) Dept of Pathology LSU Medical Ct. Shreveport, LA rla-at-mindspring.com
-----Original Message----- } From: "PostMaster {rla-at-mindspring.com} "-at-sparc5.microscopy.com [mailto:"PostMaster {rla-at-mindspring.com} "-at-sparc5.microscopy.com] Sent: Saturday, March 03, 2001 2:43 PM To: rla-at-mindspring.com
-----Original Message----- } From: Ronald Austin [mailto:rla-at-mindspring.com] Sent: Saturday, March 03, 2001 2:41 PM To: Microscopy Society of America
Email: pjp6-at-dana.nau.edu Name: Pete Polsgrove School: Northern Arizona University
Question: I'm looking for an protocol for imaging fecal pellets. The pellets are from a hydrobiid snail found in highly mineralized warm desert springs and Posos in Mexico. I am interested primarily in the bacteria found on and in them but would also like to be able to inventory the entire contents of the pellets to help get a better picture of what part they play in their isolated ecosystem. I've done some light and SEM on samples but need to get inside to determine if the bacteria that break them down are carried internally or are picked up environmentally from the water. For sample preparation and sectioning I need to consider other contents like diatoms and stromatolite pieces and the high concentration of mineral solids in the samples. To start I plan to fix with glut/OsO4, dehydrate with acetone, infiltrate with propylene oxide and go into embed 812. After sections are imaged for non organic I planned on staining with lead citrate and comparing. Being new to TEM and not having a good idea of the precise osmolarity of the sample has left me puzzling over buffer selection (sodium cacodylate vs. phosphate), concentration, and times. We have an excellent EM technician here at NAU but it seems more my responsibility to make these determinations so any suggestions or references you could offer would be greatly appreciated. Thanks Pete Polsgrove
1) The camera is of course intended for general purpose photography. Snapshots, indoors and outdoors. It has a miniature flash which is good for about ten feet distance. For microscopy, of course the flash is turned off.
2)There are some macro zoom situations where the camera does a nice job. But for compound transmitted and reflected work, I have not found it to be satisfactory. Optem has made a high quality interface for the CoolPix which interfaces to Zeiss, Olympus and many other 'scopes. I have a common optical tube with C-mount to CoolPix for Axioskop and Olympus. I cannot fault this adapter system at all. But as you point out, the native CoolPix optics are not intended for discriminatory microscopy work. At about $700 for the Optem adapter, the CoolPix package with remote release and AC adapter is not cheap.
I recently took 765 pix with the CoolPix on a trip to Australia. It did a super job. I used two 128MB AVL CF modules. Some images were in highest JPEG resolution and some were in XGA fine. All print on an Epson Stylus Photo 2000P with outstanding results. And, of course, they go to the Web without any problem.
The Pixera Penguin 600CL is quite a different camera. It is highly suitable for microscopy, but also will do excellent macro work with an inexpensive C-mount zoom lens. Optem also makes the adapter for this camera to the Zeiss and Olympus microscopes. Same high quality and fine results. I have done multiple slices and stitched them together for a final TIFF file size of 110MB.
The Pixera is non-interpolated 5.8M pixels using their Diractor prism device. At 2776x2074 pixels, it generates a 17.5MB TIFF file. In 16-bit mode, the size is twice that. As I mentioned in a prior post, their software supports multiple capture averaging and integration. It also does variable sized spot metering and average metering. The CL version has Peltier cooling and exhibits exceptionally low thermal noise. Four frame averaging reduces noise even further.
The Pixera is 10-bits per color while the Nikon MX1200 is 8-bits per color. The word on the street is that the next release of the MX1200 will be 10-bits per color. Whether this is worth anything to a particular user is their own decision.
Being a professional Nikon shooter for many years, I am no longer an intransigent fan. In fact, I have dumped nearly all of my Nikon SLR equipment in favor of Contax. This of course has nothing to do with microscopy. But the point is that in my extended experience, Nikon is struggling to develop new products, which by specifications, leapfrog their competition--while in practice, they do not.
Try before buy is a good operative in this respect. I really don't care who makes what, as long as it is good stuff. Sorting through all the marketing hype is a chore. Sometimes even working with the equipment is a chore. But the actual results of using products is much more telling than sales pitches or product brochures.
gary g.
http://photoweb.net
At 06:54 AM 3/3/01, you wrote:
} Gary, } } let me make two observations and see if you agree: } } 1) The Coolpix (and any other camera like it) is originally designed for } snapshots. That means, that the emphasis is on "nice" pictures under normal } (outside or flash) conditions. This does not necessary mean, that one can } take good scientific images. This may be the reason that you had problems } with exposure time. The camera may take a weighted measurement to calculate } exposure time, for example only from the middle. If you take a snapshot } outside, the most important feature is likely to be found somewhere around } the center of the image. So it's important to get that part right. That } might not apply to photography on a microscope. } } 2) The quality of the picture is affected by the weakest link in the chain. } My suspicion is, that the lens on the camera is far inferior to anything you } put on a microscope. For example, we spend a lot of time (and money) to } develop and make our own lenses for our TEM cameras. I don't see the point } of spending thousands of Dollars on good microscope lenses and then have all } that negated by an inferior lens on the camera. } } One word to Nikon: I am not trying to knock this camera, it is probably a } very good camera (I don't know it myself). My remarks are general and apply } to other camera manufacturers (Kodak, Canon, Fuji, etc.). This was just a } thread which mentioned the Nikon camera. Plus, all these ramblings are my } personal opinion. } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Friday, March 02, 2001 10:20 PM } To: Leonardo Lagoeiro } Cc: MSA listserver } Subject: Re: Nikon CoolPix 990 } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } } } Subject: } } } } SEM views of gold paint } } } } Content: } } } } Does anyone have any views of modern gold flake (real gold, not brass } } flake) used in making paintable substitutes for gold leaf? } } } } John Twilley } } Conservation Scientist }
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The candidate will be working in a diverse industrial R&D-environment with several links to university labs. He/she will be involved in a multidisciplinary team working on several nanostructured applications. Main focus of the work will be on microscopical and crystallographic (diffraction) techniques. In order to meet the technical requirements, the candidate should have a PhD in materials science, experience with TEM- and diffraction-techniques, and have a background in crystallography of inorganic materials. He/she should be a good teamplayer to function in a multi-disciplinary team, and be fluent in English and/or Dutch.
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For selection procedures, eligibility criteria and so on, please see homepage Marie Curie Industry Host Fellowships: http://www.cordis.lu/improving. Please note in particular that candidate fellows must be nationals of an EU Member or Associated State, or have resided in the EU for at least five years immdiately prior to their selection by Agfa.
I don't want to wake up a recent discussion (I was away in the last weeks), but I take advantage of this subject to askh a question about an other aspect of this subject. Sometimes you have so old material and maintenance services are laughing when they see what your are working with and/or on the other hand (with new materials also) it's faster and cheaper to do cleaning and (basic) maintenance yourself (and I like to do it myself !).
My question : What kind of grease/oil, where, on a LM ?
Manufacterer and maintenance services don't like to answer such a question. They say its complicate, there are much type of greases etc etc... So, does someone know something about that.
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
I was quite surprised to see the problems that are being discussed in relation to TEM focus, particularly as I was with Hitachi in the HU11 days, lets keep it simple and try to help?
When setting the image focus you should be aware of a number of problems that may complicate matters.
1. To try to focus at condenser cross over is absolutely the wrong thing to do! i) At crossover there is the poorest illumination coherence, this will lead to soft images, not out of focus but not optimised either. ii) At crossover there is a space charge effect (too many electrons in a small area and they repel each other) which may cause a false focus. (Just look at any image at crossover and then overfocus the condenser - you can see more detail!) iii) To focus at crossover and then change to a lower intensity is asking for trouble as the change in heating effect is likely to cause specimen drift and flexing which may change the contrast of material science specimens.
2. You MUST focus at the photographic magnification. Focus is the matching of the focal lengths of the objective and diffraction lenses. If you focus at a higher level and there is a diffraction lens change when dropping the magnification, you will find a matching objective correction will be required or your image will be out of focus.
3. What is best FOCUS, not always true focus? To a materials scientist it will almost certainly be true focus, no Fresnel fringes, as determined by the focus wobbler. They will set their intensity in advance as an intensity change may cause the material to flex and change in contrast. To the biological scientist it will be a degree of underfocus determined by the specimen's organelle density, its thickness, the kV, the objective aperture size and most important, the magnification; this is optimum under focus, the defocus where the (white) Fresnel fringes enhance the high contrast areas!
When taking a photograph the ideal conditions (as used in all the manufacturer's demo laboratories that I have worked with, Hitachi, JEOL and ISI) require the photograph to be taken at the same intensity level as when the focus is set. WHY? Well, once an intensity is set the operator may focus and during the focus procedure they should be concentrating sufficient to spot any image drift; it is most important to see the image under the conditions that you are going to photograph. So what will the excuses be?
1. "It is too dim to focus" - then i) increase the emission current, many people run at too low a current and make the task of optimising the instrument settings much more difficult ii) increase the kV which would increase the intensity too ii) re calibrate the photographic system to allow a focus intensity that is suitable for most operators
2. "The negatives are to dark" - i) the denser the negative the higher the contrast, so may be you could increase the kV, have a more stable specimen and an even brighter image?
3. Difficult one this - "the pathologists like a very bright image" - yes I know I guess it comes from looking down light microscopes all day? i) try 1 or 2 above - "yes but the pathologists like a high screen contrast" - yes I know but you should explain that no one publishes the screen, it it the photographic record that is the most important item(?) and we can get stacks of contrast from modern printing/publishing systems.
Regarding the reported overlap in currents between condenser and objective it is true, however the result is usually more of an image shift than a focal change in my observations, but yet another reason to focus and stigmate at the photographic intensity.
So to conclude I would
1. Always focus and stigmate at the photographic illumination level, if a biologist determine and plot out the optimum under focus for your material over your normal magnification range. 2. Always use the second condenser overfocus from crossover for high coherence and combine this with a 2 micron spot size for biology. 3. Set up the instrument's photographic procedure to suit as many operators as possible so that they too may focus at the photographic level. 4. Increase the emission current to give a higher brightness making point 2 far easier to use 5. Always use a higher kV (if 60, wow please go to 80, if 80 try 100) and work to obtain the best results on a photograph not trying to optimise for the screen. 6. To focus at low magnifications try removing the objective aperture and focus without a binocular, or a wobbler, looking for the very strong minimum contrast effect.
Well I hope this helps, its standard information on our courses, guess we do too much SEM in the States!
Regards
Steve Chapman Senior Consultant Protrain For consultancy and training by professionals World Wide Tel +44 1280 814774 Fax +44 1280 814007 www.emcourses.com
} } I have several questions for an AFM microscopist; } } 1. Can AFM really measure surface modulus?
Probably - but there are a number of factors which may have to be taken into account. These will include the nature of sample, any additional interactions between the sample and the scanning probe tip, the nature of the tip and SPM cantilever and so on.
} } 2. If yes, what is the mode of operation or measurement? } In terms of imaging modes (i.e. those where you are obtaining an image along with surface property information) there are two that may provide some surface modulus information.
i) Phase imaging (see for e.g.http://www.di.com/AppNotes/Phase/PhaseMain.html) is capable of providing a surface map based on the viscoelastic properties of the surface. Therefore the image will probably be a convolution between both the surface modulus and other surface properties (for example elasticity and whether the sample has any interactions with the tip). In this mode the probe is tapped into the surface and the lag between the voltage driving the tip oscillation and the actual tip oscillation provides this information. (It is particularly difficult, if not impossible to quantise the results obtained).
ii) Pulsed force should be able to deconvolute between sample stiffness and sample adhesion. http://www.thermomicro.com/tech/modes/pfm.htm should provide more information about this mode. However whether true values could be measured would again be a matter of some discussion.
Further mode of scanning probe microscopes are the spectroscopic ones in which the probe tip is move towards and away from the sample and the tip response measured. In AFM this mode is force-distance (see for e.g. http://www.thermomicro.com/tech/modes/fvsd.htm). This should again provide some information regarding the surface modulus. And with some tip calibration may provide something akin to a quantitative measurement.
With all of these mode the cantilever type will have an effect on the amount of information available. If the cantilever is less stiff than the surface of interest your measurement will generally be of the Young's modulus of the cantilever.
Nanoindenters may provide a more accurate measurement. http://freespace.virgin.net/micro.materials/ is an example. There are some companies that offer methods of converting commercial SPMs towards a nanoindenter.
} 3. Is it a quantitative measurement? Is it an absolute measurement } or a relative measurement?
My cynical opinion would be that most measurements available from a more qualitative than quantitative, though many would disagree. For example in this case, the area of tip-sample contact is unlikely to be able to be measured accurately but will have a major effect on your measurements, You may however be able to obtain some numbers that can be converted to an absolute measurement after careful consideration.
} } 4. Can the results be correlated to the (bulk) modulus measured by } rheometer?
This would, I presume, depend upon the material.
I hope this is of some use.
Giles
---------------------------------------------------------------------------- --------------------------- Dr. Giles Sanders Zeneca / SmithKline Beecham Centre for Analytical Sciences Chemistry Department Imperial College of Science, Technology and Medicine London SW7 2AY
I have had a request from a customer to allow remote access to our SEM in operation ( Hitachi S3500N ). The software they are proposing to use is "Timbuktoo" (?). They wish to set up three-way access across the internet along with a web-cam. I would appreciate advice from anyone who has tried this already. It would be useful to know what software has been used elsewhere and what problems have arisen. There may be better software available to carry out this function, but I have not approached this problem until now and am a bit in the dark. All advice, comments, etc. will be much appreciated.
Thanks.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
Dear Colleagues We have a CM200 (S)TEM equipped with EDAX IIs, 4pi digital imaging system and Gatan GIF. We are considering buying an integrated data acquision system for this microscope. ES Vision System from Emispec Systems, Inc. is under consideration . Comments from users of this ES Vision System would be highly appreciated. Of particular interest is operation of the Gatan Image Filter; would the autofilter functions still be available? Are there any other such systems out there? TIA Anita
To the person who is polymerizing the L.R. White at 60 degrees C:
Turn down the oven. If you want to do some sort of gold labeling on your sections try polymerizing the blocks at 45 degrees C for 2 to four days. Be sure to do this in some sort of BEEM capsule, as the stuff will not get hard if it is exposed to air. This type of plastic is not very stable in the electron beam, pick up your sections on carbon coated gold grids. Good luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
First off, I suppose that Nestor or the folks at ORNL will have much more to say about this since they have essentially developed and refined telemicroscopy. However, I have some experience with Timbuktu that I will relate.
We have been using Tibuktu for a number of years since I first heard of it several years ago at an MSA workshop on telemicroscopy. It is designed for remote observation or control of computers in general and has been borrowed for the microscopy application. There are several other products also available for the same work. Norton has PC anywhere. ATT has developed VNC which has been mentioned here before and is free.
Timbuktu (and the others) simply requires a connection to the Internet. Whatever happens on the screen is potentially available to the world once a client logs in with a password. This can slow down the host computer as it also has to transmit a copy of the screen.
I don't know about PC Anywhere, but Timbuktu is much faster on the client side than is WinVNC, even on a LAN. There can be a lot of data to transmit and it flat out takes a while to do so. Well-written code can do a lot to speed that up. You should also consider the connection between your PC and your client. If they have only a modem connection, refresh rates will be painfully slow.
I couldn't guarantee how well Timbuktu (or any other program) would work on your scope. I seem to recall that there were some windows whose contents were not available for Timbuktu to share. It may be that the SEM control programs were doing non-standard operations that circumvented the normal Windows system. Presently, I am able to view video windows whether they be from Oxford's AutoBeam, Quartz's PCI or Dazzle's video preview. But don't expect refresh rates measured in frames per second, at least not with a remote control/viewer program. Still, it could be a good alternative compared to more expensive solutions or to traveling to the site.
BTW, we also picked Timbuktu since it worked with either Macs or Windows machines.
I cannot tell you much about web cams. I know some systems are available that are stand-alone and practically plug and play. I don't know that I would want to burden my microscopy or EDS computer with the extra task. Others may have more to say.
There, that ought to be worth a couple of cents.
Warren
At 03:57 PM 3/5/2001 +0000, you wrote: } Hi, } } I have had a request from a customer to allow remote access to our SEM in } operation ( Hitachi S3500N ). The software they are proposing to use is } "Timbuktoo" (?). They wish to set up three-way access across the internet } along with a web-cam. } I would appreciate advice from anyone who has tried this already. It would } be useful to know what software has been used elsewhere and what problems } have arisen. There may be better software available to carry out this } function, but I have not approached this problem until now and am a bit in } the dark. } All advice, comments, etc. will be much appreciated. } } Thanks. } } Best wishes, } } Colin
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Timbuktu is a remote control software package made by Netopia.
http://www.netopia.com/software/
There is a free trial version which can be downloaded.
Whether it will work for you, I don't know. One of the key issues is real-time image transfer for mag and focusing. The interconnect between the SEM and the remote user is a major factor in response time. Slow analog modems are not going to be all that responsive. If running on a LAN or intranet, that would make a big difference.
A similar product is PC-Anywhere.
gary g.
At 07:57 AM 3/5/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You can do it very cheaply (i.e. free if you have Windows machines) using NetMeeting. Our LEO 1530 was easily set up to do this. You can do "over the shoulder" microscopy where the remote location observes. You need fast bandwidth for control and the instrument must be fully digitally controlled. We have tried it out with remote locations within our company and it works, but do not use it very much (i.e. never).
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Colin Reid [mailto:creid-at-truxa1.tcd.ie] Sent: Monday, March 05, 2001 10:57 AM To: MSA.Microscopy.Com (E-mail) Subject: Remote Access to SEM
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html --------------------------------------------------------------- --------.
Hi,
I have had a request from a customer to allow remote access to our SEM in operation ( Hitachi S3500N ). The software they are proposing to use is "Timbuktoo" (?). They wish to set up three-way access across the internet along with a web-cam. I would appreciate advice from anyone who has tried this already. It would be useful to know what software has been used elsewhere and what problems have arisen. There may be better software available to carry out this function, but I have not approached this problem until now and am a bit in the dark. All advice, comments, etc. will be much appreciated.
Thanks.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
to make a guess, you should can give some more information. What impurity elements did you find/expect ? Can you make any suggestion about the size of any kind of inhomogeneity - conzentration, precipitation or what ever ?
S.Guder
} We're analysing Cu wire here for purposes better known to ourselves. We } are noticing, but not able to image interesting variations in the } concentration of impurities, which suggests to us, either that the } original material was inhomogeneous, or that drawing the wire has caused } some kind of exsolution process. Have any of you encountered smilar } things in metals? Refs? } Malc. } } -- } Dr MP Roberts Phone: [+27](0)46 603 8313 } Dept of Geology Fax: [+27](0)46 622 9715 } Rhodes University Cell: 083 4060 262 (usually off) } 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za } SOUTH AFRICA
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr.-Ing. Susanne Guder Technical University Munich Department of Materials in Mechanical Engineering D - 85747 Garching Phone: + 49-(0)89-289-15308/15338 Fax: + 49-(0)89-289-15301 Email: guder-at-wm.mw.tum.de ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I own a zeiss 63X water immersion lens. If the coverslip thickness is 0, does that mean that this is meant for direct viewing of specimens in water? I purchased it with that in mind (I work in marine environments, felt it would be convenient to observe microorganisms and microanimals directly). Using a coverslip doesn't seem to work well, if at all. For some specimens it is nice, but any focusing movement will push specimens out of the way. Nice for zooxanthellae in a small sea anemone, for example, and was pretty good for a thick broth of unicellular green algae.
I am trying to get a feel for this objective. WOuld appreciate any information possible.
Alan Davis adavis-at-saipan.com
On Fri, 13 Oct 2000 08:51:07 -0400 Gary Radice {gradice-at-richmond.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I've had some experience with water immersion lenses. Their main } advantage is higher numerical aperture than dry lenses at } equivalent } magnification, so better theoretical resolution. They don't offer } quite as good resolution as oil immersion lenses, but they tend to } have longer working distances, I believe, and don't have the } messy } clean-up of oil immersion lenses. So, yes, they do have some } distinct } advantages. } } However, as others have pointed out, you may already be able to } see } everything you need to see with your current lenses. And nothing } beats a test-drive. } -- } Gary P. Radice gradice-at-richmond.edu } Associate Professor of Biology 804 289 8107 (voice) } University of Richmond 804 289 8233 (FAX) } Richmond VA 23173 http://www.science.richmond.edu/~radice
-- adavis-at-saipan.com 1-670-235-6580 Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, NMI
I have steadily endeavored to keep my mind free, so as to give up any hypothesis, however much beloved -- and I cannot resist forming one on every subject -- as soon as facts are shown to be opposed to it. -- Charles Darwin (1809-1882)
Has anyone priced the XL30 high vac FESEM and the EFESEM or variations which use LaB6? What is the general price range for these, and which Hitachi models would closely compete?
A normal (upright) epifluorescence microscope excites fluorescence in a specimen by passing the excitation light to the top of the specimen via the objective lens, which is used to focus the light into an intense spot exactly at the point on the specimen which is to be viewed. Fluoresced light from the region of interest is collected by the objective and is viewed or photographed normally after filtration to remove any excitation light and unwanted fluorescence wavelengths. The excitation illumination is commonly provided by a high-pressure mercury vapour lamp, which conveniently provides very high intensity illumination in UV, blue and green wavelengths. Other high-intensity sources are now also used, including xenon lamps, lasers (e.g. in confocal microscopes, which are a highly-derived type of epifluorescence microscope) and even tungsten-halogen lamps. The separation of excitation and fluorescence wavelengths is usually accomplished using a dichroic beam splitter which reflects green, for example, and transmits orange/red. Further fine-tuning of the excitation wavelengths may be done using dichroic filters. Many microscope manufacturers provide filter systems as beam-splitter cube assemblies with all the filters and dichroic reflectors required for a particular fluorochrome installed in one pre-aligned package for easy exchange. The fluoresced light may be further filtered for viewing or photography by "barrier" filters either of dichroic type, or of coloured glass or sometimes gelatin (e.g. Kodak Wratten).
Other than that, an epifluorescence microscope can be a normal compound microscope, but because efficiency of illumination and collection of light is important, and is limited by the objective lens, fluorescence microscopes usually employ high quality objectives with very high numerical aperture. If UV is used, it may be necessary to select ojectives with very low UV absorption.
Hope this helps Chris Date sent: Mon, 5 Mar 2001 17:25:50 -0600 To: Microscopy-at-sparc5.microscopy.com } From: ileyozerlat-at-yahoo.com ()
The FEI XL30SFEG costs in the region of £250k in UK The closest Hitachi competitor is the S4700. The FEI is thermal field emission, the Hitachi is Cold cathode field emission. There is no Hitachi ESEM, but they do a variable pressure tungsten filament SEM.
Philips / FEI service in UK is excellent. I have no first hand knowledge of the Hitachi service operation, but believe it is also first rate. Hitachi quote much lower prices for their service contracts than FEI. They tell me that their engineers generally have little to do because the instruments are so reliable! I don't know whether this is a true reflection of the cost of ownership, however.
Date sent: Tue, 06 Mar 2001 02:03:30 -0800 To: MSA listserver {Microscopy-at-sparc5.microscopy.com} } From: Gary Gaugler {gary-at-gaugler.com}
Hello,
Our Siemens Elmisjop 102 is presently out of order. We are looking for several parts of a Siemens Elmiskop 102 which must be replaced.
Parts List C73000 - E3174 - C1 - 1
Part N°335 - Objective current control fine / superfine - cat n° C72315 - A29 - A4
Part N°333 - Objective current control coarse I level - cat n° C72315 - A31 - B1
Part N°334 - Objective current control medium II level - cat n° C72315 - A31 - B2
Part N°455 - P.C. board V9 "Objective controller" LR - cat n° C72302 - A41 - A2
Many thanks in advance
Best regards
Philippe Drouillon Solvay Research and Technology Electron Microscopy and Image Analysis - Coordinateur Informatique de Division Rue de Ransbeek, 310 B-1120 Brussels (Belgium) phone : (00 32) 2 264 24 47 mailto:philippe.drouillon-at-solvay.com
We have an Emispec Vision system on our CM200FEG TEM-STEM to acquire spectrum lines and spectrum images from our Oxford EDS and Gatan GIF. We tune the GIF using the autofilter functions, and setup the GIF with ImageFilterControl, but let ESVision read out the GIF multiscan camera during acquisition. All in all, we are quite pleased with the system. If you would like to discuss this offline, please contact me directly.
} Hi, } } I am trying to distinguish between neurons and glia in } cortical tissue that has been embedded in epoxy resin } (LX112/NMA/DDSA/DMP-30). Does anyone know how to do } this on semithin sections? Toluidine-Blue staining } imparts some subtle differences in the appearance of } chromatin in the nuclei of neurons vs. glia, but i am } looking for a staining procedure that provides a more } obvious color difference (without going all the way to } immunohistochemistry).
I think you are going to have to rely on morphology alone. Get a copy of Peters, Palay and Webster, "Fine Structure of the Nervous System". You should be able to extrapolate from the low mag EMs to LM. You could probably do an immuno for GFAP (most but not all astrocytes) but that still leaves oligos and microglia. I don't know of a good immuno for these cells on plastic sections. Plus, there will be smaller neurons to contend with. Sorry!
} It is my understanding also that KMnO4 will react with } NMA (even if i disolve the resin out??), so those } procedures are out, unless I can find a substitute for } KMnO4.
And the KMnO4 is for ??
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
There is an official open position now at Genentech (the one I alluded to in an earlier post, but HR is not too speedy over there) . The posting is below and can also be found on their website: http:// www.genentech.com {http://www.genentech.com/} .
For immediate consideration please send a c.v. to Peter Schow at pschow-at-gene.com {mailto:pschow-at-gene.com} . He is not on the list, so please email him directly. I have already forwarded previous resumes I received to him, so no need to resend those.
Position: Research Associate- Cytometry Lab Requisition #: 01-0003250
Description Research Associate position available in the Research Cytometry Lab. The lab support core facility, collaborative and basic research functions. Primary responsibilities include operation and maintenance of flow cytometers and/or imaging instrumentation and may cover all aspects of data acquisition, analysis, interpretation and presentation in the contexts of basic and applied research. Lab supports Coulter and BDIS flow cytometers, Leica (confocal) and Nikon (digital fluorescence) microscopes and both Mac and PC computers. The person will work in a dynamic and interactive environment requiring a strong teamwork orientation, good communication skills, flexibility and the ability to interact with a diversity of research personnel.
Requirements Position requires a BS/MS and extensive experience with flow and/or image cytometry. Good working knowledge of cell and molecular biology necessary. Industrial experience is desirable.
Holly Aaron Head, Berkeley Imaging Center University of California, Berkeley Dept. of Molecular and Cell Biology 447 LSA Berkeley, CA 94720-2751 hollya-at-socrates.berkeley.edu
Northwestern University is drafting a complaint against a vendor of microscopy products, AMT. Has anyone else had experience with lawsuits against vendors?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
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Nestor Your Friendly Neighborhood SysOp
=========================================== Dr. Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
I am considering this, but have read about issues of contrast perception and/or low light vision. It occurred to me that microscopists, particularly EM types, depend on their eyes in low-light situations more than any other profession I can think of, with the possible exception of cat burglars.
Has anyone out there had LASIK or similar procedure, and has it been positive or negative for your work? Did you notice reduced contrast sensitivity?
Thanks for any personal experiences; since it's not a direct microscopy query, please send replies to me and (if there's interest) I'll summarize for the list.
Rick Mott (myopic, -7 or so, with some astigmatism)
We have a ten year old cryo-stage and transfer device for sale.
It is a BioRad E7400 comprising: Pump valve controller Sputtering module Evaporation module Edwards E2M5 rotary pump x2 Valve block Transfer/coating unit with dewar Stage cooling unit with dewar Specimen insertion rod ie all you need to go cryo-SEMing.
The rod and flanges suit a JEOL 6400 SEM. The unit was functioning and in good condition prior to replacement. Any offers?
Eric Hines Microscopy Centre CSIRO Entomology Canberra.
Dear all, I send you the following message written by a serious student. I hope that someone can offer a possibility to her. Thanh you very much. All the best, Elena Belluso
My name is Elisa Fornero and I doing my Degree thesis, at Turin University - Italy- (Dept. of Mineralogical and Petrologic Sciences), on "INORGANIC FIBERS IN LUNG TISSUES: MINERALOGICAL CHARACTERIZATION, COMPARISION WITH NATURAL ASBESTOS AND CALCULATION OF DIFFERENT FIBROUS MINERALOGICAL SPECIES" (the samples of tissues derived from people not professionally exsposed in order to value the environmental pollution of fibrous minerals). During my thesis I extensively worked with SEM and EDS and I learned to prepare the samples from lungs tissues filtrated. I would like to have some international experience taking part, about for two months between April and June 2001, to a stage in a foreigner University. I would like found a laboratory where develop my knowledge in this field. If there is anyone interested to host me, I can send my curriculum vitae, Thank you Elisa Fornero
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
"I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain."
It should be possible to do this much faster and without any sample preparation with X ray reflectometry.
See web site :
http://www-cxro.lbl.gov/optical_constants/
The is there the possibility to simulate some mesures.
Optical methodes could also be able to do this, by interferences in the TiO2 film.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Dear all, I send you the following message written by a serious student. I hope that someone can offer a possibility to her. Thanh you very much. All the best, Elena Belluso
My name is Elisa Fornero and I doing my Degree thesis, at Turin University - Italy- (Dept. of Mineralogical and Petrologic Sciences), on "INORGANIC FIBERS IN LUNG TISSUES: MINERALOGICAL CHARACTERIZATION, COMPARISION WITH NATURAL ASBESTOS AND CALCULATION OF DIFFERENT FIBROUS MINERALOGICAL SPECIES" (the samples of tissues derived from people not professionally exsposed in order to value the environmental pollution of fibrous minerals). During my thesis I extensively worked with SEM and EDS and I learned to prepare the samples from lungs tissues filtrated. I would like to have some international experience taking part, about for two months between April and June 2001, to a stage in a foreigner University. I would like found a laboratory where develop my knowledge in this field. If there is anyone interested to host me, I can send my curriculum vitae, Thank you Elisa Fornero
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
"I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain."
we are interested in purchasing a good software for desktop- analysis (on a PC) of X-ray EDS spectra, previously acquired on an analytical TEM and transferred to other computer.
In the book of Williams and Carter is mentioned only one such program, i.e. the DTSA, from NIST. Does anyone of you know a web address where I could get more informations about this software ? I would be delighted if there will be any possibility to check it before ordering, but where or how? Besides, does anyone know about a better program of this kind ? We would much better appreciate a free one, but we are ready to purchase even a commercial one.
Thank you for your attention.
Corneliu Sarbu, PhD Metallyrgy and Applied Materials Science Dept. Catholic University of Leuven Belgium
---------- } From: "CAROL.A.WALDER" {eencaw-at-elec-eng.leeds.ac.uk} To: mtlrmdb-at-leeds.ac.uk
Hi Listers,
I have a LKB Bromma 11800 Pyramitome whose belt has come loos and it needs to be replaced. I can't find the instruction manual or the slip of paper that my predecessor wrote the service person's name on.
Does anybody out there have a source for the instruction manual? I called Leica but they told me this machine was absolete when they acquired the LKB line.
I'll fix this thing myself if anyone can send me a copy of the manual (I'll glaly pay for the cost of copying and shipping).
If anyone knows of a repair person who still works on these ancient beasties, please let me know. It would be good to have someone who could give them tune-ups every now & then.
My screws are loose and I've got the hammer, blow torch sosme bubble gum and bailing wire....now all I need is the manual.
Pretty please??
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
An excellent write-up about the process is located at http://www.manufacturing.net/magazine/dn/archives/current/feature1.html
Cheers Peter Tarquinio peter-at-evex.com
Evex Analytical Microanalysis and Digital Imaging 857 State Road Princeton, NJ 08540 609-252-9192 T 609-252-9091 F www.evex.com info-at-evex.com ----- Original Message ----- } From: "Rick Mott" {rickmott-at-alumni.princeton.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, March 06, 2001 11:20 PM
} In the book of Williams and Carter is mentioned only one such } program, i.e. the DTSA, from NIST. Does anyone of you know } a web address where I could get more informations about this } software ? I would be delighted if there will be any possibility to } check it before ordering, but where or how? } Besides, does anyone know about a better program of this kind ? } We would much better appreciate a free one, but we are ready to } purchase even a commercial one. } } Thank you for your attention. } } Corneliu Sarbu, PhD } Metallyrgy and Applied Materials Science Dept. } Catholic University of Leuven } Belgium
Corneliu,
DTSA is no longer sold as a Standard Reference Data product. NIST has abrogated its patent on DTSA so both the executable and source code are available from the following URL free of charge:
If you find it does not meet your needs, please drop me an email. Our group welcomes suggestions for new features or improvements.
-- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
Dear all, I send you the following message written by a serious student. I hope that someone can offer a possibility to her. Thanh you very much. All the best, Elena Belluso
My name is Elisa Fornero and I doing my Degree thesis, at Turin University - Italy- (Dept. of Mineralogical and Petrologic Sciences), on "INORGANIC FIBERS IN LUNG TISSUES: MINERALOGICAL CHARACTERIZATION, COMPARISION WITH NATURAL ASBESTOS AND CALCULATION OF DIFFERENT FIBROUS MINERALOGICAL SPECIES" (the samples of tissues derived from people not professionally exsposed in order to value the environmental pollution of fibrous minerals). During my thesis I extensively worked with SEM and EDS and I learned to prepare the samples from lungs tissues filtrated. I would like to have some international experience taking part, about for two months between April and June 2001, to a stage in a foreigner University. I would like found a laboratory where develop my knowledge in this field. If there is anyone interested to host me, I can send my curriculum vitae, Thank you Elisa Fornero
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
"I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain."
You can find information about DTSA at the NIST web site
http://www.cstl.nist.gov/div837/837.02/dtsa.html
There is only a Macintosh version of this software, so it may not be of use for you if you are looking for software for a PC. NIST has officially withdrawn DTSA as a Standard Reference Data product and has abrogated its patent. NIST is now releasing the software free from charge, however no longer officially supports the software except for internal development as an internal research tool for NIST. The NIST web site I directed you to has links to download the software application and the Pascal source code.
Tyrone Daulton
} Dear fellows microscopists,
} ... } we are interested in purchasing a good software for desktop- } analysis (on a PC) of X-ray EDS spectra, previously acquired } on an analytical TEM and transferred to other computer.
} In the book of Williams and Carter is mentioned only one such } program, i.e. the DTSA, from NIST. Does anyone of you know } a web address where I could get more informations about this } software ? ...
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
I am reposting this for a colleague. If interested, please reply directly to
Ms. Linda McGuire lmcguire-at-downstate.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Experienced Histotechnologist to work in Neuropathology Research Laboratory of Department of Pathology
The person we seek will be responsible for maintaining a neuropathology research laboratory in the Department of Pathology at SUNY Downstate Medical Center. Knowledge of immunohistochemistry, special stains, and histopathology is required. Previous experience in handling central nervous system tissue preferred. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in a laboratory with hands-on experience in tissue processing, histochemistry, immunohistochemistry, and operation of a light microscope. Experience with technical aspects of neuropathology and performing special stains including silver (Bielschowsky) and myelin stains.
Two years experience working on immunohistochemistry of CNS tissue specimens with knowledge of immunoreagents and experimental protocols.
BachelorUs degree in science desirable.
Laboratory skills including communications, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, operation of computers and office equipment.
Advanced computer skills (word processing and database management) essential.
Desirable Experience
Confocal microscopy desirable.
Salary commensurate with experience
Responsibilities
Purchasing supplies and equipment, budget reports, laboratory maintenance and brain banking.
Will operate all microscopic, photographic and computer equipment, and keep accurate records of all laboratory experiments and procedures. Light and fluorescent microscopy; tissue processing for paraffin embedding, sectioning and slide stainer for immunohistochemical procedures; computer imaging (PhotoShop); general photography; and library and web searches
Familiarity with computer software for keeping records, report preparation and table/figure construction using Microsoft Office software (Word, Excel, PowerPoint) as well as Endnote and Adobe PhotoShop.
Send resume to:
Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
------------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I am reposting this for a colleague. If interested, please reply directly to
Ms. Linda McGuire lmcguire-at-downstate.edu ------------------------------------------------------------------------
Experienced Confocal Microscopy/Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for organization of a new facility that includes a confocal microscope and an electron microscope. The position includes overall management of the microscopy facility, user training, and user supervision. Requirements for the position include experience with light and transmission electron microscopy. This individual will oversee all aspects of specimen accession and processing, operation of the microscopes, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Experience with confocal and digital imaging techniques, microinjection, visualization of living cells containing fluorescent probes, photobleaching, and fluorescence in situ hybridization.
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
Excellent interpersonal and organizational skills are essential.
BachelorUs degree in science desirable.
Desirable Experience
Expertise in training in the operation of confocal microscope systems is a distinct advantage.
Familiarity with light microscopy methods, immunofluorescent staining, use of fluorescent probes for physiologic measurements and the general principles of cell biological research are critical. Significant facility with computers is desired.
Responsibilities
Serve as the technical manager of the facility and be responsible for the operation and maintenance of the confocal and EM microscope facility.
Perform routine transmission EM, including tissue processing, ultramicrotomy, and examination; do preventative maintenance on the equipment; maintain the lab, order supplies, schedule instruments, and oversee billing.
Image analysis at the light, confocal and electron microscopic levels and preparation of micrographs for publication.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I am reposting this for a colleague. If interested, please reply directly to
Ms. Linda McGuire lmcguire-at-downstate.edu
----------------------------------------------------------------------- Experienced Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for the overall operation of the EM laboratory in the Department of Pathology at SUNY Downstate Medical Center. This individual will oversee all aspects of specimen accession and processing, operation of the microscope, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
BachelorUs degree in science desirable.
Laboratory management skills including effective written/verbal communication skills to interact with a diverse group, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, and operation of computers and office equipment.
Desirable Experience
Previous experience in confocal microscopy highly desirable.
Previous experience in performing immunocytochemical staining and advanced computer skills usage (e.g. image analysis) is also desirable.
Salary commensurate with experience
Responsibilities
Maintain electron microscope in operating condition. Process clinical and research tissues for "thick" and "thin" sectioning. Darkroom management of photographic printing.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by: Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
{fontfamily} {param} Times {/param} {bigger} SCANNING ELECTRON MICROSCOPY (SEM) AND ENERGY DISPERSIVE
X-RAY SPECTROSCOPY (EDS)
WITH APPLICATIONS AT VARIOUS PRESSURES AND ATMOSPHERES
AN EDUCATIONAL SEMINAR-FRIDAY, APRIL 27, 2001
MOTOROLA GALVIN CENTER-SCHAUMBERG, IL
8:30 AM - 5:00 PM
{underline} Instructors {/underline} : Vern Robertson, JEOL-USA, and Nestor Zaluzec, Argonne National Laboratory.
{underline} Time {/underline} : Check-in at Galvin Center 8:30-9:00 am. Seminar 9:00-12:30 and 1:30-5:00.
{underline} Preregistration Required {/underline} : Deadline April 20, 2001; no walk-ins.
{underline} Cost {/underline} : Members of ASM-International and of the Midwest Microscopy and Microanalysis Society-$30; Non-members-$50; Students: $10. Registration includes lunch and refreshments at breaks. Check or money order to accompany preregistration form (below).
{underline} Exhibits {/underline} : Representatives from the following industrial co-sponsors will be present to discuss your applications and their products: EDAX, FEI, Hitachi, JEOL-USA, LEO, Noran, Oxford, and PGT.
{underline} Directions to Seminar Site {/underline} : Please see the other side.
{underline} Preregistration {/underline} : Please complete the following form. Mail it with your check or money order made out to "Chicago Regional Chapter of ASM-I" to Ms. Sheila Jungman, MSD 212, Argonne National Laboratory, Argonne, IL 60439 {bold} . {/bold} Deadline for receipt of the form is April 20, 2001. If you require a receipt for the preregistration cost, please so indicate below for pick-up at the Seminar Check-In desk. Questions? Phone 630-252-4157 or e-mail allen-at-aaem.amc.anl.gov.
{bold} SEM/EDS Educational Seminar-Friday, April 27, 2001
{/bold} Name (please print clearly)________________________________________________________
We are having problems with preparing brittle TEM sample of (single crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any suggestions beyond a kid-gloves approach?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
Mary-Jacque, Several years ago at a used book library sale, I found a copy of Forensic Geology-Earth Sciences and Criminal Investigation by Raymond C. Murray and John C.F. Tedrow. It dates from 1975 and looks interesting. I hope to read more of it. Another title which came from a special session at the MAS-MSA meeting in 1985 is titled Electron Microscopy in Forensic, occupational and environmental health sciences by Samarendra Basu and James R. Millette. I still lament that the Microbeam Analysis Society dropped Sherlock Holmes' hat and pipe from their logo.
Lawry, I don't know about magnesium orthovanadate, but sapphire will work using the small angle cleavage technique. I have done cross sections of GaN on sapphire when I taught some students at Univ. of Illinois how to do it. John McCaffrey has done YBCO on cerium oxide on sapphire. If you are only interested in the bulk, that is even easier. If the magnesium orthovanadate is as brittle as you say, then SACT will probably work on that also. If you want, I can send you an image of the GaN/sapphire.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: L. D. Marks [mailto:ldm-at-risc4.numis.nwu.edu] } Sent: Wednesday, March 07, 2001 1:22 PM } To: Microscopy List } Subject: Brittle samples } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } We are having problems with preparing brittle TEM sample of (single } crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any } suggestions beyond a kid-gloves approach? } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html } }
A quick clarification before I get deluged with responses about how to do cross-sections; we are not trying to do this. All (?) we want is a "conventional" 3mm disc sample of sapphire or magnesium orthovanadate. No glue, no mounting on a Cu ring, no nothing.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
I think a good solution may be Tripod Polishing. We have an application note on preparing samples of sapphire with a GaN film which may be useful. Let me know if you would like a copy of the note and I can email it to you.
