I would like to do imaging of live GFP expressing mammalian cells. I am concerned about regulating pH without needing a gas flow system on the stage. Is adding HEPES to media sufficient to control pH in a bicarbonate based medium? Has anyone tried the Gibco CO2 independent medium to know if it is a high riboflavin medium (proprietary formula so I can't look it up). Thanks- Dave -- Permanent Address Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
Current Address 9/01-8/02 Dr. David Knecht School of Biosciences The University of Birmingham, Edgbaston, Birmingham, B15 2TT, U.K. Lab telephone: 0121 414 2508 (44-121 414 2508 from abroad) Department Fax: 0121 414 5925 (44-121 414 5925 from abroad) Direct Fax: 0121 414 5411 (44-121 414 5411 from abroad)
Try iCal {http://www.brownbearsw.com/index.html} , it may not be completely suitable for a large shared facility (somewhat limited use of alternate logins), but we use it on several smaller pieces of shared equipment (runs on NT). You can try the software as a shareware demo.
Another possibility is {http://www.loci.wisc.edu/calendar/} , which was described in a recent article in BioTechniques.
No commercial interest in either software. Doug
At 06:38 PM 10/31/2001 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good Evening all, } } My department here is looking to acquire interactive scheduling } software, preferably for the internet, that will cover all of our } microscopes. Users should be able to create an account, manage it } (add appointments, edit them, delete them...), and be able to see } what time slots are available for the scope they wish to use. } Please feel free to give me all suggestions that come to mind as to } what software is available that will accomplish this. } Thank you all very much. } } Chris
.................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (NEW email: Cromey-at-Arizona.edu) : :...................................................................: http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
I'm searching for freeze-substitution protocols (without using of any fixative), preferably fast and slow protocols, for plant leaf specimen preparation. I'm working on localizing nickel element in the plant leaves of a hyperacumulator plant, hence using cryo preparation methods. I'll highly appreciate any contributions and help in this.
My thanks in advance.
Chris Ogomo MSc Student University of the Western Cape Rep of S. Africa
Yeah, this book is really a good book for beginners. but there is now second edition in 1999 available 670pp, ISBN 0-7637-0192-0. The content is from basic structure and principles of TEM and SEM to sample preparation with many vivid pictures, mainly for tissue samples, also some pages on FT. This book is very basic, but not advanced, and it is quite cheap($8) .
Chen Chen
Department of Biological Chemistry The Johns Hopkins University School of Medicine 725 N. Wolfe Street Baltimore, Maryland 21218
On Wed, 31 Oct 2001 akc-at-umich.edu-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } One of the particularly good ones is: Bozzola JJ, Russell LD, 1991. } Electron Microscopy: Principles and Techniques for Biologist. Jones and } Bartlett Publisher (Boston), 542 pp, ISBN 0-86720-126-6. } } Kent } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } A. Kent Christensen, Professor Emeritus } Department of Cell and Developmental Biology, Medical Science II Building } University of Michigan Medical School, Ann Arbor, MI 48109-0616 } Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 } akc-at-umich.edu http://www.umich.edu/~akc/ } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } --On Wednesday, October 31, 2001 11:15 AM -0500 Michael Jarnik } {M_Jarnik-at-fccc.edu} wrote: } } } } } Listers, } } } } I know this question was discussed in the past, but anyway. I would need } } a pretty comprehensive textbook(s)/ reference book(s) covering EM use in } } biology - possibly with stress on TEM, including sample preparation } } techniques. Any bright ideas? } } } } Thanks, } } } } Michael Jarnik } } } } } } -- } } Michael Jarnik, Ph.D. } } Fox Chase Cancer Center } } Electron Microscope Facility } } Philadelphia } } } }
I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there.
One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there".
I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques.
My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them.
Any help you can provide would be very much appreciated - before I rip all of my hair out....
Thanks in advance, listers, to all of you. I know I came to the right place.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Electron Microscopy by John Bozzola and Lonnie Russell is excellent. I use it for both my TEM and SEM class. Biological Electron Microscopy by Michael J. Dykstra is also useful. It is more compact in size so easier om students' backs, and still seems to cover the necessary material. It doesn't include the extensive ultrastructure section Bozzola's book has. Either is good, and there are also many other good books out there. I have always spent a lot of time at the library display at the MSA meetings. Joyce Craig Chicago State University
Anthrax is actually the disease caused by the spore-forming bacterium Bacillus anthracis. The bacterium in question is actually named Bacillus anthracis. I guess it is easier for the news media to say anthrax than bacillus anthracis.
Tyrone Daulton
Kristen Lennon wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Semantics because this mistake has been making me crazy - ANTHRAX IS A } BACTERIUM! This is a particularly important point because the word "virus" } scares people silly. When I think of virus, I think of AIDS and Ebolla } (sorry it that's misspelled). When I think of bacterium, I think antibiotic. } No offense intended, } Kristen } } Kristen A. Lennon, Ph.D. } Department of Plant Pathology } 351 Bessey Hall } Iowa State University } Ames, IA 50011 } } 515-294-8854 } kalen-at-iastate.edu } www.baumlab.org
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Dear Collegues. I Have a Microlumina camera in my lab and lost the EasyScan 1.2 software as a result of a virus attack. The original diskas are by now unreadable. Does anyone have a copy of this software? I would be eternally gratefull if someone could send me a copy by e-mail to the address below. I think that the camera has been descontinued and is no longer available comercially.
Thanks in advance Dr. A.P. Alves de Matos Lisbon University Dental Medical School apmatos-at-ip.pt
Dear Dee, When I looked at latex spheres made in our Pathology department, I just dried a drop out of the suspension onto a polished graphite planchet. A glass cover slip is also a good substrate, if you are going to gold-coat anyway. If the carrier is really DI, it should dry away clean. If there is stuff in the DI, you will get smut around the spheres. I believe the polystyrene is strong enough to hold up to drying. If the spheres are small enough, they stick to the graphite themselves. If they are larger and in danger of rolling off, dry them onto a sticky tab. At 06:07 PM 10/29/01 -0500, you wrote: } } Hi listers, } } I've been asked to look at some polystyrene microspherules in the SEM and } would like some advice on how to prepare them. I'm not a biomedical } person, so need some help! } } The spherules are 5.6 um size with a Lumidin (protein) bonding surface and } receptor molecules attached. They're in DI (or a DI solution?), which is } apparently the delivery system for the marker molecules. Would they need } CPD? If so, I've never CPD'd particles in solution before.... The SEM has } hivac and VP modes, plus there's a cool stage. } } Any ideas would be highly appreciated! } Many thanks, } Dee Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hi Darren, Under separate cover I am sending an Excel table with plastics, their monomers and their indices of refraction. At least this may help you look.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: dmichael-at-usc.edu } Sent: Monday, October 22, 2001 5:13 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist: high index of refraction plastic } coverslips } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (dmichael-at-usc.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } October 31, 2001 at 12:13:55 } -------------------------------------------------------------------------- } - } } Email: dmichael-at-usc.edu } Name: Darren Michael } } Organization: University of Southern California } } Education: Graduate College } } Location: Los Angeles, CA } } Question: Hi, } } I'm trying to find high index of refraction plastic coverslips. } I've read about eyeglasses with high index of refraction (1.66) } plastic lenses and have been unable to locate a manufacturer of } coverslips using the same types of materials. Have you come across } them? Do you have any suggestions as to whom to ask? } } Thanks, } } Darren } } -------------------------------------------------------------------------- } - } }
Does anyone know of a most excellent method of fixing the mucin layers of a cell culture for TEM? There is a rumor of some osmium/fluoride compound that I'm trying to track down (osmium perfluouro-something?), but no luck yet. Personal experiences, references, and sources for exotic compounds would all be most welcome.
Thanks much.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
If you are teaching biological TEM, then you might consider using as a corollary "Cell and Tissue Ultrastructure: A Functional Perspective" by Cross and Mercer, pub by W.H.Freeman & Co NY. No sample prep or EM operation, but beautifully-presented micrographs illustrating ultrastructure and tissue types.
A.L.
####################################### Ann Hein Lehman Assistant Director, EM Facility Trinity College - LSC314 300 Summit Street Hartford CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman/
-----Original Message----- } From: Joyce Craig [mailto:J-Craig-at-csu.edu] Sent: Thursday, November 01, 2001 12:07 PM To: Microscopy microscopy
Electron Microscopy by John Bozzola and Lonnie Russell is excellent. I use it for both my TEM and SEM class. Biological Electron Microscopy by Michael J. Dykstra is also useful. It is more compact in size so easier om students' backs, and still seems to cover the necessary material. It doesn't include the extensive ultrastructure section Bozzola's book has. Either is good, and there are also many other good books out there. I have always spent a lot of time at the library display at the MSA meetings. Joyce Craig Chicago State University
At 09:21 AM 11/1/01 -0500, Chen Chen wrote: } Yeah, this book is really a good book for beginners. but there is now second } edition in 1999 available 670pp, ISBN 0-7637-0192-0. The content is from basic } structure and principles of TEM and SEM to sample preparation with many vivid } pictures, mainly for } tissue samples, also some pages on FT. This book is very basic, but not } advanced, } and it is quite cheap($8) .
Not sure about mucin, but have you looked into ruthenium red????
On Thu, 1 Nov 2001, Tindall, Randy D. wrote:
} Date: Thu, 1 Nov 2001 12:45:17 -0600 } From: Tindall, Randy D. {TindallR-at-missouri.edu} } To: "'microscopy-at-sparc5.microscopy.com'" {microscopy-at-sparc5.microscopy.com} } Subject: TEM: Mucin fixation } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } Does anyone know of a most excellent method of fixing the mucin layers of a } cell culture for TEM? There is a rumor of some osmium/fluoride compound } that I'm trying to track down (osmium perfluouro-something?), but no luck } yet. Personal experiences, references, and sources for exotic compounds } would all be most welcome. } } Thanks much. } } Randy } } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
No one has mentioned this book, so I might as well: "Biomedical Electron Microscopy" by A.B. Maunsbach and B.A. Afzelius Academic Press, 1999, ISBN 0-12-480610-4 This text has many EMs comparing the effects of using different fixatives, buffers, embedding resins, and so forth. An excellent companion to Bozzola & Russell. Note: check ebay. I got my copy there for about 2/3 of the retail price.
Phil
} Electron Microscopy by John Bozzola and Lonnie Russell is excellent. I } use it for both my TEM and SEM class. } Biological Electron Microscopy by Michael J. Dykstra is also useful. It } is more compact in size so easier om students' backs, and still seems to } cover the necessary material. It doesn't include the extensive } ultrastructure section Bozzola's book has. } Either is good, and there are also many other good books out there. } I have always spent a lot of time at the library display at the MSA } meetings. } Joyce Craig } Chicago State University
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Hello Paula, How many samples you need to examine depends on the details of the experiment, i.e. nature of specimens, the types of measurements taken, and most importantly the experimental question that is addressed by the experiments. The question to whether you have significant statistics is dictated by these enumerated, and perhaps other, factors. These factors will obviously differ for different experiments which have different goals.
My advice is to sit down and examine closely the experimental questions you choose to address and decide whether the data you have collected has the necessary statistics to adequately constrain the interpretations of the data.
The only meaningful reference (of which you seek) would be one that discusses measurement statistics on similar experiments, taking similar measurements, and addressing similar questions.
I hope this is of some help, good luck. Ty
Paula Allan-Wojtas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, all, } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } Thanks in advance, listers, to all of you. I know I came to the right place. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Even statistically speaking, the number of samples that you need depends on the variability between samples in your group and the extent to which the experimental group varies from the controls.
If your experimental group consistently looks one way and your controls consistently and obviously look different from the experimental group, I don't see why you would need to run statistics.
But it really depends on what you are trying to say. If all you are trying to point out is group one looks different than normal and it is obvious in all of your samples, (by obvious, I mean obvious to a trained eye) statistics is a waste of time. However, when you start to say group one looks somewhat worse than group two which looks somewhat worse than normal, and some of the specimens in the group actually look the same as, or better than ,the ones in the better group, then you'd need statistics. There are ways to figure out how big the group actually needs to be by taking a sample and determining the variability within and between groups (say on your 4 samples already obtained). I'm betting if you really can see something with these few samples, the estimation of sample size needed for statistics will be allot less than 30-40.
Unfortunately, I don't know of any articles you can site about the use of statistics in SEM off hand. Sorry. I've personally run into people recommending these ridiculously large sample sizes because they work with people. I work with animals and have allot more control over inter subject variability than they do, therefore I can work with a much smaller sample size even for statistics.
Good luck,
Karen Pawlowski, Ph.D.
Paula Allan-Wojtas wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, all, } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } Thanks in advance, listers, to all of you. I know I came to the right place. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
We regularly use 1% osmium tetroxide with 8% dimethoxypropane (DMP) in acetone for woody plant tissues with good success. The DMP removes water from the solution. We substitute for 5 days at -80 C. Anything less that 3 days won't work at all and more than 5 days makes no difference. We have used straight acetone for immuno work but the fixation (i.e. membranes) is not so great and the images are low in contrast. Other possibilities are 0.2 to 2% glutaraldehyde in acetone, with or without tannic acid. This is good for protein preservation for immuno work. Methanol is reported to work but not as well as acetone (I haven't successfully used it).
Let me know if I can be of further help.
Kim ------- Kim Rensing Ph.D. Dept. of Botany, UBC 6270 University Blvd Vancouver, BC, Canada V6T 1Z4
} I'm searching for freeze-substitution protocols (without using of any } fixative), preferably fast and slow protocols, for plant leaf specimen } preparation. I'm working on localizing nickel element in the plant leaves of } a hyperacumulator plant, hence using cryo preparation methods. I'll highly } appreciate any contributions and help in this. } } My thanks in advance. } } Chris Ogomo } MSc Student } University of the Western Cape } Rep of S. Africa
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (griggsa-at-dbcc.cc.fl.us) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, November 1, 2001 at 14:26:59 ---------------------------------------------------------------------------
on 11/1/01 9:41 AM, Paula Allan-Wojtas at AllanWojtasP-at-EM.AGR.CA wrote:
} I am working on a multidisciplinary team of researchers on a number of very } interesting projects. I have a very large collection of top quality } micrographs from these projects which I would like to publish. The microscopy } samples have been selected and photographed within the experimental design } which has been set out for each project, so we are pretty sure they are } representative of what's there. } } One collaborator on the team is hesitant (to the point of being resistant) } about my publishing any of the microscopy as part of any of the publications } put together by the team because we have not looked at 30-40 samples of each } of the many experimental treatments ( we have done 4 samples of each } treatment) . The other researchers on the team have looked at between 30 and } 40 samples per treatment. This one collaborator says that she has no } confidence in the microscopy results because "the statistics just aren't } there". } } I have tried on numberous occasions to explain to her about why this is not } feasible to do in SEM, and is usually not done. She does not believe me and } is essentially preventing me from publishing the work. I could probably } publish this work as a stand alone ms, but it is so much more meaningful when } integrated with data from the other analytical techniques. } } My question: is there a reference (which I have somehow missed) to explain why } 30-40 samples is not feasible for microscopy and which rationalizes why this } is so? The function of microscopy in the projects has been to give pictorial } information about the samples, and to suggest trends, not for absolute } measurement of structures and changes in them. } } Any help you can provide would be very much appreciated - before I rip all of } my hair out.... } Dear Paula, As others have said, it depends on what kind of experiment you're looking at. Even if all the experimental samples look different from the controls, if you're trying to quantitate the changes--e.g., adding X to the culture shrinks the cells by Y%--you may still need sufficient data to make the numbers significant and to provide a standard deviation. Is is also important to have looked at different, independently-prepared samples, so if the few samples you observed were all from one experiment, I, too, would have doubts that they were sufficient. Then there's the problem that one selects what to photograph, and this selection always involves subjective factors. All that being said, however, if each of the other properties measured in the experiment were consistent from prep to prep, and if what you see correlates with what was measured, then, IMHO, it would be proper to publish the micrographs to illustrate what was found, regardless of how many or few you have. Good luck. Yours, Bill Tivol
Paula Most of us have grappled with this problem, but there is no generally applicable answer to the question "How many samples do I need to examine?". You are correct in saying that most electron microscopists don't attempt to examine 30-40 samples per treatment, but sometimes that is unavoidable. If the scientific question you are asking is a quantitative one - e.g. what proportion of cells have a particular morphology in normal and diseased tissues - and especially if the legal or public health or commercial cost implications of the answer are very significant - then it may be necessary to examine the number of samples and analyse the number of photographs demanded by the statisticians. If on the other hand you just want to illustrate the effect of a particular mutation on the shape of a fly's eye 2-3 representative samples are probably adequate. I feel hampered in making any really helpful suggestions about your experiment because it is not clear from your message what the nature of the experiment is, or what kind of observations you are attempting to make. Could you tell us more about that?
Best wishes Chris
} Paula Allan-Wojtas wrote: } } } -------------------------------------------------------------------- ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- ---. } } } } Hi, all, } } } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } } } Thanks in advance, listers, to all of you. I know I came to the right place. } } } } Paula. } } } } Paula Allan-Wojtas } } Research Scientist - Food Microstructure } } Agriculture and Agri-Food Canada } } Atlantic Food and Horticulture Research Centre } } Kentville, Nova Scotia Canada B4N 1J5 } } } } Tel: (902) 679-5566 } } FAX: (902) 679-2311 } } } } email: allanwojtasp-at-em.agr.ca
This article from NYTimes.com has been sent to you by vcr.group-vince-at-worldnet.att.net.
Particles Are Tiny, but Damage Can Be Great
October 30, 2001
By JAMES GLANZ
Environmental scientists have spent decades studying the physics and physiology of particles very much like those in the most dangerous forms of biological weaponry.
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Thank you to everyone who has responded, so far. It has been very helpful for me. The responses have covered the whole spectrum from "yes, do the 30-40 samples" to "3-4 is probably enough". The majority of the answers fell in between these 2 points, saying that the number of samples required to be representative really depends on what you want/need to know from the samples and the sample variability. This could be partially achieved by taking pictures randomly over different samples. I will post a summary of answers soon. Anyone who does not want me to reveal their reply/name please contact me offline to let me know.
In some other unrelated work that we have done recently, we took the approach of working with a statistician from the outset. The project was a much more focused one, and we were looking for treatment effects, and intensity of these. One of the first steps was to actually do a small study to assess the variability of the population, and factors which affected it. With this information we were able to move to the next phase, actually testing the treatments. Statistics (experimental design, proper data collection and analysis of the data) allowed us to separate the population variability from the treatment effects, even when they were subtle. The result was information which did not agree with published literature or longheld assumptions which had arrived at through studies that did not have the "statistical rigour" that our studies did. Sorry to be vague about this, but it will be some months before I can openly talk about this work. I just wanted to say, for the record, that I am not against a statistical approach. I think it's a great idea, and I think it's the way to go.
In the original work I contacted the group about yesterday, we were just making some preliminary observations about the samples and suggesting that a connection could be made between the microscopy and other analytical methods. I have published other papers of this type with other workgroups, and everyone was ok with the "representative micrographs" idea, provided the usual disclaimers were used - "possible relationship" rather than "correlation"; "preliminary observations" ;More work should be done with more samples to be certain we're seeing what we think we're seeing " a first attempt at trying to relate microstructure to..."
I really can't go into more detail about this situation for obvious reasons. Suffice it to say that I plan to do my microscopy from now on in a new frame of mind, and will take some time to speak with a statistician before I start any new projects. My quest is to get the work I originally described, and all the work that we did connected to it, published in some form (complete with all the disclaimers necessary) as soon as I can.
Thanks again for all your help and support.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
We have been looking at marine algae in the same size range for quite some time. One of the considerations is having the sample distributed well enough to see individual particles and have a uniform random distribution for statistical purposes. Drying down from a drop of liquid tends to bunch up the particles and distribute the size ranges non-uniformly. We have used polycarbonate filters (Nuclepore, Poretics) and filter from liquid solution to collect the particles on a low profile background.
There are a number of pore sizes to choose from, select the one just below the size range of interest so that smaller detritus is removed. Try a dilution series to get a useable concentration. If the particles have enough inherent rigidity so they do not collapse, air drying is sufficient. We mount the filters on SEM stubs while still somewhat damp (they will fly off with static and/or handling of the flimsy material when completely dry). We then coat with gold. The one caution here is when the vacuum is applied and released make sure that it is done slowly so that large bursts of air don't sweep over the particles. Having said that our experience is that micron sized particles tend to stick to the surface they lay on. If you are adventurous you might try looking at the particles uncoated in VP mode.
Hope this helps.