Best regards-
David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*** www.southbaytech.com ***
} RE: Brittle samples } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are having problems with preparing brittle TEM sample of (single } crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any } suggestions beyond a kid-gloves approach? } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } -------------------------------------------------------
Has anybody out there had any experiences with making your own transmitted electron detector for SEM? I'm interested in sharing ideas, references, etc. I've made one for myself and am fairly happy with its performance, but am wondering if there is more that I can do.
Thanks in advance,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I have two questions for you immunolabeling experts.
First, a client has some precious and irreplaceable histological sections of human hippocampus that have been labeled with ABC - DAB. Will it be possible for me to de-paraffinize the sections, expose them to osmium vapor and re-embed them in resin on the slide and pop them off and resection them and see the DAB precipitate? Anyone have a protocol that has worked?
Second, these researchers plan to use Vibratome sections of mouse brain and immunolabel them for light microscopy using an ABC kit. I have done on-ultrathin-section colloidal gold immunolabeling for TEM. However, for this new project we would like to try pre-embedding labeling of 50-60 micrometer Vibratome sections, then embed and resection them for TEM. Knowing what works for light microscopy, we'd like to use streptavidin/colloidal gold or whatever for TEM visualization. My question is, of course, does anyone have a favorite protocol they would be willing to share? How do I keep the sections from curling? If we stick them on a glass slide to embed in resin and (hopefully) pop off later for resectioning, when and how do I get the to adhere?
Mahalo!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
The URL I gave this morning for DTSA was correct, but not memorable. A few hours ago an alias was created to make it easier to navigate to the NIST Microanalysis Software web page. DTSA can now be accessed from:
http://www.nist.gov/dtsa
This page also contains links to Lispix, Dave Bright's excellent image processing application (now available for Win9x/WinNT/Win2k) and the NIST Monte Carlo codes. Lispix can be accessed directly at
http://www.nist.gov/lispix
-- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
Try epoxying a thin metal washer on one side of the sample, and dimple from the washer side.
Larry Thomas Pacific Northwest National Laboratory Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
---------- From: L. D. Marks Sent: Wednesday, March 7, 2001 10:21 AM To: Microscopy List Subject: Brittle samples
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
We are having problems with preparing brittle TEM sample of (single crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any suggestions beyond a kid-gloves approach?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
The purpose of gluing a metal ring to brittle samples before dimpling is to support the sample. The sample is then waxed to the dimple grinding support stub and dimpled through the washer. We use this method routinely with brittle samples and not just cross sections. It's a good idea to carefullly clean the sample and washer so the epoxy bonds well. One- or two-mil Mo hole or slot washers usually work well for us, but we sometimes use other washer materials to avoid confusing Mo in the sample with Mo deposition artifacts.
Larry
---------- From: L. D. Marks Sent: Wednesday, March 7, 2001 11:38 AM To: Microscopy List Subject: Brittle samples - NOT CROSS-SECTION
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A quick clarification before I get deluged with responses about how to do cross-sections; we are not trying to do this. All (?) we want is a "conventional" 3mm disc sample of sapphire or magnesium orthovanadate. No glue, no mounting on a Cu ring, no nothing.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
the metallurgy or materials science group in your university should be able to tell you all about grain size measurements.
Best regards
Sam Purdy Technical Center National Steel Corp. Trenton Mi USA spurdy-at-nationalsteel.com
} ---------- } From: James Clarke } Sent: Wednesday, March 7, 2001 11:45 AM } To: NIH-IMAGE-at-LIST.NIH.GOV } Subject: countig grain size in archaeological iron samples } } IMAGERS! } Hope some one can point me in the right direction. I need to be } able to measure the grain size of metal samples which are of a } heterogenous nature. I am using a PC and Scion image seems to } have some of the things I need but some advice would be nice. } } --------------------------------- } James Clarke } J.Clarke1-at-bradford.ac.uk }
Fellow Microtomists: I am seeking information regarding 2 Sorvall MT2B ultramicrotomes that still are in use and in good working condition for both semi-thin sections and ultrathin sections. However, the faster speed of the return stroke of the cutting cycle no longer engages, even though the duration and position control belts are in tact for the slow-speed (cutting phase) of the cycle. Is this a relatively simple repair, and is there a company in the Cincinnati, Ohio, area that provides such repair service? [The serial numbers of these microtomes are 7701673 and 7800584. Do the first 2 digits reflect the year of manufacture (1977 and 1978 respectively)?] What would be a fair price for microtomes of this ventage? Thanks in advance for your assistance. EL Cardell
} } } Email: de-at-payload.com } Name: De McKeown } } Organization: Payload Systems } } Education: Graduate College } } Location: Cambridge, MA, USA } } Question: I need to determine the number of TV lines my microscope is } giving me. I have a test target but it only goes to 266 l/mm and I have } better resolution than that. I can't find a 'better' target and am stuck } trying to get an answer. How can I do this or where can I go for a new } target? } } I also have to find a contrast, but haven't got enough information yet to } ask a good question. } } Thank you!! }
There is a rash of virus mail out on the net again, a number of which are using "suggestive names" in the subject line. As a precaution some people trash messages which appear out of context.
While Humorous Titles are good for the soul (especially mine) and bring a smile especially when one gets the implied joke, let me remind you to at least add in the Subject Line with some standard notation so that individuals can more easily judge source of the mail. After all, we know that my Email filtering system is not perfect and suspect mail leaks through every so often.
Some Keywords include: TEM, SEM, EDS, .Confocal, LM, SamPrep, etc... check the FAQ site if you need idea's, but you can easily see what I mean.
For example: the posting on "I've got the screws loose" could have be preceeded or prepended by an appropriate Keyword like Microtome, even if the word Microtome was in () or abbreviated as uTome.
Unfortunately getting Mo (or anything else) on the sample destroys it as far as I am concerned. So, we have to rule out anything like this unless we can safely remove the metal with an acid (and not dissolve the oxide in the process).
On Wed, 7 Mar 2001, Thomas, Larry (PNNL) wrote:
} Laurie, } } The purpose of gluing a metal ring to brittle samples before dimpling is } to support the sample. The sample is then waxed to the dimple grinding support } stub and dimpled through the washer. We use this method routinely with brittle } samples and not just cross sections. It's a good idea to carefullly clean the } sample and washer so the epoxy bonds well. One- or two-mil Mo hole or slot } washers usually work well for us, but we sometimes use other washer materials to } avoid confusing Mo in the sample with Mo deposition artifacts. } } Larry } } ---------- } From: L. D. Marks } Sent: Wednesday, March 7, 2001 11:38 AM } To: Microscopy List } Subject: Brittle samples - NOT CROSS-SECTION } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A quick clarification before I get deluged with responses about } how to do cross-sections; we are not trying to do this. All (?) } we want is a "conventional" 3mm disc sample of sapphire or } magnesium orthovanadate. No glue, no mounting on a Cu ring, } no nothing. } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html } } } }
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
A researcher has asked me to localize gluconase (a protein) in plant roots. They provided me with antibody (sera) and preimmune, which they have used in westerns. The problem is the preimmune is labelling the cell wall; the labeling is quite impressive and very specific. Now they tell me (alas I didn't ask before) that yes they have background on the westerns, but the preimmune never labels the 32 Kd protein they are interested in. What is the best approach to localize this 32 Kd protein? Should we cross absorb everything else out, or would it be better to affinity purify for the desired antibody. And why does this rabbit have such good antibodies to cell wall?
Thank you, Michelle
Michelle Peiffer ************************************************************* Electron Microscope Facility for the Life Sciences Penn State University Biotechnology Institute 001 South Frear Lab University Park PA 16802
The key to thinning brittle hard samples is to start with a doubly polished thin section 30 micrometers thick. Cut out 3 mm diameter discs with an ultrasonic probe. Glue these to a glass slide with crystalbond, a thermal plastic cement that dissolves in acetone. Mechanically thin and polish to less than 10 micrometers or thinner. Ion thin until satisfied.
Start thinning a thin sample. Don't break it. One good sample is all you need.
I have made many ion thinned samples of brittle materials, mainly shocked minerals, for over thirty years. Also this method is not for those in a hurry.
Gordon Nord USGS Emeritus Scientist
"L. D. Marks" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Unfortunately getting Mo (or anything else) on the sample destroys it } as far as I am concerned. So, we have to rule out anything like this } unless we can safely remove the metal with an acid (and not dissolve } the oxide in the process). } } On Wed, 7 Mar 2001, Thomas, Larry (PNNL) wrote: } } } Laurie, } } } } The purpose of gluing a metal ring to brittle samples before dimpling is } } to support the sample. The sample is then waxed to the dimple grinding support } } stub and dimpled through the washer. We use this method routinely with brittle } } samples and not just cross sections. It's a good idea to carefullly clean the } } sample and washer so the epoxy bonds well. One- or two-mil Mo hole or slot } } washers usually work well for us, but we sometimes use other washer materials to } } avoid confusing Mo in the sample with Mo deposition artifacts. } } } } Larry } } } } ---------- } } From: L. D. Marks } } Sent: Wednesday, March 7, 2001 11:38 AM } } To: Microscopy List } } Subject: Brittle samples - NOT CROSS-SECTION } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } A quick clarification before I get deluged with responses about } } how to do cross-sections; we are not trying to do this. All (?) } } we want is a "conventional" 3mm disc sample of sapphire or } } magnesium orthovanadate. No glue, no mounting on a Cu ring, } } no nothing. } } } } ------------------------------------------------------- } } Laurence Marks } } Department of Materials Science and Engineering } } Northwestern University } } Tel: (847) 491-3996 Fax: (847) 491-7820 } } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } } ------------------------------------------------------- } } } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } } http://xraysweb.lbl.gov/esg/phasing/index.html } } } } } } } } } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html
-- Gordon Nord Small Business Network Design and Construction Macintosh and Windows - Solutions and Conflicts
Nord Consultants 20594 Cornstalk Terrace Ashburn VA 20147
Below is the result of your feedback form. It was submitted by (linda_knoll-at-msn.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 7, 2001 at 18:43:01 ---------------------------------------------------------------------------
Email: linda_knoll-at-msn.com Name: Linda Knoll
Organization: St.Joseph'sAcademy
Education: Graduate College
Location: St. Louis, MO USA
Question: How do the cells of humans and apes differ when looked at under a microscope?
Below is the result of your feedback form. It was submitted by (georgas1-at-home.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 7, 2001 at 20:29:09 ---------------------------------------------------------------------------
Email: georgas1-at-home.com Name: Adam Georgas
Organization: UMS-Wright Preparatory School
Education: 9-12th Grade High School
Location: Mobile, Alabama, USA
Question: Dear Microscopist: I am a 10th grade student working on my science fair project. My project deals with a computer program I wrote using fractal geometry to diagnose chronic lymphocytic leukemia. I obtained 25 pictures of normal cells taken from a Phillips electron microscope and 25 pictures of leukemic cells taken with the same microscope. I obtained these photographs from my mentor, a physician/professor at the University of South Alabama. My concern with my project is that the 25 healthy cells came from only one patient, and the 25 leukemic cells came from another patient. So, I have 50 cell pictures, 25 healthy, 25 leukemic, from only 2 patients. All of the research I have read use 25 different patients with only one cell from each patient. I am wondering if my study is valid? I posed this question to a local pathologist. She told me that since the pictures were taken from an electorn microscope, it was actually better to have more cells from just 2 patients. Is this true? If it is true, what is the rationale
Here is some parameters of 2000FX with AHP2OL polepiece: --------------------------------------------- Point resolution 0.31nm Lattice resolution 0.14nm OL focal length 4.1mm Cs 3.4mm Cc 3.1mm ---------------------------------------------
As to the question of beam semi-convergence angle, Prof. O'Keef has given a perfect note. I learnt a lot there as well.
Yours, Shu-You Li ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Shu-You Li, Dr. Institut fuer Physikalische Chemie Johannes Guttenberg Universitaet Jakob-Welder-Weg 11 D-55099 Mainz, Germany
Fax: +49-6131-3923768 Tel: +49-6131-3923148(O) E-mail: syli-at-mail.uni-mainz.de; syli16-at-hotmail.com URL: http://syli.homepage.com/ (This URL contains my resume, SCI journal informations, JobList, TEMAlert, as well as other useful informations related to Transmission Electron Microscopy) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Paula: Get in touch with Helmut Patzig at MOC, Spring Valley, NY. His email is Mocleica-at-aol.com. If anybody has the know-how and parts, I am pretty sure he does. Sorry I don't know of anyone closer to DC, but I have dealt with Helmut over the years with obsolete Reichert and LKB equipment.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Wed, 07 Mar 2001 08:31:14 -0500, Paula Sicurello wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hi Listers, | | I have a LKB Bromma 11800 Pyramitome whose belt has come loos and it needs to be replaced. I can't find the instruction manual or the slip of paper that my predecessor wrote the service person's name on. | | Does anybody out there have a source for the instruction manual? I called Leica but they told me this machine was absolete when they acquired the LKB line. | | I'll fix this thing myself if anyone can send me a copy of the manual (I'll glaly pay for the cost of copying and shipping). | | If anyone knows of a repair person who still works on these ancient beasties, please let me know. It would be good to have someone who could give them tune-ups every now & then. | | My screws are loose and I've got the hammer, blow torch sosme bubble gum and bailing wire....now all I need is the manual. | | | Pretty please?? | | Paula :-) | | Paula Sicurello | George Washington Univ. Medical Center | Dept. of Pathology, Ross Hall rm 505 | Electron Microscope Lab | 2300 Eye St. | Washington, DC 20037 | 202-994-2930 phone | 202-994-2518 fax | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
The answer depends on the details of the "TV" system and microscope used. At this point I don't even know if it is an optical or electron microscope, etc... It should be noted that actual microscope resolution and the displayed resolution via TV have little in common.
NTSC (US) TV is 525 lines vertical (total, and the equevelant of 280 to over 640 horizontal (bandwidth dependent). Note that not all 525 lines are used for the image.
PAL, a standard used in many other areas of the world is slightly higher vertical resolution, but not much.
There are other "non-brodcast standard" resolutions used by custom instrumentation.
Woody
} -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } } } } } } Email: de-at-payload.com } } Name: De McKeown } } } } Organization: Payload Systems } } } } Education: Graduate College } } } } Location: Cambridge, MA, USA } } } } Question: I need to determine the number of TV lines my } microscope is } } giving me. I have a test target but it only goes to 266 } l/mm and I have } } better resolution than that. I can't find a 'better' } target and am stuck } } trying to get an answer. How can I do this or where can I } go for a new } } target? } } } } I also have to find a contrast, but haven't got enough } information yet to } } ask a good question. } } } } Thank you!! } } } } }
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Tina, Check the following reference for your answer to your second question:
Liposits, Zs., D. Sherman, C. Phelix and W.K. Paull. (1986). A combined light and electron microscopic immunocytochemical method for the simultaneous localization of multiple tissue antigens. Histochemistry 85:95-106.
We very successfully used vibraotomed sections of brain to do light and EM ICC. Becuase of the penetration problems for the gold conjugates available at that time, we used PAP-DAB for visualization on the light level followed by silver intensification of the material for EM visualization. The study involved serial sectioning of the material to locate synapses so flat embedding of the sections was manditory. The method is fully discribed in the paper but feel free to contact me if you have questions.
I would also try using the ultra small gold colloids presently on the market followed by silver intensification as an alternative to the PAP-DAB technique.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
I have two questions for you immunolabeling experts.
First, a client has some precious and irreplaceable histological sections of human hippocampus that have been labeled with ABC - DAB. Will it be possible for me to de-paraffinize the sections, expose them to osmium vapor and re-embed them in resin on the slide and pop them off and resection them and see the DAB precipitate? Anyone have a protocol that has worked?
Second, these researchers plan to use Vibratome sections of mouse brain and immunolabel them for light microscopy using an ABC kit. I have done on-ultrathin-section colloidal gold immunolabeling for TEM. However, for this new project we would like to try pre-embedding labeling of 50-60 micrometer Vibratome sections, then embed and resection them for TEM. Knowing what works for light microscopy, we'd like to use streptavidin/colloidal gold or whatever for TEM visualization. My question is, of course, does anyone have a favorite protocol they would be willing to share? How do I keep the sections from curling? If we stick them on a glass slide to embed in resin and (hopefully) pop off later for resectioning, when and how do I get the to adhere?
Mahalo!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Michelle, The answer to the last question is easy. Remember Bugs Bunny munching his carrot? Rabbits eat plants so it is not too surprising that a preimmune serum recognizes a plant antigen, if not several.
What we have done in situations like this is to incubate the serum with the protein of interest (in this case, the 32kD glucanase) and then stain with that. This removes the specific glucanse staining. It is a kind of guilt by subtraction. It is not as nice as doing affinity purification, but it is a lot faster, and conserves the serum.
Hope this helps, Tobias Baskin
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Dear Listservers, we have been recently working on structural and compositional characterization of InGaAs/GaAs quantum dots grown by MOVPE, achieving interesting results. We have tried to use CERIUS2 to obtain simulations of cross sectional HREM images of these dots, but we experienced several problems. Is there anyone who has spent time on similar matters. We would like to discuss and compare our experiences, to understand if our problems are due to a bad construction of the supercell or to limitations of the software. Thanks Massimo
Thanks for all the responses to my original query. At the suggestion of Bart Cannon, you can check out my detector design at:
http://www.mta.ca/~jehrman/ted.htm
I perhaps should have clarified originally that this is a very simple design, and does not require any additional electronics, light pipes, PMT, etc. My machinist made it for me in an afternoon. I took inspiration from a single sentence in Goldstein, et. al (SEM and X-ray Microanalysis, 2nd edition, p. 267):
"A simple, inexpensive detector can be made from a high-atomic-number scattering surface placed below the specimen and tilted so that the transmitted electrons are scattered toward the conventional E-T detector in the specimen chamber"
} From my experience with one commercially available "real" TED a number of years ago, the homemade detector seems to perform as well (or better)
than the real thing.
No reference was given in the text, so I'm rather curious who came up with the idea originally. Are there any co-authors of the text lurking nearby who could shed any light?
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Question: Dear Microscopist: I am a 10th grade student working on my science fair project. My project deals with a computer program I wrote using fractal geometry to diagnose chronic lymphocytic leukemia. I obtained 25 pictures of normal cells taken from a Phillips electron microscope and 25 pictures of leukemic cells taken with the same microscope. I obtained these photographs from my mentor, a physician/professor at the University of South Alabama. My concern with my project is that the 25 healthy cells came from only one patient, and the 25 leukemic cells came from another patient. So, I have 50 cell pictures, 25 healthy, 25 leukemic, from only 2 patients. All of the research I have read use 25 different patients with only one cell from each patient. I am wondering if my study is valid? I posed this question to a local pathologist. She told me that since the pictures were taken from an electorn microscope, it was actually better to have more cells from just 2 patients. Is this true? If it is true, what is the rationale
Dear Adam, First, there is nothing about data from an electron microscope that makes sample selection any different from that used with other forms of data. The idea is that the population in your sample--in this case the normal and leukemic cells--should be representative of the larger population--all human normal lymphocytes and all human leukemic lymphocytes. If all normal lymphocytes in any one person have identical morphology, then any one cell will represent the population; if lymphocytes vary within an individual, then you would need a selection having the same distribution of types as exist in the whole person. This is usually obtained by selecting a small sample volume "at random". You might imagine how this could go wrong; e.g., if certain types were not present in arterial blood to the same extent as in venous blood. The next consideration is whether either normal lymphocytes or leukemic lymphocytes differ from person to person. I can see no reason that this would not be the case--especially if different forms of leukemia result from different transformation processes; e.g., different mutations or different extents of gene expression. In order to be sure that your sample is relevant, you would need to know whether there were different types of normal and leukemic cells both within and among individuals. This is an area where I am completely ignorant, but there is probably an extensive literature on this. There could also be differences arising from the preparation of the cells for EM, such as variation in fixation or staining, which can give apparent differences from nomonally identical cells. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Hi, I just started to bring up an ISI-SR50A. Followed the procedure to replace the filament, saturate beam current and aligment correctly but I could not get a decent image on the CRT even at low mag, 200x. Is there a way to check the detector? I varied the applied voltage from 5KV to 20KV and WD from 7mm to 40mm without any success. Any suggestions are welcome. -Vu.
__________________________________________________ Do You Yahoo!? Get email at your own domain with Yahoo! Mail. http://personal.mail.yahoo.com/
Here are two questions that I am pondering at the moment for thin film analysis in the TEM.
If you take the integrated intensities of the center portions of the peaks (zero crossing to zero crossing) of a top hat filtered spectrum to use for quantification, will you get the same results as you would if you do a background subtraction and take the integrated intensities from the peaks themselves?
For the integrated intensities of a top hat filtered spectrum, is it more appropriate to include the two negative lobes of the filtered peak added to the center portion?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
There are two freeware software packages that can accomplish this effort.
An IBM inspired software called OpenDX (for Unix), which is quite advanced. Start here (I have just installed this software but not worked with it yet):
http://www.opendx.org/
and through scion image, which is a PC version of NIH image. Start at the link below
http://rsb.info.nih.gov/nih-image/
You must install the ImageJ plugin, which you can obtain from:
http://www.isi.uu.nl/people/michael/vr.htm
This site also has other links for related information.
I am pursuing similar work with FIB/ Auger on particles and I would love to know about your progress. I believe this area has interesting and useful applications and I have found only very limited publications in this area. I would appreciate any additional information you might have.
Good Luck !
Regards, Ed
************************************************************ Edward Principe, Ph.D. Member of Technical Staff Defect & Thin Film Characterization Laboratory Applied Materials 408-986-3882
"Richard M Langford" {richard.langford-at-materials.oxford.ac.uk} on 03/08/2001 01:51:45 AM
To: microscopy-at-sparc5.microscopy.com cc: (bcc: Edward Principe/APPLIED MATERIALS)
Dear All
Can anyone suggest suitable software for 3-D reconstruction of grains and cracks from a sequential set of focused ion beam cross-sections.
Regards
Richard -------------------------------------------------------------- Richard M Langford
Department of Materials, University of Oxford Parks Road, Oxford, OX1 3PH, UK
If you only need to do reconstruction, download our free VoxBlast Light software at www.vaytek.com. Go to VoxBlast 3D software, then VoxBlast Light 3.0.
If you need any help or want more functionality, feel free to contact us directly.
Best regards,
Steve Niemela Sales sniemela-at-vaytek.com Ph: 641-472-2227 Fax: 641-472-8131
VayTek, Inc. 305 West Lowe Avenue Fairfield IA 52556 www.vaytek.com ----- Original Message ----- } From: "Richard M Langford" {richard.langford-at-materials.oxford.ac.uk} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 08, 2001 3:51 AM
A colleague is looking for a published reference that describes cholesterol crystals using transmission electron microscopy. We have searched the web but found only light microscopy of the crystals. Any help would be appreciated.
Thank you,
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
Facility Scientist (Academic Assistant III) Flow Cytometry and Confocal Imaging Facility, Biotechnology Center
The Biotechnology Center and the Department of Molecular and Cell Biology at the University of Connecticut invite applications for the position of Facility Scientist for the Flow Cytometry and Confocal Imaging Facility. This Facility houses a Becton Dickson FACSCalibur Flow Cytometer/cell sorter, a Leica SP2 laser scanning confocal microscope and several other microscope and image processing workstations. The facility scientist will be responsible for working with University faculty and students to develop research projects that utilize these instruments and related technologies. The facility scientist will also organize periodic training sessions and workshops to familiarize new users with the instruments. This individual will also be encouraged to develop collaborative research projects with University faculty and with local biotechnology businesses. We seek an individual who is excited by the challenges of an interdisciplinary research group and who enjoys interacting with people in an active scientific environment. The preferred candidate will have a Ph.D. in the biological sciences and familiarity with one or both of the core technologies. This is a full-time, annually renewable position. Salary will be commensurate with experience and education. Submit curriculum vitae, a statement of experience and interests, and the names, addresses, telephone numbers and e-mail addresses of at least three references to: Biotechnology Center, Confocal Search Committee, University of Connecticut, 184 Auditorium Rd., Unit 3149, Storrs, CT 06269-3149. Applications may also be faxed to (860) 486-5005 or sent electronically to biotctr1-at-uconnvm.uconn.edu. Screening of applications will begin immediately and continue until the position is filled.
We encourage applications from under-represented groups including minorities, women and people with disabilities. (Search #01A372) --
************************************************************ Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
The following posting is for a colleague who is not on the listserver.
Larry Thomas Pacific Northwest National Laboratory Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
We recently experienced a fire using a 5% perchloric acid - methanol electrolyte in a commercial jet electropolisher, and request information on similar experiences.
During operation of the electropolishing unit at room temperature and moderate pump speed, when turning up the voltage towards 30 V a loud pop was heard and the reservoir was found to be in flames. The fire was quickly extinguished using a dry fire extinguisher, but the reservoir and cable shielding melted.
We would appreciate any input on similar experiences using electropolishers with this or similar electrolytes. Any explanations for this behavior that you can provide would also be appreciated.
David S. Gelles Structural Materials Research Pacific Northwest National Laboratory Richland, WA 99353 Tel: (509) 376-3141 Fax: (509) 376-0418 E-mail: ds_gelles-at-pnl.gov
There are two ways that the top-hat filter has been used for quantifying EDS spectra:
(1) One can simply apply the filter to the spectrum as you suggest -- what results looks much like the second derivative of the original spectrum and each peak is represented by a central positive lobe with a negative lobe on either side. The slowly varying (continuum) components of the spectrum are suppressed. With care, one can relate the area of the positive lobe to the area of the original peak, however, there are some issues which make this simple concept a less than satisfactory quantitative method: (1) the area of the positive lobe is proportional to the area of the original peak, but the proportionality is a function of both peak area and filter shape. Thus the proportionality varies with the peak energy since the peak width varies with energy. (2) overlapping and adjacent peaks will interfere with each other, and this interference is actually increased by the broadening effect of the filter. Oddly enough, I have seen a number of familiar reference works which seem to give me credit for inventing the "top hat filter" used in this way -- I didn't. I saw it used this way for nuclear gamma ray spectra in the early '70s and after trying it myself, realized that it really wasn't satisfactory. But this experience did lead to the "filter-fit" method which follows.
(2) The"right" way for using the top-hat filter for continuum suppression is to use it in conjunction with linear-least squares fitting. One simply filters the unknown and each of the reference spectra to be fitted to it and then applies a conventional linear fit between them. When one fits the filtered references to the filtered unknown, the fact that the spectra have been distorted by the filter is immaterial (since the filter operator is linear), the overlaps are accurately unfolded, and the continuum basically drops out of the problem. Properly implemented, this is a very good quantitiative technique and has been employed productively for almost 30 years. If interested, the technique is published in NBS 604 (P273) or interested parties could contact me for references to several other papers describing it.
Fred Schamber
"Walck, Scott D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Here are two questions that I am pondering at the moment for thin film analysis in the TEM. } } If you take the integrated intensities of the center portions of the peaks (zero crossing to zero crossing) of a top hat filtered spectrum to use for quantification, will you get the same results as you would if you do a background subtraction and take the integrated intensities from the peaks themselves? } } For the integrated intensities of a top hat filtered spectrum, is it more appropriate to include the two negative lobes of the filtered peak added to the center portion? } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax)
I've been gold immunolabelling whole retina pieces - 300um thick sheets - with great success (LM and EM) for years. I react the tissue in small vials, floating around loose. Briefly, you need a fixative like PLP or paraformaldehyde - any glut in the fix and antibody access to the tissue is decreased and you just get labelling on the cut edges. Use saponin in all solutions for penetration enhancement. Use an Fab2 anti Ig conjugated to 1 nm gold. Silver enhance then gold tone. Pop tissue in a silicon mould for embedding. If this sounds useful I can send you the full method.
As for rescuing paraffin sections, I did some a long time ago. From memory the resulting tissue looked like squashed newspaper - a mushy black mess with a hint of structure. I can't imagine even being able to distinguish the DAB from the rest, let alone see what was labelling. However, maybe methods have improved.
Cheers,
Diana --
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
In my opinion what happened was that a spark between the cathode and specimen, which often occurs at those voltages during electropolishing, ignited the electrolyte. I have used other electrolytes at voltages greater than 30V and I have also head a crackling noise which indicated to me that was sparking. Fortunately the electrolyte was not as ignitable as a 5% perchloric acid - methanol electrolyte. Nevertheless, I kept the voltage below that just in case for the electrolyte was still based on organic solvents.
You are going to be bombarded by horror stories and cautions about electrolytes containing perchloric acid. And they are right. I will not say that I have used electrolytes containing up to 30 vol.% in ethanol without problems because that would be tempting fate and I have enough Sicilian blood in me to make me a bit superstitious.
I have some recommendations.
1. As far as I know, perchloric acid electrolytes only require a voltage of 15V max. It is rare that I have used voltages in excess of 25V for any electrolyte and that was with an electrolyte of methanol, Butoxy-ethanol, magnesium perchlorate and lithium chloride (LiCl). This was the electrolyte where above 30V I was getting sparking. I have used water based electrolytes above 30V sometimes, but ignition is not an issue there.
2. Whenever making up or using an electrolyte containing perchloric acid, cool it down. When making it up, this prevents ignition due to heat of dissolution of the concentrate acid as it mixes. This may be paranoia. Cooling during electropolishing is necessary because it also reduces the risk if ignition and it improves the electropolishing process. I have found that most organic based electrolytes, especially those based on ethanol and methanol require cooling to {20°C in order to achieve the correct electropolishing conditions. So the necessity of cooling is two fold.
I hope this is useful.
"Thomas, Larry (PNNL)" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The following posting is for a colleague who is not on the listserver. } } Larry Thomas } Pacific Northwest National Laboratory } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto:Larry.Thomas-at-pnl.gov } } We recently experienced a fire using a 5% perchloric acid - methanol electrolyte } in a commercial jet electropolisher, and request information on similar } experiences. } } During operation of the electropolishing unit at room temperature and moderate } pump speed, when turning up the voltage towards 30 V a loud pop was heard and } the reservoir was found to be in flames. The fire was quickly extinguished } using a dry fire extinguisher, but the reservoir and cable shielding melted. } } We would appreciate any input on similar experiences using electropolishers with } this or similar electrolytes. Any explanations for this behavior that you can } provide would also be appreciated. } } David S. Gelles } Structural Materials Research } Pacific Northwest National Laboratory } Richland, WA 99353 } Tel: (509) 376-3141 } Fax: (509) 376-0418 } E-mail: ds_gelles-at-pnl.gov
-- Dr. Steven Celotto Department of Engineering Materials Science & Engineering University of Liverpool Brownlow Hill Liverpool L69 3BX UNITED KINGDOM
I don´t know about the ISI50A but, assuming that it has the typical Everhard-Thornley design, isn´t it just possible that the bias voltage is inverted in your detector?. If the bias voltage is set negative, it´ll be rejecting all the secondary electrons, and you would only get a generally noisy image composed of the primary electrons backscattered in the very direction of the detector´s position. Quite nice for topographic information about flat specimens, but far worse quality than in "normal" SE images. Pardon me if I raised a too obvious explanation, but it occurred to me sometimes to get puzzled with a bad functioning of my SE detector just because another user of the microscope had been playing with the knobs.
Cheers
Diego
Diego Alvarez Instituto Nacional del Carbón INCAR-CSIC Spain
Thanks to all who responded to may plea for help in finding either a manual or a service person for the wee beastie.
Slowly but surely I will tighten the screws.....
Thanks again!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Full details of Trends in Nanotechnology (TNT) 2001, Segovia, Spain, September 2001, are now available. Based on the same format as last year, the conference aims to bring together researchers from all disciplines relating to nanotechnology, in a setting with ample time for interaction. A new feature this year will be the availability of grants to enable graduate students to attend and present posters.
Full details can be found at www.cmp-cientifica.com/tnt2001
Regards
Tim
***************************************************************** Tim E. Harper CEO CMP Cientifica s.l. Space & NanoTechnology Division Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/
The NIST's web link to DTSA can be found at: http://www.cstl.nist.gov/div837/837.02/MicroscopySoftware.html
This program is free and available for the Macintosh only.
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-417-1321 100 Bureau Drive, Stop 8371 Gaithersburg, MD 20899-8371 http://www.nist.gov/cstl/div837/837.02/
TO be more specific, any advice on using formvar coated slot grids for serial sections. They seem to crinkle, or get blobby, right on the vessel I want to look at.
Ellen: Another question: are you using bare formvar or have you coated with carbon? Bare formvar will pucker, but the addition of a little carbon does wonders for stabilization and strength of the films. My experience is using 0.5% formvar, stabilized with about 10nm of evaporated carbon. These films aren't too thick, allowing good image resolution at 80kV. BTW: if the formvar films are puckered, they will show it prior to picking up sections. (Another vagrant neuron just fired.) One way to deal with the puckered formvar is to use chloroform or carbon tet. Simply pass the grid over the mouth of an open bottle, or, as a last resort, put the grid into the neck of the bottle (NOT into the fluid--just the vapors). Please be careful with this. (Exposure to these chemicals.) I may have the beginnings of interstitial pulmonary fibrosis--a not uncommon result of exposure to chemicals.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
On Sun, 11 Mar 2001 14:31:50 -0500, Ellen S. Morgan wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | TO be more specific, any advice on using formvar coated slot grids for | serial sections. They seem to crinkle, or get blobby, right on the vessel | I want to look at. |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Does anyone know of any buildings/laboratories recently constructed that house instruments (not necessarily EM's) requiring very low ( {0.5 mG) 60 Hz stray magnetic fields? I am interested in contacting someone (scientist, building manager, architect, etc.) who might know the details of how that building was constructed.
Thank you for your time and help,
Sincerely,
Mick Thomas ------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
At 5:24 PM +0100 3/9/01, Edmund Gierlik wrote: } Email: gierlik-at-delta.sggw.waw.pl } Name: Edmund Gierlik } Organization: Electron Microscopy Lab. } Education: professor } Location: Warsaw, Poland } } Question: Can anybody show the source of nice presentation of ESEM } application in a phase transition investigation (water - ice, liquid - solid)?
Trisha Rice at the ESEM applications lab might be able to provide this. She can be reached at either trice-at-feico.com or trice-at-electroscan.com (I'm not sure if this last address is still valid).
Dear Randy, I have just a few points to add to Steve's excellent advice.
1. "It is too dim to focus" - then i) increase the emission current, many people run at too low a current and make the task of optimising the instrument settings much more difficult ii) increase the kV which would increase the intensity too ii) re calibrate the photographic system to allow a focus intensity that is suitable for most operators
I often use LoDose film, which is more than an order of magnitude more sensitive than SO163, so I can barely see the beam on the screen. Fortunately, we have an intensified CCD, which is so sensitive that I can still focus. In fact, using the minimum-contrast method works very well with this system, since the contrast can be electronically enhanced.
6. To focus at low magnifications try removing the objective aperture and focus without a binocular, or a wobbler, looking for the very strong minimum contrast effect.
Furthermore, I discovered that inserting the diffraction aperture (with the objective aperture out) allows this method to be used at up to 63kx (and possibly higher). Since minimum contrast can easily be found within one second, this is well suited to radiolabile specimens.
Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
This is a different procedure from the Formvar-coated grids that you are using but it's one of my favorite journal articles/techniques - J. C. Rowley and D.T. Moran, A Simple Procedure for Mounting Wrinkle-Free Sections on Formvar-Coated Slot Grids. Ultramicrotomy 1 (1975) 151-155. They use Formvar-coated aluminum supporting racks. We use this method for slot grids all the time rather than Formvar-coated grids. good luck, Beth } TO be more specific, any advice on using formvar coated slot grids for } serial sections. They seem to crinkle, or get blobby, right on the vessel } I want to look at.
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I do quite a bit of serial sectioning and would like to offer you the following advice:
1) Cut your sections as small as possible, they can be as much as 1mm wide, but try to make them something like 0.1mm high. This will allow you to get you as many as 20 sections on a 2mm slotted grid.
2) Get a bunch of locking tweezers and from the time you pick up the grid until you put it in the microscope, do not put the grid down onto filter paper (as you would for a normal grid). So, after you pick up the ribbon of sections, leave the grid in the tweezers to dry, and then when you post stain the grids, leave them in the tweezers to dry. If you put them down onto filter paper the formvar may rupture.
3) Maintain a positive attitude, and if things aren't going your way, stop and come back to it another day (the blocks will always be there for you, but if you get burned out you won't be there for the blocks).
Best of Luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
NATIONAL INSTITUTES OF HEALTH Bethesda, Maryland Division of Bioengineering & Physical Science Supramolecular Structure & Function Lab.
Biologist GS9/GS11/GS12 trained in life sciences and/or physical sciences at BS or MS level with solid technical experience in thin-section transmission electron microscopy. Experience in advanced techniques in analytical electron microscopy and structural biology (cryosectioning, high-pressure freezing, and macromolecular preparation methods) is desirable, although on-the-job training is possible for an experienced microscopist. PROOF OF U.S. CITIZENSHIP IS REQUIRED for this appointment.
For further information please contact:
Dr. Richard Leapman Division of Bioengineering & Physical Science Bldg. 13, Rm. 3N17 National Institutes of Health Bethesda, MD 20892 Tel: (301) 496-2599 FAX: (301) 496-6608 E-mail: leapman-at-helix.nih.gov
Applications (with curriculum vitae or federal employment form SF-171) should include the reference number ORS-01-0044, and should be post marked by April 9, 2001 and sent to:
Attention: Mr. Harold Atkins National Institutes of Health ORS Human Resources Office 31 Center Drive Msc 2157 Bldg 31, Room 4b41 Bethesda MD 20892-2157
Many thanks to everyone who responded to my question about focusing. Although my actual question was answered almost immediately, the ensuing replies were very enlightening. In EM work, it seems, even the "simplest" things have dimensions that aren't always obvious.
My initial question arose from the fact that I was curious about something I had always done as a matter of course because I had been taught to do it that way. That is, I have spent years focusing TEM's with the beam at the crossover point because I believed that I'd been taught to do so because of some property of electron "optics" that made it the preferred method to ensure best focus. Turns out, though, that the reason was something else entirely, involving an earlier generation of microscopes that simply had a beam that was too dim for focusing at the intensity needed for recording an image.
It's always educational to question old assumptions. Embarrassing, too, sometimes....
Thanks again, listers.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Technically, re-embedding paraffin sections for EM is perfectly feasible. The way I have done it is to separate the sections from slides after re-hydration and before further fixation with glutaraldehyde and osmium. Since both glutaraldehyde and osmium treatment cause sections to shrink and be brittle, sections might get further torn if they are still attached to the slides. Another step you should be careful about is to keep sections flat when applying glutaraldehyde and osmium. This ensures sections not to curl or wrinkle later. You can float your sections in 0.1 M phosphate buffer in a Petri dish, and let sections lie flat on the bottom. Then you gently remove buffer and add fixative with a glass pipet without disturbing the sections. Do not shake sections for a while after adding fixative. To flat-embed the sections, you can sandwich sections that have been fully infiltrated with resin between two sheets of Aclar film, then place this sandwich between two glass slides and apply some weight on top while polymerizing the resin. As someone already pointed out, even though you can re-embed paraffin sections for EM, ultrastructural will not be adequate anymore. This is particularly a problem for examining brain tissue in which membrane outline is essential for identifying different neuronal elements. However, if the tissue is precious, I would say it is still worth trying. We work with post-mortem brain tissue often. Sometimes the brains were over twenty-four hours post-mortem, fixed with only formalin and stored in cryoprotectant in a freezer for a long time. I am often surprised how much information we can still obtain from such tissue at EM level. I would be happy to help you to evaluate your micrographs if you like.
Good luck, Tina, let me know if you need more detailed answers.
Hong
============== Hong Yi Supervisor Emory University School of Medicine Neurology Microscopy Core Laboratory Rm 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
---------- } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} } Subject: TEM - immunolabeling } Date: Wed, Mar 7, 2001, 2:55 PM }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have two questions for you immunolabeling experts. } } First, a client has some precious and irreplaceable histological sections } of human hippocampus that have been labeled with ABC - DAB. Will it be } possible for me to de-paraffinize the sections, expose them to osmium } vapor and re-embed them in resin on the slide and pop them off and } resection them and see the DAB precipitate? Anyone have a protocol that } has worked? } } Second, these researchers plan to use Vibratome sections of mouse brain } and immunolabel them for light microscopy using an ABC kit. I have done } on-ultrathin-section colloidal gold immunolabeling for TEM. However, for } this new project we would like to try pre-embedding labeling of 50-60 } micrometer Vibratome sections, then embed and resection them for } TEM. Knowing what works for light microscopy, we'd like to use } streptavidin/colloidal gold or whatever for TEM visualization. My question } is, of course, does anyone have a favorite protocol they would be willing } to share? How do I keep the sections from curling? If we stick them on a } glass slide to embed in resin and (hopefully) pop off later for } resectioning, when and how do I get the to adhere? } } Mahalo! } } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
Regarding your question about pre-embedding immunolabeling of vibratomed brain sections, the type of labeling to use depends on the questions the researchers are trying to answer. Immunoperoxidase gives greater labeling depth, but limited spatial resolution. If they simply want to know what neuronal elements are labeled, then immunoperoxidase is fine. Otherwise, imunogold should be considered because it provides much more precise localization.
The methods for pre-embedding immunogold labeling using vibratomed brain sections have been very well documented. One of the first publications was by Chan, Aoki and Pickel (1) at Cornell University in 1990, shortly after ultrasmall gold conjugates were introduced. Many brain researchers have adapted their protocol. The characteristics of this protocol are the use of acrolein in the fixative for perfusion, using little or no detergent treatment for permeabilization and using a 1 nm colloidal gold conjugated IgG secondary probe in conjunction with a light insensitive, low viscosity, but fast reacting silver enhancement reagent. I have seen many beautiful results using this protocol. However, the particle size tends to be big, and irregular in shape. The cause of this is mainly in the silver enhancement reagent. In 1999, a new EM grade silver enhancement reagent was introduced to the market. Because of the improved enhancement homogeneity in particle size and shape, this silver enhancement reagent allowed us to conduct for the first time the pre-embedding double immunogold labeling in brain tissue using only ultrasmall gold conjugates (2).