-David O'Neil Institute for Marine Biosciences National Research Council of Canada 1411 Oxford St. Halifax, NS B3H 3Z1 ph. 902-426-8258 fax 902-426-9413 david.o'neil-at-nrc.ca http://www.imb.nrc.ca/micros_e.html
At 06:07 PM 10/29/01 -0500, you wrote: } } Hi listers, } } I've been asked to look at some polystyrene microspherules in the SEM and } would like some advice on how to prepare them. I'm not a biomedical } person, so need some help! } } The spherules are 5.6 um size with a Lumidin (protein) bonding surface and } receptor molecules attached. They're in DI (or a DI solution?), which is } apparently the delivery system for the marker molecules. Would they need } CPD? If so, I've never CPD'd particles in solution before.... The SEM has } hivac and VP modes, plus there's a cool stage. } } Any ideas would be highly appreciated! } Many thanks, } Dee
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listers, } } Does anyone know of a most excellent method of fixing the mucin layers of a } cell culture for TEM? There is a rumor of some osmium/fluoride compound } that I'm trying to track down (osmium perfluouro-something?), but no luck } yet. Personal experiences, references, and sources for exotic compounds } would all be most welcome. } } Thanks much. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/
Ruthenium red, as Sara Miller wrote, works well. Osmium can be dissolved in organic compounds, the idea being not to expose the object of your study to aqueous solvents while it is being fixed. I don't know how well osmium fixes the mucin you are interested in. Glutaraldehyde and formaldehyde can also be "dissolved" or partitioned into organic solvents to accomplish the same end. This is known as phase partition fixation. I will send a reference if you like. Of course, if you are working with plastic culture dishes, the organic solvent must not react with the dish.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
} } } Dear Christopher, } } } The software that we use for scheduling is called Webevent. It is } } } fairly straightforward to use, and people can set up accounts and edit } } } them, depending on how you set it up. Their web site is: } } } http://www.webevent.com } } } Hope that helps. } } } } } } -Russell } } } } } } Christopher Gieczys wrote: } } } }
} } } } } } } } Good Evening all, } } } } } } } } My department here is looking to acquire interactive scheduling software, } } } } preferably for the internet, that will cover all of our } } } microscopes. Users } } } } should be able to create an account, manage it (add } } } appointments, edit them, } } } } delete them...), and be able to see what time slots are available for the } } } } scope they wish to use. Please feel free to give me all suggestions that } } } } come to mind as to what software is available that will accomplish this. } } } } Thank you all very much. } } } } } } } } Chris } } } } } } } } _________________________________________________________________ } } } } Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp
-- Russell McConnell Confocal Imaging Facility Technician Department of Neuroscience Tufts University School of Medicine M&V Building room #137 136 Harrison Ave. Boston, MA 02111
If you are not quantifying the microstructure with stereological measurements, then you use your qualitative judgment as to whether the micrographs that you use are representative. Typically you will get a feel for it. If on the other hand, you are using quantitative microscopy techniques, there are statistical tests that you judge your results by. These tests will tell you whether your sampling is sufficient. I recommend that you look at several books on the topic. The materials science oriented books that I am familiar with are DeHoff and Rhines "Quantitative Microscopy" and John Russ's books; try the "Image Processing Handbook" for example. John runs a short course on this topic every year and DeHoff is a co-instructor.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Tyrone L. Daulton [mailto:tdaulton-at-nrlssc.navy.mil] Sent: Thursday, November 01, 2001 5:54 PM To: Paula Allan-Wojtas; microscopy-at-sparc5.microscopy.com
Hello Paula, How many samples you need to examine depends on the details of the experiment, i.e. nature of specimens, the types of measurements taken, and most importantly the experimental question that is addressed by the experiments. The question to whether you have significant statistics is dictated by these enumerated, and perhaps other, factors. These factors will obviously differ for different experiments which have different goals.
My advice is to sit down and examine closely the experimental questions you choose to address and decide whether the data you have collected has the necessary statistics to adequately constrain the interpretations of the data.
The only meaningful reference (of which you seek) would be one that discusses measurement statistics on similar experiments, taking similar measurements, and addressing similar questions.
I hope this is of some help, good luck. Ty
Paula Allan-Wojtas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, all, } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } Thanks in advance, listers, to all of you. I know I came to the right place. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Or you could try Alcian blue/glutaraldehyde followed by osmium textroxide. See: Hear Research 1987;27(1):47-65 A newly identified surface coat on cochlear hair cells. Santi PA, Anderson CB.
Warning: This method is tricky because it can end up trapping water molecules in the mucous type tissues and it requires longer dehydration and polymerization times to set blocks hard enough for EM. But it looks great! (Actually, the molecules don't collapse as much in this as in ruthenium red, so the "coat" is closer to it's true thickness.) I only did some light level stuff with this myself, but I worked down the hall from Dr. Santi when this experiment was being done.
Karen Pawlowski
Sara Miller wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Not sure about mucin, but have you looked into ruthenium red???? } } On Thu, 1 Nov 2001, Tindall, Randy D. wrote: } } } Date: Thu, 1 Nov 2001 12:45:17 -0600 } } From: Tindall, Randy D. {TindallR-at-missouri.edu} } } To: "'microscopy-at-sparc5.microscopy.com'" } {microscopy-at-sparc5.microscopy.com} } } Subject: TEM: Mucin fixation } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Listers, } } } } Does anyone know of a most excellent method of fixing the mucin layers of a } } cell culture for TEM? There is a rumor of some osmium/fluoride compound } } that I'm trying to track down (osmium perfluouro-something?), but no luck } } yet. Personal experiences, references, and sources for exotic compounds } } would all be most welcome. } } } } Thanks much. } } } } Randy } } } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } } } } } } } } } } } } Sara E. Miller, Ph. D. } P. O. Box 3712 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-3265
Dear Randy, Sorry for the bulk of this response, but finding what you needed required two Google searches, both of which might be of use to you along with the one finding that was close to what I expected. I recalled a paper in which SEM was performed on Ab-stabilized, then fixed, brush border. I couldn't remember that so I went searching - not for you, for me. I found: http://www.uth.tmc.edu/apstracts/1995/lung/December/226l.html. My two Google searches were: "glycocalyx fixation"; and "glycocalyx antibody"
Good luck,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
I'm usually wary when someone is fixated on a particular "magic" number, 30 or 300 or whatever. Often you are dealing with a person who doesn't understand statistics nearly as well as they think they do. Since you haven't been able to convince her of your position, maybe you can ask her to justify hers? Make her really explain (and hopefully understand) where that 30-40 target comes from. I don't necessarily *recommend* this approach, since it may lead to more conflict than collaboration!
I don't know that a particular reference like the one you seek is available. As others have pointed out, it's hard to give specific advice without more specifics of your experiments. General suggestions might include 1) showing her other "comparable" microscopy examples from the literature; 2) explaining the total cost per sample in dollars and/or person-hours; 3) promising to include standard disclaimers or a brief discussion about the statistical relevance of the microscopy portion. I suspect you have already tried most or all of these. So lastly, about the only specific example I can think of is the various asbestos analysis regulations that attempt to apply statistics to microscopy (particle counting). There is a long history of trying to establish a minimum "representative" sub-sample based on sample heterogeneity. Feel free to email me if you think any of that may be helpful. Good luck!
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Thursday, November 01, 2001 9:41 AM, Paula Allan-Wojtas [SMTP:AllanWojtasP-at-EM.AGR.CA] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ------------------------------------------------------ ---- } -------------. } } } Hi, all, } } I am working on a multidisciplinary team of researchers } on a number of very interesting projects. I have a very } large collection of top quality micrographs from these } projects which I would like to publish. The microscopy } samples have been selected and photographed within the } experimental design which has been set out for each } project, so we are pretty sure they are representative of } what's there. } } One collaborator on the team is hesitant (to the point of } being resistant) about my publishing any of the } microscopy as part of any of the publications put } together by the team because we have not looked at 30-40 } samples of each of the many experimental treatments ( we } have done 4 samples of each treatment) . The other } researchers on the team have looked at between 30 and 40 } samples per treatment. This one collaborator says that } she has no confidence in the microscopy results because } "the statistics just aren't there". } } I have tried on numberous occasions to explain to her } about why this is not feasible to do in SEM, and is } usually not done. She does not believe me and is } essentially preventing me from publishing the work. I } could probably publish this work as a stand alone ms, but } it is so much more meaningful when integrated with data } from the other analytical techniques. } } My question: is there a reference (which I have somehow } missed) to explain why 30-40 samples is not feasible for } microscopy and which rationalizes why this is so? The } function of microscopy in the projects has been to give } pictorial information about the samples, and to suggest } trends, not for absolute measurement of structures and } changes in them. } } Any help you can provide would be very much appreciated - } before I rip all of my hair out.... } } Thanks in advance, listers, to all of you. I know I came } to the right place. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
} Email: griggsa-at-dbcc.cc.fl.us } Name: Alvin Griggs } } Education: Undergraduate College } } Location: Daytona Beach, FL, USA } } Question: I am looking for places that sell or lease microscope } projection oculars for use by visually impaired students. } } --------------------------------------------------------------------------- Alvin -
I would like to suggest that the current selection of add-on video cameras or direct digital input to a PC is a better way to go. You don't describe your application or budget, but simple video can be as cheap as $300. You'll find good descriptions of some possibilities at www.microscopeworld.com, but there are MANY sources.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
May 5 to 8, 2002, Palais des Congres de Montreal, Montreal, Quebec, Canada
held in conjunction with the 93nd AOCS Annual Meeting & Expo (www.aocs.org)
webaddress at the AOCS site: http://www.aocs.org/member/division/fsff/index.htm
Food Structure and Functionality Forum Bulletin Board: http://www.aocs.org/ubbcgi/ultimatebb.cgi
Tentative Technical Program Schedule (as of November 2nd, 2001)
Sunday, May 5th Short Course - Understanding structure-function relationships in food systems through specific localisation methods and microscopy. Contact: Marcel Paques (Paques-at-nizo.nl) --------------------------------------------------------------------------------------------------------------- Monday, May 6th-Morning Opening of symposium - Opening remarks
Plenary Speaker and presentation of Division Achievement Award
Dairy Applications Session. Chairs: Mark Auty, Dairy Products Research Centre, TEAGASC, Ireland (mauty-at-moorepark.teagasc.ie ) and Harjinder Singh, Massey University, NZ (H.Singh-at-massey.ac.nz)
Some Observations of a Microscopist, Author, and Reviewer on Structural Studies of Milk Products. M. Kalab, Agriculture and Agri-Food Canada, Canada
Effect of Emulsifiers and Processing Conditions on Microstructure of Milk Fat/Sunflower Oil Blends. S. Martini1, M. Cerdeira1, C. Puppo1, R.W. Hartel2, and M.L. Herrera1,3, 1CIDCA, UNLP, CONICET, Argentina; 1University of Wisconsin-Madison, USA; 3University of Buenos Aires, Argentina
Texturization of Dairy-Based Spreads. Y. Shi, B. Liang, and R.W. Hartel, University of Wisconsin-Madison, USA
Localization of Whey and Casein in Cheeses Using Microscopy and Immunochemistry Techniques. Y. Wang and D. Pechak, Kraft Foods, U.S.A
Manufacturing Yoghurt Structures with a Predicted Consumer Preference. M Langton and A. Astrom, SIK, Sweden
Dynamic Confocal Imaging of Tension and Fracture in Composite Food Materials. D.P Ferdinando1, K.P Plucknett2, and V. Normand3, 1Unilever Research, UK; 2DERA, UK; 3Firmenich SA, Switzerland
TBA - Topic: Dairy powders/caramels. C. Attapattu, University of Wisconsin, USA
Monday, May 6th - Afternoon Colloidal and Interfacial Sciences Session. Chairs: Marcel Paques (Paques-at-nizo.nl ) and David Pechak (Dpechak-at-kraft.com)
Protein Polysaccharide Interactions. C.G. De Kruif, NIZO Food Research, Netherlands (keynote speaker)
To Be Announced. B. Campbell, Kraft Foods, USA
Structure in Heat Treated Low_Fat Emulsions. R. Ofstad and V. Hoest, MATFORSK, Norway
Fatty Acid Salts-Induced Gels of Food Proteins: Their Rheological Properties and Structural Changes of the Proteins During Gelation. N. Yuno-Ohta, Nihon University, Japan
Wheat Gluten Proteins. A.S. Tatham, IACR Long Ashton research Station, United Kingdom
Interfacial Composition and Stability of Oil in Water Emulsion formed with Mixtures of Milk Proteins and Polysaccharides. H. Singh and Y. Hemar, Institute of Food, Nutrition and Human Health, Massey University, NZ
Dedicated Poster Session
Division Board Meeting -------------------------------------------------------------------------------------------------------------------- Tuesday, May 7th - Morning Agricultural Applications of Microscopy and Imaging Session/ joint with Feed Microscopy Division. Topic: Food Contamination
contacts: Mark Auty, Dairy Products Research Centre, TEAGASC (mauty-at-moorepark.teagasc.ie ) and Marge McCutcheon, West Virginia Department of Agriculture, USA (Feed Microscopy Division)
Forensic tampering. F. Platek, FDA (keynote speaker)
Contaminants in Food Processing. D. Kittleson, Pillsbury
How to approach contaminant identification. M. Auty, Dairy Products Research Centre
Growth promoters. P. Klink, South Africa
Identification of plant material. D.F. Wood, USDA
To Be Announced. J. Makowski
Quanitation of Total Fat and Fat Quality in Cheese and Dairy Products Using Membrane Separation Technology. V.C. Gordon, Safety Associates, Inc., USA
Additonal speakers to be announced.