To avoid a lengthy posting, I will send you the protocol, along with images off-line.
1. Chan J. Aoki C. Pickel VM. (1990) Optimization of differential immunogold-silver and peroxidase labeling with maintenance of ultrastructure in brain sections before plastic embedding. Journal of Neuroscience Methods. 33(2-3):113-27,
2. Hong Yi, Jan L.M. Leunissen, Ge-Ming Shi, Claire-Anne Gutekunst, and Steven M. Hersch (2001) A Novel Procedure for Pre-embedding Double ImmunogoldSilver Labeling at the Ultrastructural Level J. Histochem. Cytochem. 2001 49: 279-284.
Hong
============== Hong Yi Supervisor Emory University School of Medicine Neurology Microscopy Core Laboratory Rm 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
---------- } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} } Subject: TEM - immunolabeling } Date: Wed, Mar 7, 2001, 2:55 PM }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have two questions for you immunolabeling experts. } } First, a client has some precious and irreplaceable histological sections } of human hippocampus that have been labeled with ABC - DAB. Will it be } possible for me to de-paraffinize the sections, expose them to osmium } vapor and re-embed them in resin on the slide and pop them off and } resection them and see the DAB precipitate? Anyone have a protocol that } has worked? } } Second, these researchers plan to use Vibratome sections of mouse brain } and immunolabel them for light microscopy using an ABC kit. I have done } on-ultrathin-section colloidal gold immunolabeling for TEM. However, for } this new project we would like to try pre-embedding labeling of 50-60 } micrometer Vibratome sections, then embed and resection them for } TEM. Knowing what works for light microscopy, we'd like to use } streptavidin/colloidal gold or whatever for TEM visualization. My question } is, of course, does anyone have a favorite protocol they would be willing } to share? How do I keep the sections from curling? If we stick them on a } glass slide to embed in resin and (hopefully) pop off later for } resectioning, when and how do I get the to adhere? } } Mahalo! } } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
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A user has encountered an interesting problem . When cells are scraped and pelleted and embedded directly in epoxy they appear OK under the scope. When they are enrobed in agarose and then embedded the appearance becomes variable, although most cells appear very dense with contrast between organelles very low. This variability in appearance can be on the same grid square. Any ideas? Thanks Bruce Bruce Cutler Director, Microscopy & Electronic Imaging Laboratory University of Kansas
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id NAA02400 for dist-Microscopy; Mon, 12 Mar 2001 13:48:03 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id NAA02392 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 12 Mar 2001 13:47:33 -0600 (CST) Received: from UMKC-MAIL01.umkc.edu (email.exchange.umkc.edu [134.193.71.1]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id NAA02384 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 12 Mar 2001 13:47:22 -0600 (CST) Received: by email.exchange.umkc.edu with Internet Mail Service (5.5.2653.19) id {FCF6B0PG} ; Mon, 12 Mar 2001 13:45:30 -0600 Message-ID: {95A711A70065D111B58C00609451555C01CA7611-at-UMKC-MAIL02} "'Microscopy-at-sparc5.microscopy.com '" {Microscopy-at-sparc5.microscopy.com}
May be it's not really a phase transition, but you can find pictures of dissolution and crystallization of table salt at our web page: http://www.umkc.edu/dentistry/microscopy/Esemimg.htm
Vladimir
-----Original Message----- } From: Edmund Gierlik To: Microscopy-at-sparc5.microscopy.com Sent: 3/9/01 10:24 AM
Email: gierlik-at-delta.sggw.waw.pl Name: Edmund Gierlik
Organization: Electron Microscopy Lab.
Education: professor
Location: Warsaw, Poland
Question: Can anybody show the source of nice presentation of ESEM application in a phase transition investigation (water - ice, liquid - solid)?
Dear listers, I have a customer who wants TEM on a section of cellulose that is coated with a layer of Fe2. I have no idea about how to go about accomplishing this. He wants to see where the Fe2 is located in the cellulose matrix, and how deep it has penetrated. The sample is a layer of cellulose about 1/2 mm thick with a visible layer of Fe on the surface and it's in submerged in water. Any ideas out there? Thanks so much,
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
After several requests, I've posted a measured schematic of my transmitted electron adapter, which can be reached via a link from the page mentioned previously:
http://www.mta.ca/~jehrman/ted.htm
I had the adapter made of aluminum and brass (just because the machinist likes to work with these). The angled surface below the specimen grid is polished brass, and as mentioned by several respondents, could be plated/coated/covered with something that generates more secondaries. Signal, however, has not been a problem in our applications - more than enough even at the smallest spot sizes.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I have a LKB Ultrotome V which has given many years of faithful service to me and my students until today, when the band that raises and lowers the cutting arm snapped. I am in the middle of the semester class on EM techniques and hence am a bit desperate to get it repaired. Does anyone know where I might find a replacement part for this - and is it possible to replace and adjust it myself? There are no instructions in my manual for this procedure; are there any out there somewhere? Is there any company that will service this instrument? I am located in New England.
Many thanks in advance for any leads.
Sincerely,
Dick Briggs Biology Department Smith College Northampton, MA 01063
This notice is to invite you to attend and participate in the 28th Annual Meeting of the Microscopical Society of Canada. This three day meeting will be held at the University of New Brunswick, Fredericton, New Brunswick, Canada on 6-8 June, 2001. The Scientific Program features invited speakers from the environmental sciences, plant sciences, material sciences, EELS applications, confocal microscopy and magnetic resonance imaging (MRI). Workshops will focus on specimen preparation techniques for both the biological and material sciences, including ultramicrotomy, polishing methods, high-pressure freezing techniques and a workshop on preparation of digital images for publication. Vendors and manufacturers will be demonstrating the latest developments in the field of microscopy. Presentations can be in either platform or poster format and the Abstract Deadline is March 30, 2001. The deadline for Pre-registration is April 30, 2001. All information and required forms for registration and submission of abstracts are available at the conference website at: http://www.unb.ca/msc2001. ************ Keynote Speakers and topics include: Dr. Bruce Balcom, Department of Physics, University of New Brunswick. Title of talk: "Magnetic Resonance Imaging of Materials". Dr. Ron Gronsky, Materials Science and Engineering, University of California, Berkeley. Title of talk: TBA. Dr. Michael Hochella, Dept. of Geological Sciences, Virginia Polytechnic Institute & State University, Blacksburg, Virginia. Title of talk: "Contributions of Microscopy to the Environmental Sciences: From Bacteria to Atoms" Dr. Richard Howard, DuPont Agricultural Products, Experimental Station, Wilmington, Delaware. Title of talk: "Trends in imaging fungal pathogens for understanding the cell biology of plant disease" Dr. Peter Ottensmeyer, Medical Biophysics, University of Toronto. Title of talk: "3-D Reconstruction of Macromolecules from single-particle STEM images: Transmembrane signalling of the insulin receptor"
Invited Speakers include: Mr. Jason Davis, Medical Biophysics, University of Toronto. Title of Talk: "EM of Chromophores: Very Low Energy-Loss Imaging of Green Fluorescent Protein and other Coloured Denizens." Dr. Gianluigi Botton, Materials Technology Laboratory, Natural Resources Canada. Title of Talk: "Analytical TEM of Macrostructures and Nanostructures". Dr. Elaine Humphrey, Biosciences EM Facility, University of British Columbia. Topic: "Confocal Microscopy" Dr. John McCaffrey, Institute for Microstructural Sciences, National Research Council Canada. Title of talk: "Structural Characterisation of InAs/GaAs and InAs/InP Quantum Dots by TEM" Dr. Doug Perovic, Dept. of Metallurgy & Materials Science, University of Toronto. Topic: "EM of Semiconductors". ************ Fredericton, New Brunswick is a three hour drive north from Bangor, Maine and an eight hour drive from Boston or Montreal. June is a beautiful time in New Brunswick and you can find interesting things to do and see through the conference website tourism links.
We turned water to ice on plant surfaces using the Peltier stage. We did not take any nice pictures (I have a few ugly ones!). It was an idea re ice nucleation on plants for a student project idea that we did not follow up.
Dave
On Mon, 12 Mar 2001 13:45:29 -0600 "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } May be it's not really a phase transition, but you can } find pictures of dissolution and crystallization of } table salt at our web page: } http://www.umkc.edu/dentistry/microscopy/Esemimg.htm } } Vladimir } } -----Original Message----- } } From: Edmund Gierlik } To: Microscopy-at-sparc5.microscopy.com } Sent: 3/9/01 10:24 AM } Subject: ESEM-Need presentation of phase transition. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Email: gierlik-at-delta.sggw.waw.pl } Name: Edmund Gierlik } } Organization: Electron Microscopy Lab. } } Education: professor } } Location: Warsaw, Poland } } Question: Can anybody show the source of nice presentation of ESEM } application in } a phase transition investigation (water - ice, liquid - solid)? }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I need to fix live cells (mainly blood cells) on a microscope glass slide to prevent them from movement but to maintain their viability for 1-3 hrs. Also it should be non-stained with any dye cells in mono-layer. Material to fix cells should be optically transparent in 500-700 nm. Room temperature is asssumed.
Any input about applicable technologies (gels etc) with references about suppliers of materials that are involved and sources for description of those techniques will be highly appreciated.
Thanks in advance
Dmitri Lapotko, Ph.D.
Luikov Heat and Mass Transfer Institute 15, Brovka Street Minsk, 220072 Belarus
----- Original Message ----- } From: "Patton, David" {David.Patton-at-uwe.ac.uk} To: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} Cc: "'Edmund Gierlik '" {gierlik-at-delta.sggw.waw.pl} ; {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, March 13, 2001 4:07 AM
} I am getting confused about light sources and Ca ratio imaging. I } have talked to people who use standard Hg arc sources and seem to } have no problems. Others seem to feel that you need a stabilized } power supply or a Xenon source for stability (see below). Are we } talking about line voltage variations (which a line } conditioner/voltage regulator ought to deal with) or actual arc } source variation? Any opinions out there? Thanks- Dave
} From my observations of using both types of light source, the Xenon seems } far more stable with time. You don't notice the fluctuations that you get } with Hg bulbs. This is important for quantification, such as dual excitation } ratio methods (eg. Fura-2). } } Stephen H. Cody, } Colon Molecular and Cell Biology Laboratory, } Ludwig Institute for Cancer Research, } Post Office Royal Melbourne Hospital, } Parkville, Victoria 3050, Australia. } } Tel: 61 3 9341 3155 Fax: 61 3 9341 3104 } email: stephen.cody-at-ludwig.edu.au } www.ludwig.edu.au/confocal } } } } ---------- } } From: Tom Phillips[SMTP:PhillipsT-at-MISSOURI.EDU] } } Reply To: Confocal Microscopy List } } Sent: Wednesday, 28 February 2001 2:08 } } To: CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU } } Subject: Re: Xenon lamps } } } } All true but I believe one disadvantage is the peak illumination is } } less for some fluorochromes such as DAPI. We see a difference in our } } two systems that have Hg or Xe. } } } }
--
************************************************************ Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Greetings, Two standard points. These are not about power output but do relate to the choice of arc type (archtype??) which seemed to be on the questioner's mind. Forgive me if this is too obvious.
One is that the output of a xenon arc is much more uniform across the spectrum compared to the steep peaks of the mercury arc. For ratios with different excitation wavelengths, in principle difference in intensity between the two wavelengths doesn't matter, but in practice when the intensities differ alot, there can be problems. Therefore, if you are going to be working with a variety of wavelength pairs (lots of different dyes and applications) the continuity of the xenon will probably be helpful; whereas, if you will stick to a single pair of wavelengths, or ratio emitted light, then even spectral quality might not matter.
The other issue is that xeon arcs tend to give off lots more ozone than mercury arcs and so usually require significant venting. This can be a nuisance depdending on the lay of the land, etc.
As ever, Tobias Baskin
} } } I am getting confused about light sources and Ca ratio imaging. I } } have talked to people who use standard Hg arc sources and seem to } } have no problems. Others seem to feel that you need a stabilized } } power supply or a Xenon source for stability (see below). Are we } } talking about line voltage variations (which a line } } conditioner/voltage regulator ought to deal with) or actual arc } } source variation? Any opinions out there? Thanks- Dave } } } } } From my observations of using both types of light source, the Xenon seems } } far more stable with time. You don't notice the fluctuations that you get } } with Hg bulbs. This is important for quantification, such as dual excitation } } ratio methods (eg. Fura-2). } } } } Stephen H. Cody, } } Colon Molecular and Cell Biology Laboratory, } } Ludwig Institute for Cancer Research, } } Post Office Royal Melbourne Hospital, } } Parkville, Victoria 3050, Australia. } } } } Tel: 61 3 9341 3155 Fax: 61 3 9341 3104 } } email: stephen.cody-at-ludwig.edu.au } } www.ludwig.edu.au/confocal } } } } } ---------- } } } From: Tom Phillips[SMTP:PhillipsT-at-MISSOURI.EDU] } } } Reply To: Confocal Microscopy List } } } Sent: Wednesday, 28 February 2001 2:08 } } } To: CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU } } } Subject: Re: Xenon lamps } } } } } } All true but I believe one disadvantage is the peak illumination is } } } less for some fluorochromes such as DAPI. We see a difference in our } } } two systems that have Hg or Xe. } } } } } } } } -- } } ************************************************************ } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } 75 N. Eagleville Rd. U-125 } Storrs, CT 06269-3125 } Knecht-at-uconnvm.uconn.edu } 860-486-2200 860-486-4331 (fax) } home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html } ************************************************************
It could be a problem with penetration through the agar if cells are fixed in glutaraldehyde after the agar is added. Glut crosslinks the agar so that other solutions, including osmium and washes, can't penetrate properly. (I know this from experience.) We rinse the monolayer briefly and gently with warmed buffer (pour on, swirl, pour off). The buffer should be 300 mOsm (check with an osmometer) and pH 7.2-7.4 (check with a pH meter). Pour on the glut in buffer and let sit about 3 min (no more than 5); scrape, pellet cells in the glut and let sit for another 30 min. The pellets should be small (2 mm or so). If pellets are large, you can loosen them with the applicator stick so that the glut can get around to the underneath side. Just try not to disturb the clump too much. Remove glut with pipette. Scrape out pellet with flattened end of applicator stick. Enrobe with agar and then wash and stain as you would a piece of tissue. If there's a lot of excess agar, you can trim it away with a razor blade. This keeps them together and makes life much easier.
Some folks suspend cells in warm agar or gelatin before fixation and spin them down in the agar or gelatin and then fix. We have had variable staining this way, particularly if there is a lot of excess agar or the agar solidifies before the cells get to the bottom of the tube. If you can get a tight pellet by centrifuging fast in a warmed centrifuge (or jacketed tube) before the agar solidifies so that the layer of agar around each cell is thin, it will work. Do cut away the excess agar at the top of the tube, and make cell blocks small.
We centrifuge in a microfuge tube that looks like this:
| | | | | | | | | | \ / | | | | U
(Sorry about the graphics.) I think they come from PGC Scientific (800 424 3300). Sorry, I don't have the catelog number; our bag has been emptied, and our catelog has been "borrowed." They are about 2 " long and 3 mm in dia; the inside of the very tip is the diameter of a paper clip. This info was provided a while back, and maybe you can find it in the archives.
Centrifuge at right angles so that the pellet doesn't get stuck on the slanted neck of the tube. Then cut off the very bottom (below the cells) and then cut off the tube above the cells. Push out the cell "log" with a straightened paper clip. Chop into smaller logs.
Good luck. Address below if further questions.
On 12 Mar 2001, Bruce Cutler wrote:
} Date: 12 Mar 2001 12:58:48 -0600 } From: Bruce Cutler {bcutler-at-eureka.idl.ukans.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Agarose enrobed cells } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A user has encountered an interesting problem . When cells are scraped and } pelleted and embedded directly in epoxy they appear OK under the scope. When } they are enrobed in agarose and then embedded the appearance becomes variable, } although most cells appear very dense with contrast between organelles very low. } This variability in appearance can be on the same grid square. } Any ideas? } Thanks } Bruce } Bruce Cutler } Director, Microscopy & Electronic Imaging Laboratory } University of Kansas } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Dear Mary Gail, Do you mean a layer of metallic iron? I think you should treat the cellulose as wood or plant tissue and fix and embed, then cut a cross-section as you would a plant stem. No staining necessary. You need a TEM or STEM with an EDX on it and it will easily locate the iron to the ten nanometer scale. At 03:05 PM 3/12/01 -0500, you wrote: } } Dear listers, } I have a customer who wants TEM on a section of cellulose that is coated } with a layer of Fe2. I have no idea about how to go about accomplishing } this. He wants to see where the Fe2 is located in the cellulose matrix, } and how deep it has penetrated. The sample is a layer of cellulose about } 1/2 mm thick with a visible layer of Fe on the surface and it's in } submerged in water. } Any ideas out there? } Thanks so much, } } Mary Gail Engle
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hello, I was looking at some piping in our lab while sitting on the windowsill and noticed a sign painted over. Looking closer it was a sign for asbestos insulation that has been 80% painted over. I was told earlier that there was no asbestos pipe insulation in the lab. I collected some dust samples from near the fraying pipe insulation, on top of shelves, floor, and the windowsill and imaged them in our SEM.
Can anyone with some experience with asbestos take a look at the images at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the images looks like a candidate for asbestos? I think only fd6, fd7, and fd8 are asbestos, and the rest of the fibrous materials imaged are cellulose binding agents used in the insulation.
I will examine the material by TEM and electron diffraction, but I was wondering if someone could post the techniques they use for sampling from the dust, and any other hints and techniques I should be aware of.
Thanks for your help, Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
We have some data from a Wyko optical profilometer which we would like to work with offline & import into MATLAB for some further analysis. The lab tells me that the Wyko data can be output in "OPD" format. Does anyone know how we can covert this to a format we can work with.
Thanks
Tim
------------------------------------------------------------------------- Tim E. Harper CEO CMP Cientifica Tel. +34 91 640 7185 Fax. +34 91 640 7186
Gordon's comments concern me on several levels. The SEM, especially without EDX, is not the best tool to identify asbestos fibers. The TEM and diffraction can be absolutely definitive. However, I'm unaware of any EPA or NIOSH protocol for taking dust to the TEM grid, though one could be developed easily enough for Gordon's purposes. Gordon described friable pipe insulation with an asbestos warning. Seems wiser to bring his administration in on this situation. Does the UC system have some in-house personnel that can run a PLM for him? The lack of such in house personnel or an office dealing with asbestos abatement would be surprising for such a large university. Even a commercial lab PLM is faily inexpensive. While the amount of dust given off the pipe, barring some regular banging on the pipe, is probably minimal, the wiser course of action is bringing in someone who is well trained in dealing with asbestos pipe insulation. It doesn't have to be removed, just managed.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello, I was looking at some piping in our lab while sitting on the windowsill and noticed a sign painted over. Looking closer it was a sign for asbestos insulation that has been 80% painted over. I was told earlier that there was no asbestos pipe insulation in the lab. I collected some dust samples from near the fraying pipe insulation, on top of shelves, floor, and the windowsill and imaged them in our SEM.
Can anyone with some experience with asbestos take a look at the images at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the images looks like a candidate for asbestos? I think only fd6, fd7, and fd8 are asbestos, and the rest of the fibrous materials imaged are cellulose binding agents used in the insulation.
I will examine the material by TEM and electron diffraction, but I was wondering if someone could post the techniques they use for sampling from the dust, and any other hints and techniques I should be aware of.
Thanks for your help, Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron MicroscopeLab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id IAA07825 for dist-Microscopy; Wed, 14 Mar 2001 08:32:38 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id IAA07822 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 14 Mar 2001 08:32:08 -0600 (CST) Received: from meta.vwh.net ([192.41.39.216]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id IAA07815 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 14 Mar 2001 08:31:56 -0600 (CST) Received: from metabolix.com (cint.metabolix.vne.verio.net [199.103.134.10]) by meta.vwh.net (8.8.5) id HAA16348; Wed, 14 Mar 2001 07:30:00 -0700 (MST) X-Authentication-Warning: meta.vwh.net: Host cint.metabolix.vne.verio.net [199.103.134.10] claimed to be metabolix.com Message-ID: {3AAF8061.D94223EE-at-metabolix.com}
I am trying to develop a procedure to thin section plant leaves, stain them with Nile blue, and visualize the stained leaves by light microscopy.
I assume that a microtome would be the best way to perform the thin sectioning but have I have no experience with microtomes. Does anyone have suggestions on suitable microtomes to purchase for such a procedure? If so, where is the best place to purchase them?
Thank you.
-Kristi Snell
-- Dr. Kristi D. Snell Metabolix, Inc. 303 Third St. Cambridge, MA 02142 Fax: 617-492-1996 Phone: 617-492-0505 x 229
I want to track the performance of our EDS detector over time. We currently check: peak to background ratios via peak-to-tail of Mn k-alpha, symmetry via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha and L-alpha to known values, and resolution via Full Width Half Max. What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
A few years back when I took an EDS course put on by NORAN, they said keeping track of those K-alpha: L-alpha ratios should give you a running record of your detector sensitivity. I've been doing that ever since, and you can actually see those ratios change over time, as your detector window gets dirty. Another trick they mentioned (though I've never tried it myself) is to run a spectrum on some ordinary Teflon tape (yes, the kind you use for plumbing repairs). Teflon, apparently, contains no oxygen, so the presence of O in the resulting spectrum can only be from ice buildup. Clever, no? I also routinely do the Full Width Half Max test whenever I do a calibration. Good luck.
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
----- Original Message ----- } From: "Brian Wajdyk" {Brian.Wajdyk-at-onsemi.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 14, 2001 10:40 AM
I do not recall any problems with salt/water imaging.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----------------------------------------------------------------------. } } } We turned water to ice on plant surfaces using the Peltier } stage. We did not take any nice pictures (I have a few } ugly ones!). It was an idea re ice nucleation on plants } for a student project idea that we did not follow up. } } Dave } Like Dave, I've sometimes taken pictures of ice/water etc in our ESEM, but ice is such a good insulator, (I guess), the images never seem to come out well. So, I've never saved any. Also, salt/water interactions can be difficult too, since the salt really seems to want to charge a lot, to the detriment of image acquisition. Yes, you can play around with KeV and all that to minimize these effects, but it's still a very challenging "shoot" to get useable images.
Frank Thomas Geological Survey of Canada (Atlantic) Dartmouth, Nova Scotia
Brian, We work in a UKAS accredited laboratory and have a Pentafet light element detector with rotating turret device for windowless and thin window analysis. I perform a test with a Ni sample every month using the detector in its windowless mode. We monitor the ratio of the counts in the Ni L and Ni K peaks (NiL/NiL+NiK) (knowing what the value should be immediately after reconditioning). When the monthly value falls below a certain predetermined value, we simply perform the reconditioning routine. I suppose if you don't have a light element detector then the Cu standard you suggest is much better for a decent sized L line. I find that the above works very well for tracking the detector's sensitivity over a period of time. Best of luck Martin Roe
What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
Martin J. Roe Macaulay Land Use Research Institute Craigiebuckler Aberdeen Scotland UK Phone 01224 318611 e-mail m.roe-at-mluri.sari.ac.uk
I'd say some of those fibers are definitely "candidates". They are also large enough that you could analyze by PLM. If you want to do the TEM for fun or for the experience: dust samples are usually suspended in water and filtered onto a PC or MCE filter. I'm sure someone will post the appropriate reference, I can't seem to find it here.
Matt
On Tuesday, March 13, 2001 10:37 PM, Gordon Vrololjak [SMTP:gvrdolja-at-nature.Berkeley.EDU] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I was looking at some piping in our lab while sitting on the windowsill } and noticed a sign painted over. Looking closer it was a sign for } asbestos insulation that has been 80% painted over. I was told earlier } that there was no asbestos pipe insulation in the lab. I collected some } dust samples from near the fraying pipe insulation, on top of shelves, } floor, and the windowsill and imaged them in our SEM. } } Can anyone with some experience with asbestos take a look at the images } at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the } images looks like a candidate for asbestos? I think only fd6, fd7, and } fd8 are asbestos, and the rest of the fibrous materials imaged are } cellulose binding agents used in the insulation. } } I will examine the material by TEM and electron diffraction, but I was } wondering if someone could post the techniques they use for sampling from } the dust, and any other hints and techniques I should be aware of. } } Thanks for your help, } Gordon. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } }
This is a general site for Design News magazine; search that site for the term "lasik" to find an uncritical article featuring the inventor of the laser-vision machine now marketed by Visx. Even though it's a puff piece, it contains a great deal of info on the process.
2 recommended against the surgery, citing the risk of bad results for small gain if you have good correction now with glasses or contacts. Here are sites they cited with good info and extensive warnings of potential problems, and some additional sites I found in my own searches:
http://www.americaneye.com/bbs3/
This is a bulletin board where you can read many personal comments.
http://www.surgicaleyes.com/
This is a site founded by people who had unsuccessful laser vision correction resulting in complications. Its major point is that success defined by Snellen eye chart acuity may be inadequate.
http://wakeup.to/lasik
The site's name should tell you its slant on the topic.
http://www-psy.ucsd.edu/~mm/eyeknowwhy/index.htm
Very well-organized site, balanced viewpoint, but a bit out of date (last updated 1999). Well worth a careful reading anyway.
Detailed study results comparing PRK and LASIK outcomes. 3 years old (1998, therefore procedures done at least a year earlier). 220 eyes in the study. Interesting point (for me): 11.8% of PRK eyes and 3.2% of LASIK eyes reported a loss of 2 Snellen lines or more of best spectacle-corrected visual acuity (in English, you can't see as well as before even with glasses).
A more recent study (1999) with better outcomes. Equipment and experience makes a significant difference; be aware the technology changes fairly rapidly. 2-line loss rates from 2% to 0.6% in larger groups (1000+ and 700+ eyes).
A very good Feb. 2000 presentation of FDA pre-market approval data for several laser systems. 2-line loss is down to 0.3% with one system, but they show glare and night-vision problems reported by ~10% of patients. Remarkable Snellen acuity results: 87% 20/20 post-op without lenses.
5 respondents to my query had actually had laser surgery. All ultimately reported being satisfied with the results. 4 had no side effects at all. 3 reported good to excellent results immediately with no side effects. One (57 years old, farsighted) reported better post-op vision than previous lens-corrected vision. 1 respondent cited trouble for almost 1 year after surgery (couldn't see TEM images properly, trouble adjusting astigmatism), and said he was "initially devastated". However, he said he adjusted and/or the problems corrected themselves after a year. He can now handle all imaging tasks, and was happier overall than with glasses.
My personal decision has been to wait and see. I'm still not satisfied with the fairly high reported incidence of glare and night-vision problems, which I think are related to limited correction diameters compared to the dilated pupil of a larger-than-average eye (mine are). The changes I saw over just a few years of clinical trial results convinced me this is still evolving technology. The risks, while small, are still too high for me to accept in a purely elective procedure.
Thank you for your patience with this long-winded summary. Hope the info is useful to some of you.
Rick Mott (still reaching for his specs in the morning)
Dear Gordon, As someone in an old building with asbestos floor tiling, asbestos fume cupboard lining and old technicians who like to machine asbestos, I have looked at lots of samples of asbestos insulation. First, when asbestos is used, it is usally 30 % of the materials by volume, so it is very evident. Second it is characterized by very long, fairly straight fibres that divide down to smaller and smaller fibres, through and beyond the range of the SEM magnification. By that critierion, none of your pictures look like they contain asbestos. It helps to have looked at a sample so you can recognise it. Third, I check all suspicious fibres by EDX. The crysotile asbestos used in most insulation has a distictive Mg-Si spectrum, with a little Ca and Fe, that isn't found in most minerals. If it has that signature, then I would suspect asbestos and do the TEM. At 07:37 PM 3/13/01 -0800, you wrote: } Hello, } I was looking at some piping in our lab while sitting on the windowsill } and noticed a sign painted over. Looking closer it was a sign for } asbestos insulation that has been 80% painted over. I was told earlier } that there was no asbestos pipe insulation in the lab. I collected some } dust samples from near the fraying pipe insulation, on top of shelves, } floor, and the windowsill and imaged them in our SEM. } } Can anyone with some experience with asbestos take a look at the images } at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the } images looks like a candidate for asbestos? I think only fd6, fd7, and } fd8 are asbestos, and the rest of the fibrous materials imaged are } cellulose binding agents used in the insulation. } } I will examine the material by TEM and electron diffraction, but I was } wondering if someone could post the techniques they use for sampling from } the dust, and any other hints and techniques I should be aware of. } } Thanks for your help, } Gordon. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hello All, I have the opportunity to obtain a Seiko SMI8300 focused ion beam machine. I can't find out very much about it (I didn't even know Seiko made them - I suspect they are usually native to Japan). Does anyone have any useful information or experience? Could I use one to make TEM specimens?
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
One last note: of the 4 respondents who enthusiastically recommended the procedure, all were in agreement on the importance of finding a highly experienced surgeon. Complication rates are inversely related to experience.
I've never tried this either, but doubt it should work for the stated reason. A layer of ice on the detector (actually the oxygen in the ice) will act mostly as an x-ray absorber rather than a source of O x-rays. In fact, the ice will be a poor absorber for O-K x-rays because the oxygen x-ray energy is below its absorption edge.
Larry Thomas Pacific Northwest National Laboratory Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
---------- From: Frank Thomas Sent: Wednesday, March 14, 2001 7:30 AM To: Brian Wajdyk; Microscopy-at-sparc5.microscopy.com Subject: Re: EDS detector chracterization: sensitivity/efficiency
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Brian -
A few years back when I took an EDS course put on by NORAN, they said keeping track of those K-alpha: L-alpha ratios should give you a running record of your detector sensitivity. I've been doing that ever since, and you can actually see those ratios change over time, as your detector window gets dirty. Another trick they mentioned (though I've never tried it myself) is to run a spectrum on some ordinary Teflon tape (yes, the kind you use for plumbing repairs). Teflon, apparently, contains no oxygen, so the presence of O in the resulting spectrum can only be from ice buildup. Clever, no? I also routinely do the Full Width Half Max test whenever I do a calibration. Good luck.
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
----- Original Message ----- } From: "Brian Wajdyk" {Brian.Wajdyk-at-onsemi.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 14, 2001 10:40 AM Subject: EDS detector chracterization: sensitivity/efficiency
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Fellow Microscopists, } } Fellow Microscopists, } } I want to track the performance of our EDS detector over time. We } currently } check: peak to background ratios via peak-to-tail of Mn k-alpha, } symmetry } via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha } and } L-alpha to known values, and resolution via Full Width Half Max. What I } don't have is a good way to determine sensitivity of the detector to } determine when to remove ice/hydrocarbon build up. In the past I used a } 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha } because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. } Unfortunately I no longer have this standard or the means to acquire a } new } one thanks to the slow down in the e semiconductor industry. My best } guess } would to be to clean the detector then use the k-alpha to l-alpha ratio } of } Cu. However I know that we have the best microscopists in the world at } our } disposal. Any suggestions for sensitivity tracking, or other useful } detector characterizations I may have missed, would be greatly } appreciated. } } Sincerely, } } Brian Wajdyk } Sr. Electron Microscopist } On Semiconductor } brian.wajdyk-at-onsemi.com } 602-244-4883 } }
Perhaps a Macintosh, computer guru could figure this one out.
We are capturing JPEG images from a standard (Sony, analog 470 line, NTSC) video camera using a program called Reel Eyes. The images are captured on a Mac 8500 via the built-in S-Video port and saved as Quicktime JPG's (as mandated by the Reel Eyes program). When we put these images on our server they now appear with ".bin" appended to the file name so that the new image is described as a "Application/x-macbinary". Mac folks can open the files after processing through Stuffit but PC folks are unable to deal with the files. Anyway, this is an additional step that should not be present.
We can remove this problem by re-opening each of the images in Photoshop and saving them without the Thumbnail (or preview). So, I conclude that Quicktime is always saving with the Thumbnails in place. Does anyone know of a way that we can prevent the thumbnails from occurring in Quicktime?
Any other suggestions would be most welcome.
Many thanks,
John B.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
The problem is that your files are encoded as Macbinary (.bin) before being stuffed. Windows machines can't decode .bin files unless they have the windows version of Stuffit expander. Encourage them to get it.
To correct this you need to change a preference setting.
In Magic Menu or Dropstuff preferences, go to the "Cross-Platform" or "MacBinary" icon and choose "Never" (instead of "Smart"). There is no way to change this preference in the StuffIt Deluxe application.
The URL for this information is {http://www.aladdinsys.com/support/techsupport/mac/dlx/dlx611.html}
Happy capturing. Gordon Nord "My real job is Mineralogy"
"John J. Bozzola" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Perhaps a Macintosh, computer guru could figure this one out. } } We are capturing JPEG images from a standard (Sony, analog 470 line, } NTSC) video camera using a program called Reel Eyes. The images are } captured on a Mac 8500 via the built-in S-Video port and saved as } Quicktime JPG's (as mandated by the Reel Eyes program). When we put } these images on our server they now appear with ".bin" appended to } the file name so that the new image is described as a } "Application/x-macbinary". Mac folks can open the files after } processing through Stuffit but PC folks are unable to deal with the } files. Anyway, this is an additional step that should not be present. } } We can remove this problem by re-opening each of the images in } Photoshop and saving them without the Thumbnail (or preview). So, I } conclude that Quicktime is always saving with the Thumbnails in } place. Does anyone know of a way that we can prevent the thumbnails } from occurring in Quicktime? } } Any other suggestions would be most welcome. } } Many thanks, } } John B. } } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ##############################################################
-- Gordon Nord Small Business Network Design and Construction Macintosh and Windows - Solutions and Conflicts
Nord Consultants 20594 Cornstalk Terrace Ashburn VA 20147
Yes, I did use Hitach 4500S and 3500N to study some coated fibers in the recent past years. The coating layers were up to three.
General speaking, I think this kind of preparation needs patient and do it properly at every step as the processing goes.
The steps I did for preparing the coated fibers for SEM are 1. Cut the fibers carefully into about 1 cm length of pieces 2. Arrange them in a longitudinal direction (do not hurt them) 3. Hold one end of the fibers by using a pair of tweezers 4. Apply thin M-Bond (16) on to the fibers Since the M-bond is so thin, it will go along with the fibers and surround almost every fiber very well. 5. Leave the fibers in the air (room temperature) overnight. Do not let the wet part touch any solid surface such as metal or microscope slide. Otherwise when it cures, it is very difficult to separate them. Another way to cure it is putting the fiber with the tweezers on to a hot plate at 130 C for one hour. Do not let the wet part touch the hot plate. 6. When the fibers are cured, embed them into epoxy to make a big enough piece so that you can hold and polish it for SEM. At this step, you have to avoid trapping air in the epoxy. Especially when a fabric of fibers is going to be embedded into epoxy, you have to use a pump to get rid of air bobbles before curing. You may choose a kind of epoxy which can be cured in couple of hours in air (room temperature) or on a hot plate for about 1 to 2 hours at 130 C. The epoxy I used is a kind of mixture of two parts. 7. Then you may follow the routing procedures for polishing SEM samples, i.e., grinding first then polishing, from coarse to fine carefully.
Finally if you are going to study a fabric, then you can cut a size of 1 x 1 cm2, and hold it with a tweezers at a corner, and apply a layer of M-bond (16) on both sides, put it onto a hot plate for curing. Then embed this piece of fabric into epoxy. Again, you have to pump the air out before curing.
Good luck.
Zhenquan Liu
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Zhenquan Liu (Dr.)
------------------------------------ Zhenquan Liu (Dr.) Arizona State University Center for Solid State Science Room PSA213 Tempe, AZ 85287 Tel (480) 965 4544 (o) (480) 775 7428 (h) Fax (480) 965 9004 (o) Email zhenquan.liu-at-asu.edu ------------------------------------
Hello, We have a Gatan Duomill IBT Model 600. We have been having some problems with it lately. When I turn on the IBT, the vacuum in the main chamber works fine. When pumping down the specimen chambers, the left side does not pump at all. The right side pumps to the proper level, but when you lower the sample into the chamber, it takes a while for the vacuum to recover. There is no vacuum instability during sample rotation. There is no arcing in the guns. The argon flow is at the proper level. We have done the following: -cleaned, regreased and changed o-rings in the Whisperlock Assembly, sample viewing port, sample chamber, vent/vac buttons, and ion guns -changed the cathode tubes -dusted out the main sample chamber -check the vacuum pumps and topped off oil in the roughing pump -DP seems to be running fine
Still it doesn't seem to help. We are lost. Any suggestions about what might be going wrong here? Thanks. Prad
Pradyumna Prabhumirashi Department of Materials Science Northwestern University Phone: (847)-491-7798 Fax: (847)-491-7820 http://vpd.ms.northwestern.edu/prad.htm
We have used the k:alpha:l-alpha ratio for copper and nickel for many years as a measure of ice buildup on our detectors. Either ratio is very sensitive to ice buildup. With single element standards you don't have to worry if the mixture of two elements in your standard has changed. For sources we use copper or nickel TEM grids which are cheap and available from all the EM vendors.
Regards, Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Brian.Wajdyk-at-onsemi.com on 03/14/2001 10:24:35 AM To: Microscopy-at-sparc5.Microscopy.Com cc:
Fellow Microscopists,
Fellow Microscopists,
I want to track the performance of our EDS detector over time. We currently check: peak to background ratios via peak-to-tail of Mn k-alpha, symmetry via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha and L-alpha to known values, and resolution via Full Width Half Max. What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
Just got a call from a researcher on campus looking for service on a Reichert Frigo-Cut type 2800, s/n 0110117 microtome. I am not familiar with this piece of equipment (we don't do light level microtomy) and I do not know this gentleman but he had already contacted one northern California service and was quoted $130/hr p to p. He balked. I explained that I had been quoted as much from services in New Hampshire. If anyone has a resource for repairing one of these microtomes in the Northern California region I will pass the info back to this person.
Thanks in advance.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Thanks for all of your replies, We had someone from our physical plant come in, and they revealed that the colour of the painted over sign indicates asbestos - free - insulation. Wierd scare for me since the painter covered the most important part of the label - that the insulation was asbestos free, not asbestos. They'll come back to tape back up all of the loose insulation tomorrow.
Since one of the specimens I looked at is very similar in morphology to asbestos, they'll be doing some wipe tests and counts by polarized light microscopy for asbestos particles to be sure. It likely was left behind from when the original asbestos insulation was removed about 10 years ago or so.
I appreciate all of the replies on the thread and supportive messages. Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Wed, 14 Mar 2001, Mary Mager wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Gordon, } As someone in an old building with asbestos floor tiling, asbestos fume } cupboard lining and old technicians who like to machine asbestos, I have } looked at lots of samples of asbestos insulation. First, when asbestos is } used, it is usally 30 % of the materials by volume, so it is very evident. } Second it is characterized by very long, fairly straight fibres that divide } down to smaller and smaller fibres, through and beyond the range of the SEM } magnification. By that critierion, none of your pictures look like they } contain asbestos. It helps to have looked at a sample so you can recognise } it. Third, I check all suspicious fibres by EDX. The crysotile asbestos used } in most insulation has a distictive Mg-Si spectrum, with a little Ca and Fe, } that isn't found in most minerals. If it has that signature, then I would } suspect asbestos and do the TEM. At 07:37 PM 3/13/01 -0800, you wrote: } } Hello, } } I was looking at some piping in our lab while sitting on the windowsill } } and noticed a sign painted over. Looking closer it was a sign for } } asbestos insulation that has been 80% painted over. I was told earlier } } that there was no asbestos pipe insulation in the lab. I collected some } } dust samples from near the fraying pipe insulation, on top of shelves, } } floor, and the windowsill and imaged them in our SEM. } } } } Can anyone with some experience with asbestos take a look at the images } } at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the } } images looks like a candidate for asbestos? I think only fd6, fd7, and } } fd8 are asbestos, and the rest of the fibrous materials imaged are } } cellulose binding agents used in the insulation. } } } } I will examine the material by TEM and electron diffraction, but I was } } wondering if someone could post the techniques they use for sampling from } } the dust, and any other hints and techniques I should be aware of. } } } } Thanks for your help, } } Gordon. } Regards, } Mary } } Mary Mager } Electron Microscopist } Metals and Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchg.ubc.ca } }
A few years ago, a EDS reparer explain me a test, which seems to be interresting, to check the "health" of a detector with UTW, but I never have tried it.
With a fluorine (CaF2) sample at 10 keV, You should have a ratio of one betwween F K alpha and Ca K alpha. When not, or when it diminish in the time (less F), you have ice (absorbtion effect of F by O2). (Why 10 kev? Is 10 keV not a bit high for F. It think it's a compromise between the window caracteristics and the primary energy).
Same thing with quartz (SiO2), at 5 keV, and than, if O diminish, you have oil on the window (absorbtion effect of O2 by C).
As I said, I never tried this test, the CaF2 and SiO2 samples are in my drawer, waiting for polishing. I suppose that geologist should have an advice about that, but of course they don't often work at 5 keV for EDS.
I usually use a aluminum sample holder, with a piece of carbon adhesive tape and a copper foil, and I take a spectrum with a quarter of the SEM screen on the copper, a quarter on the carbon, and the half on the aluminium,at low mag, and at 5 keV. I adust the probe current to a preset value (typically a few nA, and each time the same value) mesured on the carbon tape (less SE and BSE, better would be a Faraday cup). So I see if the sensitivity on C K, Cu L and Al K seems to diminish from on mesure to an other. In such condition you'll see that the count rate (and of course, the spectrum shape) is very sensitive to window or detector contamination (with allways the same probe current, time constant, etc.). You can imagine similar test with for exemple, Vandium L and silicon K, etc.
best regard
Jacques
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
I was asked to perform double labelling of surface antigens on leukaemia cell cultures and cord blood progenitors.
In the past I used a pre-embedding protocol for single labelling which worked quite well.
I would like to continue with a pre-embedded protocol, but both primary antibodies are raised in the same animal and I wonder how I can block unspecific labelling in that case.
(I have a protocol for post-embedded double staining with primary AB raised in the same animal, but would prefer to use that as a last resort)
So far I used a 10 nm gold F(ab)2- AB - if I use a 5nm gold F(ab)2 AB will I be able to distinguish them in the TEM?
I would appreciate your input very much.
Many thanks
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
I have a technique that I use with clients on a wide range of fibres which takes very little time.
1. Drill two or three holes through two SEM stubs placed face to face. 2. Pass the fibres through the holes. 3. Fix in place with a water based carbon solution, make sure the fibre/stub interface is well wetted. 4. When fully dry place in liquid nitrogen CARE!!. 5. When the bubbling stops lift out CARE!! 6. Place on an insulating surface and with a knife strike the interface between the two stubs - the system will fracture. 7. When dried off you have two sets of surfaces to look at in LM or SEM.
It works great on a number of differing media other than fibres, the only snag is they must not be affected by the water base.
Steve Chapman Senior Consultant Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
I imagine that the problem arises when you transfer the files to the server. I'll assume that you are using some FTP program such as Fetch. In the upload preferences, make sure that you are transferring non-text data in the "raw data" format, as opposed to MacBinary, etc. Also, make sure that the FTP doesn't append a ".bin" to raw data.
Remember you still need to have a ".jpg" at the end of the filename or else the Windows boxes still won't know how to handle the file (sigh...). There are numerous utilties on the Mac to batch process the renaming of files for cross-platform compatibility.
Hope this helps, John B.
} } Perhaps a Macintosh, computer guru could figure this one out. } } We are capturing JPEG images from a standard (Sony, analog 470 line, } NTSC) video camera using a program called Reel Eyes. The images are } captured on a Mac 8500 via the built-in S-Video port and saved as } Quicktime JPG's (as mandated by the Reel Eyes program). When we put } these images on our server they now appear with ".bin" appended to } the file name so that the new image is described as a } "Application/x-macbinary". Mac folks can open the files after } processing through Stuffit but PC folks are unable to deal with the } files. Anyway, this is an additional step that should not be present. } } We can remove this problem by re-opening each of the images in } Photoshop and saving them without the Thumbnail (or preview). So, I } conclude that Quicktime is always saving with the Thumbnails in } place. Does anyone know of a way that we can prevent the thumbnails } from occurring in Quicktime? } } Any other suggestions would be most welcome. } } Many thanks, } } John B. } } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ##############################################################
At 01:26 PM 3/14/01 -0600, John J. Bozzola wrote: } The images are captured on a Mac 8500 via the built-in S-Video port and saved as Quicktime JPG's (as mandated by the Reel Eyes program). When we put these images on our server they now appear with ".bin" appended to the file name so that the new image is described as a "Application/x-macbinary". Mac folks can open the files after processing through Stuffit but PC folks are unable to deal with the files. Anyway, this is an additional step that should not be present. } We can remove this problem by re-opening each of the images in Photoshop and saving them without the Thumbnail (or preview). So, I conclude that Quicktime is always saving with the Thumbnails in place. Does anyone know of a way that we can prevent the thumbnails from occurring in Quicktime?
The Mac's file system is slightly different than that on a PC or a Unix server. Mac files have two parts, known as forks. The icon or thumbnail image resides in the "resource fork". The regular JPEG image data that a PC expects is in the "data fork." The resource fork also holds the Mac's association between the file and the program that made it, while the PC uses the file extension for that purpose.
MacBinary files are a way of wrapping up the data and resource forks into a single file, such as for transmission via an e-mail attachment, ftp site, etc. or in your case, handy for putting on a server that doesn't speak with forked tongue.
You didn't say much about your server. If it's a WinNT server, your admin can install some AppleTalk extensions that can make PC {-} Mac file exchange a bit easier. They automatically manage the resource fork, hiding it for PC programs but allowing networked Macs to store and access it.
You also didn't say much about which PC programs you were using. I don't know of any that handle MacBinary files. (I've written a few that did, once upon a time, but they're obscure.) You may be forced to either strip the files on the Mac, or use a utility on the PC side to convert the MacBinary files to data-only files.
I don't think Quicktime thumbnails are the problem, I think it's a MacBinary problem.
Question: I have been analyzing stress around STI (shallow trench isolation) in DRAM. In general, stress is higher at bottom corner and top corner of STI, which is probed by my data. But, the strange thing is that HOLZ line split and blurring occurs in CBED pattern at the bottom of FOX (field oxide). I know that the split and blurring is the sign of high stress. And I don't think that stress is higher at the bottom of FOX. So, my question is what makes line split and blurring except stress. Have you seen that kind of phenomenon? Thank you for your answer.
If you go to our new website at www.southbaytech.com, you will find a listing of technical papers that you can request. There are several there dealing with coated fibers. Please take a look. If any look interesting, submit a request and we will send them out to you at no charge.
When you get to the site, click on Applications support then technical papers. There are a lot of papers there, so the easiest way to find the ones you are interested in is to use the "find on page" function typically found under your edit command. If you type in "fiber", you'll find some relevant papers.
I hope this helps.
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
The ion getter pump on the CM10 (12 and 20) does have a finite life and if your machine is 10 years old with the original IGP then I suspect that the pump could be finished. I assume that it will not pump down properly on the IGP but has a good penning gauge (P3) pressure.
The pump is not a standard Edwards pump as it has an extra port so that it can pump both the column and the gun. As I understand it the pump body is cut open to replace the plates and then rewelded before being leak tested and baked.
There are companies that will do this for you but you are without the pump while they do it, this may take two weeks or more. FEI should hold replacement pumps that they would swop in a day, more expensive but faster and more convenient. It depends on your budget and facilities.
Good luck, Ron
On Thu, 15 Mar 2001 14:08:46 +0100 Marco Arienti {marienti-at-tiscalinet.it} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have no big experience with Philips microscopes, but I need some } information. } } A CM10 installed about 10 years ago have some vacuum problems. } } Looks like the Ion Getter Pump is not working anymore. } } As far as I understood the problem may be that there is no more Titanium in } the pump. } } Is possible In this one to change the only grids (like in the Leibold pumps) } or the all pump have to be exchanged? } } It is a pump manufactured from Edwards, any info about the model? } } Thanks a lot. } } } } Marco Arienti } } www.electronrescue.com } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Could you recommend a water based carbon solution?
Dave
On Thu, 15 Mar 2001 04:56:05 -0500 Steve Chapman {PROTRAIN-at-CompuServe.COM} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi } } I have a technique that I use with clients on a wide range of fibres which } takes very little time. } } 1. Drill two or three holes through two SEM stubs placed face to face. } 2. Pass the fibres through the holes. } 3. Fix in place with a water based carbon solution, make sure the } fibre/stub interface is well wetted. } 4. When fully dry place in liquid nitrogen CARE!!. } 5. When the bubbling stops lift out CARE!! } 6. Place on an insulating surface and with a knife strike the } interface between the two stubs - the system will fracture. } 7. When dried off you have two sets of surfaces to look at in LM or } SEM. } } It works great on a number of differing media other than fibres, the only } snag is they must not be affected by the water base. } } Steve Chapman } Senior Consultant Protrain } For professional training in SEM, TEM and EDX world wide } www.emcourses.com }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I need some info on liquid nitrogen dispensing devices. Our EM lab has four 20 liter dewars for liquid nitrogen storage. A piece of styrofoam (2 x 6) is attached to the lid. We would like to purchase a dispensing device because it would be easier to use and less waste of liquid nitrogen. Thanks in advance Karen Schlueter PWA, ARS, USDA EM Lab 1636 East Alisal St. Salinas, CA 93905 (831) 755-2847 FAX: (831) 755-2814 kschlueter-at-salinas.ars.usda.gov
As a materials scientist, I haven't had any experience doing TEM sample prep of biological samples, so I need some assistance in this.
I've been asked to look at in the TEM some dehydrated beef looking for the changes in sarcomere length etc due to the dehydration process.
I have at my disposal staining solutions of 0.5% RuO4, and 2% methanolic UA and need some procedures on how to go about preparing these specimens. (Things like cut the dehydrated sample into cubes, stain with this for how long at this temp then embed and microtome or such.)
Thanks in advance.
dz
David Ziegler U.S. Army, SBCCOM AMSSB-RSS-MS(N) Materials Science Team, SS&T Natick, MA 01760-5020 TEL: (508) 233-6484 FAX: (508) 233-5521 Email: David.Ziegler-at-Natick.Army.Mil
I am looking to purchase a sputter coater ~$5000. I currently have a Polaron E5200 that is inoperable. I do Failure Analysis on semiconductors, probably a 6 inch chamber & Au/Pd target. Can anyone in the list give me some good tools that work for them.
Anyone familiar with BOC Scancoat Six?
Thanks in advance
Jim Arnold Senior Quality Technician / Failure Analysis Honeywell - Microelectronics and Technology Center Columbia, MD 21045 email: jim.arnold6-at-honeywell.com
For organic contamination, monitoring the ratio of TiKa to TiLa is good because TiLa is absorbed heavily by organic layers on the detector window. For ice, a spectrum from say calcium fluoride is good because again fluorine Ka is heavily absorbed by oxygen containing layers such as ice.
Carl Miller ---------------------- Forwarded by Norman C Miller/RES/Raytheon/US on 03/14/2001 02:00 PM ---------------------------
"Brian Wajdyk" {Brian.Wajdyk-at-onsemi.com} on 03/14/2001 09:40:51 AM
To: Microscopy-at-sparc5.microscopy.com cc: (bcc: Norman C Miller/RES/Raytheon/US)
Fellow Microscopists,
Fellow Microscopists,
I want to track the performance of our EDS detector over time. We currently check: peak to background ratios via peak-to-tail of Mn k-alpha, symmetry via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha and L-alpha to known values, and resolution via Full Width Half Max. What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
I am looking to replace my target on my sputter coater and would like to find out if there is an optimal thickness for the target. Is it material dependant? I had requested 2.5mil from the thickness of my old target but the vendor I spoke with will only source a minimum 4mil thick target. I am looking at Pt and Au/Pd. Will I have to increase current to get the same deposition rate for a thicker target?
TIA
David B Rose W. L. Gore and Associates 297 Blue Ball Road Elkton, MD 21921 410-506-2958
Many thanks to all of you who offered advice, assistance, consolation and even a loan of the part or a used instrument for parts. I was able to find a replacement ($100 - overpriced perhaps but worth every penny!) and will get it installed next week with the directions from another microscopy list reader.
Again my thanks to all and if anyone wants the names and addresses of some LKB parts dealers and/or service folks, I will gladly provide you with my new list off-line.
Some years ago I had Melins (510-234-5749) repair our Reichert 2800. They insisted on taking it to their shop and it needed a new compressor from Europe. The bill was over $2000. $130/ hour is a typical charge for this type of work in this area. When I the Reichert unit was destroyed in a water flood I purchased a Microm cryostat. It uses standard easily available refrigeration components. The cost and repair time is gresatly reduced. Good luck.
At 03:35 PM 3/14/01 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
No, thicker target = longer life, but it makes no difference to deposition rate, current etc. Chris
----- Original Message ----- } From: {"drose-at-wlgore.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 15, 2001 7:23 PM
No, thicker target = longer life, but it makes no difference to deposition rate, current etc. Chris
----- Original Message ----- } From: {"drose-at-wlgore.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 15, 2001 7:23 PM
G'day
I have an aging Jeol 840 and the monitor image is getting a bit dim and noisy to the extent that I have to use large spot sizes that are damaging some samples! Jeol tell me they cannot supply new tubes. Does anyone know of compatible tubes that could be utilised? Is there anything special about SEM monitor tubes?
Thanks Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
I have some difficulties to stick my protein (Mojor Sperm Protein, a basic protein from Nematode Sperm ) to the microscope slide, therefore, it is hard to perform further perfusion experiment. When I try to do so, I lose everything. Does anyone have experience in such a manipulation? Any suggestion is highly appreciated. Long
The CRTs can be rebuilt for about $600.00 to $800.00 USD by Richardson Electronics in Georgia.
I have their number & can give it to you offline.
Earl weltmer
----- Original Message ----- } From: "Dave Phelan" {emudp-at-mail.newcastle.edu.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 15, 2001 4:35 PM
Try Richardson Electronics http://www.rell.com/ . Have CRT information handy (labels on the CRT) when calling Richardson Electronic. Good luck.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
G'day
I have an aging Jeol 840 and the monitor image is getting a bit dim and noisy to the extent that I have to use large spot sizes that are damaging some samples! Jeol tell me they cannot supply new tubes. Does anyone know of compatible tubes that could be utilised? Is there anything special about SEM monitor tubes?
Thanks Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
Hi: We are lookng to purchase a Zeiss 10C TEM, preferable in working condition but would consider one needing some work. Please contact us direct via e-mail or call 909 301-9130 (Los Angeles area) EMSI Peter Jordan
Dear All, I'm looking for circuit diags for J-TEC X-ray counting electronic units XM-XCU. These were supplied with some JEOL 733 'probes although most seem to have gone out with ORTEC. Can anyone help? Thanks, MAlc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
Both display and photo-monitor tubes can be recoated. We had our photo CRT recoated by Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if they're still in business, but they did a good job. JSIII
} Try Richardson Electronics http://www.rell.com/ . Have CRT information handy } (labels on the CRT) when calling Richardson Electronic. Good luck. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } } This message is made of 100% recycled electrons. } -----Original Message----- } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Thursday, March 15, 2001 10:28 PM } Subject: Jeol 840 viewing screen replacement } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Julian P.S. Smith III Dept. of Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x6427 (vox) 803-323-3448 (fax)
Does anyone use this resin and use it for antibody staining? We will be using this resin for embedding. Also any suggestions on etching. Thanks. Caroline Miller
I have a Tracor Northern TN-5402 EDS system mounted on a JEOL SEM. The monitor just went up in smoke and I need to a) find a replacement or b) try to replace the hardware. Since I don't have much money and we would like to replace the whole system in a couple of years I can't spend much to repair the monitor.
Does anyone have a monitor for sale/offer? Does anyone know of a replacement monitor available on the market? A standard BNC type of monitor does not work because of a proprietary modulation. Has anyone successfully upgraded the hardware minus the detector?
Thanks, Louie
-- Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
VISIT OUR WEB SITE: http://www.mbl.edu/ http://www.courses.mbl.edu/
Our Sorvall MT2B is having some problems, to say the least. I would very much appreciate any info. concerning service of this machine. We are located in Milwaukee, WI. I have contacted Boeckeler (new owners of RMC) regarding service. They informed me they are no longer providing service on these models. What other options do I have?
TIA Craig M. Klotz, BS,CT(ASCP) EM Tech II Neuromuscular Lab Medical College of Wisconsin cklotz-at-mcw.edu
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
Claudia, I would direct you to the March issue of journal of histochemistry and cytochemistry, they have published a procedure to double-label with gold at the pre-embedding stages, good luck Marge
Margaret Springett IEM Specialist Electron Microscopy Core Facility Mayo Foundation email: springett.margaret-at-mayo.edu
} ---------- } From: Claudia Hayward-Costa } Sent: Thursday, March 15, 2001 2:49 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: IEM: Double labelling of cells } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear Microscopists, } } I was asked to perform double labelling of surface antigens on } leukaemia cell cultures and cord blood progenitors. } } In the past I used a pre-embedding protocol for single labelling } which worked quite well. } } I would like to continue with a pre-embedded protocol, but both } primary antibodies are raised in the same animal and I wonder how } I can block unspecific labelling in that case. } } (I have a protocol for post-embedded double staining with primary } AB raised in the same animal, but would prefer to use that as a } last resort) } } So far I used a 10 nm gold F(ab)2- AB - if I use a 5nm gold F(ab)2 } AB will I be able to distinguish them in the TEM? } } I would appreciate your input very much. } } Many thanks } } Claudia } } } Dr. C. Hayward-Costa } School of Life Sciences } Kingston University } Penrhyn Road, Kingston upon Thames } Surrey KT1 2EE, UK } 44(0)208 547 2000 x 2240 } Email: c.hayward-at-kingston.ac.uk } }
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
As many of you know, I maintain the WWW pages for Project Micro which is dedicated to Educational Outreach to pre-college students. One of their projects is sand from around the world which is used in the classroom activities section.
Today I uploaded their latest web page documenting their sand collection administered by Joe Neilly of Abbott Labs . You can see this page at
I could not help to notice that their supply of sands from outside the USA has become sorely lacking. In particuliar
Europe, Africa and South America
are now all completely depleted. And
Australia and Asia
only have supplies from only one location left.
Can I suggest to members of this list particularly if you are not from the USA, that if you have the opportunity this would be an excellent and simple way that you can contribute to the education of our next generation of microscopist's. The sand is available to any educator, regardles of their affiliation and location.
All the information you need to contribute "sand" from your local beach is listed on that WWW page as well as the end of this message.
It will cost you a few $$ out of your pocket to send the sand , but remember, it is for a good cause. So spend a few minutes next time you drive past a beach and fill up a small plastic bag, and send it to Joe.
Cheers.... Nestor Your Friendly Neighborhood SysOp.
--------------------------- Here is an exerpt from the Sands WWW page
To Request Sand
Select the sands you would like from the list below and e-mail your request to joe.neilly-at-abbott.com. Include your selection (limit 6 per request), where the sands should be sent, and how you plan to use them. This collection was created for Microscopic Explorations but how you ultimately use them is up to you. Great stories and photos about how you used the sands are always welcome!
To Donate Sand
Sand donations are what keeps this collection going. All donations are appreciated and will be shared with many educators. To send your sand, fill a Ziplock sandwich bag half full and place it in another Ziplock bag. Mail your donation to:
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Dear Listservers, recently there was a discussion about the stability of LR-White in the EM-beam. One distribtion mentioned a polymerization of LR-White by microwave treatment. I don´t remember who sent this mail. I would be interested to test this procedure. Who knows how to do this kind of LRW polymerization? What is the set-up? How long does it take, how much Energy (Watt) is needed? Maybe the colleague who posted this mail remembers the discussion and is able to give some information.
Thanks in advance, with best regards, Michael
Michael Reiner University of Cologne, Germany Dept. of Anatomy I _______________________________________________________________________________ Alles unter einem Dach: Informationen, Fun, E-Mails. Bei WEB.DE: http://web.de Die große Welt der Kommunikation: E-Mail, Fax, SMS, WAP: http://freemail.web.de
Dear Michael: The following procedure for polymerization of LR-White has worked of me in my microwave oven: I will start with the infiltration phase: The wattage setting I used was about 700-750 watts.
1:1 95% ETOH / LR-White 50'C temperature restriction setting (trs) x2 for 10 minutes each.
100% LR-White 50'C (trs) x3 for 10 minutes each.
Polymerization in the oven: In this phase be sure to fill the beem capsule to the very top and cap off with parafilm to make a tight seal to prevent water from getting into the resin. Submerge the capsules in a water bath and place the temperature probe in the bath to monitor the temp.
The blocks will be firm and will cut very well. For more information on this technique you can contact Rick Giberson at TED PELLA CORP. USA using their toll free number, I don't know what that is in Germany! He can give you many more details about LR-White and the microwave oven. I understand that lesser wattage setting can be used to infiltrate and polymerize LR-White but I have not experimented with them as yet!
Good Luck
Ron Austin (Research Associate) Dept. of Pathology L.S.U. Medical Ct. Shreveport, LA 318-675-4775 rla-at-mindspring.com
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by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id LAA21224 for dist-Microscopy; Sun, 18 Mar 2001 11:59:42 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id LAA21217 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Sun, 18 Mar 2001 11:59:11 -0600 (CST) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id LAA21210 for {microscopy-at-msa.microscopy.com} ; Sun, 18 Mar 2001 11:59:00 -0600 (CST) X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {v03130303b6daa5368faf-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Colleagues
A number of people have reminded me about the growing problems with hoof and mouth disease around the world and that sending possible organic material from country to country should not be considered at this time.
I humbly acknowledge that you are all absolutely correct!
It simply did not occur to me to connect sand with this problem. I guess my physics background is just showing it's tunnel vision, since I think of sand as inorganic compounds and neglect to think of organic "contaminants" which might be included.
For the moment it would therfore certainly be prudent to hold off collecting and/or send and sand samples for the Micro Project, even if it can be documented that the samples have been properly sterilized. Let's wait until this potential problem is controlled. It is better to error on the safe side.
Thanks again, to everyone that pointed out my error.
Dear Microscopists, I am working with Ilex paraguariensis seeds. After dissection, with mechanical extraction of the endocarp, the micropylar endosperm is enveloped by a lignified cap. This one do not allow the cellular visualization, in clarified material (Herr´s method), of the endosperm because the yellow color of the lignin. Is there a non aggressive protocol for lignin extraction? Thank´s.
Rinaldo Pires dos Santos
Dr. Rinaldo Pires dos Santos - e-mail: rinaldop-at-uol.com.br Lab. of Plant Anatomy - Dept. of Botany - UFRGS Porto Alegre - RS - Brazil
Well, I stepped in it this time. I agreed to do some "gee-whiz" shots for my dentist of a tooth with a rather large amalgam filling. Only later did it sink in that the mercury might be a problem under vacuum and the beam. Has anybody out there done this? I'd be looking at the thing with gold coating an a fairly light beam (say 10 kV), but unfortunately, not with a cold stage. I've checked the archives and found one brave soul who said that he's done this on old amalgams. I can't really say how old this one is, but I'm pretty sure it wasn't made yesterday. Call me paranoid, but I'd like some more opinions before I do something really stupid!
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I am trying to evaluate some tools that perform automated TEM sample prep prior to FIB thinning. I am looking for some one that has experience using Sela microcleaving tools in conjuction with the TEMstation. I work with failure and material analysis in the IC industry so submicron accuracy and relaibility are important concerns of mine. Also, we will soon be facing chips with low K dielectrics which from what I have heard are extreme soft and hard to polish. I am not sure if the sawing technique they employ will be able to section this type of material.
If anyone on this list has any experience with these tools and can give me insight into their worth as a prep technique please get in touch with me.
We are interested in buying a liquid helium TEM cold stage. Has anyone had experience with it and how was it? Is there any other companies selling this apart from Gatan?
I would appreciate very much for any input on this.
Best regards Yan Xin ======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
Does anyone have a step by step protocol for decalcifying bone for transmission electron microscopy using EDTA?
Thanks,
Kenn
Dr. Corazon D. Bucana, Ph.D. Mr. Kenneth Dunner, Jr. Department of Cancer Biology U.T. M.D. Anderson Cancer Center High Resolution Electron Microscopy Facility 7777 Knight Road, Box 173 Room SRB 1.660 Houston, Texas 77054 PH: 713-792-8106 FAX:713-792-8747
I am in need of a gas flow proportional counter (GFPC) for a JEOL 733 microprobe. Alternately, I may also need some W wire of the appropriate size to restring an existing unit. Thanks for any help/advice.
Ed Holdsworth General Mgr. SEMTEC Laboratories, Inc.
I have had good luck decalcifying bone for TEM using this EDTA protocol:
Fix 1-2 mm sized bone pieces for two days in 2.5 percent glutaraldehyde in 0.1 M Sorensen's buffer.
Decalcify on a shaker for 3-7 days in 7.5 % disodium EDTA, 2.5 % glut. in 0.1 M Sorensen's buffer. (pH the decal solution to physiological pH with NaOH.)
Change to fresh decal solution every couple of days.
Check for complete decalcification by taking before and after X-rays of the tissue.
Rinse twice in buffer
Post fix in 1 % osmium in buffer.
Rinse once with buffer, then once with ddH2O
en bloc stain for 1 hour with aqueous 3% uranyl acetate.
Dehydrate in a graded series of EtOH, infiltrate, and embed in epon.
Good luck, and I hope this helps.
Dotty
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Mon, 19 Mar 2001 semcore-at-audumla.mdacc.tmc.edu-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have a step by step protocol for decalcifying bone for } transmission electron microscopy using EDTA? } } Thanks, } } Kenn } } } } } } } Dr. Corazon D. Bucana, Ph.D. } Mr. Kenneth Dunner, Jr. } Department of Cancer Biology } U.T. M.D. Anderson Cancer Center } High Resolution Electron Microscopy Facility } 7777 Knight Road, Box 173 } Room SRB 1.660 } Houston, Texas 77054 } PH: 713-792-8106 } FAX:713-792-8747 }
I'm looking for a part no longer available from the manufacturer. We have a TMC Micro-g air table, and one of the black plastic parts that holds together the leveling valve (at the ends of the table) has broken.
I can't get one of these from TMC anymore -- the whole valve kit has to be bought for $120, and the bloody things were special-made for TMC.
Does anyone perchance have any of these things that they no longer need?
Thanks to all.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Hi all, I need to have TEM screens recoated and heard that Grant Scientific is the place to contact. I called 803-799-6716 (a # I found in my address archives) and got the lovely fax machine scream sound. any help? thanks, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I have examined amalgams in the SEM in the past. There is little problem with the mercury as it is amalgamated with silver. I have even used pure mercury in the SEM as a standard for WDS with no problem. I believe mercury is used in high vacuum diffusion pumps.
Good luck!
Sam O. Mancuso Group Leader, Physical Metallurgy Special Metals Corporation New Hartford NY 13413 phone: (315) 798-2920 fax: (315) 798-2001
I've got 2 really basic questions. First, I'm trying to determine the actual magnification power of my objectives. I'm using a calibration slide with 0.01 mm (I think that's the increments). What are the procedures for using this slide? I'm assuming I need to find out the diameter of the field and then do a reverse calculation. For example, if I have 36 lines (0.36 mm), using a 40x objective coupled with a 10x ocular, how do I calculate the actual mag power of my 40x obejctive. My second question is about fluorescent microscopy. I'm using a Zeiss fluorescence microscope with 40x and 63x Zeiss "neofluro" objectives, when I'm in focus, there are halos around the fluorescence image. What does this mean? Are my lens dirty? Oh, and one last thing. Does anyone know any good websites that teaches the basics about microscope maintenance and general troubleshooting faq?
I have a wonderful old Leitz Ortholux which I use to look at diatoms in my off time. Unfortunately, the coating on one of the prisms in the trinocular head has degraded with time, and shows up as a mottled shadow when viewing specimens.
Does anyone have recommendations and/or know of resources for reconditioning optics? Any suggestions concerning parts suppliers for old microscopes would be appreciated as well.
Thanks so much for your time!
Todd ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Todd A. Clason, Imaging Coordinator Department of Zoology University of Washington Box 351800 Seattle, WA 98195-1800
Grant Scientific Corp 1385 Rock Island Rd, Gilbert, SC 29054-8821 Phone: (803) 892-2841
Hope that's the right one.
Regards,
Rick Powell
} Hi all, } I need to have TEM screens recoated and heard that Grant Scientific is the } place to contact. I called 803-799-6716 (a # I found in my address } archives) and got the lovely fax machine scream sound. } any help? } thanks, } Beth } } ************************************** } Beth Richardson } EM Lab Coordinator } Botany Department } University of Georgia } Athens, GA 30602 } } Phone - (706) 542-1790 } FAX - (706) 542-1805 } Email - beth-at-dogwood.botany.uga.edu } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } **************************************
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
Question: We recently bought a home microscopy kit and some "GUM MEDIA" was included in the set but was not explained in the manual. What is gum media and how would we use it? HELP!?
Osram Sylvania (http://www.sylvania.com) is a major manufacturer of tungsten wires. If you contact their local sales branch and request a small sample of the appropriate wire or range of wires, you'll get a lifetime supply. I've used their wire myself on XRF detectors and found them at least as good as the manufacturer's.
I can't think of many reasons why the whole detector should ever have to be replaced. You may need to clean it real well and change windows and wire. Any other parts that are damaged should be fairly easy to replace or duplicate.
On Monday, March 19, 2001 1:54 PM, "Edsworth-at-aol.com"-at-sparc5.microscopy.com [SMTP:"Edsworth-at-aol.com"-at-sparc5.microscopy.com] wrote: } } } I am in need of a gas flow proportional counter (GFPC) for a JEOL 733 } microprobe. Alternately, I may also need some W wire of the appropriate size } to restring an existing unit. Thanks for any help/advice. } } Ed Holdsworth } General Mgr. } SEMTEC Laboratories, Inc. } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
We have a liquid helium cold stage made by Oxford Instruments. It is used rarely because the safe handling precautions needed when using liquid helium (not to mention the cost) put many people off. However, if temperatures as low as 6K are a requirement in your experiment and the relevant safety precautions are taken, the thing works well. We also have Gatan's liquid nitrogen cold stage, which is much easier to use although only capable of minus 196C(?).
Hope this helps.
Kind regards,
Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
We have an Oxford Instruments double tilt He holder that fits both out JEOL 3000F and our 2010. We bought it about 2 years ago but Oxford Instruments have since been bought out by Gatan. We use the holder for a few days every couple of months and provided that you have common sense the hazards associated with handling liquid He are not a concern.
We are happy with the performance of the holder. Before buying we tested both the ultra-low temperature (5K) and low temperature (15K) versions and bought the 15K version as it looked more robust and we did not think that there was any real advantage to us to go to 5K. The holder we have gets below 10K.
It takes approximately 45 minutes to fill from room temperature and 30 minutes to refill when cold. If we operate at 20K it will hold the temperature for about 2 hours.
The tilt drives still work well at 10K and the image is stable enough to see gold fringes (0.2nm), after the cooling drift has stopped.
Since Gatan bought out Oxford Instruments I can only think of Fischione as a possible alternative supplier. It really is a shame that competition has reduced in the EM accessory field.
Good luck
Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
} Investigations of biomolecular interactions and mechanics (4 studentships) } These projects will employ SPM and state-of-the-art computational } methods to investigate the origins of biomolecular recognition and } protein folding or RNA folding pathways. Specifically, SPM will be } employed to characterize interaction forces between complementary } biological molecules e.g. individual coiled-coil proteins or } RNA-protein interactions (2 studentships). In parallel, two } studentships will aim to develop complementary computational methods } to further investigate the molecular origins of the obtained forces, } and also to explore the energetics of RNA/protein folding pathways. } } Ultra-high resolution studies of surface-chemistry and protein } structure (2 studentships) } In collaboration with Professor Richard James (Clinical Laboratory } Sciences), one project will utilize state-of-the-art SPM methods to } investigate, in-situ, the structure-function relationships of the } colicin class of bacterial membrane proteins. A second project will } aim to image foliate surfaces with high-resolution, and to } investigate the influence and surface distribution of various } agrochemicals (in collaboration with Syngenta-agrochemicals) } } The development and analysis of surface engineered biomimetic } systems for drug-delivery & tissue engineering (3 studentships) } In collaboration with the School's drug-delivery and tissue } engineering group, these projects will aim to develop novel } biomimetic polymeric surfaces for drug-delivery. In addition the } projects will employ state-of-the-art analytical techniques for } their surface-chemical characterization of polymer systems, } including SPM, contact angle, X-ray photoelectron microscopy and } time-of-flight secondary ion mass spectrometry. } } Single molecule optical microscopy using a single-molecule light } source (1 studentship) } This project will aim to address the current limitations of } near-field scanning optical microscopy, by replacing the physical } aperture by a small number of fluorescent molecule at the probe (as } proposed by W Heckl). The developed technology will be employed for } the high-resolution analysis of microfabricated biomolecular arrays. } } ________________________________________ } Phil Williams } Laboratory of Biophysics and Surface Analysis } School of Pharmaceutical Sciences and The Pharmacy School } University of Nottingham NG7 2RD } tel: +44 (0)115 9515025 } fax: +44 (0) 115 9515110 } http://www.nottingham.ac.uk/lbsa }
I am looking for quotes/sources for an anitivibration table for a TEM. The table will have to be larger than normal to accomodate some other instrumentation.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
I have imaged OLD amalgam without problem. FWIW, A copy of the image can be seen on my web site. No experience with "fresh" material.
http://woody.white.home.att.net
Woody -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi listers, } } Well, I stepped in it this time. I agreed to do some "gee-whiz" shots } for my dentist of a tooth with a rather large amalgam filling. Only } later } did it sink in that the mercury might be a problem under } vacuum and the } beam. Has anybody out there done this? I'd be looking at the } thing with } gold coating an a fairly light beam (say 10 kV), but } unfortunately, not } with } a cold stage. I've checked the archives and found one brave soul who } said } that he's done this on old amalgams. I can't really say how } old this one } is, but } I'm pretty sure it wasn't made yesterday. Call me paranoid, } but I'd like } some } more opinions before I do something really stupid! } } Cheers, } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/~jehrman } } }
Email: ken.andrew-at-bigpond.com.au Name: Ken ANDREW
Education: Graduate College
Location: Melbourne Australia
Question: Can I convert a Zeiss Jena polarized microscope from monocular to binocular. I have a student model from late-70's-early 80's which happens to also have metallographic attachment. E-bay often features binocular eyepieces. Is the barrel fitting common to all brands or just zeiss jena ? Does being a polarized scope make any difference ? I wear spectacles.
Hi there I am looking for a high resolution image of the MSA Logo in either Freehand, Illustrator or high resolution TIFF or JPEG, dows anyone have one or does anyone know who does have one? Many thanks.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
I would like to inform you that the new version of the homepage of the 5th Multinational Congress on ELectron Microscopy, that will be held in Lecce, Italy, from September 20-25 2001, is now ready and available at the URL: www.MCEM5.unile.it. In the "satellite events" section of the page, you will also find details about the "International School on Advances in Electron Microscopy in Materials Science" that will be held before the Congress.
The second circular/call for papers will be distributed to all who pre-registered very shortly.
Up to now we have 300 preregistrations from all over the world, and we really hope to have a nice and succesfull meeting.
Hope to see many of you in Lecce.
Best regards
Massimo
Dr. Massimo Catalano President of MCEM5 CNR-IME Via Arnesano 73100 Lecce - ITALY massimo.catalano-at-ime.le.cnr.it
Hi, I am looking for a new sputter coater to coat specimens for our FEG SEM, I was thinking of a Chromium coater but they are expensive and the films don¹t last very long (apparently). Is there a coater available that gives you small grain size and film longevity? Also are there brands who I should avoid for poor quality? Please reply off line as such opinions could be considered hurtful and libelous (even if preceded by IMHO!) ADVthanksANCE
-
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
I'd appreciate hearing from users of Disk Inspector software in the ISIS platform by Oxford. I'm especially interested in manual analysis of particles detected that are smaller than the minimum size established in the particle detection setup menu. Hope to share experiences and weed out some potential problems.
TIA
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
Hi Frank, Lets take your questions in reverse. It is likely that your 63x lens is an oil objective. The halo you see is most likely left over oil immersion media used on the lens. It is obligatory to use the oil for correct image formation. You indicate the 40x is presenting with the same symptoms, it doesn't necessarily have to be an oil objective however. It is possible it was designed as a dry lens but was accidentally smeared with oil. Specifications for each objective are on the body of the objective. oil lenses usually have a black colored indicator marking. Cleaning with non-abrasive lens paper (not chemwipes) and Windex wouldn't hurt either objective. You also need to make sure your light source, most likely mercury, is properly aligned. You may need help with this step, Zeiss manuals usually mention the steps required, but I strongly urge you seek out an experienced microscopist for this step. Actual magnification is a slightly tricky subject. Ocular x objective is a good range. It is further complicated by additional light gathering / magnification devices, i.e. field lenses, and the final form of the image. If you use 35mm film, print the micrograph, and measure your stage micrometer with a standard ruler to get a ratio which will indicate your final magnification. This figure will include any incidental magnification due to any other in line processes. If you are capturing video or digital, assign xx number for pixels to the stage micrometer. Good luck Ramin
-----Original Message----- } From: Frank Lee [mailto:flee-at-uhnres.utoronto.ca] Sent: Monday, March 19, 2001 6:55 PM To: Microscopy-at-sparc5.microscopy.com
Hi,
I've got 2 really basic questions. First, I'm trying to determine the actual magnification power of my objectives. I'm using a calibration slide with 0.01 mm (I think that's the increments). What are the procedures for using this slide? I'm assuming I need to find out the diameter of the field and then do a reverse calculation. For example, if I have 36 lines (0.36 mm), using a 40x objective coupled with a 10x ocular, how do I calculate the actual mag power of my 40x obejctive. My second question is about fluorescent microscopy. I'm using a Zeiss fluorescence microscope with 40x and 63x Zeiss "neofluro" objectives, when I'm in focus, there are halos around the fluorescence image. What does this mean? Are my lens dirty? Oh, and one last thing. Does anyone know any good websites that teaches the basics about microscope maintenance and general troubleshooting faq?
At 11:28 PM 3/19/01 -0600, you wrote: } I've never heard of "Gum Media" can someone answer this question?
It must be "gum arabic", the water-soluble plant-based gum, for mounting specimens or even slide labels.
There are several mixtures that can be made with it. One comes to mind that involves chloral hydrate - but perhaps they don't let high school students work with that.
http://www.nmnh.si.edu/iz/copepod/techniques.htm has some recipes.
- John
} --------------------------------------------------------------------------- } } Email: JNDKepnerLovers-at-aol.com } Name: Josh Kepner } } Education: 9-12th Grade High School } } Location: City, State, Country } } Question: We recently bought a home microscopy kit and some "GUM MEDIA" was } included in the set but was not explained in the manual. What is gum media } and how would we use it? HELP!? } } ---------------------------------------------------------------------------
We are looking for a source for either an actual gold sputter target or plating service for our gold target for our Edwards 306A coater. Alternatively a source for gold foil would do. As usual time is of the essence. Vendors welcome. Thanks.
David O'Neil Institute for Marine Biosciences National Research Council 1411 Oxford St. Halifax , NS B3H 3Z1 Canada ph. 902-426-8258 fax. 902-426-9413 david.o'neil-at-nrc.ca
Mercury was indeed used in diffusion pumps, but as far as I know, that has been replaced by turbo pumps in most cases, especially due to the health hazards of mercury vapor.
As far as I know (and I am not a doctor!) mercury is fairly harmless in liquid form. It's the vapor that is dangerous. The problem is, that tiny drops of mercury tend to evaporate (due to surface tension), so you don't want to spill mercury. Keeping it in a bottle is not such a problem, as the vapor pressure is very low and it does not evaporate much.
Amalgam, however, is a different material. As Sam says, it's mercury amalgamated with silver. To find out what happens upon heating, you should probably consult some tables.
I heard, that the biggest health problem with Amalgam in teeth did not occur in patients, but in dentists. They had to handle the mercury and amalgamate it with silver all the time. Of course, there have been reports about health risks to patients also, which is probably why I haven't received any Amalgam fillings for a number of years (?).
If I had to look at a tooth with a filling, I would probably try to avoid heating it too much by keeping the beam current low.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: "SMancuso-at-specialmetals.com"-at-sparc5.microscopy.com [mailto:"SMancuso-at-specialmetals.com"-at-sparc5.microscopy.com] Sent: Monday, March 19, 2001 3:06 PM To: Microscopy Listserv
I have examined amalgams in the SEM in the past. There is little problem with the mercury as it is amalgamated with silver. I have even used pure mercury in the SEM as a standard for WDS with no problem. I believe mercury is used in high vacuum diffusion pumps.
Good luck!
Sam O. Mancuso Group Leader, Physical Metallurgy Special Metals Corporation New Hartford NY 13413 phone: (315) 798-2920 fax: (315) 798-2001
I need some ideas on how to reembed kidney tissue that has been embedded in Spurr's. I've heard of a method of using propylene oxide. Are there any other methods out there in TEM land???
Thanks in advance,
Donald G. Awbrey, HT(ASCP) QIHC Image Analysis / Electron Microscopy donaldawbrey-at-texashealth.org
It appears that Barnes and Noble (www.bn.com), Fatbrain (www.fatbrain.com) and Borders (www.borders.com) have it in stock. However, Amazon.com lists it as out of print.
Bookfinder.com is usually a good place to look for new and used books. However, for this book there were very few hits and it appears to list the author as Oliver Flint, but the ISBN number is the same as the book by Olga Flint.
Hope you find the book. I have it sitting on my desk now on interlibrary loan. I was wanting to look at it to determine if I would find it useful.
Judy Bowen Buckman Laboratories Memphis, TN USA
Previous message: I have been looking for a copy of "Food Microscopy", by Olga Flint. It is one volume of the Royal Microscopy Society handbook series. I saw copies available during the last MSA meeting, but cannot recall who the vendor was. Can somebody point me towards a company or individual who may have a copy of this book available for sale?
Karl Hagglund, Researcher P&G Food and Beverage Analytical and Microbiology 6071 Center Hill Ave., Box 117 Cincinnati, OH 45224 (513)634-0146
I have done extensive work on amalgams using SEM and TEM. There is no problem in the vacuum. Mercury is completely consumed/ reacted with alloy particles and the mercury containing phases are stable to 100C.
Tejpal Kaur Hooghan Agere Systems ---------- From: White, Woody N. [SMTP:nwwhite-at-mcdermott.com] Sent: Tuesday, March 20, 2001 8:24 AM To: 'James M. Ehrman'; 'Microscopy-at-MSA.Microscopy.Com' Subject: RE: dental amalgam in SEM?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
I have imaged OLD amalgam without problem. FWIW, A copy of the image can be seen on my web site. No experience with "fresh" material.
http://woody.white.home.att.net
Woody -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi listers, } } Well, I stepped in it this time. I agreed to do some "gee-whiz" shots } for my dentist of a tooth with a rather large amalgam filling. Only } later } did it sink in that the mercury might be a problem under } vacuum and the } beam. Has anybody out there done this? I'd be looking at the } thing with } gold coating an a fairly light beam (say 10 kV), but } unfortunately, not } with } a cold stage. I've checked the archives and found one brave soul who } said } that he's done this on old amalgams. I can't really say how } old this one } is, but } I'm pretty sure it wasn't made yesterday. Call me paranoid, } but I'd like } some } more opinions before I do something really stupid! } } Cheers, } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/~jehrman } } }
Hello, I'm having trouble finding a comprehensive source of info on what wavelengths to use with a multiphoton (Ti:sapphire) laser attached to a confocal to excite fluorochromes (if there is one yet...). Specifically, I'm having trouble with the alexa dyes, but a database of the usual and general dyes would help as well. I already have a cheat sheet from the confocal manufacturer listing the usual (FITC, TRITC, DAPI, etc..) and have "discovered" some not listed (Nile red, aniline blue, alexafluor 594).