Division Luncheon and round table (expert) discussion. Topic to be announced
Tuesday, May 7th - Afternoon Microbiology and Food Session Chairpersons: Judy Arnold and Ida Yates, USDA, ARS, Russell Research Center, USA
Prevention of Bacterial Fouling on Food Equipment Surfaces. J.W. Arnold, USDA, ARS, Russell Research Center, USA
Probiotics and Their Use in Food Animal Production. R. Droleskey, USDA, ARS, SPARC, USA
The Effect of High Pressure Steriliztion on Listeria Inoculated Seafood. K.R.S. Schneider and M.V.W. Wood, University of Florida, USA
Food Microstructure Investigations by Atomic Force Microscopy. J. Thornton, Digital Instruments/Veeco Metrology Group, USA
Controlling Growth of the Toxigenic Fungus, Fusarium Verticillioides. I. Yates and J. Arnold, Russell Research Center, ARS, USDA, USA
Division Members Meeting (immediately following the afternoon session) -------------------------------------------------------------------------------------------------------------------- Wednesday, May 8th- Morning Ingredients and Food Processing Session- Chairpersons: Diana Kittleson, Pillsbury Co, TPC Labs, USA; and Bernhard Tauscher, Federal Research Center for Nutrition, Germany
Non-Invasive Quality Determination of Fruit and Vegetables: Application of a Multi-Wavelength NIR-Diode Laser Array. B. Tauscher, Federal Research Center for Nutrition, Germany
Characterisation of the Application of Novel Oil Structurants. E. Floeter, F. Gandolfo, and W. Hogervorst, Unilever Research Vlaardingen, The Netherlands
High Pressure Processing. E. Ting and E. Raghubeer, Flow International, USA
Microstructure of Rice Starch Isolates. D.F. Wood1, A.M. Ibanez_Carranza2, and C.F. Shoemaker2, 1USDA, ARS, WRRC, USA; 2University of California, USA
To Be Announced. F. Escher, B. Conde-Petit, ETH, Switzerland
To Be Announced. M. Michel, Nestec Ltd., Nestle Research Center, Switzerland
To Be Announced. M. Salmenkallio-Marttila , VTT Biotechnology, Finland
Wednesday, May 8th - Afternoon New Methods and Techniques for Food Structure and Functionality Analysis Session Chairpersons: Kathy Groves, Leatherhead Food Research Association, England; and Maud Langton, SIK, Sweden
Quantifying Microstructures through Image Analysis. G.M.P. van Kempen1, M. van Ginkel2, C.L. Luengo Hendriks2, L.J. van Vliet2, and S. Singleton3, 1Unilever Research Vlaardingen, Netherlands; 2Delft University of Technology, The Netherlands; 3Unilever Research Colworth House, Great Britain
Cryo-TEM of Biopolymers in Comparision with Other TEM-techniques. M. Langton, A. Altskar, and A.-M Hermansson, SIK, Sweden
Freeze-substitution and low temperature embedding of dairy products for electron microscopy. A.K. Smith and H.D. Goff, Department of Food Science, University of Guelph, Canada
Recent Advances in our Understanding of the Relationship Between Crystallization Behavior, Microstructure and Rheological Properties of Fat Crystal Networks. A. Marangoni, University of Guelph, Canada
Spectroscopic Prediction of Rheological Properties in Grains. F. Meadows and F. Barton USDA, ARS, QARU, USA
Staining Techniques for Detection of Components in Fish Muscle. K. Hanneson Eggen and G. Enersen, Matforsk, Norway
Closure of Symposium ---------------------------------------------------------------------------------------------------------------------- Posters
Relationships between Microstructure and Rheological Properties of Model Lipid Systems. B. Liang, Y. Shi, and R.W., Hartel Univ. of Wisconsin-Madison, USA
Minor Biomolecules from the Olive Drupe to Olive Oil: The Technology and the Well-being Effects. N. Uccella, CIRASAIA-Mediterranean Agrifood Research Centre, Calabria University, Italy
Formation and Physical Properties of ?-Fat Gel. I: Macroscopic and Microscopic Observations. K. Higaki1, Y. Sasakura1, I. Hachiya1, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. Ii: In situ Observation of Gel-Formation Processes. K. Higaki1, Y. Sasakura1, I. Hachiya1, S. Ueno2, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. III: Rheological Properties. K. Higaki1, T. Koyano1, I. Hachiya1, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. IV: Why and How? K. Sato1, K. Higaki2, T. Koyano2, and I. Hachiya2, 1Faculty of Applied Biological Science, Hiroshima University, Japan; 2Meiji Seika Kaisha Ltd., Japan
} Date: Fri, 02 Nov 2001 07:40:47 -0800 } To: "Paula Allan-Wojtas" {AllanWojtasP-at-EM.AGR.CA} } From: Gary Gaugler {gary-at-gaugler.com} } Subject: Re: SEM - how many samples are enough? } } Coming from a semiconductor background, I have a } somewhat different take on this topic. } } In evaluating the quality of IC wafer runs, there are } two methods I use. One way is to check each wafer } while the other is to check only one wafer. Either } way, only four die off of the wafer are examined via } SEM. This is in accordance with MIL-STD-883 Method } 2018, which can be found at: } } http://www.dscc.dla.mil/Downloads/MilSpec/Docs/MIL-STD-883/std883_2018.pdf } } In particular, look at paragraph 3.1.1 and Figure 2018-4. } } This area discusses die sampling procedures for wafer lots. } Considering that one wafer may contain upwards of 5,000 die } (depending on wafer size and die size), examination of } all die, or even a large number of die is unrealistic. The issue } is whether the sample is representative of the rest of the wafers } and the die therein. There are indeed strange things that can } and do happen to wafers as they are processed such that } the results of examining a randomly-pulled wafer may not } be correct about the whole wafer lot--or some of the other } wafers in the lot. } } If there is a reasonable expectation of repeatability from } wafer-to-wafer, then examination of one wafer and the } prescribed number of die will pass or fail the whole wafer } lot. perhaps this same reasoning might apply to your } situation? I certainly don't SEM 40,000 die! There are } in-process control wafers which are examined during } the course of fabrication. Failure at any of these steps } fails the whole wafer. But the small sample at the end } of fabrication is a pas/fail for the whole wafer lot. } } If you were to be examining E. coli for example and } imaged say ten individuals, and all were rod-shaped. } How many more specimens would you have to image } to find one that was round? The issue seems to me } to be more of the characteristics of the specimens } being examined. 100% coverage will always be } someone's desired approach. But of course, this } is not practical nor realistic for most situations. Thus, } the sampling criteria is applied. Which one is } applied is the key issue. How much or how many } is sufficient to draw a conclusion? } } gary g. } } } At 06:41 AM 11/1/2001, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We did a lot of freeze substitutions of plant leaf tissues, but with fixatives. However, I do believe the principle would be the same though without fixatives you would not clearly see the ultrastructure of the cells. Following are some references you might like to check.
Xu and Mendgen (1994) Planta 195:282 Xu and Mendgen (1997) MPMI 10:87
and some publications in Dr. K. Mendgen's lab.
Hope it helps.
Haixin Xu (Ph D)
Biological Sicences University od Maryland Baltimore County 1000 Hilltop Circle Baltimore, MD 21250
On Thu, 1 Nov 2001, Christopher Ogomo wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Greetings fellow listers. } } I'm searching for freeze-substitution protocols (without using of any } fixative), preferably fast and slow protocols, for plant leaf specimen } preparation. I'm working on localizing nickel element in the plant leaves of } a hyperacumulator plant, hence using cryo preparation methods. I'll highly } appreciate any contributions and help in this. } } My thanks in advance. } } Chris Ogomo } MSc Student } University of the Western Cape } Rep of S. Africa } } } } } } } }
Thanks to all the generous folks who sent me their microspherule protocols - The info is most appreciated! Dee
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
I think this is a very interesting thread. I agree with most of the other comments contributed. It is the sort of problem that is not always clear but always important. In certain situations there are clear quantitative ways to arrive at the number of samples (measurements) required for a given confidence interval (key term), of say 95%, applied to a t-distribution assumption (usually). This type of formula can be plucked directly from statistical texts and there are even nifty little sample size calculators on the web. But, it is not always obvious how to apply that to a given measurement, and as others have said, we don't know exactly what you are trying to quantify. Within a given type of measurement, there are also ways to establish the precision of a given measurement method and then knowing that value, apply it to a single independent measurement acquired under conditions your accuracy/precision estimate was valid. There are very commonly scenarios where your final calculation also has errors that are cumulative and generally these add in quadrature. So, say you measure something and then calculate a result that involves ratios of this intensity or that, and/or multiplication of one length with another to get your final answer. Errors associated with division, multiplication and exponents are also known. In related schemes, say your calculation involves some intensity measurement and you know your the level of "noise" in that intensity measurement with a given RMS level, then that error may, in the most simple approximation, be given as 3X the rms and however that may be related to your final results would possibly be part of the quadrature-based calculation of your total error. I guess my point is there should be a way to estimate error within a given confidence interval in a single measurement if you know what the errors involved in the fundamental measurement, be it intensity, distance, etc. But, you need to break down that calculation and pick out the individual errors to determine the final error. Accuracy, versus precision, is a distinction I am sure you can appreciate.
I should say, I am definitely not a statistical expert but like you I have had to deal with this situation on several occaisons, as I am sure others have. Let's face it, it is also more work to generate error estimates for everything we do. Given the option we'd all like to plot and quantify with error bars and confidence limits, it is just not always clear how to "get there from here". My overall opinion is it is quite likely you have a valuable contribution and you need to determine some way to express qualitatively the limit in the interpretable information from the image (assuming it is an image) or quantitatively estimate that will satisfy your collegues. Good luck and please share any enlightment you might find as a results of this quest!
Regards, Ed Principe
} Paula Allan-Wojtas wrote: } } } -------------------------------------------------------------------- ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- ---. } } } } Hi, all, } } } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } } } Thanks in advance, listers, to all of you. I know I came to the right place. } } } } Paula. } } } } Paula Allan-Wojtas } } Research Scientist - Food Microstructure } } Agriculture and Agri-Food Canada } } Atlantic Food and Horticulture Research Centre } } Kentville, Nova Scotia Canada B4N 1J5 } } } } Tel: (902) 679-5566 } } FAX: (902) 679-2311 } } } } email: allanwojtasp-at-em.agr.ca
Hi listers, I am looking for an ER specific fluorophore that can be excited using visible wavelength lasers. The cells that this would be used in are cultured fibroblasts. If possible, either the excitation or emission (one or the other, it needn't be both) would be different from those of Cy3 and Cy5. If anyone knows of any good kits available for this, I would greatly appreciate it (Mol. probes has a kit, but it requires UV excitation, a laser that I unfortunately do not have).
Thanks in advance, Russell -- Russell McConnell Confocal Imaging Facility Technician Department of Neuroscience Tufts University School of Medicine M&V Building room #137 136 Harrison Ave. Boston, MA 02111 Tel. (617) 636-3795
I am forwarding the email from van Heel (sent to 3dem). It is a good summary of scheduling/booking softwares. I personally like iCal (http://www.brownbearsw.com/). Hope it'll help--Zhaojie Zhang
************************************** Zhaojie Zhang, Ph.D. Manager of Microscopy Facility University of Wyoming http://www.uwyo.edu/microscopy **************************************
-----Original Message----- } From: van Heel, Secretary T P M [mailto:vanheel.office-at-ic.ac.uk] Sent: Thursday, October 25, 2001 5:36 AM To: '3dem-at-sdsc.edu'
Paula writes ...
} ... I have a very large } collection of top quality micrographs from these projects which I } would like to publish. The microscopy samples have been selected } and photographed within the experimental design which has been } set out for each project, so we are pretty sure they are } representative of what's there. } } One collaborator on the team is hesitant (to the point of being } resistant) about my publishing any of the microscopy as part of } any of the publications put together by the team because we have } not looked at 30-40 samples of each of the many experimental } treatments ( we have done 4 samples of each treatment) . The } other researchers on the team have looked at between 30 and 40 } samples per treatment. This one collaborator says that she has no } confidence in the microscopy results because "the statistics just } aren't there". } ...
As others have implied ... how many samples depends on the variation you see. If you had 3 samples and absolutely no variation, you may be justified in stopping there and drawing a conclusion ... however, this is generally never the case. Statistical tools, such as the "chi-square distribution", are sometimes used for drawing conclusions from a small set, but even generalized chi-square formulas don't begin to yield meaningful conclusions unless the # of samples is ~30. This "rule-of-thumb" is probably at the root of your colleagues criticisms ... as if we remembered anything from "statistics 101".
The National Institute of Standards & Technology (NIST) and the Materials Science and Engineering Laboratory have post-doctoral positions open. These positions are offered competitively through the National Research Council (NRC). The Materials Structure and Characterization Group at NIST has many scientists utilizing our research facilities including TEMs (HRTEM, S/TEM, EELS, EDS) and SEMs (FEG, LaB6 equipped with OIM/EBSD).
We employ microscopy techniques to fundamental and challenging problems in materials science, semiconductors and optoelectronics. Recent topics have included semiconductor interconnections, nanometer scale multilayered materials, quantum dot materials, spintronic devices, etc.
The NIST/NRC Post Doc program offers a two-year position at an annual salary of approximately $53,200 with an additional $5,500 for research expenses. The applications are due to the NRC by Jan 15, 2002. The application must include a brief proposal and several recommendations.
A candidate must be a U.S. citizen and start work (with their PhD in hand) at NIST between July 2002 and January 2003. Students graduating this spring through next fall and others that have received their degree within the last five years are encouraged to apply. The NIST/NRC Post Doctoral program only selects positions once a year, unlike some other institutions.
You may contact me or visit our web site for further information at
Paula writes ... "I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there.
One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment). The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". "
} From Paul Webster: I am getting into this subject a little late so forgive me if I repeat something that has already been said.
The answer to the question, as posted by Michael S. a little earlier, is "... how many samples depends on the variation you see." This is exactly the solution to the problem. However, to get to a meaningful answer requires us to know what is the variation within the population. This may take a little more experimental planning. This point has been taken up in the rest of Michaels answer and also in previous replies concerning the examination of homogeneous materials samples. Homogeneous sample populations will require less sampling that heterogeneous populations.
To get a "representative" estimate of normal human blood temperature should be pretty easy as we all hover around the same temperature. Of course, a bias will be introduced if we include hospital patients in our sample. This is only a bias if we want an estimate for healthy individuals.
Conversly, obtaining an estimate for the mean height of a population will depend on who is being measured and their location. There will be many different, and correct answers for this one. Of course, the limitations of the estimation should be included with the final estimate (e.g. estimated mean height of adult males, age 35, attending a Yankees baseball game). Hwo many males should be measured so that we obtain a meaningful result? This will depend on the total population, their habits, and the sampling protocol applied. Selecting males going to the game who are seen coming out of "Big and Tall" stores may not be a representative sample!
Representative images in microscopy, selected to be "representative" are actually biased samples in that they have been selected by the operator. To get a truly "representative" result implies that unbiased estimators must be applied to the samples under investigation. An unbiased estimation should then be in the form of a quantitative result and not a "representative" image.
There are many powerful, efficient and easily applied protocols available for estimating number, length, surface area and volume. However, to be applied correctly, they must be applied to truly representative samples of the population. To obtain this unbiased sample requires the application of unbiased sample collection.
I saw in an earlier posting the cautious warning not to confuse accuracy with precision. This is an important warning. It is easy to be precise and still get the wrong answer. Imagine, for example, the result of a target practice using a gun with crooked sights but in the hands of an exceptional marksman. The holes will probably all be clustered together. The bias due to the crooked sight may cause the holes to be way of center. However, in the hands of a novice, the gun will fire off-target, but the operator may possibly be so inaccurate that a stray shot will hit the bulls-eye.
In the words of the sterologists "do more - less well". In biology, most of the variation occurs at the level of the cell or the animal. There will be less variation between samples taken from the same animal or cell population. It is far more reliable to measure small samples from more animals or cells than to take lots of images from one sample.
There are many unbiased sampling protocols available to use. However, most of them are designed to be applied to thin sectioning. It is possible that they can be adapted for use in the SEM. Estimating number and particle size should be possible using ramdom fields.
To obtain more information on these protocols I suggest looking up papers by John Lucocq and Terry Mayhew, who have both written very readable reviews of the challenges of random sampling. In particular, look for descriptions of Systematic Random Sampling. You will also see references to uniform isotropic sampling to eliminate bias.
You may come to the conclusion that taking one or two random images from your 30-40 samples, and then estimating the paramenters of interest from them is the way to go.
Microscopy has become a powerfully quantitiative branch of science. As such it is possible to express the qualitative results we see every day in unambiguous ways. We should make use of these tools to remove the fuel for argument.
In setting up the monthly archives for October of 2001 I got moderatley curious about the number of Email messages/ deliveries have run through the system running out of my home/office these past 8 years (Oct 1, 1993 was the Listserver's start date).
According to my extremely precise calculations ;-) the 100 Millionth Email message was delivered to a Microscopy Listserver subscriber sometime this year.
In case your curious, there have been 58, 280 postings to the Microscopy Listserver over the last 8 years, all of which are stored in the searchable archives. The subscriber base varies (daily), but as of this morning we had 2562 subscribers from at least 62 countries on 6 Continents. There is no one from Antarctica that I can find, or shall we say that I know about, nor is anyone from the International Space Station subscribed. :-( We did have at least one person subscribed whose mail is being forwarded to an Oceanographic survey vessel via a satellite link.
Not counted in the above totals were 6415 "suspect" messages caught by the Email filter and were rejected. Just under 3000 of these had attachments, 700+ had forged IP addresses, another ~2000ish were offers for credit/mortages/loans/viagra/porn etc... but at the same time number of valid postings { 3 % were inadvertently caught and these are generally passed through after I get contacted by the authors asking what is wrong.
I also noticed that the very first subscriber to the Listserver (Arthur Day from ANSTO just outside of Sydney Australia) is still receiving his daily dose of Microscopy Email. Well done, Arthur!
Now if I only had a beer for each of those emails, I'd be set for life.
} } Now if I only had a beer for each of those emails, I'd be set for life. { {
Actually Nestor, by my extremely precise calculations, someone should buy you your 100,000,000th beer about MSA time next year...something to look forward to in Quebec City!
Larry :-)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Listserverites... } } In setting up the monthly archives for October of 2001 } I got moderatley curious about the number of Email messages/ deliveries } have run through the system running out of my home/office } these past 8 years (Oct 1, 1993 was the Listserver's start date). } } According to my extremely precise calculations ;-) the 100 Millionth Email } message was delivered to a Microscopy Listserver subscriber } sometime this year. } } In case your curious, there have been 58, 280 postings to the Microscopy } Listserver over the last 8 years, all of which are stored in the } searchable archives. The subscriber base varies (daily), but as of this } morning we had 2562 subscribers from at least 62 countries on 6 } Continents. There } is no one from Antarctica that I can find, or shall we say that I know about, } nor is anyone from the International Space Station subscribed. :-( } We did have at least one person subscribed whose mail is being } forwarded to an Oceanographic survey vessel via a satellite link. } } Not counted in the above totals were 6415 "suspect" messages caught } by the Email filter } and were rejected. Just under 3000 of these had attachments, 700+ } had forged IP addresses, another ~2000ish were offers for } credit/mortages/loans/viagra/porn etc... but at the same time } number of valid postings { 3 % were inadvertently caught and these are } generally passed through after I get contacted by the authors asking } what is wrong. } } I also noticed that the very first subscriber to the Listserver } (Arthur Day from ANSTO just outside of Sydney Australia) } is still receiving his daily dose of Microscopy Email. Well done, Arthur! } } Now if I only had a beer for each of those emails, I'd be set for life. } } Cheers... } } Nestor } Your Friendly Neighborhood SysOp
-- Dr. Lawrence F. Allard Distinguished Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
for symmetry I think Arthur Day should be the one to buy Nestor his 100,000,000th beer, and the 17th Australian Conference on Electron Microscopy in Adelaide this coming February would be the perfect place. Adelaide is quite warm that time of year , perfect beer drinking conditions. Why don't all you listservers come and join us. Cheers,
Mark Blackford Materials Division, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} ---------- } From: Larry Allard } Sent: Monday, November 5, 2001 7:59 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Administrivia: 100,000,000 and counting } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Nestor states: } } } } Now if I only had a beer for each of those emails, I'd be set for } life. { { } } Actually Nestor, by my extremely precise calculations, someone should } buy you your 100,000,000th beer about MSA time next year...something } to look forward to in Quebec City! } } Larry :-) } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Listserverites... } } } } In setting up the monthly archives for October of 2001 } } I got moderatley curious about the number of Email messages/ } deliveries } } have run through the system running out of my home/office } } these past 8 years (Oct 1, 1993 was the Listserver's start date). } } } } According to my extremely precise calculations ;-) the 100 Millionth } Email } } message was delivered to a Microscopy Listserver subscriber } } sometime this year. } } } } In case your curious, there have been 58, 280 postings to the } Microscopy } } Listserver over the last 8 years, all of which are stored in the } } searchable archives. The subscriber base varies (daily), but as of this } } morning we had 2562 subscribers from at least 62 countries on 6 } } Continents. There } } is no one from Antarctica that I can find, or shall we say that I know } about, } } nor is anyone from the International Space Station subscribed. :-( } } We did have at least one person subscribed whose mail is being } } forwarded to an Oceanographic survey vessel via a satellite link. } } } } Not counted in the above totals were 6415 "suspect" messages caught } } by the Email filter } } and were rejected. Just under 3000 of these had attachments, 700+ } } had forged IP addresses, another ~2000ish were offers for } } credit/mortages/loans/viagra/porn etc... but at the same time } } number of valid postings { 3 % were inadvertently caught and these are } } generally passed through after I get contacted by the authors asking } } what is wrong. } } } } I also noticed that the very first subscriber to the Listserver } } (Arthur Day from ANSTO just outside of Sydney Australia) } } is still receiving his daily dose of Microscopy Email. Well done, } Arthur! } } } } Now if I only had a beer for each of those emails, I'd be set for life. } } } } Cheers... } } } } Nestor } } Your Friendly Neighborhood SysOp } } -- } Dr. Lawrence F. Allard } Distinguished Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory } 1 Bethel Valley Road } Bldg. 4515, MS 6064 } PO Box 2008 } Oak Ridge, TN 37831-6064 } } 865-574-4981 } 865-576-5413 Fax } allardlfjr-at-ornl.gov } }
Does anybody have the information about the using possibility of image recording systems (like PIXsysTEM from JEOL or the same from Fuji) and CCD cameras (for example from Gatan or another companies) in 1,000 kV TEM?