Thanks! john shields university of georgia jshields-at-cb.uga.edu
I'm trying to prepare cryoTEM specimens of oil emulsions for one of my users, and it's clear that I don't have plunge-freezing, using ethane, figured out. I start with a Quantifoil grid, which I've glow-discharged, held at the edge with a pair of fine forceps. There's an o-ring keeping the forceps closed. I use a Pipetperson to place 4 microliters of oil emulsion on one side of the grid. I suspend the forceps/grid over a tiny cup of liquid ethane, centered within liquid nitrogen, using a guillotine apparatus with a foot-operated trigger. I blot the grid carefully with a piece of filter paper. At the instant in which the paper and grid separate, I trigger the guillotine. (The timing as I've described it differs from what I've read, but this is how I was shown to do it, and it seems to have worked -- once or twice. The people who showed me how to do this freeze virus suspensions, not oil emulsions.)
Now I have my forceps tip/grid in liquid ethane. I detach the forceps from the guillotine, keeping the grid suspended in the ethane, and then with a quick movement I immerse the grid in a small cup of liquid nitrogen that's suspended a few cm away in the same glass dewar. This is where I get a shallow dome of frozen ethane on the face of my grid. How do I prevent this from happening? Do I need to blot better, or longer, or blot and then leave the grid in air briefly before I dunk it? Or is there a trick to getting the ethane off at this point without damaging the grid?
Scott Robinson
Microscopy Suite Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana IL 61801 217 265-5071 (office); 217 244-6219 (fax) sjrobin-at-uiuc.edu
Awhile back I went to an art opening of Stefan Eberhard's photomicrographs. He's a researcher here at UGA at the Complex Carbohydrate Research Center. He does photomicroscopy using polarized light on commonplace chemicals (crystal images) such as cystine, niacin, vitamin C and my fav image Saccharin. I thought some of you might enjoy seeing the images. His website - http://home.att.net/~seberhard Something to do in your spare time;-) Beth
Disclaimer: I have no financial interest in this work. Just thought it was nice.
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I'm trying to prepare cryoTEM specimens of oil emulsions for one of my users, and it's clear that I don't have plunge-freezing, using ethane, figured out. I start with a Quantifoil grid, which I've glow-discharged, held at the edge with a pair of fine forceps. There's an o-ring keeping the forceps closed. I use a Pipetperson to place 4 microliters of oil emulsion on one side of the grid. I suspend the forceps/grid over a tiny cup of liquid ethane, centered within liquid nitrogen, using a guillotine apparatus with a foot-operated trigger. I blot the grid carefully with a piece of filter paper. At the instant in which the paper and grid separate, I trigger the guillotine. (The timing as I've described it differs from what I've read, but this is how I was shown to do it, and it seems to have worked -- once or twice. The people who showed me how to do this freeze virus suspensions, not oil emulsions.)
Now I have my forceps tip/grid in liquid ethane. I detach the forceps from the guillotine, keeping the grid suspended in the ethane, and then with a quick movement I immerse the grid in a small cup of liquid nitrogen that's suspended a few cm away in the same glass dewar. This is where I get a shallow dome of frozen ethane on the face of my grid. How do I prevent this from happening? Do I need to blot better, or longer, or blot and then leave the grid in air briefly before I dunk it? Or is there a trick to getting the ethane off at this point without damaging the grid?
Dear Scott, Since the ethane is drawn up as you transfer the grid to the LN2, it would have to be removed during the time it is briefly in the N2 vapor (over the LN2 and ethane) before dunking into the small cup. This would expose your grid to higher temperatures and, possibly, water vapor, either of which will be far worse than the ethane. When the cryo-grid is inserted in the EM, the vacuum in the airlock should be good enough to cause the ethane to evaporate, so, unless the ethane affects the structure of the oil droplets, you should be better off just leaving the ethane in place. I would also doubt that there is a good cryogen which is hydrophyllic enough not to affect the oil, and is, at the same time, hydrophobic enough not to affect the water. If examination of the grids shows there to be a problem, you might try to withdraw the grid rapidly from the ethane--to try to draw up as little as possible--and move the grid quickly through a volume of dry N2 (the vapor over the LN2 should qualify) to try to dislodge or evaporate the ethane. I do not think that this procedure will be easy, and it could well raise the temperature of the specimen to unacceptable levels if the N2 vapor is not cold enough. Ideally you would want the temp to be just above the ethane boiling point. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Could this be gum arabic? It is a mounting medium for permanently mounting specimens under coverslips, and in another form (perhaps more dilute?) it can be used as a general adhesive.
} } Colleagues } } I've never heard of "Gum Media" can someone answer this question? } } Nestor } } --------------------------------------------------------------------------- } } Email: JNDKepnerLovers-at-aol.com } Name: Josh Kepner } } Education: 9-12th Grade High School } } Location: City, State, Country } } Question: We recently bought a home microscopy kit and some "GUM MEDIA" was } included in the set but was not explained in the manual. What is gum media } and how would we use it? HELP!? } } ---------------------------------------------------------------------------
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Scott Robinson asked: ..I get a shallow dome of frozen ethane on the face of my grid. How do I prevent this from happening?
The answer depends upon how much ethane is retained and where it is located. A small covering of solid ethane actually serves as an additional frost shield. This will pump off in the airlock of the microscope. On our Philips CM-20 we actually insert the holder into the airlock and pump for a while before dumping the LN2 and inserting the holder. This will work on a standard airlock with increased pumping time by the cryo software. Our airlock is a special turbo-pumped system that presumably gives us a cleaner vacuum.
The problem comes when there is so much ethane film that it interferes with loading in the holder. When a solid ethane film splits off the grid, it usually takes the sample with it. This is very annoying (to say the least). We have blotted the grid on a dry piece of filter paper in the cryochamber prior to plunging in the liquid nitrogen. This works ok if you work fast. It is another opportunity to get frost contamination on the sample so we only do this as a last resort...
I'd be interested in hearing what problems you have with oil emulsions. Our work, along with that of Ishi Talmon, suggests when you have high concentration of oil phase in contact with ice (vitreous or otherwise) that radiation damage (visible as bubbling) is worse than in most aqueous based systems.
Best Regards,
John Minter Eastman Kodak Co. Analytical Technology Division Rochester, NY 14650-2152 Phone: (716) 722-3407 FAX: (716) 477-9303
I'm passing on a question from a colleague. Does anyone know of a technique for etching LR White from sections on a slide? I gave him all the info I had on epoxy removal using ethoxide and so on, but I've been unable to locate any references specific to LR White.
Thanks. Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
This course has been designed to teach microscopists the preparation of high quality SEM and Light Microscopy specimens using new broad ion beam techniques and instrumentation. The two-day practical course intended to offer end-users an alternative/replacement method to the traditional "wet" chemical etching technique, used as final step in the sample preparation process.
Please visit http://www.gatan.com for details.
To register, please e-mail info-at-gatan.com and request a Gatan Schools Registration Kit.
Olympus POS for sale. Good monocular PLM student scope. Comes with Mahogany case, 10x, 20x and 40x stage, Leitz spindle. All offers welcomed. Lou Solebello microls1297-at-mindspring.com
Dear Scott, Why don't you try pure grade propane? The freezing procedure is exactly the same as with the ethane. You will not get frozen ethane, but may have contamination from comercial propane if you do not find pure one. svetla
Scott Robinson asked: .. Do I need to blot better, or longer, or blot and then leave the grid in air briefly before I dunk it? Or is there a trick to getting the ethane off at this point without damaging the grid?
You should use a small cryochamber with enough space for the ethane cup, the specimen holder, some small tools and a piece of filter paper all located above a boiling liquid nitrogen bath. The evaporating nitrogen prevents all parts from contamination with ice. After immersing the grid into the ethane the grid should be blotted on the dry filter paper where the excess of liquid ethane is drained off. When the specimen holder is located near by the transfer from liquid ethane to the holder via the filter paper can be done very fast. In the humidity free atmosphere of the chamber the intermediate storage in liquid nitrogen can be omitted.
Best regards, Markus
******************************************* Dr. Markus Drechsler Friedrich-Schiller-Universitaet Jena Institut fuer Pharmazie Lehrstuhl fuer Pharmazeutische Technologie Lessingstr.8 D-07743 Jena
You can order 'Food Microscopy', and indeed any other Royal Microscopical Society Handbook, from the publisher, Bios Scientific.
Contact sales-at-bios.co.uk to order directly, or visit www.bios.co.uk or our website, www.rms.org.uk/handbook.html if you want more information on any of the RMS handbooks.
Best wishes
Sue ************************************************************************** Sue Betteridge, Publications Officer, Royal Microscopical Society 37/38 St Clements, Oxford OX4 1AJ, UK. Tel +44 (0)1865 248768, fax +44 (0)1865 791237, email jmicrosc-at-rms.org.uk, website http://www.rms.org.uk **************************************************************************
My sugestion would be to either build or buy a carbon evaporator. While it may not be impossible to sputter carbon with more exotic sputtering devices, a Hummer V will not work.
Wooody White ------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------. } } } I have a Technics sputter coater- Hummer -V. I'd like help finding a } carbon source for it. } Any ideas? } Thank you. } } }
This is a discussion archived at "Tips & Tricks" There might be more than one, but this is the first I hit with the search.
The rest of the archive can be found at:
www.biotech.ufl.edu/~emcl
Follow the Tips & Tricks link.
I will begin adding material to the site soon. Anything you have found useful in the course of your studies would be welcome. Simply forward it to whittaker.scott-at-nmnh.si.edu. You will of course be given the credit.
Scott Whittaker SEM Lab Manager National Museum of Natural History Smithsonian Institution 10th & Constitution Ave, NW Washington DC 20560-0104 202-357-1651 {*^-at-)~~~ It was recently discovered that research causes cancer in rats. ~~~(-at-^*}
} } } "Tindall, Randy D." {TindallR-at-missouri.edu} 03/20/01 05:45PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
I'm passing on a question from a colleague. Does anyone know of a technique for etching LR White from sections on a slide? I gave him all the info I had on epoxy removal using ethoxide and so on, but I've been unable to locate any references specific to LR White.
Thanks. Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
I would like to hearing from you opinions about Scanning microscopes. We have a limited fund to buy a new scanning electron microscope and we have just received a proposal from a Japanese company named SHIMADZU. They offered us a scanning electron microscope model SS-550. I confess that I don't know much about this company and neither about the quality of its instruments (particularly SEM). I would appreciate if someone out there could give me an opinion about this equipment. Right now we can buy a unit without the EDS analitical system which we are planning to buy later. Then I also would like to know if we would buy only the image unit first we could buy later the EDS system (from the same company) separately to upgrade the same model (SS-550). The vendor said that it is possible, but sometimes it is hard to trust in sellers especially when you live in South America.
Thank all of you
Best wishes,
Leonardo -- --- Leonardo Lagoeiro Departamento de Geologia Universidade Federal de Ouro Preto Ouro Preto, MG, 35400-000 Brazil E-mai: lagoeiro-at-degeo.ufop.br
I am trying to find a procedure to detect cellular and membrane cholesterol at the EM level in RBC's. Using the standard EM processing techniques, cholesterol is dissolved out leaving familiar clefts. Does anyone out there know of a protocol where I can preserve the cholesterol at the EM level and still have decent ultrastructure.
Howard Mulhern Children's Hospital Dept. of Pathology Surgical Path EM Facility Boston, 02115, Ma.
I'm looking for a simple DLS program, freeware and preferably source code. This would be "distance least squares" for cleaning up atomic positions (for example those input by hand from a model), minimizing the mean squared deviation from expected bond angles and spacings.
I'm particularly interested in code which would do this for silicates (SiO2), using typical ~1.6 Angstrom Si-O distance and 109 degree Si-O-Si angle.
I actually found something of this kind, ostensibly free for downloading, at
http://www.kristall.ethz.ch/LFK/software/xrs/
However, I'm consistently refused the ftp connection listed there, and have gotten no response to queries.
Does anyone know of other sources of such software?
Greetings, If you look up protocols for doing PAS staining to show polysaccharides (in plant and I suppose any tissue), they start with an aldehyde quench, with either of two chemicals. Years ago we discovered that for our purposes we don't need that step, and now I have forgotten the name of the chemicals. The names are something like DNPH and Dimedone but I can't remember exactly. In any book of botanical microtechnique that gives the PAS procedure, they should be there.
Sorry about my flawed memory.
Tobias
} } } I'm looking for a method to remove aldehydes from tissue before } processing? Thanks, Caroline Miller
Bodycote Ortech is seeking an experienced electron microscopist for our physical characterization group. The successful candidate will provide hands-on microscopy services to our clients in the healthcare, general manufacturing and materials sectors. You will characterize a wide variety of man-made and naturally occurring materials, and investigate materials breakdown and failure in support of manufacturing processes. You will work with our clients to define project scope, write quotations, perform analytical work and prepare project reports.
Your expertise lies in analytical scanning electron microscopy, including low temperature SEM. You are adept at using light microscopy and image analysis techniques. You enjoy problem solving and research activities, manage your time well and are able to work effectively in a team atmosphere. Ideally you have a graduate degree in chemistry, biology or a related field and more than three years experience working in an electron microscopy laboratory.
To join our team, please forward your resume to Human Resources Department, Bodycote Ortech, 2395 Speakman Drive, Mississauga, Ontario, L5K 1B3, email bruce.a-at-bodycote.ca Fax: (905) 823-1446
Ammonium chloride (50-100 mM) in PBS or glycine (sorry, don't remember concerntration--probably 1 or 2 % ??) in PBS have been reported. We use NH4Cl.
Sara Miller
On Wed, 21 Mar 2001, Caroline Miller wrote:
} Date: Wed, 21 Mar 2001 12:24:55 -0600 } From: Caroline Miller {camiller-at-creighton.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Aldehyde Removal } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm looking for a method to remove aldehydes from tissue before } processing? Thanks, Caroline Miller } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I'm afraid that I have never heard of Shimadzu SEM's or EDS's. Moneywise, I evaluated JEOL, Hitachi, and KLA/Tencor (Amray) SEM's and found that JEOL was by far the most economical for equivalent systems. (We also purchased an EDAX EDS system to go with it.)
I have no interest in JEOL or EDAX (other than being a happy customer.)
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 978-470-1620 jlagoy-at-bodycote-imt.com
Here are the replies I have received so far on my question about sputter coaters. They are as I have received them except I have removed the senders names. Some are obviously from vendors. These are NOT my opinions I am simply reflecting what I have learned. YMMV.
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Dear Sir, I saw your posting on the MSA list server and wanted to know if you were aware of Cressington products. You can view our web site at www.cressington.com {http://www.cressington.com} . Also I would offer to come to your lab to demonstrate our 208HR sputter coater. As far as price being an issue we are always tailoring quotes around tight budgets. Please feel free to contact me if you would like more information.
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We have a KMC Ion-Tech, which I've been told has been taken over by South Bay Technologies. This is good. KMC was everything bad in a company. If South Bay has redesigned -- extensively -- the ion-coater, then this might be a good machine. Otherwise, I'd avoid it like the plague. Even though I generally think good thoughts about South Bay.
When the thing works right, we use it for platinum-coating specimens for FESEM down to 1 or 2 nm (measured to 0.7 nm, but that really requires the right specimen). The Pt coat is very fine. I saw grain structure once at something over 200,000X (might have been 400,000, this was a while ago), but typically I see no structure.
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We used a sputter coater with a chromium target for awhile, with good results. You are correct, however, in that the chromium begins oxidizing immediately and will quickly form an insulating oxide layer. Samples should be viewed as quickly as possible after coating and stored in the best vacuum possible if repeated viewings are required. Chromium is not a good way to go for specimens that need to be kept for repeated viewings.
Our compromise was to purchase a platinum target, which provides somewhat larger grain size than chromium, but smaller than gold or gold-palladium. Platinum, of course, does not oxidize. Platinum and other targets are available at very reasonable cost (relative to the sputter coaters' manufacturer's prices) from Abe Dayani at Refining Systems, Inc. He can be reached at P.O. Box 72466, Las Vegas, NV 89170. Phone is (702) 368-0579 and fax is (702) 368-0933. I have dealt with him before and found him to be very helpful and knowledgeable.
We are using an EMITECH sputter coater currently, but I cannot in good faith recommend them at this time. The coater works well for the most part, but the company is recovering from some internal problems and their service is spotty (to be kind). I think that if or when the company is over its present difficulties that this will again be a wonderful line of equipment.
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I can't address relative grain size and longevity, nor Cr coats specifically, but the carbon coater we opted for last year is the Emitech 950C. It also has an easily interchangeable metal head. We felt this turbo unit was the best value by a considerable stretch. So far we've not done metal [though may do some Au-Pd fairly soon], only carbon, but the carbon coats have been very consistent. URL:
http://www.emitech.demon.co.uk/K950.htm
US contact: Linda Dailey {emitech-at-earthlink.net}
If you feed power to your roughing pump through the K950, allowing automatic control of the rotary by the coater, there is a fuse/circuit issue we had that is solved by using the auto control for a relay switch.
Best of luck.
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The 681 and the 682 BOTH provide an amorphous coating. The 682 also does ecthing. The 681 is coating ONLY.
If you want to talk with someone about applications, please call Dick Mitro at 925-224-7319.
If you don't mind seeing the coating(s), then a cheap $5-$10K coater will suffice.
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I recently went through this decision process for a HR coater for our new LEO 1550. I decided on the Cressington 208HR. It can do Cr, but also Au or Pd or Pt or Au/Pd. It has several nice features such as a rotary/planetary/tilting stage to allow for a thin 2nm coating even on rougher samples. My research led me to this coater or one by Emitech. I think both are good coaters and both were in the $25,000 range. We are taking delivery of the Cressington in about X weeks. If you are still looking at that point I can tell you what I think once I get a chance to use it on several of our samples. Of course I had them coat some samples for us before I made my decision but lets see how it works in the lab. We run a wide range of samples, polymer, metal, ceramic. Good luck.
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I saw this posted on the listserver and thought you might have an interest in our IBS/e Ion Beam Sputter Deposition and Etching System. It is on the pricey side compared to a sputter coater ($50-60k), but can deposit very thin, uniform, small grain films using chromium, iridium, carbon etc. It has many advantages over magnetron systems.
If it is of interest, I would be happy to send you more detailed information. Please let me know.
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IMHO! I know it is not the least expensive choice, but I recently purchaced an SBT IBSS (Ion Beam Sputter/etch System). It does a fantastic job with Ir. We have a LEO 1530 (FEG) and routinely image up to } x200,000 with no evidence of the coating structure. It's durability is fine for us, we have specimens in us often for several days and up to a week or more, without significant deterioration in conductivity. Vince Carlino is with them (South Bay) now and this coater is basically his previous system in a new and improved system/package. Of course you can use Cr or Pt or virtually any other metal to "coat", but we have had tremendous success with Ir. It is a very flexible, durable, and effective instrument for us.
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For DC sputtering the next best option is Pt, better than Au or Pd or Au/Pd mix.
However Cr is best but has all of the inherent issues that you have mentioned, We are currently evaluating using Irridium for DC sputtering, all the advantages of Cr in grain size but none oxidizing.
However very hard to obtaining targets of sufficient purity and very expensive.
Have a look at our site in the US this shows our equipment and has some tech information that you may find useful.
www.empdirect.com
The Emitech K575X turbo sputter coater is designed with Cr and FE sem in mind, the unit may also be used to coat with noble metals as mentioned above.
Please call or email if you have any questions or would like price information on any of our products.
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I too am interested in purchasing another sputter coater for use with our JEOL FEG-SEM. For the last 8 years or so we have been using a Plasma Sciences CrC-100 coater with a chromium target. Generally the unit has served us well, however, I have had two nagging concerns with respect to this coater, 1) its age - it's becoming tempermental of late and 2) Plasma Sciences is no longer in business, service and parts have been taken over by Torr International. I don't know where or from whom you were informed regarding problems associated with chromium. We have not had that experience.
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Denton desk II did a good job while I was at university of xxxx with just gold/palladium on a hitachi s-4000 FESEM.
I now have a Cressington Scientific 108 SE automatic but it is so new I have yet to really put it through the paces. They say it does a wonderful job and for the price it better. It is also what the FEI applications lab has for the FEG demonstrations. Those pics were impressive
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Take a look at URL http://www.2spi.com/catalog/osmi-coat.html
It is now almost impossible to sell a chromium coater in Japan; the Japanese learned over the past few years that nothing comes even close to what the osmium coater will do. I know that sounds almost too good to be true but consider a) the metal layer is completely amorphous, there is absoulutely no grain size, and of course, with Cr, there is some point where you do start to see the grain, and b) the metallization has the same inertness as gold or Pt, so unlike Cr which starts to oxidize almost immediately, the osmium metal coating lasts almost forever.
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I've used gold-palladium targets with good results in ordinary sputter coaters. Emitech and Anatech/Hummer are two brands we've had good luck with.
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Pertaining to your recent question posted to Microprober list, perhaps you may want to try Polaron sputter coater SC series at reasonable price, I have been using few brands of sputter coater for FESEM, I think this is recommended.
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Would appreciate hearing what you learn about sputter coaters.
Our 15-year-old Au-coater (Denton) has been reliable and user-friendly, but considering its age, I could be faced with replacing it before too much longer.
Topographic artifacts produced by the Au-sputter have been of interest because of the 'nannobacteria' problem. With our little garden-variety W-filament SEM, a 30 sec coating does not produce detectable artifacts (below about 50,000x) whereas longer coating times may (See Fig 7, p. 588 in Folk and Lynch, JSR v. 67).
In the FE-SEM, samples coated with our sputter for 45 sec begin to show potentially troubling textures (artifacts?) around 80,000x.
Thanks for any info you can share!
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I purchased a Cressington 208 HR Sputter Coater for use with our FEG SEM. It has worked very nicely. I decided that the Cr target was too much of a hassle for routine prep work, so for the majority of work, I use Au/Pd and apply a coating of from 1 to 8 nm in thickness, depending on my samples. The system has been very reliable, pumps down quickly, and releases the vacuum quickly. The little shield that swings in and out works to protect the sample from the initial oxidized Cr from the target surface, but when the oxide layer is sputtered off, you swing the small shield out of the way to coat the sample with Cr. The rotating, tilting stage allows for good coverage of the sample with the target material. I know there are other good brands out there, but the Cressington works very well.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
SHIMADZU is definitely a small player here in the US.
In the 27 yeatrs I have been involved in SEMs, I have never heard of them.
Regards,
Earl
----- Original Message ----- } From: "Leonardo Lagoeiro" {lagoeiro-at-degeo.ufop.br} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 21, 2001 6:57 AM
I didn't know that Shimadzu (who are a large and reputable scientific equipment manufacturer who afew years ago bought Kratos)) made an SEM, and a search of their website www.shimadzu.com revealed none. However, I see that their regional Brasil website lists not only the SEM 550, but also the EPMA 1600! I remember hearing, years ago, that they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that may not have been so.
I'd be very interested to hear more about either instrument.
Anybody know anything?
cheers
rtch
} } Hi there, } } I would like to hearing from you opinions about Scanning } microscopes. We have a limited fund to buy a new scanning electron } microscope and we have just received a proposal from a Japanese } company named SHIMADZU. They offered us a scanning electron } microscope model SS-550. I confess that I don't know much about this } company and neither about the quality of its instruments } (particularly SEM). I would appreciate if someone out there could } give me an opinion about this equipment. Right now we can buy a unit } without the EDS analitical system which we are planning to buy } later. Then I also would like to know if we would buy only the image } unit first we could buy later the EDS system (from the same company) } separately to upgrade the same model (SS-550). The vendor said that } it is possible, but sometimes it is hard to trust in sellers } especially when you live in South America. } } Thank all of you } } Best wishes, } } } Leonardo } -- } ---
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
G'day Caroline, You need to carry out an "aldehyde blockade" process, This will convert all aldehyde groups. Including tissue aldehyde groups. Method: A saturated solution of 2,4-DNP(2,4dinitrophenyl hydrazine) in 15% acetic acid. Time 0.5Hrs. Wash thoroughly in tap water. Time 0.5Hrs
If you are going on to do a PAS reaction you then begin your oxidation step with 1% Periodic acid. Regards JVN
Caroline Miller wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm looking for a method to remove aldehydes from tissue before } processing? Thanks, Caroline Miller
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
Applied Molecular Evolution (AMEV), is an early stage biotech company with secure financial funding and high growth potential. We focus on novel high throughput approaches to drug discovery, screening compound libraries against thousands of variants of G protein coupled receptors using highly advanced instrumentation.
We are looking for an entry level Scientist in the department of Fluorescence Imaging. The candidate is expected to have a Ph.D. and be experienced in fluorescence imaging. Experience with Ca2+-sensitive dyes and G Protein coupled receptors is a plus.
We are willing to recognize the added value of highly skilled and motivated individuals. Novasite employees receive excellent benefits and compensation, including equity participation and generous performance-linked incentives.
Please Reference Job Code: NR0321A jobs-at-novasite.com Novasite Pharmaceuticals 3520 Dunhill Street San Diego CA 92121 Fax: (858) 597-4950
San Diego based Novasite Pharmaceuticals, Inc., a subsidiary of Applied Molecular Evolution (AMEV), is an early stage biotech company with secure financial funding and high growth potential. We focus on novel high throughput approaches to drug discovery, screening compound libraries against thousands of variants of G protein coupled receptors using highly advanced instrumentation.
We are looking for an entry level Scientist in the department of Fluorescence Imaging. The candidate is expected to have a Ph.D. and be experienced in fluorescence imaging. Experience with Ca2+-sensitive dyes and G Protein coupled receptors is a plus.
We are willing to recognize the added value of highly skilled and motivated individuals. Novasite employees receive excellent benefits and compensation, including equity participation and generous performance-linked incentives.
Please Reference Job Code: NR0321A jobs-at-novasite.com Novasite Pharmaceuticals 3520 Dunhill Street San Diego CA 92121 Fax: (858) 597-4950
We are preparing to embark on an in-depth project with outside funding that will require, among other things, 3-D reconstruction of series' of transmission electron micrographs of sections of biological material. We don't have a great deal of recent experience in this area and would welcome (off list if you are a commercial concern) suggestions for the "best" or most capable, or most flexible or useful 3-D reconstruction software including what computer platform is required and what configuration is considered adequate or better for this type of work. In addition, what digital data collection system currently available would be best for this type of work? We presently have a Philips CM12S STEM with a slow scan CCD camera with a 1k x 1k chip that has done good service for an extended period, but it seems that it might be a bit difficult to use for serial section recording and image orientation in preparation for 3-D reconstruction. If something new in a hardware-software package is available, we would be interested to hear how it works for others, or to hear directly from vendors off list.
Separately, it is possible that TEM tomography will also be in our future - It is our understanding that either at least IVEM or an energy filtered TEM are required to do useful tomography of thicker sections (0.25 - 0.5 micron thick). What are others in this area of interest using for microscopes, computers, and software? Has something of a "standard" come about, or are people still writing their own code and putting systems together from scratch?
Thanks in advance for any and all information and advice.
Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
I do not know this company either, but why do you have proposal just from one company? Ask other vendors and compare their proposals. As for EDS it is better to by system directly from manufacturer and ask them in advance proposals too - if SEM you choose is not in their "standard microscopes" list they may charge you extra for an adapter.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Leonardo Lagoeiro [mailto:lagoeiro-at-degeo.ufop.br] } Sent: Wednesday, March 21, 2001 8:57 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM inquiry } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi there, } } I would like to hearing from you opinions about Scanning microscopes. } We have a limited fund to buy a new scanning electron microscope and } we have just received a proposal from a Japanese company named } SHIMADZU. They offered us a scanning electron microscope model } SS-550. I confess that I don't know much about this company and } neither about the quality of its instruments (particularly SEM). I } would appreciate if someone out there could give me an opinion about } this equipment. Right now we can buy a unit without the EDS } analitical system which we are planning to buy later. Then I also } would like to know if we would buy only the image unit first we could } buy later the EDS system (from the same company) separately to } upgrade the same model (SS-550). The vendor said that it is possible, } but sometimes it is hard to trust in sellers especially when you live } in South America. } } Thank all of you } } Best wishes, } } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br }
Caroline didn't say whether she wanted the aldehyde procedure for LM or EM, but I suspect the 15% acid wouldn't be to good for ultrastructural studies.
On Thu, 22 Mar 2001, John V Nailon wrote:
} Date: Thu, 22 Mar 2001 09:14:34 +1000 } From: John V Nailon {J.Nailon-at-mailbox.uq.edu.au} } To: Caroline Miller {camiller-at-creighton.edu} } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: Aldehyde Removal } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } G'day Caroline, } You need to carry out an "aldehyde blockade" process, This will convert } all aldehyde groups. Including tissue aldehyde groups. } Method: A saturated solution of 2,4-DNP(2,4dinitrophenyl hydrazine) in } 15% acetic acid. Time 0.5Hrs. } Wash thoroughly in tap water. Time 0.5Hrs } } If you are going on to do a PAS reaction you then begin your oxidation } step with 1% Periodic acid. } Regards } JVN } } Caroline Miller wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } I'm looking for a method to remove aldehydes from tissue before } } processing? Thanks, Caroline Miller } } -- } John Nailon } Operations Manager } The Centre for Microscopy and Microanlaysis } The University of Queensland } St Lucia QLD 4072 } Tel: +61-7-33654214 } Fax: +61-7-33654422 } WWW: http://www.uq.edu.au/nanoworld } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Could it be that Shimadzu are now the manufactures/distributors of the Topcon/Akashi/ISI range of instruments?? Regards JVN
Ritchie Sims wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I didn't know that Shimadzu (who are a large and reputable scientific } equipment manufacturer who afew years ago bought Kratos)) made an } SEM, and a search of their website www.shimadzu.com revealed none. } However, I see that their regional Brasil website lists not only the } SEM 550, but also the EPMA 1600! I remember hearing, years ago, that } they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that } may not have been so. } } I'd be very interested to hear more about either instrument. } } Anybody know anything? } } cheers } } rtch } } } } } Hi there, } } } } I would like to hearing from you opinions about Scanning } } microscopes. We have a limited fund to buy a new scanning electron } } microscope and we have just received a proposal from a Japanese } } company named SHIMADZU. They offered us a scanning electron } } microscope model SS-550. I confess that I don't know much about this } } company and neither about the quality of its instruments } } (particularly SEM). I would appreciate if someone out there could } } give me an opinion about this equipment. Right now we can buy a unit } } without the EDS analitical system which we are planning to buy } } later. Then I also would like to know if we would buy only the image } } unit first we could buy later the EDS system (from the same company) } } separately to upgrade the same model (SS-550). The vendor said that } } it is possible, but sometimes it is hard to trust in sellers } } especially when you live in South America. } } } } Thank all of you } } } } Best wishes, } } } } } } Leonardo } } -- } } --- } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
I was searched Shimadzu web site at http://www.shimadzu.com, but there is no
any information about scanning electron microscope.
Henrik
Ritchie Sims wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I didn't know that Shimadzu (who are a large and reputable scientific } equipment manufacturer who afew years ago bought Kratos)) made an } SEM, and a search of their website www.shimadzu.com revealed none. } However, I see that their regional Brasil website lists not only the } SEM 550, but also the EPMA 1600! I remember hearing, years ago, that } they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that } may not have been so. } } I'd be very interested to hear more about either instrument. } } Anybody know anything? } } cheers } } rtch } } } } } Hi there, } } } } I would like to hearing from you opinions about Scanning } } microscopes. We have a limited fund to buy a new scanning electron } } microscope and we have just received a proposal from a Japanese } } company named SHIMADZU. They offered us a scanning electron } } microscope model SS-550. I confess that I don't know much about this } } company and neither about the quality of its instruments } } (particularly SEM). I would appreciate if someone out there could } } give me an opinion about this equipment. Right now we can buy a unit } } without the EDS analitical system which we are planning to buy } } later. Then I also would like to know if we would buy only the image } } unit first we could buy later the EDS system (from the same company) } } separately to upgrade the same model (SS-550). The vendor said that } } it is possible, but sometimes it is hard to trust in sellers } } especially when you live in South America. } } } } Thank all of you } } } } Best wishes, } } } } } } Leonardo } } -- } } --- } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
-- Henrik Kaker, Ph.D. Metal Ravne d.o.o. SEM-EDS Lab Koroska cesta 14, 2390 Ravne Slovenia Phone: +386 02 82 21 131 Fax: +386 02 82 20 436 http://www.kaker.com
Some questions arising: 1) The use of iridium for sputtering is news to me, and looks interesting. I would be interested to see a comparison of iridium film structure with platinum etc. Can a garden variety gold or gold/palladium unit sputter iridium, or are more exotic conditions required?
2) Does anyone have experience of the practical and safety issues arising from osmium coaters? Is the release of osmium tetroxide vapour to room atmosphere well controlled? How economical are they to run? Would they be practical in an environment where there could be several days or even weeks between coating runs? Presumably there are no safety issues with the coated specimens if the metal is inert. True?
Chris
snip
IMHO! I know it is not the least expensive choice, but I recently purchaced an SBT IBSS (Ion Beam Sputter/etch System). It does a fantastic job with Ir. We have a LEO 1530 (FEG) and routinely image up to } x200,000 with no evidence of the coating structure. It's durability is fine for us, we have specimens in us often for several days and up to a week or more, without significant deterioration in conductivity. Vince Carlino is with them (South Bay) now and this coater is basically his previous system in a new and improved system/package. Of course you can use Cr or Pt or virtually any other metal to "coat", but we have had tremendous success with Ir. It is a very flexible, durable, and effective instrument for us.
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For DC sputtering the next best option is Pt, better than Au or Pd or Au/Pd mix.
However Cr is best but has all of the inherent issues that you have mentioned, We are currently evaluating using Irridium for DC sputtering, all the advantages of Cr in grain size but none oxidizing.
However very hard to obtaining targets of sufficient purity and very expensive.
Have a look at our site in the US this shows our equipment and has some tech information that you may find useful.
www.empdirect.com
The Emitech K575X turbo sputter coater is designed with Cr and FE sem in mind, the unit may also be used to coat with noble metals as mentioned above.
Please call or email if you have any questions or would like price information on any of our products.
___________________
I too am interested in purchasing another sputter coater for use with our JEOL FEG-SEM. For the last 8 years or so we have been using a Plasma Sciences CrC-100 coater with a chromium target. Generally the unit has served us well, however, I have had two nagging concerns with respect to this coater, 1) its age - it's becoming tempermental of late and 2) Plasma Sciences is no longer in business, service and parts have been taken over by Torr International. I don't know where or from whom you were informed regarding problems associated with chromium. We have not had that experience.
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Denton desk II did a good job while I was at university of xxxx with just gold/palladium on a hitachi s-4000 FESEM.
I now have a Cressington Scientific 108 SE automatic but it is so new I have yet to really put it through the paces. They say it does a wonderful job and for the price it better. It is also what the FEI applications lab has for the FEG demonstrations. Those pics were impressive
___________________
Take a look at URL http://www.2spi.com/catalog/osmi-coat.html
It is now almost impossible to sell a chromium coater in Japan; the Japanese learned over the past few years that nothing comes even close to what the osmium coater will do. I know that sounds almost too good to be true but consider a) the metal layer is completely amorphous, there is absoulutely no grain size, and of course, with Cr, there is some point where you do start to see the grain, and b) the metallization has the same inertness as gold or Pt, so unlike Cr which starts to oxidize almost immediately, the osmium metal coating lasts almost forever.
___________________
I've used gold-palladium targets with good results in ordinary sputter coaters. Emitech and Anatech/Hummer are two brands we've had good luck with.
___________________
Pertaining to your recent question posted to Microprober list, perhaps you may want to try Polaron sputter coater SC series at reasonable price, I have been using few brands of sputter coater for FESEM, I think this is recommended.
___________________
Would appreciate hearing what you learn about sputter coaters.
Our 15-year-old Au-coater (Denton) has been reliable and user-friendly, but considering its age, I could be faced with replacing it before too much longer.
Topographic artifacts produced by the Au-sputter have been of interest because of the 'nannobacteria' problem. With our little garden-variety W-filament SEM, a 30 sec coating does not produce detectable artifacts (below about 50,000x) whereas longer coating times may (See Fig 7, p. 588 in Folk and Lynch, JSR v. 67).
In the FE-SEM, samples coated with our sputter for 45 sec begin to show potentially troubling textures (artifacts?) around 80,000x.
Thanks for any info you can share!
___________________
I purchased a Cressington 208 HR Sputter Coater for use with our FEG SEM. It has worked very nicely. I decided that the Cr target was too much of a hassle for routine prep work, so for the majority of work, I use Au/Pd and apply a coating of from 1 to 8 nm in thickness, depending on my samples. The system has been very reliable, pumps down quickly, and releases the vacuum quickly. The little shield that swings in and out works to protect the sample from the initial oxidized Cr from the target surface, but when the oxide layer is sputtered off, you swing the small shield out of the way to coat the sample with Cr. The rotating, tilting stage allows for good coverage of the sample with the target material. I know there are other good brands out there, but the Cressington works very well.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
As is mentioned by other members you are unable to use a carbon target with your coater but you may possibly be able to obtain a "carbon string" head and power pack for it?
This package usually contains a new top plate with through terminals, a power pack (not much different to adding a car battery) and some safety interconnections. Most sputter coater manufacturers have this type of accessory.
You are basically using a carbon fibre which you "burn" at high temperature to evaporate the carbon. The coating is not from a point source (as would the conventional carbon electrodes used in a high vacuum coater) so it is better for SEM samples. However the carbon is very soft and easily damaged even when evaporated under (the desired) best sputter coating vacuum level.
In short it is a quick and easy SEM technique but it does not provide a coat as efficient as a sputtered metal, or provide a coating that would allow the production of free floating carbon films here, a high vacuum coater is essential .
Good luck but talk to the manufacturer.
Steve Chapman Senior Consultant Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
I have undertaken several SEM tests with EDS analysis of minute "Black Spots" that we are having problems with in the Ultra High Molecular Weight polyethylene components that we manufacture at our plant. We thought that identifying the spots would help us eliminate the problem. To a certain extent it did. i.e. any Fe,Cr& Ni that was found together is almost certainly stainless steel splinter.
However, it is the other results that are proving difficult to identify. Fe is present along with S, Si & Na? Another spot yielded Si, K ,Na and Ca together. A further example of a spot would be; Fe,Ca,Si,Ti,K and Na.
Would anyone have any ideas on how to identify what these particles are?
Any information or help would be gratefully appreciated.
It's difficult to ID material without knowing how the spectra were collected and what elements were dominant. You list Si in the three spots that you describe. If it was the major foreign element, then you might have some silicate material.
If available it would be good to try Micro IR to further characterize the spots.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027
paulfinnegan-at-ireland.com on 03/22/2001 07:27:09 AM To: Microscopy-at-sparc5.Microscopy.Com cc:
Hi all,
I have undertaken several SEM tests with EDS analysis of minute "Black Spots" that we are having problems with in the Ultra High Molecular Weight polyethylene components that we manufacture at our plant. We thought that identifying the spots would help us eliminate the problem. To a certain extent it did. i.e. any Fe,Cr& Ni that was found together is almost certainly stainless steel splinter.
However, it is the other results that are proving difficult to identify. Fe is present along with S, Si & Na? Another spot yielded Si, K ,Na and Ca together. A further example of a spot would be; Fe,Ca,Si,Ti,K and Na.
Would anyone have any ideas on how to identify what these particles are?
Any information or help would be gratefully appreciated.
There exists a homepage of 3-Dimensional Microscopy Labs (http://3dem.sdsc.edu/) in which are listed various packages for 3D-EM Image Analysis. Additionally a mailing list exists.
BTW: A nice animation can be found in a Web-Page of the University of Utrecht (The Netherlands): http://emsaserv.bio.uu.nl/3dem/ANIMATED_INTRODUCTION/animated_introduction_1.html
I hope this helps a little bit. Of course, you are welcome to contact me off-line.
Best wishes, Ingo
+++++++++++++++++++++++++++++++++++++++++++++++++++++ Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH Herbststrasse 7 D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: www.tvips.com Email: ingo.daberkow-at-tvips.com
-------- Original Message --------
Dear List -
We are preparing to embark on an in-depth project with outside funding that will require, among other things, 3-D reconstruction of series' of transmission electron micrographs of sections of biological material. We don't have a great deal of recent experience in this area and would welcome (off list if you are a commercial concern) suggestions for the "best" or most capable, or most flexible or useful 3-D reconstruction software including what computer platform is required and what configuration is considered adequate or better for this type of work. In addition, what digital data collection system currently available would be best for this type of work? We presently have a Philips CM12S STEM with a slow scan CCD camera with a 1k x 1k chip that has done good service for an extended period, but it seems that it might be a bit difficult to use for serial section recording and image orientation in preparation for 3-D reconstruction. If something new in a hardware-software package is available, we would be interested to hear how it works for others, or to hear directly from vendors off list.
Separately, it is possible that TEM tomography will also be in our future - It is our understanding that either at least IVEM or an energy filtered TEM are required to do useful tomography of thicker sections (0.25 - 0.5 micron thick). What are others in this area of interest using for microscopes, computers, and software? Has something of a "standard" come about, or are people still writing their own code and putting systems together from scratch?
Thanks in advance for any and all information and advice.
Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
Never heard of them either but a quick internet search showed the following three sites:
1. Shimadzu Scientific Instruments, Inc. We provide solutions to analytical laboratory science by producing an extensive range of products, software and unrivaled customer service. In the United States, Shimadzu
http://www.ssi.shimadzu.com/
2. Shimadzu - Solutions for Science Shimadzu Scientific Instruments - Solutions for Science since 1875 http://www.shimadzu.com/
3. Shimadzu Solutions for Science We provide solutions to analytical laboratory science and medical science by producing an extensive range of products, software and customer service. In Europe, Shimadzu has a
http://www.sel.shimadzu.com/
====} As well as a specific South American Local site:
http://www.shimadzu.com.br
===================================
And since none of us seem familiar with Shimadzu I thought this site was interesting . .
4. Shimadzu SUCKS homepage Shimadzu specializes in lies, denials and cover-ups!
http://shimadzu.wxs.org/
(Now, take it for whatever its worth, but someone did go the effort and expense of setting up this site, eh?)
================================
On 21 Mar 2001, at 11:57, Leonardo Lagoeiro wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi there, } } I would like to hearing from you opinions about Scanning microscopes. } We have a limited fund to buy a new scanning electron microscope and } we have just received a proposal from a Japanese company named } SHIMADZU. They offered us a scanning electron microscope model } SS-550. I confess that I don't know much about this company and } neither about the quality of its instruments (particularly SEM). I } would appreciate if someone out there could give me an opinion about } this equipment. Right now we can buy a unit without the EDS } analitical system which we are planning to buy later. Then I also } would like to know if we would buy only the image unit first we could } buy later the EDS system (from the same company) separately to } upgrade the same model (SS-550). The vendor said that it is possible, } but sometimes it is hard to trust in sellers especially when you live } in South America. } } Thank all of you } } Best wishes, } } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
} ... } we have just received a proposal from a Japanese company named } SHIMADZU. They offered us a scanning electron microscope model } SS-550. I confess that I don't know much about this company and } neither about the quality of its instruments (particularly SEM). } ...
Any reputable SEM manufacturer should be able to provide you with a list of facilities with their SEM in use. If not, you should be wary about being the first.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
I use 1% glycine in PBS. Publish data : 0.15 M glycine or lysine.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } Sara Miller {saram-at-duke.edu} 03/21 4:19 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ammonium chloride (50-100 mM) in PBS or glycine (sorry, don't remember concerntration--probably 1 or 2 % ??) in PBS have been reported. We use NH4Cl.
Sara Miller
On Wed, 21 Mar 2001, Caroline Miller wrote:
} Date: Wed, 21 Mar 2001 12:24:55 -0600 } From: Caroline Miller {camiller-at-creighton.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Aldehyde Removal } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm looking for a method to remove aldehydes from tissue before } processing? Thanks, Caroline Miller } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
In my continuing quest to morph my lab from materials-oriented to bio-oriented I may finally get a chance to get a critical point drier or freeze drier. Could someone with more bio experience than I please help me out with some guidance? Or primary efforts will be intact bacteria and viruses although we may ultimately get into some minor amounts of tissue work. If I can only get either CPD or freeze drier which do I go with? is there any strong justification for both? what accessories are "must haves"? how much should I budget? and finally is there anything beyond the drier and accessories that I should be arranging?
I will not object if any suppliers reading choose to privately send a budgetary quote with the understanding that I am a US Govt. employee and am in no way authorized to negotiate purchases or commit resources.
Thank-you! Erica Valdes US Army SBCCOM Edgewood Chem Bio Center APG, MD 410-436-2608
To find them, you have to go to the Brasilian site, where there the SEM 550, the EPMA 1600, and an AFM are pictured on www.shimadzu.com.br/analitica/portugues/microscopia.htm
There's a page for the EPMA, seems to say 5 spectrometers www.shimadzu.com.br/analitica/portugues/microsonda.htm
Does anybody recognise it as a different brand?
Why only in Brazil?
Strange.
cheers
rtch
} Date: Thu, 22 Mar 2001 07:53:51 +0100 } From: Henrik Kaker {Henrik.Kaker-at-guest.arnes.si} } Reply-to: Henrik.Kaker-at-guest.arnes.si } Organization: Metal Ravne d.o.o., SEM-EDS Lab } To: Ritchie Sims {r.sims-at-auckland.ac.nz} , } "microscopy-at-sparc5.microscopy.com" {microscopy-at-sparc5.microscopy.com} } Subject: Re: SEM inquiry
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } ---. } } } Hello, } } I was searched Shimadzu web site at http://www.shimadzu.com, but } there is no } } any information about scanning electron microscope. } } Henrik } } Ritchie Sims wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } I didn't know that Shimadzu (who are a large and reputable scientific } } equipment manufacturer who afew years ago bought Kratos)) made an } } SEM, and a search of their website www.shimadzu.com revealed none. } } However, I see that their regional Brasil website lists not only the } } SEM 550, but also the EPMA 1600! I remember hearing, years ago, that } } they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that } } may not have been so. } } } } I'd be very interested to hear more about either instrument. } } } } Anybody know anything? } } } } cheers } } } } rtch } } } } } } } } Hi there, } } } } } } I would like to hearing from you opinions about Scanning } } } microscopes. We have a limited fund to buy a new scanning electron } } } microscope and we have just received a proposal from a Japanese } } } company named SHIMADZU. They offered us a scanning electron } } } microscope model SS-550. I confess that I don't know much about this } } } company and neither about the quality of its instruments } } } (particularly SEM). I would appreciate if someone out there could } } } give me an opinion about this equipment. Right now we can buy a unit } } } without the EDS analitical system which we are planning to buy } } } later. Then I also would like to know if we would buy only the image } } } unit first we could buy later the EDS system (from the same company) } } } separately to upgrade the same model (SS-550). The vendor said that } } } it is possible, but sometimes it is hard to trust in sellers } } } especially when you live in South America. } } } } } } Thank all of you } } } } } } Best wishes, } } } } } } } } } Leonardo } } } -- } } } --- } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } } The University of Auckland email : r.sims-at-auckland.ac.nz } } Private Bag 92019 } } Auckland } } New Zealand } } -- } Henrik Kaker, Ph.D. } Metal Ravne d.o.o. } SEM-EDS Lab } Koroska cesta 14, 2390 Ravne } Slovenia } Phone: +386 02 82 21 131 } Fax: +386 02 82 20 436 } http://www.kaker.com } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I've been asked to look into XRD equipment for our lab here, and was wondering if any of you know who the major players are in this area. We deal mostly with polymers and polymer fibers, and want to be able to do both small and wide angle work.
Thanks for the help. dz
David Ziegler U.S. Army, SBCCOM AMSSB-RSS-MS(N) Materials Science Team, SS&T Natick, MA 01760-5020 TEL: (508) 233-6484 FAX: (508) 233-5521 Email: David.Ziegler-at-Natick.Army.Mil
Dear Member of the Microscopy/Microanalysis Listserver,
following the very successful workshops at Lake Tahoe, Leukerbad and Port Ludlow, the next international workshop on electron energy loss spectroscopy and imaging will be held in Guadeloupe from May 5 - 9, 2002.
The abbreviated title will be
SALSA 2002
which stands for
'Strategies and Advances in Atomic Level Spectroscopy and Analysis'.
We think that this title very nicely summarises the central themes that will be discussed at the workshop and also suggests the stimulating environment in which the workshop will be held. You can find further announcements about the workshop on the new homepage
http://www.energyloss.com/
where you will also find a form to express your interest in SALSA 2002 (} This Meeting } Interested?).
May we kindly ask you to fill out the form with as much information as possible. This will help us to demonstrate the level of interest in the meeting in order to attract possible sponsors, including local authorities! If you fill in your e-mail address on the form, you will automatically be placed on our email distribution list and informed about future updates. Your e-mail address will only be used for information directly related to the workshop and will not be given to any other parties. If you prefer not to receive any future e-mails, please help us nonetheless by filling out the form and simply entering 'no' in the e-mail address field.
We hope to see you all at SALSA 2002!
The organisers:
Andrew Bleloch Christian Colliex Bernd Kraus Ondrej Krivanek Richard Leapman Jean-Louis Mansot Joachim Mayer David A. Muller Stephen J. Pennycook
Chris Jeffree wrote the following: ============================================================ Some questions arising: 1) The use of iridium for sputtering is news to me, and looks interesting. I would be interested to see a comparison of iridium film structure with platinum etc. Can a garden variety gold or gold/palladium unit sputter iridium, or are more exotic conditions required?
2) Does anyone have experience of the practical and safety issues arising from osmium coaters? Is the release of osmium tetroxide vapour to room atmosphere well controlled? How economical are they to run? Would they be practical in an environment where there could be several days or even weeks between coating runs? Presumably there are no safety issues with the coated specimens if the metal is inert. True? ============================================================== Chris raised two good points, one I can comment about with greater certainty than the other:
1] When sputtering precious metals with a typical ("garden variety") "SEM lab coater", with or without a turbo pump, there is a certain physics going on, and it is basically the same for all of the precious metals. Without going into a discussion of the nucleation and growth of a layer, the process is fundamentally the same, be it Au, Pt, alloys of these, or even Ir.
The mention of Ir in a previous posting was not in the context of a conventional SEM coater, and presumably there could be a different physics going on so the conclusion when coating is done in that equipment could be quite different (e.g. a much finer grain for example).
So while Pt in a conventional SEM coater does give a slightly smaller grain size, there are some trade offs (e.g. longer coating times) and in any case, it still does not satisfy one with a FESEM seeking to get the most out of their equipment.
2] We at SPI, in our demo lab have had one of the original OPC-40 Osmium Plasma Coaters operating for more than one year and then that was upgraded to the OPC-60 which is described on our website. Yes, the source of the osmium is osmium tetroxide, in 0.1g ampoules. For those not used to looking at osmium tetroxide in ampoules, a 1g amount of the material is described as being "1/3 of a teaspoon". So we are talking about pretty tiny amounts to begin with (e.g. 10% of 1/3 of a teaspoon). On the rotary vane mechanical pump pumping out the unit, in addition to the normal oil mist filter, on top of that is another filter, we call it an "osmium filter", probably a misnomer but it is designed to be a further filter of any mist. At the top is a white paper that serves as an indicator paper: the slightest amount of tetroxide getting out would turn the paper black.
So the first point is that this white paper is still the original paper that has been in continuous operation for more than two years. Translated this means that no tetroxide whatsoever is exiting the pumping system. Even when the unit is vented to atmosphere, there is a procedure where the chamber is first pumped out, and then the chamber can be opened. But one does not get a whiff of tetroxide vapor when the chamber is opened. Those seeing the demos at the shows would attest to that fact.
Of course, at some point there is some tetroxide being pumped out of the chamber, and if indeed some of the OsO4 did indeed get to the pump oil as tetroxide, it would be instantly reduced to the dioxide, something fairly innocuous, which ends up as a colloid in the pump fluid. So other than the apparent need to change the pump oil a bit more often, that seems to be the only consequence we have ever seen from the presence of the tetroxide.
With regard to the unit itself, it is completely safety interlocked, we have done literally hundreds of demonstrations at M&M, PITTCON, and other meetings and safety is the number 1 issue that comes up. But once anyone sees such a demonstration, there is agreement that safety no longer is an issue. It really is impossible for someone to get a whiff of the bad stuff when the chamber is opened.
Further information on this point would be the following: a) It is CE tested (passed the tests) for sale in Europe; these tests are among the most serious in the world in terms of safety. b) I have been in laboratories in Japan and they run the units out in the open although if one really was concerned about that, there is no reason why the unit could not be run in a fume hood.
Since there are well over 100 of the Nippon Laser coaters installed in Japan alone, I trust that there must be at least a few users of this equipment in Japan who might also feel qualified to comment on this approach to the coating of SEM samples. The unique technology embodied in the OPC-60 Plasma Coater is covered by US Patent #5855682; it makes interesting reading for one wanting to understand better the operation of this system.
3] The other issues raised had to do with cost and practicality. While the number of runs one could get depends of course on the thickness of the coatings being applied, from our own experience, using 0.1g ampoules, we typically get 20-40 runs per ampoule, so at a current cost of $8.00 per ampoule, the cost per run is indeed quite low, and in the ball park of cost one would associate with coating with gold. The capital cost is comparable to or less than most chromium coaters.
When the system is turned on, including the pump, the pump down is literally just a few minutes to coating. If the unit has sat unused for several months, we would expect that, as with any vacuum system, a bit longer for the pump down would be needed.
For the long term storage of osmium coated SEM samples, as with any delicate SEM sample, once prepared, we would recommend storing dry, not but not necessarily oxygen free.
Disclaimer: SPI Supplies is a distributor of the Osmium Plasma Coating equipment so we have a vested interest in seeing more coaters being sold. We would be happy to hear what others have to say about this system, pro or con.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Question up front: I¹d like to know how many labs go straight from 100% ethanol (no propylene oxide)to epoxy resin (both Spurr and Epon substitutes, or any other epoxies).
For years, we have deleted the propylene oxide step a.) to decrease the processing time and b.) to prevent having to deal with this toxic chemical. This works fine as long as you use dry ethanol, which we did by keeping drying beads (molecular sieves) in a small (200-500 ml) bottle of ethanol and oven-drying the beads when this absolute EtOH stock was depleted. We also made 3 changes of 100% ethanol (10-15 min each for small blocks, or longer for bigger pieces).
I have other folks in my realm that have routinely used propylene oxide; understandably, they don¹t want to change because it works. It would be nice to have one set of rules that always works. What I¹d like to know from you is how many of you have eliminated the PO, and are there instances where you must use it? Are there other procedures, such as using absolutely dry ethanol or extended times, that you follow if you use only ethanol?
Reply to me directly to save the net from hundreds of answers, and I will tabulate and send out a summary.
Thanks, Sara
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Hi All, I worked as a service engineer for Kratos after they became a subsidiary of Schimadzu. I installed a surface analysis system in Japan at the same time as a Schimadzu SEM with WDX detector was being installed in the room opposite. The system looked very well put together. Unfortunately since the installation engineer and customers spoke no English I have no specific information about the quality of the SEMs they make, but I can tell you they are a multibillion dollar corporation with headquarters in Kyoto, Japan and a significant presence in the US in facilities in Columbia, MD. Some of their products are only offered in Japan but they have an extensive range of analytical instruments that they do offer in the US.
Steve Buckingham Excellatron Solid State LLC 1640 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 617 812 5920
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-muohio.edu] Sent: Thursday, March 22, 2001 9:51 AM To: microscopy-at-sparc5.microscopy.com
Leanardo etc.:
Never heard of them either but a quick internet search showed the following three sites:
1. Shimadzu Scientific Instruments, Inc. We provide solutions to analytical laboratory science by producing an extensive range of products, software and unrivaled customer service. In the United States, Shimadzu
http://www.ssi.shimadzu.com/
2. Shimadzu - Solutions for Science Shimadzu Scientific Instruments - Solutions for Science since 1875 http://www.shimadzu.com/
3. Shimadzu Solutions for Science We provide solutions to analytical laboratory science and medical science by producing an extensive range of products, software and customer service. In Europe, Shimadzu has a
http://www.sel.shimadzu.com/
====} As well as a specific South American Local site:
http://www.shimadzu.com.br
===================================
And since none of us seem familiar with Shimadzu I thought this site
was interesting . .
4. Shimadzu SUCKS homepage Shimadzu specializes in lies, denials and cover-ups!
http://shimadzu.wxs.org/
(Now, take it for whatever its worth, but someone did go the effort and expense of setting up this site, eh?)
================================
On 21 Mar 2001, at 11:57, Leonardo Lagoeiro wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi there, } } I would like to hearing from you opinions about Scanning microscopes. } We have a limited fund to buy a new scanning electron microscope and } we have just received a proposal from a Japanese company named } SHIMADZU. They offered us a scanning electron microscope model } SS-550. I confess that I don't know much about this company and } neither about the quality of its instruments (particularly SEM). I } would appreciate if someone out there could give me an opinion about } this equipment. Right now we can buy a unit without the EDS } analitical system which we are planning to buy later. Then I also } would like to know if we would buy only the image unit first we could } buy later the EDS system (from the same company) separately to } upgrade the same model (SS-550). The vendor said that it is possible, } but sometimes it is hard to trust in sellers especially when you live } in South America. } } Thank all of you } } Best wishes, } } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
} From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } Organization: Dept of Geology, Univ of Auckland } To: Henrik.Kaker-at-guest.arnes.si, microscopy-at-sparc5.microscopy.com } Date: Fri, 23 Mar 2001 07:34:08 GMT+1200 } Subject: Shimadzu } Priority: normal
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } ---. } } } } Yes, that's what I said. } } To find them, you have to go to the Brasilian site, where there the } SEM 550, the EPMA 1600, and an AFM are pictured on } www.shimadzu.com.br/analitica/portugues/microscopia.htm } } There's a page for the EPMA, seems to say 5 spectrometers } www.shimadzu.com.br/analitica/portugues/microsonda.htm } } Does anybody recognise it as a different brand? } } Why only in Brazil? } } Strange. } } cheers } } rtch } } } } } } } } Date: Thu, 22 Mar 2001 07:53:51 +0100 } } From: Henrik Kaker {Henrik.Kaker-at-guest.arnes.si} } } Reply-to: Henrik.Kaker-at-guest.arnes.si } } Organization: Metal Ravne d.o.o., SEM-EDS Lab } } To: Ritchie Sims {r.sims-at-auckland.ac.nz} , } } "microscopy-at-sparc5.microscopy.com" {microscopy-at-sparc5.microscopy.com} } } Subject: Re: SEM inquiry } } } -------------------------------------------------------------------- } } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } } ---. } } } } } } Hello, } } } } I was searched Shimadzu web site at http://www.shimadzu.com, but } } there is no } } } } any information about scanning electron microscope. } } } } Henrik } } } } Ritchie Sims wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } I didn't know that Shimadzu (who are a large and reputable scientific } } } equipment manufacturer who afew years ago bought Kratos)) made an } } } SEM, and a search of their website www.shimadzu.com revealed none. } } } However, I see that their regional Brasil website lists not only the } } } SEM 550, but also the EPMA 1600! I remember hearing, years ago, that } } } they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that } } } may not have been so. } } } } } } I'd be very interested to hear more about either instrument. } } } } } } Anybody know anything? } } } } } } cheers } } } } } } rtch } } } } } } } } } } } Hi there, } } } } } } } } I would like to hearing from you opinions about Scanning } } } } microscopes. We have a limited fund to buy a new scanning electron } } } } microscope and we have just received a proposal from a Japanese } } } } company named SHIMADZU. They offered us a scanning electron } } } } microscope model SS-550. I confess that I don't know much about this } } } } company and neither about the quality of its instruments } } } } (particularly SEM). I would appreciate if someone out there could } } } } give me an opinion about this equipment. Right now we can buy a unit } } } } without the EDS analitical system which we are planning to buy } } } } later. Then I also would like to know if we would buy only the image } } } } unit first we could buy later the EDS system (from the same company) } } } } separately to upgrade the same model (SS-550). The vendor said that } } } } it is possible, but sometimes it is hard to trust in sellers } } } } especially when you live in South America. } } } } } } } } Thank all of you } } } } } } } } Best wishes, } } } } } } } } } } } } Leonardo } } } } -- } } } } --- } } } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } } Department of Geology Fax : 64 9 3737435 } } } The University of Auckland email : r.sims-at-auckland.ac.nz } } } Private Bag 92019 } } } Auckland } } } New Zealand } } } } -- } } Henrik Kaker, Ph.D. } } Metal Ravne d.o.o. } } SEM-EDS Lab } } Koroska cesta 14, 2390 Ravne } } Slovenia } } Phone: +386 02 82 21 131 } } Fax: +386 02 82 20 436 } } http://www.kaker.com } } } } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Ir as an ion beam sputtering target began in'92 after a scientist at Lucent Technology, formerly Bell Labs, was looking at silicon by STM. Before AFM, STM was the only type of SPM. In STM the tip contacts the specimen for tunneling. To increase specimen conductivity without introducing artifacts, Pt was ion beam deposited in an IBS, instrument developed by VCR GROUP now called IBS/e manufactured by SBT. Different metals were compared to Pt for minimum film resistivity and smoothness including Cr, Ti, W & Ir. Surprisingly Ir films prevented specimen surfaces from STM tip damage, were as conductive as Pt films, sputtered like Pt ie sputter rate & material distribution were the same and Ir does not oxidize.
After learning Bell Labs results, we hypothesized that if Ir films minimized STM tip damage, there may be a benefit to delicate specimens from electron beam interaction in FESEMs. Therefore thin films of Ir & Pt were deposited on 200 nm polystyrene spheres dispersed on mica. Several specimens with different metal thickness of Ir & Pt were examined in a Hitachi in-lens FESEM. It was observed that the PS spheres with Ir films showed less beam damage than those with Pt films. During separate TEM studies we learned that 5 to 100 angstrom thick Ir films had virtually identical structure as Pt films.
Further tests were done in Hitachi & JEOL FESEMs of thin Ir films on various substrates. No Ir structure, grains, could be seen below 200kX. Since '93, we have recommended Ir targets for the IBS since most FESEM work is carried out below 200kX. We still recommend Cr or other refractory metal targets for examination above 200kX.
Vince Carlino SBT, Inc.
Some questions arising: 1) The use of iridium for sputtering is news to me, and looks interesting. I would be interested to see a comparison of iridium film structure with platinum etc. Can a garden variety gold or gold/palladium unit sputter iridium, or are more exotic conditions required?
2) Does anyone have experience of the practical and safety issues arising from osmium coaters? Is the release of osmium tetroxide vapour to room atmosphere well controlled? How economical are they to run? Would they be practical in an environment where there could be several days or even weeks between coating runs? Presumably there are no safety issues with the coated specimens if the metal is inert. True?
vince thank you for this information. Iridium suppliers: Johnson Matthey Noble Metals Division supply it in various forms from facilities in Royston, Cambridge UK and West Chester, Pennsylvania USA. Sheet of 99.99% purity up to 10" wide by 0.1 to 5mm thick is available, and discs, wire etc. http://www.noble.matthey.com/product/detail.asp?id=34 An information sheet with contact addresses is available from the web site as a 47k .pdf file
Chris
----- Original Message ----- } From: {"VCRVINCE-at-aol.com"-at-sparc5.microscopy.com} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} Cc: {microscopy-at-sparc5.microscopy.com} ; "John F. Mansfield" {jfmjfm-at-umich.edu} Sent: Friday, March 23, 2001 1:00 AM
Folks, this is for a colleague of mine. If you have such a document please mail it to me at your earliest convenience Simon
I am seeking information appropriate to preparing an SOP to handle the eventuality that a mercury lamp explodes in one of our fluorescent microscopes. I am looking for sepcific information about potential personnel risks associated with the mercury vapor and what, if any, decontamination procedures are necessary/appropriate for potentially exposed equipment or personnel. I appreciate that the possibility of an explosion is remote but, I am required to address this eventuality.
Thank you again for sharing your findings with me.
------------------------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 3500 Terrace St University of Pittsburgh Pittsburgh PA 15261 Tel: 412-648-3051 Fax: 412-648-8330 URL: http://www.cbi.pitt.edu --------------------------------------------
I am in the market for a digital camera upgrade to my LaB6 2010. At the risk of receiving more information than I want to know about, venders are welcome to contact me off line to bring me up to date on the technology. Invitations to bid are expected to follow a review of the technology.
Thanks, Bruce Brinson Optical Analyst Rice University
Dear colleguages, I'm seeking a controller for a JSM840 whicsh is reliable and can control the SEM beam in X-Y direction at least with 4096 pixel resolution. And it would be nice if the stage and the controller would be optically coupled to the computer. I saw such a controller in some older letter, but I don't remember where? I thank any kind of help LAjos Pogany dr. Pogány Lajos Senior Research Fellow, Metals Research Department, Research Institute for Solid State Physics and Optics, Hungarian Academy of Sciences Office Address: H-1121 Budapest, Konkoly-Thege ut 29-33 Letters: H-1525 Budapest, P.O.B. 49, Hungary Phone: (00)-36-1-392-2222/17-25; Fax: (00)-36-1-392-2215 e-mail:pogany-at-szfki.hu
Announcement and Call for Papers TENTH CROATIAN-SLOVENIAN CRYSTALLOGRAPHIC MEETING Lovran, Croatia, June 21-24, 2001
The Meeting is open for the contribution dealing with all aspects of crystallography and its applications (Chemical Crystallography, Industrial Crystallography, Biological Crystallography, Physical Crystallography, Applied Crystallography and Mineralogy) and its techniques (XRD, TEM, SEM, STM)
please visit the web site
http://www.chem.pmf.hr/~hkz/lovran01/
Prof. A. Tonejc Department of Physics Faculty of Science Zagreb Croatia
In a message dated 03/23/2001 6:26:00 AM US Mountain Standard Time, swatkins+-at-pitt.edu writes:
{ { I am seeking information appropriate to preparing an SOP to handle the eventuality that a mercury lamp explodes in one of our fluorescent microscopes. I am looking for sepcific information about potential personnel risks associated with the mercury vapor and what, if any, decontamination procedures are necessary/appropriate for potentially exposed equipment or personnel. I appreciate that the possibility of an explosion is remote but, I am required to address this eventuality.
Thank you again for sharing your findings with me.
} }
Simon,
I know the manufacturers of the bulbs have this info. I have seen the precautionary statements on the flyers which accompany Osram bulbs, and Ushio probably has a similar document. If you don't hear from someone who already has this info, you could probably get it from Osram.
In a nutshell, Osram recommends that all personnel immediately leave the area for at least 30 minutes to avoid inhaling mercury vapor, and then any mercury residue, debris, envelope shards, etc. should be cleaned up with a mercury cleanup kit, and the debris properly disposed of as biohazardous material with mercury clearly identified as the hazard. Alternatively, if you have a hazmat team, they could be called in to do the cleanup, I suppose.
Hi! I am preparing a sample for TEM. The sample is GaN on sapphire substrate. I am supposed to make a sample in plane-view. Its needless to say how hard sapphire is. Grinding it down to 150microns was itself a tough job. Now I have put it on the dimpler, and using a diamond slurry to grind it. Its been 10 hrs and I have been successful in taking off only 70 microns. Does anyone have any bright ideas to speed up this process?
Praveena bhaskara Department of Chemical Engineering. University of Massachusettes, Lowell Lowell _________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
One of my clients needs to do immunocytochemistry on mouse bone marrow, and needs to use frozen sections. I haven't got a clue where to begin (she will harvest the bone marrow). Any ideas?
Thanks as always, Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I'd just like to say "Thank You" to all of you who replied to my question on the EDS analysis of the contamiants. you answers proved to be most helpful.
Yep! The small angle cleavage angle works well for GaN on sapphire. I prepared about 8 samples in about 2 hours while teaching the technique to students and staff at the Univ. of Illinois. We only looked at three samples. Two were very very good. I will send you an image off-list. You can obtain information about South Bay Technology's Micro-Cleave kit at their web site, http://www.southbaytech.com
For grinding down the samples, use a 30 or 45 um diamond lapping film with a hand grinder. (I used 30um.) Works well.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Praveena Bhaskara [mailto:bubbyp-at-hotmail.com] } Sent: Friday, March 23, 2001 1:00 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM sample preparation } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Hi! } I am preparing a sample for TEM. The sample is GaN on } sapphire substrate. I } am supposed to make a sample in plane-view. Its needless to } say how hard } sapphire is. Grinding it down to 150microns was itself a } tough job. Now I } have put it on the dimpler, and using a diamond slurry to } grind it. Its been } 10 hrs and I have been successful in taking off only 70 } microns. Does anyone } have any bright ideas to speed up this process? } } Praveena bhaskara } Department of Chemical Engineering. } University of Massachusettes, Lowell } Lowell } ______________________________________________________________ } ___________ } Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
} Dear Praveena: } } We have an application note on preparing GaN on Sapphire using our } Tripod Polisher. You can download it from our webiste at } www.southbaytech.com. Go to Applications Support then click on } Applications Notes. } } I hope this helps. } } David } } } ----------------------------------------------------------------------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hi! } } I am preparing a sample for TEM. The sample is GaN on sapphire substrate. I } } am supposed to make a sample in plane-view. Its needless to say how hard } } sapphire is. Grinding it down to 150microns was itself a tough job. Now I } } have put it on the dimpler, and using a diamond slurry to grind it. Its been } } 10 hrs and I have been successful in taking off only 70 microns. Does anyone } } have any bright ideas to speed up this process? } } } } Praveena bhaskara } } Department of Chemical Engineering. } } University of Massachusettes, Lowell } } Lowell } } _________________________________________________________________________ } } -- } David Henriks TEL: +1-949-492-2600 } South Bay Technology, Inc. FAX: +1-949-492-1499 } 1120 Via Callejon } San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com } } *** www.southbaytech.com ***
Hg clean up kits are available from Lab Safety Supply, Box 1368, Janesville, WI 53547 (800 356 0783).
I have no commercial interest in this company. That's just where we got our kit. Fortunately, we haven't had to use it.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
---------- Forwarded message ----------
In a message dated 03/23/2001 6:26:00 AM US Mountain Standard Time, swatkins+-at-pitt.edu writes:
{ { I am seeking information appropriate to preparing an SOP to handle the eventuality that a mercury lamp explodes in one of our fluorescent microscopes. I am looking for sepcific information about potential personnel risks associated with the mercury vapor and what, if any, decontamination procedures are necessary/appropriate for potentially exposed equipment or personnel. I appreciate that the possibility of an explosion is remote but, I am required to address this eventuality.
Thank you again for sharing your findings with me.
} }
Simon,
I know the manufacturers of the bulbs have this info. I have seen the precautionary statements on the flyers which accompany Osram bulbs, and Ushio probably has a similar document. If you don't hear from someone who already has this info, you could probably get it from Osram.
In a nutshell, Osram recommends that all personnel immediately leave the area for at least 30 minutes to avoid inhaling mercury vapor, and then any mercury residue, debris, envelope shards, etc. should be cleaned up with a mercury cleanup kit, and the debris properly disposed of as biohazardous material with mercury clearly identified as the hazard. Alternatively, if you have a hazmat team, they could be called in to do the cleanup, I suppose.
I'm in another lab now, and in the hood here I found about 50 ml of DMF (or DMFA) - see below - and wondering what EMer's might use it for. Here's what the Merck Index says about it:
--------------------------- from the Merck Index:
DMF = N,N - Dimethylformamide (also abbreviated DMFA); C3H7NO Colorless to very slightly yellow liquid. Faint amine odor.
Human toxicity: Vapor harmful. Irritating to skin, eyes and mucous membranes. Liver injury has been produced in experimental animals by prolonged inhalation of 100 ppm.
Use: Solvent for liquids & gases. In the synthesis of organic compounds. Solvent for Orlon and similar polyacrylic fibers. Wherever a solvent with a slow rate of evaporation is required. Has been termed the universal organic solvent. ----------------------------
Any thoughts will be appreciated, thanks.
Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
To the person who was wondering weather or not P.O. was needed after dehydration and before embedding in Spurrs:
For the past 15 years I have been successfully embedding in Spurrs by using graded steps of acetone and going from my third change in 100% acetone to one third Spurrs and two thirds 100% acetone with out the use of P.O.. Of course then I follow that with a 1:1 mixture and then a 2:3 mixture and then a 100% Spurrs before embedding in 100% Spurrs at 65 degrees Celsius for 12 hours or more. Best of luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
I have been receiving conflicting reports concerning Amray service on the FE SEMs. The "official" word has been that Amray will continue to support the FESEMs "for some time" but I have had several customers call me & request service for their FESEMs as Amray will not offer a contract for the next year.
Has anyone had a similar experience or give me a striaght answer on this subject?
I was wondering if anyone has had any experience with lead aspartate "en Block" staining. I have tried "en block" staining before with disappointing results, but I have never tried it with lead asparate. The catalog describes it such that with Lead Aspartate you can then skip the staining step after sectioning.
Soft Imaging's ADDA is optically coupled between controlling computer and physical electrical interface to SEM.
http://www.soft-imaging.com
gary g.
At 08:18 AM 3/23/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Everyone, I want to make my own lacey carbon films for cryo-tem and was wondering if anyone had some good methods, experiences, insights, or references?
I found references for lacey formvar, pure carbon films, and a few others... But nothing explicitely for lacey carbon.
All comments greatly appreciated, Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Email: dfvillamar-at-acuabiotec.com Name: Daniel F. Villamar
Organization: AcuaBiotec LLC
Education: Graduate College
Location: Damascus, MD USA
Question: We are interested in SEM of samples of marine zooplankton, i.e., larval and postlarval shrimp. We would like to have a ventral view of the mouth parts and a view of the inside surface of the gut wall. The latter would require sectioning of the specimens along the longitudinal axis and scanning the inside surface starting at the mouth and working down through the foregut, midgut and hindgut of the specimens.
1. Does enyone provide such a service? 2. How should we fix and preserve the specimens for best result.
If I remember correctly, you make holey carbon films by first making your holey formvar (or other plastic), coating this film with a thin layer of carbon to make it continuous, and then dissolving the support plastic with a suitable solvent. This is off the top of my head. A few years ago, Mr. Pella suggested some literature to me on how to do it.
It's easier and more cost-effective to buy them from any of the EM supply houses!
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] } Sent: Friday, March 23, 2001 6:49 PM } To: microscopy-at-sparc5.microscopy.com } Subject: ? how to make lacey carbon films? } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Hello Everyone, } I want to make my own lacey carbon films for cryo-tem and was } wondering if } anyone had some good methods, experiences, insights, or references? } } I found references for lacey formvar, pure carbon films, and a few } others... But nothing explicitely for lacey carbon. } } All comments greatly appreciated, Thanks. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } \/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 } Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } }
The word I have received so far is that later model FESEMs will be covered under contracts. The distinction of "later model" is a perhaps nebulous term. The 1800 series instruments will probably not qualify. My 1910FESEM is based on 1800 electronics but uses the 305FE gun--and so far, is covered under contract. I am good for the rest of CY 2001. If Amray KLA stopped supporting FESEMs, something major would have to occur to keep the guns (at a minimum) available to established systems. The guns seem to last about 2.5-4 years. During that time, they are rock solid.
It seems to me that the more that a microscopist can do in regards to maintenance is better in the near term. This is despite a maintenance contract. If simple things go wrong and you can fix them, doing this will keep your system below the cost accountant's radar screen. The key for having the contract is if something really major goes wrong. An FE gun under contract is about $2K. Otherwise, it is at least $18K. How about a turbo pump? No cost under contract. Otherwise???
what about changing final apertures? I have not done a good job of doing this. But now I know more about how to do it correctly. And cleaning the holder is not a trivial process, nor one to be ignored.
gary g.
At 01:56 PM 3/23/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for TEM images for two reasons: 1. to become part of a digital image database for cell biology courses here at the University of British Columbia; 2. to be part of an image database used on CDs accompanying the new edition of a popular cell biology textbook.
I am looking for high quality images of a variety of cell types. These images can be either digitized or the original negatives. If they are digitized then they must be scanned-in to standards set by ourselves and the publisher. If the images are in the form of the original negatives we can arrange to have them sent here, scanned-in and returned.
In return contributors to the UBC collection can have access to other images in the collection and those who's images are used by the publisher will receive $50/US per image chosen. The publisher does not want to hold the copyright on the images used, rather they have proposed a generous licence agreement.
Interested parties can contact me (Brian Oates) at:
brian.oates-at-ubc.ca -- Dr Brian Oates Department of Botany University of British Columbia Vancouver, B. C. V6T 1Z4 604.822.4630 brian.oates-at-ubc.ca
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Snoring is the audible symptom of a {/FONT} {FONT color=3D"#0000A0"} {B} blocked airway during sleep. {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} The noise is caused by a vibration in the soft palate as the lungs pull ha= rd to take in a {/FONT} {FONT color=3D"#0000A0"} {B} weakened current of incoming air. {/B} {/FONT} {FONT color=3D"#0000A0"} This blockage may result from any number of circu= mstances, and these offer clues to get rid of the problem. {BR} {BR} Snoring is also more than likely to occur among people who {/FONT} {FONT color=3D"#0000A0"} {B} sleep on their backs {/B} {/FONT} {FONT color=3D"#0000A0"} ; the tongue falls back toward the throat and par= tly closes the airway. {BR} {BR} {/FONT} {FONT color=3D"#800040"} {B} The hidden dangers of snoring? {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {B} {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {UL} {LI} Snoring could increase the chances of a h= eart attack if not treated {BR} {LI} Hypertension and increased anxiety are associated with snoring {BR} {LI} Snoring causes sleep apnea a serious treatable medical condition {BR} {/LI} {/LI} {/LI} {/UL} {/FONT} {FONT color=3D"#0000A0"} {BR} {/FONT} {FONT color=3D"#800040"} {B} Snoring decreases sexual performance {/B} {/FONT= } {FONT color=3D"#800040"} {/FONT} {FONT color=3D"#0000A0"} by half, due to lack of oxygen to the brain which= decreases sexual responsiveness. {BR} {BR} Most people who snore have a {/FONT} {FONT color=3D"#800040"} {B} shortage of oxygen {/B} {/FONT} {FONT color=3D"#0000A0"} for a significant portion of that person's life.= In recent studies, daytime fatigue and cardiovascular disorder are relate= d to snoring. In a few cases, heavy snoring is a sign of potentially life = -threatening problem. Sleep apnea in which breathing stops for a second or= even minutes at a time and finally resumes with snorting and tossing arou= nd. This pattern may be repeated hundreds of times all night long. The res= ult is chronic oxygen shortage, which leads to abnormal heart rhythms, hig= h blood pressure, and heart strain. {BR} {BR} {/FONT} {FONT color=3D"#800040"} {B} How the Snore Eliminator works! {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} Snore Eliminator offers a unique break through wi= th its all {/FONT} {FONT color=3D"#800040"} natural enzymes {/FONT} {FONT color=3D"#0000A0"} which metabolize the secretions, allowing the bo= dy to absorb them into the back of the throat. The herbs reduce tissue swe= lling. The result is to open the airway and smooth the airflow. This in tu= rn eliminates the snoring for hours at a time. It also {/FONT} {FONT color=3D"#800040"} {B} reduces the noise {/B} {/FONT} {FONT color=3D"#800040"} made by breathing through the mouth while sleepi= ng {/FONT} {FONT color=3D"#0000A0"} . By simply spraying the back of the throat, tong= ue and uvula with our special formula, manufactured using our patented bre= ak through process, the soft tissue is coated. This allows the {/FONT} {FONT color=3D"#0000A0"} {B} {/B} {/FONT} {FONT color=3D"#800040"} {B} throat a chance to relax {/B} {/FONT} {FONT color=3D"#0000A0"} thus a reduced amount of snoring. In most cases = snoring is eliminated. Snore Eliminator coats the throat area with a uniqu= e patented process of all natural lubricants. {BR} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} TESTIMONIALS from several of our satisfied cus= tomers: {BR} {BR} {/B} {/FONT} {FONT face=3D"Times New Roman"} {FONT color=3D"#FF0000"} {B} Lorenzo writes... {BR} {BR} I am having a good nights sleep, finally. I used to wake myself up during= the night which was a real bummer as I would not always go back to sleep.= Sleeping through the night is great. Thanks a lot Snore Eliminator. {BR= } PS: My girlfriend loves it as much or more than I do. {BR} {BR} Alex writes... {BR} {BR} My husband uses it for another reason. He has a problem with blocked nasa= l passages. While he's asleep the passages close, he stops breathing and = he wakes up gasping. When he takes Snore Eliminator his nose stays cleare= r and he's able to sleep more restfully. {BR} Thank you for a product that has really helped us! {BR} {BR} Glen writes... {BR} {BR} In February 1995, I was diagnosed with severe sleep apnea. A CPAP was the= suggested treatment for my affliction, but it did not help my condition. = I heard your advertisement and ordered a bottle of Snore Eliminator and m= y sleep improved immediately. I continue to use it and my sleep has been = better than it had been in years. Thanks. {BR} {BR} Steve writes... {BR} {BR} Let me take this opportunity to thank you for your wonderful product and t= ell you how much my wife, Tracy, and I appreciate the quiet nights we have= enjoyed since discovering it. It's very rare when a product actually del= ivers what it promises but Snore Eliminator truly does. We have tried most= of the other advertised remedies without any satisfaction and I am very p= leased to be able to write and say that this product is as close as you ca= n get to an absolute "Miracle". {BR} {BR} {/B} {/FONT} {FONT face=3D"MS Sans Serif"} {FONT color=3D"#800040"} If you have a snoring problem what can you do? {/F= ONT} {FONT color=3D"#0000A0"} Who would you turn to? Look no further, the ans= wer to your snoring problems are over. Let our patented {/FONT} {FONT color=3D"#0000A0"} {B} Snoring Eliminator {/B} {/FONT} {FONT color=3D"#0000A0"} help you get through those sleepless nights. We = guarantee it. In fact, we are so confident in Snore Eliminator that if you= are not 100% satisfied with it you can return the unused portion for a fu= ll refund, no questions asked* {BR} {BR} It is estimated that 90 million Americans, over the age of 18,suffer from = habitual snoring. {BR} {BR} {/FONT} {FONT color=3D"#800000"} {B} How to Use Snore Eliminator: {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} Open your mouth {BR} Hold the 4oz bottle about four inches from your open mouth {BR} Spray one or two application onto the back of the throat. {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} {BR} Snore Eliminator is safe to use nightly because of its' natural ingredient= s. {BR} Please note as with any health-related product, it is recommended that you= consult with your physician prior to use if you have any medical conditio= ns. {BR} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} A Money Back Guarantee! * {BR} {/B} {/FONT} {FONT face=3D"Times New Roman"} {BR} {FONT face=3D"MS Sans Serif"} {FONT color=3D"#0000A0"} {B} We are so confident in Snore Eliminator that w= e offer a 15-day money back guarantee. Merely return the unused portion o= f Snore Eliminator within 15 days and we will refund your purchase price w= ith no questions asked! {BR} {BR} Our normal price for Snore Eliminator is $29.95, but for a limited time on= ly, you can purchase it for only {/B} {/FONT} {FONT color=3D"#800040"} {B} $19.49 plus $3.95 {/B} {/FONT} {FONT color=3D"#0000A0"} {B} for shipping handling. We will also continue t= o offer you Snore Eliminator at this low price as long as you purchase fro= m us in the future. To take advantage of this savings you must order with= in the next 10 days. {BR} {BR} Here's How to order {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} You can order by mailing us a Check, Cash or a Money Order. You can also = order by faxing us a copy of your Check. Faxing your order to us speeds y= our order up by 3-5 days. {BR} {BR} To order by Check, Cash or Money Order merely send $19.49 plus $3.95 for a= total of $23.44 per bottle. 1 bottle of Snore Eliminator will last you 4-= 6 weeks. Order TWO bottles for $19.49 and we do not charge you any shippin= g and handling. This means you will receive an 8 to 12 week supply of Sno= re Eliminator for only $38.98, including shipping and handling. This is a = 17% savings. {BR} {BR} Mail your check, cash or Money Order along with your name address, phone n= umber and email address filled out on the following form: {BR} {BR} Your Name:___________________________________________(please print) {BR} {BR} Street: ______________________________________________ {BR} {BR} City:______________________State/Province:____________________ {BR} {BR} Zip/Postal Code:_________________Country:_________________________ {BR} {BR} Phone number:______________________________________________ {BR} {BR} FaxNumber:_________________________________________________ {BR} {BR} Email address:_______________________________________ {BR} (if there is a problem with your order this is where we will notify you) {B= R} {BR} To: {BR} {BR} John Taylor {BR} CEO {BR} Internet Information Services, Snore Eliminator Division {BR} PO Box 21442 {BR} Billings, MT 59104 {BR} {BR} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} {BR} To Fax us your Order (Fill out and fax us every page which follows): {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} MAKE YOUR CHECK PAYABLE TO {/B} {/FONT} {FONT color=3D"#0000A0"} : Internet Information Services, Snore Eliminator= Division, tape your check to the following form and Fax to {/FONT} {FONT color=3D"#0000A0"} {B} 1-775-599-3092 {/B} {/FONT} {FONT color=3D"#0000A0"} . If this line is busy, please try faxing to {BR= } {/FONT} {FONT color=3D"#0000A0"} {B} 1-801-383-9082. {/B} {/FONT} {FONT color=3D"#0000A0"} Faxing your check to us speeds your delivery up = by 3-5 days. {BR} {BR} Please ship me 1 bottle of Snore Eliminator for $19.49 plus $3.95 shipping= and handling for a total of $23.44 per bottle. Order TWO bottles for $19= .49 and we do not charge you any shipping and handling. This means you wi= ll receive an 8 to 12 week supply of Snore Eliminator for only $38.98, inc= luding shipping and handling. This is a 17% savings. {/FONT} {FONT face=3D"Times New Roman"} {FONT color=3D"#0000A0"} {BR} {/FONT} {FONT face=3D"MS Sans Serif"} {FONT color=3D"#0000A0"} {BR} Your Name:___________________________________________(please print) {BR} {BR} Street: ______________________________________________ {BR} {BR} City:______________________State/Province:____________________ {BR} {BR} Zip Code:_________________Country:_________________________ {BR} {BR} Phone number:______________________________________________ {BR} {BR} Fax Number:_________________________________________________ {BR} {BR} Email address:_______________________________________ {BR} (if there is a problem with your order this is where we will notify you) {B= R} {BR} ************************************************************** {BR} {BR} Internet Information Services Check Authorization and Order Form {BR} {BR} I wish to authorize the purchase of the Snore Eliminator from Internet Inf= ormation Services using this order and check authorization form. I hereby= authorize Internet Information Services to duplicate the attached check i= n bank draft form for the amount shown on this attached check. I will ret= ain my original copy for my record of this transaction. {BR} {BR} This Authorization is valid for this transaction only. No other bank draf= ts may be created without my direct written authorization. {BR} {BR} Dated:_________________________________ {BR} {BR} Signed:________________________________________ {BR} {BR} {BR} .Tape your check here and Fax it to us... {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} Fax this form to: {/FONT} {FONT color=3D"#0000A0"} {B} 1-775-599-3092 {/B} {/FONT} {FONT color=3D"#0000A0"} . If this line is busy, please try faxing to {/FO= NT} {FONT color=3D"#0000A0"} {B} 1-801-383-9082. {BR} {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} Thank you for your business, {BR} {BR} John Taylor {BR} CEO {BR} Internet Info Service, Snore Eliminator Division {BR} PO Box 21442 {BR} Billings, MT 59104 {BR} {BR} {BR} {BR} {BR} {BR} {/FONT} {FONT face=3D"Times New Roman"} {FONT color=3D"#000080"} {BR} {BR} {BR} {BR} {BR} {/FONT} {FONT face=3D"Courier New"} {FONT size=3D3} {FONT color=3D"#000000"} +++++++++++++++++++++++++++++++++++++++++++++++++= +++++++ {BR} This ad is produced and sent out by: {BR} Universal Advertising Systems {BR} To be removed from our mailing list please email us at {BR} vanessagreen-at-freeze.com with remove in the subject line or {BR} call us toll free at 1-888-605-2485 and give us your email address or writ= e us at: {BR} Central DB Removal, PO Box 1200, Oranjestad, Aruba {BR} ++++++++++++++++++++++++++++++++++++++++++++++++++++++++ {/FONT} {FONT face=3D"Times New Roman"} {FONT size=3D3} {BR} {FONT face=3D"MS Sans Serif"} {FONT color=3D"#000000"} {BR} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FO= NT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/BODY} {/HTML}
Michael Reiner University of Cologne, Germany Dept. of Anatomy I
} Hi, } } I'm passing on a question from a colleague. Does anyone know of a technique } for etching LR White from sections on a slide? I gave him all the info I had } on epoxy removal using ethoxide and so on, but I've been unable to locate } any references specific to LR White. } } Thanks. } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } }
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I am negotiating with my college to purchase a block of time on their new variable pressure SEM (Hitachi S-3000N with EDX). We have agreed to abide by the 'average' cost per hour charged by universities and colleges. I seem to remember that there was a discussion along these lines some time ago but I cannot find it in the archives. If you pay per-hour fees for SEM usage I would be much obliged if you would let me know what they are.