Thank you in advance, Victor Sidorenko, ANTRON Co., Ltd., scientific service, Moscow, Russia.
Do any of the more geologically-inclined persons have an e-address for the company that makes the Franz magnetic separator?
thanks
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Dear Chris, If you check our new website, em-preparation.com you can download our manual, 'AFS Recipe book'. Here you will find a chapter on the freeze substitution of plant tissue including leaves. I hope you find it useful. Regards,
Ian Lamswood Marketing Manager, EM Products Leica, Vienna
Having run a listserv I can appreciate the stable service that Nestor provides and appreciate the quality assistance the list provides. I have seen it found many things that I though were made of pure unobtainium with ease and help the list provides people world wide.
This is the best all around list I follow and I follow a lot. I don't have a lot to do being retired and not to mobile. I have a broad range of interests in science and agriculture and there is no question that Nestor and you members provide the best product out there. Be it a Nobel laureate or a 6th grader you treat their question with consideration and usualy present several approaches.
You should be presented a pin at the next meeting and clusters with each additional billion messages or a pitcher of beer your choice.
} From all of us the appreciate your work
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
} From: {zaluzec-at-sparc5.microscopy.com} Sent: Sunday, November 04, 2001 12:32 PM
Hi,
CMP Cientifica today published the first Nanotechnology Overview white paper, attempting to give a global overview of nanotechnology. Although written with a business audience in mind, there is probably plenty in there to interest anyone working at the nanoscale.
The white paper (1.5 Mb) is available for free download at www.cmp-cientifica.com & other locations.
Any feedback from the microscopy community would be much appreciated, feel free to contact me directly.
Regards
Tim
Disclaimer: The full version of the Report is a commercial product, but the White Paper is freely available at no charge.
***************************************************************** Tim E. Harper CEO CMP Cientifica s.l. Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/
I'd like to add a suggestion to the one's by Matt listed below:
} } I'm usually wary when someone is fixated on a particular } "magic" number, 30 or 300 or whatever. Often you are } dealing with a person who doesn't understand statistics } nearly as well as they think they do. Since you haven't } been able to convince her of your position, maybe you can } ask her to justify hers? Make her really explain (and } hopefully understand) where that 30-40 target comes from. } I don't necessarily *recommend* this approach, since it } may lead to more conflict than collaboration!... } } Matt
} } } } My addition; If this person is so hung up on doing 30-40 } } } } samples, peerhaps you can give him'her instructions n sample prep } } } } and then tell him'her that s/he should feel free to run up all } } } } the samples on his/her time and that you would then gladly 'scope } } } } them. I have found that when people learn by personal } } } } experience how long it takes to do even a "standard' TEM } } } } embedding they suddenly readjust their "n" requirement to a more } } } } realistic goal.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
A customer of mine has a Kevex Delta II analyzer (circa 1989) equipped with the 8 inch/10 Mb Bernoulli disk drives. He would like to continue using this media for the time being but is unable to locate any of these disks for data storage. Thermo Noran is apparently unable to upgrade his drives to something more current.
If anyone has these disks laying around, and would like to sell them, please contact me off-line. Thank you.
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 480.967.3946 "Home of the 2001 World Series Champion Arizona Diamondbacks"
Don't see what you need? Don't worry! We'll make it!
GaAs Stock (More Available, including 4") 1103SACON 76.2 mm x 635 um (100)SI,undp R} 3x107 EPD {7 x 104 2Pol Epi=09 1104SACON 76.2 mm x 635 um (100)SI,undp R} 2x107 EPD {7x104 2Pol Epi E-J Fl= ts=09 A0244 20x25mmx381um Zn HB cc=3D1018, (111) 1pol/1etch, EPI, EPD {104=09 AR250E 76.2 mm dia. (100) Semi Insul, 2 x 108 ohm-cm, As-Cut=09 A0432 2=94dia x 450 um (100) N/Te, cc=3D1-5 x 1017, 2pol, EPD {8x104=09 A0437 2=94dia x 450 um (100) P/Zn, cc=3D1-5 x 1017, 2pol, EPD {8x104=09 A0860 76.2 mm x 600 um (100) N/Te cc=3D3 x 1018, 1 Pol, EPI=09 A1291 50.8 mm x 450 um (111) P/Zn, cc=3D1-2 x 1017 EPD:3-5x104 , 2 pol Ep= i=09 A1397 76.2 mm x 600 um (100) SI, Undp R: 1-2 x 108 EPD {8x104, 2 pol, Epi=09 A1583 76.2 mm x 600 um (111) N/Te, cc} 2 x 1018, EPD {8x104, 1 pol, Epi=09 A1595 76.2 mm x 600 um (111) N/Te, cc} 1 x 1018, EPD {8x104, 1 pol, Epi=09 A1784 50.8 mm x 375 um (111) N/Si, cc} 1 x 1017, EPD {5x104, 1 Pol, Epi=09 L6049 76.2 mm x 600 um (100) P/Zn, cc=3D1-5 x 1017, EPD { 8 x 104, 2 pol,= Epi=09 L6097 50.8 mm x 400 um (110) SI, Undp R} 1x108, EPD {8x104, 2pol, Epi=09 L6133 50.8 mm x 400 um (111)Ga, SI, Undp R} 1x107, EPD {8x104, 1pol, Epi=09 L6165 50.8 mm x 400 um (100), SI, Undp R} 1x107, EPD {8x104, 2 pol, Epi=09 L6223 50.8 mm x 450 um (100) P/Zn, cc=3D2-4 x 1018, EPD { 8 x 104, 1 pol,= Epi=09 ---------
GaP L6114 50.8 mm x 300 um (111) N/S, cc: 1 - 5 x 1017 EPD { 2 x 105 2 pol EP= I=09 L6208 50.8 mm x 300 um (100),N/Undpd,cc { 1 x 1016,EPD { 2 x 105,2Pol EPI= =09
------------
GaSb 11108SA 2=94 2 Polished=09 1.75=94 (100), P-Type =09 2=94 x 4 mm (100), N/Te, Blanks, R { 1017, EPD { 104, Unpol=09 L5467 50.8 mm x 500 um (100), N/Te, cc =3D 6-8x1017, EPD { 1x103, 1 pol=09 L6116 50.8 mm x 500 um (100) P/Undpd, cc:1-2 x 1016,EPD {5 x 103, 1 Pol Ep= i=09
------=20 InAs A0254 50.8 mm x 450 um Zn/P-Type (100) 4o to(111), cc:2x1018, 1 pol/1 etc= h=09 A0300 50.8 mm x 450 um S/N-Type (100) 4o to(111)A, cc: 1018, 1 pol/1 etch= =09 A0352 50.8 mm x 450 um Zn/P-Type (100)2o to(110), cc:5-7x1018, 1pol, Epi=09 A0358 50.8 mm x 450 um S/N-Type (100) 4o to(111)A, cc: 3-5x1018, 1 pol=09 A0359 50.8 mm x 450 um S/N-Type (100) 4o to(111)A, cc:3x1018, 1 pol/1 etc= h=09
------- =09 InSb L5728 50.8 mm x 450 um (111)A, N/Undpd cc: 4-7x1014, EPD {200, 2 pol, Epi= =09 L5885 50.8 mm x 450 um (100), N/Te, cc: 3-5x1017, EPD {200, 1 pol, Epi=09 L6207 50.8 mm x 450 um (100), N/Undpd, cc: 4-7x1014, EPD {200, 2 pol, Epi= =09
-------- =09 InP 50.0 mm x 500 um Prime, SI/Fe,(100),R} 1x107,EPD {5x104,E-J,1pol,Epi=09 50.0 mm x 500 um Test, SI/Fe,(100), R} 1x107,EPD {5x104,E-J, 1pol,Epi=09
InP Indium Phosphide, LEC (100) +/- 0.2 degrees Orientation Semi Insulating, Iron doped Resistivity } 1 x 10^7 Ohm cm EPD { 5 x 10^4 cm^-2 Euro-Japanese Flats 1 Side Polished, Back Lapped and Etched 50.0 mm diameter x 500 microns thick
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InGaAs Wafer 2"=D8 with InGaAs EPI on SI InP {100} , by MOCVD deposition=20 Substrate: Semi-Insulating InP (eg. InP:Fe), Resistivity: } 1 x 10E7 Ohm = cm, EDP { 1 x 10E4/cm2)EPI: L attice matched In/Ga alloy layer of n-type InGaAs Nc } 2 x 10E18/cc (usin= g Si as dopant) Thickness: 0.5 um (+20%)
Ultra-Thin wafers now in-stock as is Float Zone and Ultra-flat wafers.
We also have fresh stocks of 2"-6" GaAs, GaSb, GaP Germanium, InP, InAs, = InSb, Al203, ZnO, ZnSe and more! They're ready to ship!
I'll forward this to the Classic Computer Collector's mailing list. Someone there certainly will have a lead on some of these.
- John
At 10:18 AM 11/5/01 -0700, Bob Roberts wrote: } A customer of mine has a Kevex Delta II analyzer (circa 1989) equipped with the 8 inch/10 Mb Bernoulli disk drives. He would like to continue using this media for the time being but is unable to locate any of these disks for data storage. Thermo Noran is apparently unable to upgrade his drives to something more current. } } If anyone has these disks laying around, and would like to sell them, please contact me off-line. Thank you. } } Bob Roberts } EM Lab Services, Inc. } 2409 S. Rural Rd Suite C } Tempe, Arizona 85282 } 480.967.3946 } "Home of the 2001 World Series Champion Arizona Diamondbacks"
----- Original Message ----- } From: "Leona Cohen-Gould" {lcgould-at-med.cornell.edu} To: {mlynn-at-miami.edu} ; {microscopy-at-sparc5.microscopy.com} } } } } Hi Paula, } } } I'd like to add a suggestion to the one's by Matt listed below: } } } } } I'm usually wary when someone is fixated on a particular } } "magic" number, 30 or 300 or whatever. Often you are } } dealing with a person who doesn't understand statistics } } nearly as well as they think they do. Since you haven't } } been able to convince her of your position, maybe you can } } ask her to justify hers? Make her really explain (and } } hopefully understand) where that 30-40 target comes from. } } I don't necessarily *recommend* this approach, since it } } may lead to more conflict than collaboration!... } } } } Matt } } Have someone that really understands statistics look at the paper. There is no magic number it all depends on the varibility of the data and what you are saying about the data.
I doesn't sound like either of you can make a strong case for your position. At least not one that convinces the other.
Good luck Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
----- Original Message ----- } From: "John Foust" {jfoust-at-threedee.com} To: "Bob Roberts" {bobrobs-at-earthlink.net} ; {Microscopy-at-sparc5.microscopy.com} Cc: {classiccmp-at-classiccmp.org} Sent: Monday, November 05, 2001 7:49 PM
I might be willing to share some of my 10 MB Bernoulli cartridges. Since I'll be holding onto some of the drives, I expect I'll not let them all go.
I do have 20 MB cartridges and drives as well, and those drives read the 10 MB media, yet write only the 20's. At one time it was convenient to put a bernoulli box at each of several clients' locations in order simply to move the cartridges along with me, yet be able to accomplish equivalent useful work at home, which has always been my primary work location.
regards,
Dick
----- Original Message ----- } From: "John Foust" {jfoust-at-threedee.com} To: "Bob Roberts" {bobrobs-at-earthlink.net} ; {Microscopy-at-sparc5.microscopy.com} Cc: {classiccmp-at-classiccmp.org} Sent: Monday, November 05, 2001 7:49 PM
Maybe..
} } } Arthur Day {ard-at-ansto.gov.au} 11/06/01 02:50PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It would be easier to send Nestor to where the beer is. Less postage that way...
} } I say we all mail Nester one beer, something regional or unique. Just } picture the scene :-) } } } -Ed
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
Tell you what - If all the List subscribers come here I can promise the Hawaii Visitors Bureau will buy you ALL a beer! And 2,562 for Nestor ...
Mahalo nui loa, Nestor!
Tina
http://www.pbrc.hawaii.edu/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Ok Sally I admit that last message was a bit ambiguous. I'll blame the Melbourne Cup BBQ for that (beer). So how do I weasel out of it?
Nestor wrote: "Now if I only had a beer for each of those emails, I'd be set for life."
If nature abhors a vacuum/empty stomach, and if there is no such thing as one beer, then each of us may have to contribute up to 100,000,000 beers each -- one per message. That's up to 100,000,000 times 2562. So, therefore, it would be easier to send Nestor to each exotic location to sample the beer in person?
Bit of an extrapolation I suppose.
If anybody needs some tips for their next grant proposal.... ;-(
} Maybe.. } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I saw a hand full of this media that VCF East...Did they get sold? I'm not sure who the seller was...maybe Tom Owad? -Chandra
-----Original Message----- } From: owner-classiccmp-at-classiccmp.org [mailto:owner-classiccmp-at-classiccmp.org] On Behalf Of John Foust Sent: Monday, November 05, 2001 9:50 PM To: Bob Roberts; Microscopy-at-sparc5.microscopy.com Cc: classiccmp-at-classiccmp.org
Having mis-spent my youth at Lehigh University, I suggest we all HAVE a beer FOR Nestor!
Fred Monson
} ---------- } From: "Edward_Principe-at-amat.com"-at-sparc5.microscopy.com } Sent: Monday, November 5, 2001 7:59 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Beer fund } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I say we all mail Nester one beer, something regional or unique. Just } picture the scene :-) } } } -Ed } } }
The web page for Franz is http://www.sgfrantz.com/
Sincerely, James Talbot
K/T GeoServices, Inc. Bulk and Clay Mineralogy by X-ray Diffraction (214) 403-6342 web site: http://www.ktgeo.com/
Ritchie Sims wrote:
} } Hello world } } Do any of the more geologically-inclined persons have an e-address } for the company that makes the Franz magnetic separator? } } thanks } } rtch } } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
} Kristen Lennon wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Semantics because this mistake has been making me crazy - ANTHRAX IS A } } BACTERIUM! This is a particularly important point because the word "virus" } } scares people silly. When I think of virus, I think of AIDS and Ebolla } } (sorry it that's misspelled). When I think of bacterium, I think antibiotic. } } No offense intended, } } Kristen } } } } Kristen A. Lennon, Ph.D. } } Department of Plant Pathology } } 351 Bessey Hall } } Iowa State University } } Ames, IA 50011 } } } } 515-294-8854 } } kalen-at-iastate.edu } } www.baumlab.org } } -- } _____________________________________________________ } Tyrone L. Daulton } Director: Marine Geosciences - Electron Microscopy Center } Physicist and Senior Electron Microscopist } } Naval Research Laboratory } Marine Geosciences Division (Code 7400) } Stennis Space Center, MS 39529-5004 } } Voice (228)-688-4877 } Fax (228)-688-4093 } email tdaulton-at-nrlssc.navy.mil } tld-at-howdy.wustl.edu }
Does anyone know what type of microscope the JEOL 100 CX TEMSCAN is/was? I've come across a reference to it in a 1982 paper by Eddy and Koehler and need to describe the SEM imaging technology for something I'm writing.
Thanks in advance for any help.
Best,
Angela
Angela V. Klaus
Director, Core Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
I think someone had once posted a URL to a website with an HTML presentation that moved the observer from the sub-atomic viewing level to one outside of our solar system using metric increments. Could someone please send me that URL again?
I still say it would be easier to have a sampling of beer, and, Oh, alright, tell Nestor how good it (OR THEY!!!!) WAS!!!!
Fred Monson I'm heading out now to start!!! See all of you who are down-time from me, later!!! } ---------- } From: Arthur Day } Sent: Tuesday, November 6, 2001 2:43 AM } To: Sally Stowe } Cc: microscopy-at-sparc5.microscopy.com; Edward_Principe-at-amat.com } Subject: Re: Beer fund } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ok Sally I admit that last message was a bit ambiguous. I'll blame } the Melbourne Cup BBQ for that (beer). So how do I weasel out of it? } } Nestor wrote: "Now if I only had a beer for each of those emails, } I'd be set for life." } } If nature abhors a vacuum/empty stomach, and if there is no such } thing as one beer, then each of us may have to contribute up to } 100,000,000 beers each -- one per message. That's up to 100,000,000 } times 2562. So, therefore, it would be easier to send Nestor to each } exotic location to sample the beer in person? } } Bit of an extrapolation I suppose. } } If anybody needs some tips for their next grant proposal.... ;-( } } } } Maybe.. } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It would be easier to send Nestor to where the beer is. Less postage } } that way... } } } } } } } } I say we all mail Nester one beer, something regional or unique. Just } } } picture the scene :-) } } } } } } } } } -Ed } } }
Comments to comments: } } I'm usually wary when someone is fixated on a particular } "magic" number, 30 or 300 or whatever. Often you are } dealing with a person who doesn't understand statistics } nearly as well as they think they do. Since you haven't } been able to convince her of your position, maybe you can } ask her to justify hers? Make her really explain (and } hopefully understand) where that 30-40 target comes from. } I don't necessarily *recommend* this approach, since it } may lead to more conflict than collaboration!... } } Matt
} } } } My addition; If this person is so hung up on doing 30-40 } } } } samples, peerhaps you can give him'her instructions n sample prep } } } } and then tell him'her that s/he should feel free to run up all } } } } the samples on his/her time and that you would then gladly 'scope } } } } them. I have found that when people learn by personal } } } } experience how long it takes to do even a "standard' TEM } } } } embedding they suddenly readjust their "n" requirement to a more } } } } realistic goal. Lee
Reply: Unfortunately (or fortunately), the actual sample number is not arrived at in an arbitrary manner, it is dictated by the nature of the specimen under study.
Nobody makes up these numbers, but if you look at the article by John Lucocq, Trends in Cell Biology 1993 Unbiased 3-D quantitation of ultrastructure in cell biology 3:354-358, he suggests using 3-5 experiments/animals/cultures, only 1 or more blocks/dishes, one or more sections but 10-40 micrographs and between 100-200 probe counts. I am pretty sure this is a common sampling scheme for stereologists.
Paraphrasing slightly form this manuscript, Sampling is "..an inherent part of any quantitative electron micrscopy study since only a portion of all the possible blocks, sections and micrographs are analyzed. It is essential therefore that at each level in a sampling scheme items are selected in a way that gives all parts of the specimen the same chance of being included."
The actual amount of sampling that is performed will be directly related to the variance occurring within the population. Samples with low variance will require a lower sample number to give you an answer with an error of 5% or less. High variance between individual samples will require more sampling to be performed.
These are all experimental design considerations that have to be applied much before all the clever statistical analysis is performed. Working out probabilities on poorly sampled data is not very useful.
It can be said that proper sampling techniques should also be applied to morphological data too. Where would that put all those text-book images?
AIDS is a name of the illness caused by VIRUS, I believe. Virus itself has a name: HIV, Human Immuno-deficit Virus. Sergey
At 10:36 AM 11/6/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Yes, Ebola is a virus, AIDS is caused by a virus and Anthrax is caused by a bacterium. How come I, as a lowly physicist, know this but others do not, although they take great delight in trumpeting their ignorance for all of us to see!
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } AIDS is a name of the illness caused by VIRUS, I believe. Virus } itself has a name: HIV, Human Immuno-deficit Virus. } Sergey } } At 10:36 AM 11/6/01, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Note new cell phone number!
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
I'm a bit of a novice with antibody staining, but so are my colleagues and so they've all got conflicting and inconsistent results from trying the Alkaline Phosphatase staining.
We're trying to stain sea urchin larvae whole-mounts with the 2 antibody-alkaline phosphatase system(larvae are formalin fixed and rehydrated from 70% EtOH cold storage). Our AkPh substrate is a few months old and has a small amount of blackish precipitate at the bottom--is this OK? My colleagues have had different results with using the substrate undiluted on the animals vs. diluted in some small amount of PBS. I notice that with the latter, there is a slight whitish precipitate that occurs--is this due to the pH differential between the AkPh substrate(ph 10 or more) and PBS(7.4)? What are the ideal staining conditions for this to work? Please note--I'm getting problems with the preliminary staining where I know I'm supposed to get endogenous gut AkPh staining with just substrate and nothing else, but I'm getting almost imperceptible staining.
All suggestions and admonishments will be taken into consideration. Thanks in advance.