Thanks
Michael
Michael McInerney Rose-Hulman Institute, Terre Haute, IN.
Amray customers, as well as the rest of us, are scrambling to figure out a lot of things. I've had loyal Amray customers running to me because they've had their service dropped, they've had implied threats that their service would be dropped in the near term and because they are just sick and nervous of the way the whole thing was handled by the manufacturer. I am quite sure that there are many who are just plain confused over whether they have been dropped or not.
On the other hand, a new customer of mine had to field a question from an Amray rep over whether they were renewing a contract on a 3300 FESEM. When told they were not, the rep apparently went over the line about the 'impossibility' of getting other service and the 'proprietary' nature of their service.
I think that, having punched a hole in their own boat, Amray is now surprised that they are taking on water.
There are a lot of long-time, dedicated people at Amray who feel betrayed by their own company now. I know, having been through a very similar business turn with ETEC when they were bought out by Perkin-Elmer and their SEM business was left to suffer an ignoble death. It's hard to believe that anyone could possibly have made such decisions without having a long term desire to shutdown the business.
The only question that seems to remain is - what business is being shut down? Amray has always been a major player in the semi-conductor field. These days, the FESEM is the most attractive portion of that business. My guess would be that they have chosen to still pursue the semi manufacturers with FESEMs but close down their general purpose SEM business. In the mean time, they want to still provide the lucrative service contracts on the general purpose FESEMs out there. Cash is king, and the cash in their operation is in the selling and service of FESEMs, primarily to those customers who are willing and able to pay a premium for these instruments.
Overall, this move could only be construed as an attempt to increase the profit margin of their service operation. The general purpose SEMs are spread out geographically around the country, where the semi business is far less so. The older instruments represent an additional training cost for new service people. By limiting the number of instruments being serviced, the geographical spread of those instruments and the learning curve of their service staff, they can drastically cut back their service staff, weather attrition and defections of service staff by having new hires online quicker and in the end, probably have a staff that is paid less to do more. The same could also be said of their sales force.
Don't sweat, Gary - cathode emitters are available for the Amray FESEMs in the aftermarket for around $2,500 and turbopump exchanges cost about the same from the manufacturer. I can't really address the pricing strategies of other service providers, but in my case, whatever type of contract you choose, or even on a billable basis, you would save at least a few thousand a year over the Amray service costs. I'll cover the cathode by tacking on the $2,500 per year to the contract and simply change it once a year, and still beat Amray's cost by $4,000 - $6,000 per year. But I suggest that the customer cover it themselves - buy a new emitter when they first take out a contract and keep it on hand. In this way, the customer can themselves better control the cost through proper care and operation of the instrument, and deciding for themselves when a replacement is needed. This also gives you that warm fuzzy feeling knowing that you have one standing by, ready to use if needed. Third party service providers have great flexibility in writing contracts - in the case above, I would normally cover the cost of installation of the emitter even though you may have chosen to be responsible for the emitter.
Your mention of those using the instrument having some ability to take care of many problems they run across is right on the mark. I always prefer customers who have a real interest and knowledge of the instrumentation they use. Not because it may make my life easier (although it does because I know I'm talking to someone knowledgeable and am more willing to take what they say at face value), but because it's fun to share with, and learn from, those with similar interests.
I'm all for having a customer watch over my shoulder and understand what I'm doing, because it protects their better interests as well as mine. I'm not perfect, and sometimes (perhaps often) I'll get off on the wrong direction on a problem. The better the description of the problem I get, the better I'll do. It also helps sometimes to have a back-seat driver to let you know when you've taken a wrong turn, as damaging to the ego as that can be. Of course, for the customer, having a better knowledge of the instrument and its service means some peace of mind in finding someone to service it.
Kind of like knowing something about the various systems and construction methods in a house before hiring a remodeling company.
On Friday, March 23, 2001 8:32 PM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } } The word I have received so far is that later model FESEMs will be } covered under contracts. The distinction of "later model" is } a perhaps nebulous term. The 1800 series instruments } will probably not qualify. My 1910FESEM is based on 1800 } electronics but uses the 305FE gun--and so far, is covered } under contract. I am good for the rest of CY 2001. If Amray } KLA stopped supporting FESEMs, something major would have } to occur to keep the guns (at a minimum) available to established } systems. The guns seem to last about 2.5-4 years. During } that time, they are rock solid. } } It seems to me that the more that a microscopist can do } in regards to maintenance is better in the near term. This } is despite a maintenance contract. If simple things go } wrong and you can fix them, doing this will keep your } system below the cost accountant's radar screen. The key } for having the contract is if something really major goes } wrong. An FE gun under contract is about $2K. Otherwise, } it is at least $18K. How about a turbo pump? No cost } under contract. Otherwise??? } } what about changing final apertures? I have not done a good } job of doing this. But now I know more about how to do it } correctly. And cleaning the holder is not a trivial process, } nor one to be ignored. } } gary g. } } } At 01:56 PM 3/23/2001, you wrote: } } } } } Hi Listers, } } } } I have been receiving conflicting reports concerning Amray service on the FE } } SEMs. } } The "official" word has been that Amray will continue to support the FESEMs } } "for some time" but I have had several customers call me & request service } } for their FESEMs as Amray will not offer a contract for the next year. } } } } Has anyone had a similar experience or give me a striaght answer on this } } subject? } } } } } } Thank You, } } } } Earl Weltmer } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
} [snip] } } The only question that seems to remain is - what business is being shut } down? Amray has always been a major player in the semi-conductor field. } These days, the FESEM is the most attractive portion of that business. My } guess would be that they have chosen to still pursue the semi manufacturers } with FESEMs but close down their general purpose SEM business. In the mean } time, they want to still provide the lucrative service contracts on the } general purpose FESEMs out there. Cash is king, and the cash in their } operation is in the selling and service of FESEMs, primarily to those } customers who are willing and able to pay a premium for these instruments.
Amray, as a company, no longer exists. Amray is a division of KLA-Tencor. For some reason, Amray apparently could not compete in the general purpose (lab) SEM marketplace. Or, other forces came into play which caused the change.
I do know that Amray had developed very good FESEM systems--optics, control software, stages, etc. KLA-Tencor is in the semiconductor defect analysis business. They lacked the type of SEM technology that Amray had. Consequently, KLA-Tencor purchased Amray, ostensibly I believe, to acquire Amray's SEM technology. This is similar to the relationship between FEI and Philips, wherein Philips acquired FEI for its ion beam expertise. To wit, FEI now produces a fine dual beam focused ion beam system which combines ion beam technology and SEM imaging (e.g., FEI 830).
KLA-Tencor produces a top line semiconductor defect analysis tool set (8100xl, 8300, 8500, etc.) which combines KLA's best attributes along with the FESEM qualities of Amray. The defect analysis tools cost about $2M each, compared to a typical lab FESEM at say $400K. The earlier Amray systems were priced lower than this. It seems to me that KLA, with Amray in tow, had made a decision to focus on the high cost (high profit) business area of semiconductor defect analysis, at the exclusion of lab SEMs. That is why we are seeing what is happening now with Amray service contracts.
In some respects, this was a myopic decision. Currently, the semiconductor industry is in a decline (albeit not monumental). There surely was and is a market for lab SEMs. KLA obviously made a business decision to focus on semiconductor defect analysis tools. From what I am seeing, the older Amray systems are no longer being supported by Amray. Since the thermionic systems are rather simple in design and easier to maintain than FESEMs, this may not be too bad. But, if one had a bad PC-10, that is of course a different matter.
So far as I can tell, Amray will support the FESEMs. Why they would not support a 3300 eludes me. Perhaps that is the bow wave of non-support for all Amray systems. I don't know. But I do know that a new Philips, Jeol, or Hitachi system at say $400K+ is about triple what I have invested in my current Amray FESEM. As long as my Amray system does what I need and can be maintained to do it, it makes no economic sense for me to dump it for a horribly expensive replacement. I suppose that what may happen is that the few lab SEM makers will offer about the same compliment of systems at about the same price mix. And the prices will be painful.
I have seen this same type of consolidation in the UK and other countries. For us Amray owners, it is a real problem....conundrum?
----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'Gary Gaugler'" {gary-at-gaugler.com} ; "Earl Weltmer" {earlw-at-pacbell.net} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, March 24, 2001 3:31 PM
Didn't mean to leave you with any misconceptions. I'm aware of the purchase by KLA and as far as I am aware, any decisions made were theirs. To date, as far as I know, sales have still been under Amray brand name, which is why I referred to them that way. I believe that there were some non-general purpose SEM technologies that KLA was primarily interested in and will roll those into its own production. Perhaps Amray was poised to take a firmer stance in those areas? As far as Amray, it has always been more of a family run business and perhaps the offer was right and the time was right. I don't think there were any particular problems in the operation or bottom line of Amray.
I also didn't mean to leave anyone with the impression that they are not supporting the 3300. On the contrary, what I wrote was that they were upset that anyone would consider a third-party service provider for one. They were quite upset that they had lost one.
On Saturday, March 24, 2001 9:34 PM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } At 03:31 PM 3/24/2001, you wrote: } } } [snip] } } } } The only question that seems to remain is - what business is being shut } } down? Amray has always been a major player in the semi-conductor field. } } These days, the FESEM is the most attractive portion of that business. My } } guess would be that they have chosen to still pursue the semi manufacturers } } with FESEMs but close down their general purpose SEM business. In the mean } } time, they want to still provide the lucrative service contracts on the } } general purpose FESEMs out there. Cash is king, and the cash in their } } operation is in the selling and service of FESEMs, primarily to those } } customers who are willing and able to pay a premium for these instruments. } } Amray, as a company, no longer exists. Amray is a division of KLA-Tencor. } For some reason, Amray apparently could not compete in the general } purpose (lab) SEM marketplace. Or, other forces came into play which } caused the change. } } I do know that Amray had developed very good FESEM systems--optics, } control software, stages, etc. KLA-Tencor is in the semiconductor defect } analysis business. They lacked the type of SEM technology that Amray } had. Consequently, KLA-Tencor purchased Amray, ostensibly I believe, } to acquire Amray's SEM technology. This is similar to the relationship } between FEI and Philips, wherein Philips acquired FEI for its ion beam } expertise. To wit, FEI now produces a fine dual beam focused ion beam } system which combines ion beam technology and SEM imaging (e.g., FEI 830). } } KLA-Tencor produces a top line semiconductor defect analysis tool set } (8100xl, 8300, 8500, etc.) which combines KLA's best attributes along } with the FESEM qualities of Amray. The defect analysis tools cost about } $2M each, compared to a typical lab FESEM at say $400K. The earlier } Amray systems were priced lower than this. It seems to me that KLA, } with Amray in tow, had made a decision to focus on the high cost (high } profit) business area of semiconductor defect analysis, at the exclusion } of lab SEMs. That is why we are seeing what is happening now with } Amray service contracts. } } In some respects, this was a myopic decision. Currently, the semiconductor } industry is in a decline (albeit not monumental). There surely was and } is a market for lab SEMs. KLA obviously made a business decision } to focus on semiconductor defect analysis tools. From what I am seeing, } the older Amray systems are no longer being supported by Amray. } Since the thermionic systems are rather simple in design and easier } to maintain than FESEMs, this may not be too bad. But, if one had } a bad PC-10, that is of course a different matter. } } So far as I can tell, Amray will support the FESEMs. Why they } would not support a 3300 eludes me. Perhaps that is the bow wave } of non-support for all Amray systems. I don't know. But I do know } that a new Philips, Jeol, or Hitachi system at say $400K+ is about } triple what I have invested in my current Amray FESEM. As long } as my Amray system does what I need and can be maintained to do it, } it makes no economic sense for me to dump it for a horribly } expensive replacement. I suppose that what may happen is that } the few lab SEM makers will offer about the same compliment of systems at } about the same price mix. And the prices will be painful. } } I have seen this same type of consolidation in the UK and other } countries. For us Amray owners, it is a real problem....conundrum? } } gary g. } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Question: I recently purchased a microscope used in forensics with a state agency. I would like to study microscopy so I could show this fascinating subject to my grandchildren. As a youngin' I enjoyed biological and petrology studies with the microscopes. I'm looking for a course that would allow me to use the different features of the microscope in light and dark field applications. I'm retired and would appreciate any help in this endeavor that you could offer. I am suffering from a physical disability and erudition is how I go about studies these days. THANX again.
For a fairly comprehensive list try http://www.xraysite.com/index.html?producers.html
I have experience with diffractometers made by Rigaku, Scintag and Siemens - all have good systems.
I have also dealt with Jim Stauver at JSTechnical. He sells and services Siemens systems and can probably get you a good deal on a used system. His web site is http://www.jstechnicalservices.com/
If you need any more info or help, contact me off-list.
Sincerely, James Talbot
K/T GeoServices, Inc. Bulk and Clay Mineralogy by X-ray Diffraction (214) 403-6342 visit my web site at http://www.ktgeo.com/
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} } Hello listers } } I've been asked to look into XRD equipment for our lab here, and was } wondering if any of you know who the major players are in this area. We } deal mostly with polymers and polymer fibers, and want to be able to do both } small and wide angle work. } } Thanks for the help. } dz } } David Ziegler } U.S. Army, SBCCOM } AMSSB-RSS-MS(N) } Materials Science Team, SS&T } Natick, MA 01760-5020 } TEL: (508) 233-6484 } FAX: (508) 233-5521 } Email: David.Ziegler-at-Natick.Army.Mil
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Larry -
You're not lilely to find a "course" that will meet your needs. Try starting with this book; this listing is taken from the Project MICRO bibliography, which you will find at http://www.msa.microscopy.com/ProjectMICRO/PMBooks.html
Nachtigall, W. 1995 Exploring With the Microscope 160 pp. hardbound. 6.5x9.5". $19.95. ISBN 0-8069-0866-1 Sterling Publishing Co., NY. Although this book is intended for adult amateur microscopists, it is well written and will provide teachers and classroom volunteers with much useful information on "serious" light microscopy; it's unequalled as a basic reference for beginners. Almost half of the book is devoted to simple preparation methods for biological specimens and descriptions (with good illustrations) of commonly encountered organisms. Adult. RECOMMENDED
If you want a more sophisticated, interactive (and more expensive) approach, try this CD-ROM (from the same bibliography):
Pagliaro, l., Murray, C., Curran, G., Orkand, A., and Astion, M. 1997 Microscopy-Tutor. ISBN 0-7817-1217-3 $195.00 plus shipping; distributed by Lippincott-Raven Publishers, 12105 Insurance Way, Hagerstown, MD 21740, 800-638-3030. For Macintosh and Windows 3.1 or 95/NT; easy installation. Developed by the Department of Laboratory Medecine and the Center for Bioengineering at the University of Washington, Seattle, this is a college-level introduction to the use of a research-quality compound microscope. Its extensive use of QuickTime animation to illustrate alignment steps and optical principles make it much more than a "book on a CD"; moving ripples on a pond really do make it easier to understand wave theory! Kohler illumination is emphasized and explained. Most, but not all, terminology is defined; a glossary would help beginners. Proper care of lenses is covered well, but there's no instruction on how to use immersion oil properly. There is a brief concluding self-test that is more of a review of important information than a quiz. Although it's a good CD, it's definitely not appropriate for precollege microscopists. Adult.
And there are dozens of things listed for the grandkids...
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Having advised one of our customers to try the above resin (glutaraldehyde/carbohydrazide mixture) to embed her particularly sensitive material for LM (it was dissolving using other mainstream embedding media), she has now successfully got to the sectioning stage. Next problem is getting the sections to stick to the microscope slide. They curl and crack and disappear Lemming like into the abyss.
Having cut resin sections for many years I have now exhausted all the tricks that I have used to keep them on the slide from poly-L-lysine to silane to drying in an oven overnight (instead of using a hotplate or slide dryer). That last advice has surprisingly saved many desperate microscopists but not this time. Maybe one of the adhesive sticks which lays down a sticky membrane will work. We will even try double sided tape (or maybe not), but does anyone out there have experience with sections cut from this material? Help,
Regards
Terry Cooper TAAB Laboratories Equipment Ltd England
Thanks to all who offered help on the subject of the high resolution image if the MSA logo. Most thanks go to Bev Maleef who provide the goods.
I did have someone offer to take a micrograph of it with an SEM! I didn¹t know we actually had a copy ofthe MSA logo that small, has any one been playing the STM tricks with the individual atoms a la IBM?
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
Greetings, I have found that polyethlyene-imine (PEI) is much stickier than poly-lysine. I have no idea about the resin you mentioned, but PEI is sticky stuff. It comes as a liquid. I make a 0.1% soloution in ddwater, which I freeze in aliquots. Then I keep a working one in the fridge. I coat coverslips in the stuff by floating them on a drop of the PEI solution for about 10 sec, and then blotting off the excess and letting them air dry. In that conditions, the coated 'slips are good for at least months.
Hope this helps, Tobias Baskin
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Hi all, Can anyone in the Atlanta area help with this request? Please send your replies to Ulrika - E-mail: Ulrika.Egertsdotter-at-ipst.edu. thanks for your help, Beth Here's the request: } I am an } assistant scientist currently working at the Institute of Paper science and } technology in Atlanta and I am trying to find a cryomicrotome that I could } use for my project. Within my project, we are broadly speaking trying to } reveal information about the biological mechanisms of wood formation. We } are using different types of arrays, and for the sample preparations we are } trying to get as thin sections as possible. For this purpose I am now } looking for a cryomicrotome that I could access to make thin sections of } wood samples that would then be processed for RNA to probe the arrays, and } also hopefully in situ hybridisation. I am planning to go to Sweden to } learn the techniques from the group that has already been doing this type } of work in poplars. I will however need to find an accesible cryomicrotome } to work with that are somewhat closer to Atlanta. Do you think it would be } possible at all for me to make use of the cryomicrotome in your department, } or would you perhaps know of any other available equipment? } } Thank you. } } Sincerely, } Ulrika Egertsdotter } E-M Ulrika Egertsdotter, Ph.D. } Institute of Paper Science and Technology } 500 10th Street, N.W. } Atlanta, GA 30318 } } Direct dial: +1 404 894 7842/4807 } Telephone: +1 404 894 5700 } Fax. +1 404 894 4778 } E-mail: Ulrika.Egertsdotter-at-ipst.edu
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I am very new to electron microscopy, so I ask your indulgence in some basic questions.
I have an old JEOL SEM that has been sitting at atmospheric pressure for some time now and will need a thorough baking. It has a plain old tungsten wire filament, so the vacuum requirements aren't all that stringent. The SEM has rubber o-rings (I don't know how to tell if they are Viton). I doubt that I want to go above 100 oC.
1) What characteristics are important in choosing a heat tape suitable for baking the instrument?
2) Where would I find a suitable heat tape (my web searches on heat tapes got me lots of information about ice buildup on my roof, but very little about getting water out of an SEM)?
3) How do I determine how much to buy - how much column on each side of the tape can I assume to be heated properly by the tape - what is an appropriate center-to-center distance between adjacent wraps of tape?
4) Is there a more practical method than heat tapes for baking the instrument?
5) What else may I be missing here?
Thank you,
Bruce Girrell in snowy northern Michigan with a few more basic questions waiting in the wings
The Sudan dyes are those most commonly used for suberin, and perhaps cutin. The lignin component of both can be stained with phloroglucinol (or observed as autofluroescence with blue excitation in unstained material). If the autofluroescence is quenched by the Sudan, this supports the interpretation that the substance is suberin or cutin rather than lignin.
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R. Howard Berg, Ph.D. Director, Integrated Microscopy Facility, Associate Member Donald Danforth Plant Science Center/Nidus Center 893 North Warson St. Louis, MO 63141
Would anyone know of a general protocol I could use to stain a nylon 6,6 sample for TEM analysis? I'm studying cross-sections of nylon fibers. I've read that tin chloride is a good stain for nylons, I've also seen reference to work using iodine.
I would not recommend baking out that system if it has not been designed for it. The heat tapes will give you hot spots and other places may not take the heat well. If you are not going above 100C, it is not going to help you much anyhow. At least 200C is required for modest bakes. The purpose of a bakeout is to get water off the walls and you have to overcome the heat of chemsorption.
A better way is to rig a leak of nitrogen from a liquid nitrogen container (open to atmosphere) so that you get flow in the viscous flow range. You have to throttle your vacuum pump so if you do not have a way to do this with your high vacuum pump, do it with your mechanical pump and throttle your leak. I think that if your run at about 0.1 Torr into a typical 1.5 inch vacuum line that you will be in the viscous range.
If you use clean stainless steel or copper tubing that is coiled and put your heat tape around that, you can heat the nitrogen gas going into your system for added benefit. Run this leak overnight and it will take out all the water off of the walls.
There is a commercially available system to do this and more. Contact Ron Vane at XEI.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Bruce Girrell [mailto:bigirrell-at-microlinetc.com] } Sent: Monday, March 26, 2001 10:47 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Bake out } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Greetings, } } I am very new to electron microscopy, so I ask your } indulgence in some basic } questions. } } I have an old JEOL SEM that has been sitting at atmospheric } pressure for } some time now and will need a thorough baking. It has a plain } old tungsten } wire filament, so the vacuum requirements aren't all that } stringent. The SEM } has rubber o-rings (I don't know how to tell if they are } Viton). I doubt } that I want to go above 100 oC. } } 1) What characteristics are important in choosing a heat tape } suitable for } baking the instrument? } } 2) Where would I find a suitable heat tape (my web searches } on heat tapes } got me lots of information about ice buildup on my roof, but } very little } about getting water out of an SEM)? } } 3) How do I determine how much to buy - how much column on } each side of the } tape can I assume to be heated properly by the tape - what is } an appropriate } center-to-center distance between adjacent wraps of tape? } } 4) Is there a more practical method than heat tapes for baking the } instrument? } } 5) What else may I be missing here? } } Thank you, } } Bruce Girrell } in snowy northern Michigan with a few more basic questions } waiting in the } wings } }
We have recently decommissioned a Fullam closed circuit TV imaging system, purchased in the late 80's-early 90's for use on a JEOL-1200EX. The system is in excellent condition, includes a side-port camera mount and detector, Dage MTI 70 camera, Nikkor lens, etc., and is available for donation to a school or university.
If interested, please respond directly to me and not to the list.
Regards, Bev Maleeff
GlaxoSmithKline Pharmaceuticals Safety Assessment 709 Swedeland Road M/C UE 0462 King of Prussia, PA 19406
I think a bigger question here is why Bruce thinks he needs a bake out? If he "heard" from someone that is what you have to do for a vacuum system then we should give him some easier advice.
First, we must all understand that most SEMs are not ultra high vacuum system that require baking. SEMs have specimen chamber with a typical vacuum between 10-5 to 10-6 Torr. High vacuum at best. An old JEOL SEM was never baked. Water vapor does not bother the electron beam or the secondary electrons at these pressures. I would clean the vacuum system with IPA, check and change as needed the pump oil, and try pumping down. Then I would fix any leaks.
Bruce, have you tried to pump it down? What make you think you should bake it? If oily dirt is a problem, then nitrogen purging like Scott Walck suggests might be a solution after you wipe down the chamber wall with IPA.
I have a long discussion of oil and hydrocarbon contamination and removal methods at www.SEMCLEAN.Com.
Ronald Vane XEI Scientific Notice: XEI makes anti-contamination system for SEMs.
Bruce have you tried pumping it down?
-----Original Message----- } From: Walck, Scott D. {walck-at-ppg.com} To: 'bigirrell-at-microlinetc.com' {bigirrell-at-microlinetc.com} Cc: 'Microscopy' {microscopy-at-sparc5.microscopy.com}
I appreciate the many responses that I have gotten already.
Several have questioned the need for a bake out. Well, I did say I was new to this...
The reasons I felt that the instrument should be baked are: 1) Wilbur Bigelow, in _Vacuum Methods in Electron Microscopy_ suggests that, short of pumping endlessly, baking is almost a necessity to remove adsorbed water. 2) The instrument has been sitting at atmospheric pressure for an untold period of time, at least several months. The E-T detector had been removed, leaving a nice 6" dia. hole in the system. God knows what else besides water vapor is in there. 3) The logbook that came with the unit recorded routine baking. Along with item 1), I concluded that baking was simply just a matter of course.
What I am hearing so far is that 1) for the relatively low vacuum required for this instrument, baking may not be necessary and 2) short of going to about 200C, baking may not be very effective.
Baking won't be necessary, you're biggest problem will be the o-ring seals. I assume this is a diffusion pump system.
O-ring seals have a tendency to dry out and shrink, if they've been in the system for some time, they will also have been shaped by the pressure of the flanges. The result will be poor mating with the flanges and lots of leaks.
You'll need to take each one out of the system and inspect it for drying and cracking. I do mean each one - if the system has been at atmosphere for a long time, you'll want to get every o-ring that seals the sample stage controls, vacuum valves and the gun translation mechanism, if so equipped.
If the o-rings are not too badly out of round and not cracking, then they can generally be reconditioned by cleaning them off with acetone and applying a very light layer of vacuum grease evenly on the surface. The grease is only to seal the surface, prevent drying and increase flexibility. I use acetone, although many will probably warn you not to, because it slightly swells the o-ring. When you're doing a general reconditioning like this, that helps the o-rings seal up right away, but it will still take a day or two at high vacuum to get them to fully seal.
This is also a good time to get in and clean the column and wipe down the sample and gun chambers. It's hard to say what will work here for you, as it depends largely on the pump oils that have been used in the past and the accumulated contamination from samples. Try some standard solvents like methanol, ethanol, acetone, etc. Hopefully one or a combination can remove some of the build-up.
The outgassing rates of adsorbed gasses at the vacuum levels you'll attain in the instrument are not really a problem. You'll want to leave the instrument in high vacuum for at least 24 hours. Over the first day or two, you will notice that the vacuum level slowly improves, primarily due to the o-rings being sucked into better contact with the flanges. Over this time, the system will also reach equilibrium with outgassing products.
On Monday, March 26, 2001 9:47 AM, Bruce Girrell [SMTP:bigirrell-at-microlinetc.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings, } } I am very new to electron microscopy, so I ask your indulgence in some basic } questions. } } I have an old JEOL SEM that has been sitting at atmospheric pressure for } some time now and will need a thorough baking. It has a plain old tungsten } wire filament, so the vacuum requirements aren't all that stringent. The SEM } has rubber o-rings (I don't know how to tell if they are Viton). I doubt } that I want to go above 100 oC. } } 1) What characteristics are important in choosing a heat tape suitable for } baking the instrument? } } 2) Where would I find a suitable heat tape (my web searches on heat tapes } got me lots of information about ice buildup on my roof, but very little } about getting water out of an SEM)? } } 3) How do I determine how much to buy - how much column on each side of the } tape can I assume to be heated properly by the tape - what is an appropriate } center-to-center distance between adjacent wraps of tape? } } 4) Is there a more practical method than heat tapes for baking the } instrument? } } 5) What else may I be missing here? } } Thank you, } } Bruce Girrell } in snowy northern Michigan with a few more basic questions waiting in the } wings } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
We are relatively new to atomic force microscopy and need some estimate of costs for routine maintenance. I don't mean expendible items but repairs.
For example, we have been facing some (in my opinion) rather high costs to repair hardware: broken scanner $3,900, electronic module $400 (twice). Are AFM's prone to such frequent, high repair costs?
Has anyone ever had to replace a scanner that "broke" for no apparent reason? BTW, none of these repairs were caused by operator error.
Just wondering.......
Thank you,
John
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
In conducting RGA studies of polymer outgassing inside hermetic electronic packages some years ago we found that hydrogen seems to have the effect of displacing moisture adsorbed onto metal surfaces. It was a problem that amounted to a "memory effect" within the RGA* for us but the effect can be used to advantage in displacing water. You might try Scott's suggestion substituting bottled, high purity hydrogen for the liquid nitrogen bleedoff. Obviously, hydrogen is flammable and requires caution when in use.
(* e.g. if a hermetic package punctured in the RGA sample compartment contained moisture, some of the released water vapor would adsorb onto the chamber wall. If the next hermetic package contained hydrogen, it would displace the moisture from the previous package causing it to be attributed to the second unit when, in fact, it was left over from the first. During pumping out of the sample compartment in between samples, the background level of moisture would be achieved even though it was lurking on the metal surface waiting to be liberated.)
John Twilley Conservation Scientist
Walck, Scott D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would not recommend baking out that system if it has not been designed for it. The heat tapes will give you hot spots and other places may not take the heat well. If you are not going above 100C, it is not going to help you much anyhow. At least 200C is required for modest bakes. The purpose of a bakeout is to get water off the walls and you have to overcome the heat of chemsorption. } } A better way is to rig a leak of nitrogen from a liquid nitrogen container (open to atmosphere) so that you get flow in the viscous flow range. You have to throttle your vacuum pump so if you do not have a way to do this with your high vacuum pump, do it with your mechanical pump and throttle your leak. I think that if your run at about 0.1 Torr into a typical 1.5 inch vacuum line that you will be in the viscous range. } } If you use clean stainless steel or copper tubing that is coiled and put your heat tape around that, you can heat the nitrogen gas going into your system for added benefit. Run this leak overnight and it will take out all the water off of the walls. } } There is a commercially available system to do this and more. Contact Ron Vane at XEI. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } } } -----Original Message----- } } From: Bruce Girrell [mailto:bigirrell-at-microlinetc.com] } } Sent: Monday, March 26, 2001 10:47 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Bake out } } } } } } -------------------------------------------------------------- } } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } } } } } -------------------------------------------------------------- } } ---------. } } } } } } Greetings, } } } } I am very new to electron microscopy, so I ask your } } indulgence in some basic } } questions. } } } } I have an old JEOL SEM that has been sitting at atmospheric } } pressure for } } some time now and will need a thorough baking. It has a plain } } old tungsten } } wire filament, so the vacuum requirements aren't all that } } stringent. The SEM } } has rubber o-rings (I don't know how to tell if they are } } Viton). I doubt } } that I want to go above 100 oC. } } } } 1) What characteristics are important in choosing a heat tape } } suitable for } } baking the instrument? } } } } 2) Where would I find a suitable heat tape (my web searches } } on heat tapes } } got me lots of information about ice buildup on my roof, but } } very little } } about getting water out of an SEM)? } } } } 3) How do I determine how much to buy - how much column on } } each side of the } } tape can I assume to be heated properly by the tape - what is } } an appropriate } } center-to-center distance between adjacent wraps of tape? } } } } 4) Is there a more practical method than heat tapes for baking the } } instrument? } } } } 5) What else may I be missing here? } } } } Thank you, } } } } Bruce Girrell } } in snowy northern Michigan with a few more basic questions } } waiting in the } } wings } } } } } } }
I am looking for a X-ray radiation detector used on TEMs. Currently we have a Geiger counter that is only sensitive to low energy X-ray. The purpose of the new detector is to sense high energy X-ray leakage from a 200 kV scope.
Any suggestion or clue will be greatly appreciated. Please contact off-line.
There's a whole chapter (#4) on support films in Hayat MA, Miller SE. 1990. Negative Staining. Pages 201-214 are on perforated films and discuss making of films with different size holes.
On Fri, 23 Mar 2001, Gordon Vrololjak wrote:
} X-Sieve: cmu-sieve 1.3 } Return-Path: {Microscopy-request-at-sparc5.microscopy.com} } Received: from jeter.acpub.duke.edu (jeter.acpub.duke.edu [152.3.233.10]) } by moorcock.acpub.duke.edu (8.9.3/8.9.3/Duke-5.0.0) with ESMTP id UAA15529 for {saram-at-moorcock.acpub.duke.edu} ; } Fri, 23 Mar 2001 20:14:59 -0500 (EST) } Received: from ultra5.microscopy.com ([206.69.208.10]) } by jeter.acpub.duke.edu (8.9.3/8.9.3/Duke-5.0.0) with ESMTP id UAA05887; } Fri, 23 Mar 2001 20:11:33 -0500 (EST) } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id RAA13779 } for dist-Microscopy; Fri, 23 Mar 2001 17:51:15 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id RAA13774 } for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 23 Mar 2001 17:50:45 -0600 (CST) } Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.175.19]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id RAA13763 } for {microscopy-at-sparc5.microscopy.com} ; Fri, 23 Mar 2001 17:50:34 -0600 (CST) } Received: by nature.Berkeley.EDU (Postfix, from userid 7458) } id DFB298B64; Fri, 23 Mar 2001 15:48:36 -0800 (PST) } Received: from localhost (localhost [127.0.0.1]) } by nature.Berkeley.EDU (Postfix) with ESMTP id D3AB54DB35 } for {microscopy-at-sparc5.microscopy.com} ; Fri, 23 Mar 2001 15:48:36 -0800 (PST) } Date: Fri, 23 Mar 2001 15:48:36 -0800 (PST) } From: Gordon Vrololjak {gvrdolja-at-nature.Berkeley.EDU} } To: {microscopy-at-sparc5.microscopy.com} } Subject: ? how to make lacey carbon films? } Message-ID: {Pine.GSO.4.33.0103231546140.28537-100000-at-nature.Berkeley.EDU} } MIME-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } Errors-to: Microscopy-request-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Everyone, } I want to make my own lacey carbon films for cryo-tem and was wondering if } anyone had some good methods, experiences, insights, or references? } } I found references for lacey formvar, pure carbon films, and a few } others... But nothing explicitely for lacey carbon. } } All comments greatly appreciated, Thanks. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I would appreciate hearing how the experts out there prepare their microscope slides for measuring a PSF. We got a couple of prepared slides from Molecular Probes and were quite disappointed with the quality of the Blue and Green beads (the red ones were fine). This was somewhat surprising since they usually have great quality. We bought the beads and are making our own but I would be interested in any protocols or alternative sources for prepared slides. Thanks in advance, Tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
A W gun system doesn't need a particularly high vacuum to operate. But a better vacuum will extend the life of the filament dramatically. While I am not familiar with your JEOL instrument, I have worked with Amray 1610T and 1830 systems which had an ion pump so a LaB6 cathode could be used. Each system had separate guns for W and LaB6. I only ran W in the 1610T but only LaB6 in the 1830. In this respect, the ion pump had to be working and the vacuum had to be quite good.
When the gun chamber was opened, either for column liner cleaning, liner aperture change, anode aperture change or for cathode change and Wenhelt change, I baked out the gun chamber. If the ion pump indicated an inability to pump down quickly, I baked it out too. I never baked the chamber. Usually, when a system would sit around without being under vacuum, a long pumping after internal wipe down with methanol would bring the chamber vacuum back. If not, that is unchartered territory for me.
I baked out the gun chamber and the ion pump at 175F. All pumps (except ion) were on during the bake. I used flexible heating tape made by BH Thermal http://www.bhthermal.com and their control thermostat. The thermostat control is PN TS0991-550 and has a fluid bulb sensor. I used two silicone rubber extruded heating tapes, PN BS0101060L (313 Watts, 1" x 72") for gun and PN BS0201040L (418 Watts, 2" x 48") for pump. The first tape cost $95 and the second was $135 list. The controller was about $175.
The tape is wound tightly around the chamber or pump and taped down with high temperature fiberglass tape. This is PN PSAT36 (1/2" x 180ft) and costs $15 a roll. PN AAT260 (2" x 180ft) is also handy...$61 per roll. Wrapped around the heating tape is thick aluminum foil (cooking variety) and taped tight to the heating tape. The sensor bulb is taped down to the top of the gun assembly and should be enclosed by the aluminum foil. This makes it a set and forget deal for overnight baking.
The gaskets in my systems were Viton and copper. At 175F, I suspect rubber components would be OK. As far as I know, new replacements are Viton these days. But I can't say for certain if your system has rubber or Viton or even if replacements are Viton. Try heating a spare gasket or O-ring enclosed in aluminum foil in an oven at 175F and see if it is destroyed. That ought to tell the story. If the logs say that the system was baked, 175F ought to be safe.
The difference baking made was quite remarkable. If I did not bake, the ion pump would draw about 100-120uA and get worse as the cathode was on for much time. After baking, it idled at 50-60uA and would stay there for a long time.
The problem with a W-only system is that the gun chamber is typically connected to the specimen chamber via a stand pipe or some other vacuum connection. This means that the gun area is basically at the vacuum level of the chamber. Thus, my discussion of baking the gun chamber may be useless to you. But you could apply the same baking principles to the chamber and the gun chamber. I would not recommend at all baking or heating the column. This would surely destroy the scan coils.
If you have a turbo pump system, it is probably worth baking the system to purge the trapped moisture. If not, then it may not be of any value to bake. What kind of pumps does the system have?
Hope this is a good starting point for you.
gary g.
At 07:47 AM 3/26/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone know where one could get a microprocessor of the type that was used in the Polaron E5200 automatic coater. This was their first version of an automatic coater and the replacement part is no longer available via the manufacturer. Please respond to me off-line since this topic is probably not of interest to many followers of this list. If anyone is interested in the responses, let me know and I will send a summary of anything I learn.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Thanks to everyone who sent me their ideas for adhering sections to slides. We have had lots of suggestions and I will certainly post any successful results we have from the input,
As sometimes happens, some people find they can't get money to attend courses at the last minute while others don't hear about the courses until it is too late to apply.
Now may be your chance!
Several late cancellations at the UBC Live-Cell course mean three opportunities for those of you who have ten days free this June. In addition, the presence of more microscopy systems allows us to set up one more group-of-three.
We expect to have an international faculty of 15, and on-site equipment that includes 2-photon systems from Zeiss and Bio-Rad. There will also be single-photon confocals from Bio-Rad, Olympus, Nikon, Yokogawa/Solamere and Yokogawa/EG&G as well as deconvolution set-ups from Applied Precision Instruments, AutoQuant/Universal-Imaging, Improvision, Intelligent-Imaging- Innovations and Zeiss, all set up for "live-cell" work. In addition, there will be a laser tweezers/scissors from Cell-Robotics, microinjection from Eppendorf and sundry other delights including a fully equipped cell biology lab (with tireless technician!!) just for students.
These microscopy systems are not just to look at but to use for over 45 hours of organized 2D and 3D live-cell laboratory sessions, plus 10 hours of evening sessions for group live-cell projects.
Although the eight "Groups-of-3" have already been set up and have chosen their "individual projects," we are able to accept 3-2 more students as long as their backgrounds will fit into one of these groups, and we may even be able to set up one additional group.
which has the rest of the story, including a preliminary program.
Then fill out the Registration Form from the site to tell us about you, Fax it to Dr. Elaine Humphrey ASAP and we will see if we can fit you in. (I will be away April 1-14)
Hope that you can join us in Vancouver this June 18-28.
Jim Pawley, Organizer -- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
As pointed out by many listers, it may not be necessary to bake your instrument. Since the baking part has been well covered, I'll just add a very effective way to tell Viton o-rings from rubber ones. If you can find some Freon TF just put some in a beaker and drop in the questionable o-ring. If the o-ring floats, it is rubber and if it sinks, it is Viton.
Good luck, Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone 512/282-5507 FAX 512/280-0702
Sustaining Member - MICROSCOPY SOCIETY OF AMERICA
-----Original Message----- } From: Bruce Girrell {bigirrell-at-microlinetc.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Greetings, I am taking overlapping video images of a growing root through a horizontal light microscope. For analysis, I need to put the images back in register. I have tried incorportating various high contrasty things into the background but this introduces various problems ranging from death to the root to requiring a large overlap between successive images. As an alternative, I thought of measuring the displacement of the stage. A typical displacement between images is around 300 microns and I need to measure this displacement to within around 1 micron. I only need to work in one dimension, which can be parallel to one of the edges of the stage. I have in mind something like a "giant" afm detection set up, with a laser bouncing off the edege of stage and picked up by a detector. But this is just wild imagination on my part. Anyone out there have any *knowledge* about how this might be accomplished?
Thanks in advance, Tobias -- _ ____ __ ____ Tobias I. Baskin / \ / / \ / \ \ 109 Tucker Hall / / / / \ \ \ Biological Sciences /_ / __ /__ \ \ \__ University of Missouri / / / \ \ \ Columbia, MO USA / / / \ \ \ 65211-7400 / / ___ / \ \__/ \ ____ voice: 573-882-0173 fax: 573-882-0123
I was a student intern with Dr. Judi Walton when she was first using lead aspartate in 1977. I don't remember if we used it with tissue, but I know we used it with cell culture monolayer samples for TEM. We used it without poststaining. She developed this as Walton's Lead Aspartate. I don't know if it is still on the market....
Barbara L. Plowman Univ. of the Pacific School of Dentistry 2155 Webster Rm 632 San Francisco, CA 94115 email: Bplowman-at-sf.uop.edu
Thank you for the answers to my inquiry regarding 3D reconstruction of TEM serial sections and / or EM tomography. They were very useful and we have a much better idea of how to proceed with our project. I have, at the request of a couple of list subscribers, summarized the answers in a Word file and sent the file to them. It is available to anyone who emails me and asks - it is kind of long, but there is good advice and contact information included for anyone who is beginning in this area.
Again - Thanks to all who helped. I wouldn't know how to cover the bases for those who depend on me without this list! Thanks to Nestor, too!
Regards, Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
Dear Readers, I have a Nikon 63x oil objective that is used on an inverted scope. It is a multi user facility and I just noticed today that oil has seeped into the objective making it almost useless to use.
(i) Should I tackle the job of cleaning this lens myself or (ii) Does anyone know of a facility that would clean this lens at a reasonable price?
I used to have silicon around the top of the lens to prevent this problem but it came off and I forgot to replace it.
Thanks for any information on this topic.
Sean M. Ward Associate Professor Department of Physiology and Cell Biology University of Nevada School of Medicine Manville Medical Sciences Building Reno NV 89557 Tel: (775) 784-6061 Fax:(775) 784-6903
I've been asked to look at the microscopic distribution of a mineral oil used in plant sprays on a plant surface. The spray is dispersed in water at 1-2% so the final volumes on the surface can be quite low. Most of the techniques I have seen are concerned with the total distribution of the spray and use either fine particles or dyes in the aqueous phase and do not track the oil. I have had only limited success with the natural autofluorescence of the oil, cathodoluminescence in the SEM and with common chromatic dyes such as oil red. Visualization is just possible if we deposit pure oil on the surface but not the suspension. Has anyone any suggestions??
Thanks in advance
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
Yes, an interferometer could be used to measure the stage movement, but it's an expensive option. If you truly need 1 micron placement, there are few methods available. If you can accept somewhat less resolution, a machinist's dial micrometer with a stand can give you 1/10,000 of an inch measurement rather inexpensively, but with limited travel. You didn't mention the overall length of your samples.
Being based at a university, try your local campus machine shop. Machinists tend to covet those precision instruments, but perhaps you could borrow one for a short time (just don't sign in blood).
On Tuesday, March 27, 2001 2:33 PM, Tobias Baskin [SMTP:BaskinT-at-missouri.edu] wrote: } } Greetings, } I am taking overlapping video images of a growing root } through a horizontal light microscope. For analysis, I need to put } the images back in register. I have tried incorportating various high } contrasty things into the background but this introduces various } problems ranging from death to the root to requiring a large overlap } between successive images. As an alternative, I thought of measuring } the displacement of the stage. A typical displacement between images } is around 300 microns and I need to measure this displacement to } within around 1 micron. I only need to work in one dimension, which } can be parallel to one of the edges of the stage. I have in mind } something like a "giant" afm detection set up, with a laser bouncing } off the edege of stage and picked up by a detector. But this is just } wild imagination on my part. Anyone out there have any *knowledge* } about how this might be accomplished? } } Thanks in advance, } Tobias } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 Tucker Hall } / / / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ University of Missouri } / / / \ \ \ Columbia, MO USA } / / / \ \ \ 65211-7400 } / / ___ / \ \__/ \ ____ voice: 573-882-0173 } fax: 573-882-0123 } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Dear collegues has anyone out there has any experiences with imaging bacteriophages in a FESEM and does anyone know of some published pictures. Thank's in advance. Manfred
Ian I have never tried this, but one possibility would be to add a fluorochrome to the system. Nile red is an oil-soluble fluorochrome, only sparingly soluble in aqueous phase, strongly fluorescent only when it partitions into an apolar environment but non-fluorescent in aqueous phase. It should be possible to image Nile-red labelled oil droplet footprints by fluorescence LM with FITC-type filter systems and possibly also by cathodoluminescence in SEM. If you are working on leaves the red autofluorescence of the chloroplasts should be filter out with a green barrier filter. The Leica GFP Plant filter sets may therefore be suitable. see Greenspan, P., Mayer,E.P. and Fowler,S.D. (1985) Nile red: a selective fluorescent stain for intracellular lipid droplets. Journal of Cell Biology 100, p965-973.
Hope this helps Chris ----- Original Message ----- } From: "IAN HALLETT" {ihallett-at-hortresearch.co.nz} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 28, 2001 5:04 AM
1 micron is a pretty demanding tolerance. Do you really need it to be this good? Stages used to be provided with x and y vernier scales, but today a stepper motor stage drive is often used controlled by a computer which counts the steps. Mitutoyo make electronic measuring devices accurate to about 5 or 10 microns which are used in electronic calipers. Similar modules are available for mounting on engineering x,y stages, for example of milling machines. Electronic supplies companies like Farnell, RS Components and others supply these.
----- Original Message ----- } From: "Tobias Baskin" {BaskinT-at-missouri.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, March 27, 2001 9:33 PM
The Department of Engineering Metals, Chalmers University of Technology (Göteborg, Sweden) is seeking:
DOCTORAL STUDENT Microstructure and Chemistry of Nanostructured Materials using Transmission Electron Microscopy
Any technological application of nanostructured materials is only possible if their properties and their microstructure are well understood so that they can be manufactured reproducibly. Concentrating on Ni- and Co-based materials analytical transmission electron microscopy will be applied to characterize the microstructure of the material as well as to monitor the microstructural evolution upon heating. The work will be performed in close collaboration with colleagues at the University of Toronto, Canada. The experiments will be carried out on a Zeiss 912 Omega with inbuilt imaging energy filter, however, also a Philips CM 200 FEG equipped with EDS and Gatan imaging filter (GIF) is accessible. A special heating holder is available to perform the in-situ heating experiments.
The appointment applies to full time for a maximum period of five years. In addition to the research activities departmental duties, i.e. a limited amount of educational, research and/or administrative work up to 20% of full time has to be performed. The initial monthly salary is SEK 18.000, however depending on the academic degree. The applicant must have an academic degree (MSc, BSc or equivalent) in materials science/materials engineering, physics or a similar subject area. Experience in (analytical) transmission electron microscopy is a merit. The position is available immediately.
For further information about the position, please contact: Prof. Uta Klement Chalmers University of Technology, Department of Engineering Metals Tel: +46 31-772 1264, Fax: +46 31-772 1262 E-mail: Klement-at-em.chalmers.se
A written application including the reference number 23/2001, with (attested) personal record, brief description of research interests, name and e-mail addresses to three reference persons, possible day for taking possession of the position, and the documents that you want to refer to should have arrived at the Registrar, Chalmers University of Technology, SE-412 96 Göteborg, Sweden no later than April 27, 2001.
I can't visualize exactly how you are doing this, but I assume you are using reflected rather than transmitted light.
In the dim and distant past I have used a 'traveling microscope' which consists of a lens system with a cross-hair which can be moved along a metal rail (at least 200mms long). The rail had divisions marked off accurately in mms and a sliding Vernier scale so I would guess that it would be accurate to 10um or maybe better. I suppose that it may be possible to attach a camera system instead of the lens but it would be best to try and borrow one to find out. I assume that a stage micrometre (slide marked off in fractions of 1mm or 10mm) could not be adapted to your purposes but it might be a useful way of calibrating any system you adopt.
Malcolm
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Tobias Baskin wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Greetings, } I am taking overlapping video images of a growing root } through a horizontal light microscope. For analysis, I need to put } the images back in register. I have tried incorportating various high } contrasty things into the background but this introduces various } problems ranging from death to the root to requiring a large overlap } between successive images. As an alternative, I thought of measuring } the displacement of the stage. A typical displacement between images } is around 300 microns and I need to measure this displacement to } within around 1 micron. I only need to work in one dimension, which } can be parallel to one of the edges of the stage. I have in mind } something like a "giant" afm detection set up, with a laser bouncing } off the edege of stage and picked up by a detector. But this is just } wild imagination on my part. Anyone out there have any *knowledge* } about how this might be accomplished? } } Thanks in advance, } Tobias } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 Tucker Hall } / / / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ University of Missouri } / / / \ \ \ Columbia, MO USA } / / / \ \ \ 65211-7400 } / / ___ / \ \__/ \ ____ voice: 573-882-0173 } fax: 573-882-0123
Hi, All, I take some diffraction contrast pctures. As the thickness of the samples changes sharply the brightness of the images is not uniform. Does someone know any software to adjust the brightness for the whole image? Thanks inadvance. Best regards. Feng ********************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
A good tool maker should be able to make you someting the will do an order of magnitued beter than 1/10,0000 by using a standard micrometer head and setting to drive a wedge at about 10 degrees. I don't remember the exact angle but i used it to get .00001 on a lathe wiht 0001 devisions. I dont expect you would get all the precision but you sould get .00022 to .00004
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'Tobias Baskin'" {BaskinT-at-missouri.edu} ; {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 28, 2001 12:03 AM
Hi folks, If anyone has experience with and opinions about the product Auto- Montage by Syncroscopy, please contact me or Mark Farmer off- listserve. Thanks in advance,
John Shields EM Lab University of GA Athens jshields-at-cb.uga.edu
Hi Sean, We had the same problem with one of our objectives on the Nikon inverted. The lens should not allow oil in if it is sealed correctly, even without the "seal" you provided. Your lens is either older and the seal is worn, or you've been cleaning the oil off with a harsh solvent (I've seen some people using xylene regularly). Somewhere in the back of my mind (a dusty and unreliable source) I think silcon is not good for the seal. Another solution is to take finger cots or make them by cutting the finger tips from gloves and pull them down over the lens if you are still unsure of your seal. We ended up sending the lens to Nikon for repair. I seem to remember it cost around $200 or so to have them clean it and replace the seal, but you can call to get a quote. good luck
On 27 Mar 2001, at 17:39, Sean Ward wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Dear Readers, } I have a Nikon 63x oil objective that is used on an inverted scope. } It is a } multi user facility and I just noticed today that oil has seeped into } the objective making it almost useless to use. } } (i) Should I tackle the job of cleaning this lens myself or (ii) Does } anyone know of a facility that would clean this lens at a reasonable } price? } } I used to have silicon around the top of the lens to prevent this } problem but it came off and I forgot to replace it. } } Thanks for any information on this topic. } } } Sean M. Ward } Associate Professor } Department of Physiology and Cell Biology } University of Nevada School of Medicine } Manville Medical Sciences Building } Reno NV 89557 } Tel: (775) 784-6061 } Fax:(775) 784-6903 } }
John P. Shields, PhD Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080 FAX 706-542-4271 jshields-at-cb.uga.edu
Hi Tobias, What you suggest may not be worth the hassle. Potential error introduced by temp. variations on the stage would be problematic. Is it not possible to include an internal standard, for example an embedded stage micrometer, ruler, calibrated cover plate for the incubation vessel,... Ramin
-----Original Message----- } From: Tobias Baskin [mailto:BaskinT-at-missouri.edu] Sent: Tuesday, March 27, 2001 3:33 PM To: Microscopy-at-sparc5.microscopy.com
Greetings, I am taking overlapping video images of a growing root through a horizontal light microscope. For analysis, I need to put the images back in register. I have tried incorportating various high contrasty things into the background but this introduces various problems ranging from death to the root to requiring a large overlap between successive images. As an alternative, I thought of measuring the displacement of the stage. A typical displacement between images is around 300 microns and I need to measure this displacement to within around 1 micron. I only need to work in one dimension, which can be parallel to one of the edges of the stage. I have in mind something like a "giant" afm detection set up, with a laser bouncing off the edege of stage and picked up by a detector. But this is just wild imagination on my part. Anyone out there have any *knowledge* about how this might be accomplished?
Thanks in advance, Tobias -- _ ____ __ ____ Tobias I. Baskin / \ / / \ / \ \ 109 Tucker Hall / / / / \ \ \ Biological Sciences /_ / __ /__ \ \ \__ University of Missouri / / / \ \ \ Columbia, MO USA / / / \ \ \ 65211-7400 / / ___ / \ \__/ \ ____ voice: 573-882-0173 fax: 573-882-0123
I have heard mention of replicating surfaces with carbon for analysis w/ EM, it seems like it may be history now, but I was wondering if someone could "clue me in" with when it was required.
At 5:39 PM -0800 3/27/01, Sean Ward wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You have my deepest sympathies! We had the same experience with a Zeiss lens on a multi-user Axiovert a few years ago. I would NOT recommend trying this on your own. High power lenses are complexes of many lens elements and, in my humble opinion, should only be disassembled by those who know what they are doing. We ended up sending our lens back to Germany. 6 weeks and many $$$ later, it was like new.
Ask you Nikon dealer what s/he would suggest.
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Is this correct? There isn't such a tight seal at the telescoping part of the objective barrel - this (in my experience) is where oil can get in unless a lens condom is used - or have I been woefully misled?
Tamara
On Wed, 28 Mar 2001 jshields-at-cb.uga.edu-at-sparc5.microscopy.com wrote:
} } Hi Sean, } We had the same problem with one of our objectives on the Nikon } inverted. The lens should not allow oil in if it is sealed correctly, } even without the "seal" you provided. Your lens is either older and } the seal is worn, or you've been cleaning the oil off with a harsh } solvent (I've seen some people using xylene regularly). Somewhere } in the back of my mind (a dusty and unreliable source) I think } silcon is not good for the seal. Another solution is to take finger } cots or make them by cutting the finger tips from gloves and pull } them down over the lens if you are still unsure of your seal. } We ended up sending the lens to Nikon for repair. I seem to } remember it cost around $200 or so to have them clean it and } replace the seal, but you can call to get a quote. } good luck }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
You might try an ordinary dial gauge, a device widely used for measuring small movements in machine shops. Check with your friendly machinist, or try your local hardware store. Dial gauges will easily measure 0.0005" (0.01 mm), and are simple, inexpensive, and easy to use. I am sure you can order one from Small Parts Co. I don't happen to have their address here, but you can probably find them on the internet.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Prior to the introduction of techniques for preparing and examining thin metal specimens by transmission electron microscopy, which came into general use in the late 1950s, virtually all electron microscopic studies of non-biological materials was done by examining exposed surfaces (treated in various ways to reveal the characteristics of the internal structure) using replical techniques. Early on, these replicas were made of various polymeric materials (e.g. formvar, colloidion, etc.). There were various shortcomings of these replicas, mainly their resolution was limited to something of the order of 50 Angstroms. In 1954 D. E. Bradley introduced a convenient method for depositing very thin films of carbon. These films were proven to be much better than the polymer films for producing surface replicas. They were stronger, they were electrically conductive, they could be made much thinner, but most of all could provide resolution below 20 Angstroms. Therefore they quickly became the most widely used type of replica film. You can find a rather complete discussion of the many replica techniques that were in use back then in the book 'Techniques for Electron Microscopy', D. E. Kay, Editor, Blackwell Scientific Pub., 1965. These techniques are hardly ever mentioned now-a-days, because there are so many newer techniques now available for producing thin specimens for direct examination.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Listers, I received this inquiry recently. Can anybody help this person? Hi. I am a biology student and I have to do an assignment on electron or light microscopy an it's impact on society. For example, in forensic science or medical research. If you have any information that would help with this, me please email it to me as soon as possible. It would be greatly appreciated. Thankyou, Nicole.
Nicole Forzan {phindola16-at-hotmail.com} Thanks, Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
} Date: Wed, 28 Mar 2001 13:20:14 -0800 } To: "Smartech" {smartech-at-javanet.com} } From: Mary Mager {mager-at-interchange.ubc.ca} } Subject: Re: EM, Why did they replicate surfaces w/ carbon? } Cc: Microscopy } } Dear Ric, } We routinely do carbon replicas of steel surfaces to study the very fine precipitates by TEM, free of the steel matrix. The method is: etch the steel lightly, carbon coat in the evaporator, score the carbon coat into 3 mm. squares with a razor blade, place the steel in the etchant until the carbon squares float off. Scoop up the floating carbon squares with a TEM grid. } For SEM of surfaces that are too big to go into the SEM chamber or that the cops won't let you take away, a cellulose acetate replica is used. It also removes surface material, allowing you to analyse it free of the matrix and cleaning the matrix. It is often used for fractures. The cellulose acetate film is softened in acetone and the sample surface is wetted with acetone. The acetate is placed on the acetone-wetted surface and left for about 0.5 hour to solidify. It conforms exactly to the surface, so you can study the fracture surface without putting the sample in the SEM, after coating the acetate piece to make it conductive. } That is very brief, you can contact me for more information. } At 10:17 AM 3/28/01 -0500, you wrote: } } } I have heard mention of replicating surfaces with carbon for analysis w/ EM, } } it seems like it may be history now, but I was wondering if someone could } } "clue me in" with when it was required. } } } } Thanks } } } } Ric } } } } SMARTech } } 860-491-3299 } } www.semguy.com } } 19 Cornwall Drive } } Goshen CT 06756 } } } Regards, } Mary } } Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Can someone tell me what the mains supply voltage is in Sweden?
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
----- Original Message ----- } From: "Wil Bigelow" {bigelow-at-engin.umich.edu} } You might try an ordinary dial gauge, a device widely used for measuring } small movements in machine shops. Check with your friendly machinist, or } try your local hardware store. Dial gauges will easily measure 0.0005" } (0.01 mm), and are simple, inexpensive, and easy to use. I am sure you can } order one from Small Parts Co. I don't happen to have their address here, } but you can probably find them on the internet. } Wil,
Thanks for mentioning this it solves a problem I have on how to tell how much I move the stage for some experments I am doing trying to enhance the quality of video images by stacking multiple images. I need to get a little shift of the view for better averageing. I never thought using the test indicator laying 6 feet from the scope.
Looking through my machine tool catalog what you want if you go this route is called a test indicator. It is an order of magnitude more accrute than most dial indicators. They can be purchased with calimed accurcies down to 0.00012 or .003 mm. They are avalible in either mecanical dial or digital and range in cost from $100 to $500USD. They are built to be used in machine shops and should give very long service in a labrotory. The accurcy ranges from .001 to .00012 over the price range.
A lever arangement could be built to extend the resolution. It would require careful work to retain the accurcy.
If you need help in finding a vendor or how one would work I have spent countless hours using one.
good luck Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Panel components, Inc. info-at-panelcomponents.com has a great catalog with info on power,plugs, receptacles, etc. for most countries.
I have no $ interest...... but have purchased items from them.
Don Marshall
} From Microscopy-request-at-sparc5.microscopy.com Wed Mar 28 21:07:09 2001 From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } Organization: Dept of Geology, Univ of Auckland } To: Microscopy-at-sparc5.microscopy.com } Date: Thu, 29 Mar 2001 14:07:04 GMT+1200
} Hi } } Can someone tell me what the mains supply voltage is in Sweden? } } thanks } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to dmrelion-at-aol.com. Thank you.)
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all those who responded to my question regarding enrobing of cells in agarose. Bruce Bruce Cutler Director, Microscopy & Electronic Imaging Laboratory University of Kansas
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id MAA03114 for dist-Microscopy; Thu, 29 Mar 2001 12:22:45 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id MAA03110 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 29 Mar 2001 12:22:14 -0600 (CST) Received: from srvr20.engin.umich.edu (srvr20.engin.umich.edu [141.213.75.22]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id MAA03103 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 29 Mar 2001 12:22:03 -0600 (CST) Received: from [141.213.21.161] ([141.213.21.161]) by srvr20.engin.umich.edu (8.9.3/8.9.1) with ESMTP id NAA02225; Thu, 29 Mar 2001 13:19:58 -0500 (EST) User-Agent: Microsoft-Entourage/9.0.2509
Let me actually try and answer your question.
In keeping with most of the rest of Europe I think it is 220/240.
On 3/29/01 8:47 AM, "donald j marshall" {dmrelion-at-world.std.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } ritch, } } } Panel components, Inc. info-at-panelcomponents.com has a great } catalog with info on power,plugs, receptacles, etc. for most countries. } } I have no $ interest...... but have purchased items from them. } } } } Don Marshall } } } } } From Microscopy-request-at-sparc5.microscopy.com Wed Mar 28 21:07:09 2001 } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } } Organization: Dept of Geology, Univ of Auckland } } To: Microscopy-at-sparc5.microscopy.com } } Date: Thu, 29 Mar 2001 14:07:04 GMT+1200 } } } } Hi } } } } Can someone tell me what the mains supply voltage is in Sweden? } } } } thanks } } } } rtch } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } } The University of Auckland email : r.sims-at-auckland.ac.nz } } Private Bag 92019 } } Auckland } } New Zealand } } } } Donald J. Marshall } Relion Industries } P.O. Box 12 } Bedford, MA 01730 } Ph: 781-275-4695 } FAX: 781-271-0252 } email dmrelion-at-world.std.com } } Cathodoluminescence, mass spectroscopy, electron beam technology } } } "A weed is a flower out of place." } } (Please note: Do not send email with attachments to this address. Instead, } send it to dmrelion-at-aol.com. Thank you.) } }
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
} From http://www.sverigeturism.se/smorgasbord/smorgasbord/service/electricity.html:
The voltage is 230V in Sweden. All new buildings erected from January 1, 1994, will be in accord with EU standards. That is, all sockets are earthed. In every public milieu there will only be earthed sockets. In all other buildings erected before the date of January 1, 1994, earthed sockets will be found in bathrooms, as well as in kitchens. Otherwise there are simple sockets. In case one needs an adapter it can be acquired in an electricity store. There are no transformers for 130 volt to 230 volt to be found.
"John F. Mansfield" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Let me actually try and answer your question. } } In keeping with most of the rest of Europe I think it is 220/240. } } On 3/29/01 8:47 AM, "donald j marshall" {dmrelion-at-world.std.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } ritch, } } } } } } Panel components, Inc. info-at-panelcomponents.com has a great } } catalog with info on power,plugs, receptacles, etc. for most countries. } } } } I have no $ interest...... but have purchased items from them. } } } } } } } } Don Marshall } } } } } } } } } From Microscopy-request-at-sparc5.microscopy.com Wed Mar 28 21:07:09 2001 } } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } } } Organization: Dept of Geology, Univ of Auckland } } } To: Microscopy-at-sparc5.microscopy.com } } } Date: Thu, 29 Mar 2001 14:07:04 GMT+1200 } } } } } } } Hi } } } } } } Can someone tell me what the mains supply voltage is in Sweden? } } } } } } thanks } } } } } } rtch } } } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } } Department of Geology Fax : 64 9 3737435 } } } The University of Auckland email : r.sims-at-auckland.ac.nz } } } Private Bag 92019 } } } Auckland } } } New Zealand } } } } } } } Donald J. Marshall } } Relion Industries } } P.O. Box 12 } } Bedford, MA 01730 } } Ph: 781-275-4695 } } FAX: 781-271-0252 } } email dmrelion-at-world.std.com } } } } Cathodoluminescence, mass spectroscopy, electron beam technology } } } } } } "A weed is a flower out of place." } } } } (Please note: Do not send email with attachments to this address. Instead, } } send it to dmrelion-at-aol.com. Thank you.) } } } } } } -- } } Dr. John Mansfield CPhys MInstP } North Campus Electron Microbeam Analysis Laboratory } 417 SRB, University of Michigan } 2455 Hayward, Ann Arbor MI 48109-2143 } Phone: (734) 936-3352 FAX (734) 763-2282 } Cellular Phone: (734) 358-7555 } (Leaving a phone message at 936-3352 is preferable to 358-7555) } Email: jfmjfm-at-engin.umich.edu } URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html } Location: Lat. 42° 16' 48" Long. 83° 43' 48"
-- Henrik Kaker, Ph.D. Metal Ravne d.o.o. SEM-EDS Lab Koroska cesta 14, 2390 Ravne Slovenia Phone: +386 602 21 131 Fax: +386 602 20 436 http://www.kaker.com
I would like to know what computer programs are out there for taking serial sections and creating a 3 D image. Can any one help? Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
{html} {font face="Times, Times"} Dear Listers, {br} {br} Immunogold Workshop Anouncement {br} {br} The Electron Microscopy Facility for the Life Sciences, a shared technology laboratory in the Life Sciences Consortium at Penn State University is hosting a three-day workshop on immunogold techniques from May 21-23, 2001. Dr. Jan Luenissen from the Aurion Immunogold Reagent & {br} Accessories, an internationally known expert in the field, will be the instructor for the workshop. The workshop will include lectures, hands-on training, round table discussions, and presentations on applications. Also, participants of the workshop will be able to work on their own samples during the workshop. The following is the workshop main curriculum. If you are interested in attending or need more information about the workshop, please contact the workshop technical coordinator Hong Yi by phone (404-727-8692) or email (hyi-at-emory.edu). {br} {br} {br} MAIN CURRICULUM {br} {br} The properties of gold particles and their protein conjugates. {br} Theories underlying immunogold labeling protocols. {br} Silver enhancement of gold particles {br} Immunogold labeling on a variety of sample preparations for LM. {br} Immunogold labeling for EM {br} a. Pre-embedding immunogold labeling using ultrasmall gold conjugates and silver enhancement. {br} b. Post-embedding immunogold labeling using conventional colloidal gold conjugates and ultrasmalll gold conjugates. {br} Pre- and post-embedding double immunogold labeling. {br} Background minimization in immunogold labeling {br} Signal amplification in immunogold labeling. {br} {br} {br} Rosemary Walsh, Manager {br} The Electron Microscope Facility for the Life Sciences {br} A Shared Technology in The Life Sciences Consortium {br} 1 South Frear Lab {br} Penn State University {br} University Park, PA 16802 {br} (815) 865-0212 {br} {a href="http://www.lsc.psu.edu/stf/em/home.html" eudora="autourl"} http://www.lsc.psu.edu/stf/em/home.html {/a} {br} {br} {br} {br} {br} {/font} {/html}
There exists a homepage of 3-Dimensional Microscopy Labs (http://3dem.sdsc.edu/) in which are listed various packages for 3D-EM Image Analysis (including reconstruction of serial secions). Additionally a mailing list exists.
BTW: A nice animation can be found in a Web-Page of the University of Utrecht (The Netherlands): http://emsaserv.bio.uu.nl/3dem/ANIMATED_INTRODUCTION/animated_introduction_1.html
I hope this helps.
Best wishes, Ingo
+++++++++++++++++++++++++++++++++++++++++++++++++++++ Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH Herbststrasse 7 D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: http://www.tvips.com/ Email: ingo.daberkow-at-tvips.com
-------- Original Message --------
Dear Listers:
I would like to know what computer programs are out there for taking serial sections and creating a 3 D image. Can any one help? Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Dear collegues, I need to perform an autoradiographic study in electron microscopy. From some papers in the literature it seems usuful to employ a phenidon developer and a gold latensification solution; since I did not find any commercial product available, I need to prepare it in my lab. I would be very grateful to you if you can provide me with some information about the composition and detailed method of preparation and storage of the different solutions. Thank you very much. Maria Meli
I am pleased to announce the completely revised and updated edition of "Microscopy & Imaging Resources on the WWW" is now available at a new URL. The goal of this site has always been to provide resources for University students, staff and faculty who want to learn more about microscopy and imaging. The site includes K-12 educational links, information on Light Microscopy, Histology, Confocal Microscopy, Fluorescence Techniques, Electron Microscopy, and Digital Imaging.
http://swehsc.pharmacy.arizona.edu/exppath/
"Microscopy & Imaging Resources on the WWW" is an outreach of the Southwest Environmental Health Sciences Center, which is funded by the National Institute of Environmental Health Sciences (NIEHS is one of the National Institutes of Health, part of the U.S. Department of Health and Human Services).
Yours, Doug Cromey Manager, Experimental Pathology Core, SWEHSC .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (NEW email: Cromey-at-Arizona.edu) : :...................................................................: http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
Listerservers, I have heard that there are a set of fonts called 'Fraser fonts' with over bars etc. that are useful for indices. Does it exist. Where can such a font be obtained?
Thanks in advance, Steven
-- Dr. Steven Celotto Department of Engineering Materials Science & Engineering University of Liverpool Brownlow Hill Liverpool L69 3BX UNITED KINGDOM
ok taking a little survey. i am in a diagnostic EM lab. we mount out sections on formvar coated slotted grids, so he can shoot pics of the whole glomerlus. ok my question. how may of you out there doing diagnostis EM on renals do this? john
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} ok taking a little survey. i am in a diagnostic EM lab. we mount out sections {BR} on formvar coated slotted grids, so he can shoot pics of the whole glomerlus. {BR} ok my question. how may of you out there doing diagnostis EM on renals do {BR} this? {BR} john {/FONT} {/HTML}
We have just acquired a used microscope made by ISI Inc. (their model alpha - 1980). Would anyone please have information on how we can obtain spare parts, filaments etc., for this machine. Many thanks for your help...
John: I did renal biopsies by E-M for 12 years and knew the other two people in town who were also doing diagnostic E-M of renal biopsies. We all used grids with grid bars ( I always used naked grids). I chose grids with slender bars for all my work to maximize viewing without using slotted grids. We did not view "whole" glomeruli slices (quite low power I would think). I don't feel diagnoses were compromised versus using slotted grids. Since several sections are mounted on the 200 mesh grids, one should be able to view areas of interest just fine.
} } } {"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com} 03/30 11:20 AM } } } ok taking a little survey. i am in a diagnostic EM lab. we mount out sections on formvar coated slotted grids, so he can shoot pics of the whole glomerlus. ok my question. how may of you out there doing diagnostis EM on renals do this? john
will add this link to the upcoming SMECC website. thanks Ed Sharpe
} Subj: Web site announcement } Date: 3/30/01 11:16:39 AM US Mountain Standard Time } From: Cromey-at-Arizona.edu (Doug Cromey) } To: microscopy-at-sparc5.microscopy.com, confocal-at-listserv.acsu.buffalo.edu } } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues, } } I am pleased to announce the completely revised and updated edition of } "Microscopy & Imaging Resources on the WWW" is now available at a new } URL. The goal of this site has always been to provide resources for } University students, staff and faculty who want to learn more about } microscopy and imaging. The site includes K-12 educational links, } information on Light Microscopy, Histology, Confocal Microscopy, } Fluorescence Techniques, Electron Microscopy, and Digital Imaging. } } http://swehsc.pharmacy.arizona.edu/exppath/ } } "Microscopy & Imaging Resources on the WWW" is an outreach of the Southwest } Environmental Health Sciences Center, which is funded by the National } Institute of Environmental Health Sciences (NIEHS is one of the National } Institutes of Health, part of the U.S. Department of Health and Human } Services). } } Yours, } Doug Cromey } Manager, Experimental Pathology Core, SWEHSC } .................................................................... } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } : Research Specialist, Principal University of Arizona : } : (office: AHSC 4212A) P.O. Box 245044 : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } : (FAX: 520-626-2097) (NEW email: Cromey-at-Arizona.edu) : } :...................................................................: } http://swehsc.pharmacy.arizona.edu/exppath/ } Home of: "Microscopy and Imaging Resources on the WWW" } } } } } ----------------------- Headers --------- }
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} will add this link to the upcoming SMECC website. {BR} thanks Ed Sharpe {BR} {BR} {BR} {BLOCKQUOTE TYPE=CITE style="BORDER-LEFT: #0000ff 2px solid; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px; PADDING-LEFT: 5px"} Subj: {B} Web site announcement {/B} {BR} Date: 3/30/01 11:16:39 AM US Mountain Standard Time {BR} {I} From: Cromey-at-Arizona.edu (Doug Cromey) {BR} To: microscopy-at-sparc5.microscopy.com, confocal-at-listserv.acsu.buffalo.edu {BR} {/I} {BR} {BR} {BR} {BR} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {BR} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {BR} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} -----------------------------------------------------------------------. {BR} {BR} {BR} Dear Colleagues, {BR} {BR} I am pleased to announce the completely revised and updated edition of {BR} "Microscopy & Imaging Resources on the WWW" is now available at a new {BR} URL. The goal of this site has always been to provide resources for {BR} University students, staff and faculty who want to learn more about {BR} microscopy and imaging. The site includes K-12 educational links, {BR} information on Light Microscopy, Histology, Confocal Microscopy, {BR} Fluorescence Techniques, Electron Microscopy, and Digital Imaging. {BR} {BR} http://swehsc.pharmacy.arizona.edu/exppath/ {BR} {BR} "Microscopy & Imaging Resources on the WWW" is an outreach of the Southwest {BR} Environmental Health Sciences Center, which is funded by the National {BR} Institute of Environmental Health Sciences (NIEHS is one of the National {BR} Institutes of Health, part of the U.S. Department of Health and Human {BR} Services). {BR} {BR} Yours, {BR} Doug Cromey {BR} Manager, Experimental Pathology Core, SWEHSC {BR} .................................................................... {BR} : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : {BR} : Research Specialist, Principal University of Arizona : {BR} : (office: AHSC 4212A) P.O. Box 245044 : {BR} : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : {BR} : (FAX: 520-626-2097) (NEW email: Cromey-at-Arizona.edu) : {BR} :...................................................................: {BR} http://swehsc.pharmacy.arizona.edu/exppath/ {BR} Home of: "Microscopy and Imaging Resources on the WWW" {BR} {BR} {BR} {/XMP} {/FONT} {FONT COLOR="#0f0f0f" SIZE=2 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BR} {BR} ----------------------- Headers --------- {BR} {/BLOCKQUOTE} {BR} {/FONT} {/FONT} {FONT COLOR="#000000" SIZE=2 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BR} {/FONT} {/HTML}
ISI became Topcon, and then dropped out of the American market directly awhile ago. Any of the SEM supply houses can provide supplies or you can address your requests to Aspex, which was R. J. Lee until recently. They currently represent the Topcon lines in North America and can be found at http://www.rjleeinst.com/. Confused yet? It gets better. Good luck finding anything about ISI or Topcon products on their website. Try the phone lines.
Frankly, I hate having to trace the providence of this and other companies that give up well known brand names for reasons known only to them. ISI made inroads in this country primarily a s a low-bidder, thus many instruments were originally in government labs. Had they made some attempt to produce a product for the middle of the pack they may have done better and may still be a player.
I'm not saying that they didn't produce a decent instrument - I still have many that perform quite well. But they cut their own throats by playing to an audience that has suffered devastating budgetary cutbacks in an era when other segments were willing to pay vastly more for instruments that offer little basic operational improvements.
On Friday, March 30, 2001 2:53 PM, Rishi Raj [SMTP:Rishi.Raj-at-Colorado.edu] wrote: } ------------------------------------------------------------------------ } } We have just acquired a used microscope made by ISI Inc. (their model } alpha - 1980). Would anyone please have information on how we can obtain } spare parts, filaments etc., for this machine. Many thanks for your } help... } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
-----Original Message----- } From: "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com [mailto:"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com] Sent: Friday, March 30, 2001 2:20 PM To: microscopy-at-sparc5.microscopy.com
-----Original Message----- } From: Garrison, Becky Sent: Saturday, March 31, 2001 8:33 AM To: '"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com'; microscopy-at-sparc5.microscopy.com
John: We do around 500 renal biopsies per year and all the sections are mounted on 200 mesh uncoated copper grids. We have an 8 year old Hitachi 7100 and use 60kv. The majority of the glomerulus can be viewed with the 3-4 serial sections lying randomly across the grid bars. We do not need a picture of the whole glomerulus, rather most pictures are between 3,000 and 10,000X. Dr. Tibor Nadasdy is the renal pathologist and decided last year that all our renal biopsies would be captured with the digital camera onto a computer and sent up to him via a network to his computer. So, at the present time we use very little EM film. He diagnoses each biopsy and e-mails representative digitized images to the nephrologists.
Karen L. Jensen, M.S. Project Manager & Associate Scientist Electron Microscopy Research Core
-----Original Message----- } From: "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com [mailto:"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com] Sent: Friday, March 30, 2001 2:20 PM To: microscopy-at-sparc5.microscopy.com
Hello -
Has anyone out there tried to adhere xylem sections to pre-coated microscope slides like Fisher Probe-On Plus slides? I've tried and had a zero percent section retention. I can imagine that this is because of the scarcity of live cells (and plasma membranes). I've tried slow air drying and various temperatures on the slide warmer. The sections are 15 microns, and are from fixed (buffered paraformaldehyde) but not embedded samples. I'm about to try Poly-L-lysine and amino-acyl silane treated slides, but I'm not too hopeful because they (at least Poly-L) rely on the same positively-charged surface principle. I want to avoid gelatin or albumin subbing because I'm treating sections with protease, and also want to minimize background staining. Any tips would be greatly appreciated.
Rachel
****************************************** Rachel Spicer Biological Laboratories 3119 Organismic and Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138
Rishi Raj wrote: ============================================================ We have just acquired a used microscope made by ISI Inc. (their model alpha - 1980). Would anyone please have information on how we can obtain spare parts, filaments etc., for this machine. Many thanks for your help... ============================================================ The business of the ISI SEM's is now being handled by
Aspex Instruments LLC Formerly: RJ Lee Instruments Ltd. 175 Sheffield Drive Delmont, PA 15626 USA Tel: 1-724-468-5400 Fax: 1-724-468-0225 E-mail: pssales-at-rjleeinst.com
The former manager of the SEM operation when it was still ISI, Michael McCarthy, is now with Aspex. Michael might be the single most knowledgeable person in terms of spare parts for the column and vacuum system. Several of the main suppliers of consumables, like SPI Supplies, also offer filaments, new and retipped, apertures, and the other items of that nature you would be needing to maintain the microscope.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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