Pauline Yu pcy-at-usc.edu Manahan Research Laboratory http://www.usc.edu/manahanlab University of Southern California
There's a high school teacher in Hull, Massachusetts with 20 or so microscopes that need cleaning and (hopefully minor) repair. She's willing to work on them herself, but doesn't know how to begin. Is there a lister who can visit and help her get started? If so please contact me.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
on 11/4/01 4:39 PM, Victor Sidorenko at antron-at-space.ru wrote:
} } Does anybody have the information about the using possibility of } image recording systems (like PIXsysTEM from JEOL or the } same from Fuji) and CCD cameras (for example from Gatan or } another companies) in 1,000 kV TEM? } Dear Victor, A few years ago (when the 1250 kV scope had been installed at Stutgart, but I don't remember the exact year) there was a poster at MSA which gave the quantum efficiency for image plates as a function of voltage up to 1250 kV. Since I was interested in quantitating ED data at 1200 kV, and since I had heard great things about IPs--single-electron sensitivity, linearity for a range which would include all the expected intensities in an ED pattern, etc.--I remember the graph well. The QE for somewhat more than 100 kV was 100% (giving the required sensitivity), but it dropped off at intermediate voltages--I don't remember whether it was still 100% at 200 kV, but definitely lower at 400 kV--and was only ~25% at 1000+ kV. I don't know what improvements may have been made; a thicker active layer will give more sensitivity, but lower spatial resolution (there is no free lunch). The manufacturers of the plates would be able to tell you whether they can sell you what you would need. Good luck. Yours, Bill Tivol
ACEM 17 The 17th Australian Conference on Electron Microscopy Adelaide Convention Centre, Adelaide, South Australia February 4 - 8, 2002 Hosted by Australian Society for Electron Microscopy (Inc.)
This meeting will provide an opportunity to explore the many applications of Electron, Scanned Probe and Confocal Microscopies, to keep abreast of the latest developments and to hear and interact with many of the leaders in the field. Sessions on Cryotechniques, Virology/Microorganisms, Cell Tomography, Nanotechnology, Emerging EM Technologies, Materials and Microstructure, Superresolution and an Industry Forum are being planned, among many others.
Invited and Keynote Speakers (to date) Martin Bootman; Confocal (biological), David Cockayne; Nanostructure, Patrick Echlin; Analytical Microscopy of Biomaterials, Mark Fricker; Confocal (biological), David Jefferson; Emerging Techniques, Jan Leunissen; Immunolabelling, Ohad Medalia; Cellular Tomography, Greg Meeker; Microbeam Analysis, Sara Miller; Virology, Daniel Mueller; SPM - (biological), David Williams; Microanalysis, Nestor Zaluzec; Analytical Microscopy/Telepresence Jamie Chapman; Reproductive Biology, Jacques Dubochet; Cryotechniques, Alex Hyatt; Virology/Cryotechniques, Sergei Magonov; SPM, Mary, Mah-Lee Ng; Virology/Confocal
A series of pre-conference workshops will be held to allow participants to gain hands-on experience in the latest techniques and applications. Many of the invited and keynote speakers will participate as presenters, providing a wonderful opportunity for interaction with these noted scientists. Be sure to book soon, as places in some of the workshops are limited.
Workshops Cryo Scanning Electron Microscopy Environmental Scanning Electron Microscopy, Techniques and Applications Image Analysis Leica-Aurion Workshop on Cryotechniques and Immunolabelling Light Element Analysis: Problems and Solutions Quantitative Confocal Microscopy Scanning Probe Microscopy Trace and Major Quantitative X-ray Mapping (a Hands on Approach) Virology Using Electron & Light Microscopy
During the week, delegates will have the chance to mingle and relax after hours in several of Adelaide's noted heritage locations. A range of accommodation choices, from five-star to budget, ensures choices suitable for all.
The Trade Exhibition will be a comprehensive and exciting display of the latest equipment and accessories for microscopy, imaging and analysis. We are most grateful to our sponsors and exhibitors for their very generous support of ACEM17.
As is traditional at all ACEMs, Poster and Micrograph Competitions will be run. Categories for the Poster Competition will be Best Biological Poster, Best Physical Poster and Best Student Poster. Categories for the Micrograph Competition are SEM, TEM, SPM and Confocal. Entries will be judged on aesthetic, photographic and scientific merit. Entry details are available on the website.
With the deadline for abstract submission fast approaching, (November 30), make plans now to join us in February! (Early bird registration deadline - November 16). Abstracts should be submitted electronically, and the registration form can be downloaded from the website - visit often for regular updates.
We look forward to seeing you at ACEM17.
Full details can be obtained at http://www.adelaide.edu.au/CEMMSA/acem17
The JEOL 100CX TEMSCAN is a 100kV transmission electron microscope (TEM) with the attachment for scanning imaging device (ASID) that provides scanning electron microscope (SEM) facilities (the SCAN part of TEMSCAN). It could image with secondary electrons (SE) or backscattered electrons (BSE) in a standard SEM mode or use brightfield or darkfield detectors for scanning transmitted electron microscopy (STEM). With an energy dispersive x-ray (EDX) specrometer fitted it could also provide chemical analysis, x-ray maps and line scans.
Undoubtably there are still some out there being used and their users could give you more information but your local JEOL office could also give you any information you want (ask for an engineer over 40). If you don't know the local JEOL office look on the web, although they maybe rushing out to you now in case you want to buy something!!!
Regards, Ron
On Tue, 6 Nov 2001 14:00:30 -0500 (EST) Angela Klaus {avklaus-at-amnh.org} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all... } } Does anyone know what type of microscope the JEOL 100 CX TEMSCAN is/was? } I've come across a reference to it in a 1982 paper by Eddy and Koehler and } need to describe the SEM imaging technology for something I'm writing. } } Thanks in advance for any help. } } Best, } } Angela } } Angela V. Klaus } } Director, Core Imaging Facility } American Museum of Natural History } Central Park West at 79th Street } New York, NY 10024 USA } } Email: avklaus-at-amnh.org } Tel: 212-769-5977 } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
The 100CX TEMSCAN is a JEOL 100kV TEM fitted with a scanning attachment which allows imaging by way of "in lens" surface studies (SEM) in addition to transmitted electron scanning (STEM). The microscope was a second generation 100C hence the X.
The instrument was designed by Yanaka and Shirota the scanning system interface was made by Koike (the man who invented the in lens SEM system). Koike went on to design the ISI DS130 SEM and Yanaka and Shirota the ISI 002B TEM.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
The 100CX TEMSCAN is a JEOL 100kV TEM fitted with a scanning attachment which allows imaging by way of "in lens" surface studies (SEM) in addition to transmitted electron scanning (STEM). The microscope was a second generation 100C hence the X.
The instrument was designed by Yanaka and Shirota the scanning system interface was made by Koike (the man who invented the in lens SEM system). Koike went on to design the ISI DS130 SEM and Yanaka and Shirota the ISI 002B TEM.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
Angela Klaus wrote: Does anyone know what type of microscope the JEOL 100 CX TEMSCAN is/was?
If we're talking about the same instrument, the JEOL 100 CX that I spent years on was a TEM (100 kV with a booster switch to 120 kV accelerating voltage). Mine had the SAED (selected area electron diffraction) accessory and Scanning EM functions.
Coincidentally, I know where just such an instrument is available for the cost of tear down and shipping to any federal institution OR an institution with an active NSF grant using electron microscopy. It includes a full-sized, pneumatic anti-vibration table, chiller, compressor, film carriers, etc.
Contact me off-line if interested and I'll see if it's still available.
S. Frank Platek Forensic Chemistry Center U.S. Food and Drug Administration 6751 Steger Drive Cincinnati, OH 45237-3097 (513) 679-2700 X254 (513) 679-2761 FAX fplatek-at-ora.fda.gov
We are interested in doing EELS work with a LaB6 filament TEM. Can anyone tell us how best is the energy resolution with this type of filament?
Thanks
Best Regards Yan Xin ======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
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That would be "Powers of 10", by Eames. I know it as a video, book, and flip book (see the MICRO bibliography, URL below), but not as a website. Will you please post the URL on the list when you get it?
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Absolutely correct John! And, since you know, I'll ask.
Which physicist is most responsible for "nucular" physics [did I spell it right???], and which Computer Scientist for "data is"?
I know the biologists are responsible for "dIsection"!
Blessings upon the lexicographers who say, "Cut it any way you want! DIsect it with a cleaver or DISsect it with fine forceps. Your choice, whether you know the difference or not!"
As Grandfather used to say when Grandmother was near, "Oh! For the love of words!!!"
Fred Monson, who hath some affection for the silly!
P.S. By the way, Nestor, I stopped on the way home last evening, tried a few different brands to be found in Eastern Pennsylvania, just Northwest of Philadelphia, and most were worth the try. Every one went down as a toast to your good works, and that was good too! So, you are taken care of here, and I hope you appreciate the effort on your behalf. FCM
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: John F. Mansfield } Sent: Tuesday, November 6, 2001 9:12 PM } To: microscopy-at-microscopy.com } Subject: Virus and Bacterium } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Yes, Ebola is a virus, AIDS is caused by a virus and Anthrax is } caused by a bacterium. How come I, as a lowly physicist, know this } but others do not, although they take great delight in trumpeting } their ignorance for all of us to see! } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } AIDS is a name of the illness caused by VIRUS, I believe. Virus } } itself has a name: HIV, Human Immuno-deficit Virus. } } Sergey } } } } At 10:36 AM 11/6/01, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } AIDS isn't a virus either. More semantics. } } } } } } } } } } Kristen Lennon wrote: } } } } } } } } } } ------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } Semantics because this mistake has been making me crazy - ANTHRAX IS } A } } } } } BACTERIUM! This is a particularly important point because the word } "virus" } } } } } scares people silly. When I think of virus, I think of AIDS and } Ebolla } } } } } (sorry it that's misspelled). When I think of bacterium, I think } } } antibiotic. } } } } } No offense intended, } } } } } Kristen } } } } } } } } } } Kristen A. Lennon, Ph.D. } } } } } Department of Plant Pathology } } } } } 351 Bessey Hall } } } } } Iowa State University } } } } } Ames, IA 50011 } } } } } } } } } } 515-294-8854 } } } } } kalen-at-iastate.edu } } } } } www.baumlab.org } } } } } } } } -- } } } } _____________________________________________________ } } } } Tyrone L. Daulton } } } } Director: Marine Geosciences - Electron Microscopy Center } } } } Physicist and Senior Electron Microscopist } } } } } } } } Naval Research Laboratory } } } } Marine Geosciences Division (Code 7400) } } } } Stennis Space Center, MS 39529-5004 } } } } } } } } Voice (228)-688-4877 } } } } Fax (228)-688-4093 } } } } email tdaulton-at-nrlssc.navy.mil } } } } tld-at-howdy.wustl.edu } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } -- } Note new cell phone number! } } Dr. John Mansfield CPhys MInstP } North Campus Electron Microbeam Analysis Laboratory } 417 SRB, University of Michigan } 2455 Hayward, Ann Arbor MI 48109-2143 } Phone: (734) 936-3352 FAX (734) 763-2282 } Cellular Phone: (734) 834-3913 } (Leaving a phone message at 936-3352 is preferable to 834-3913) } Email: jfmjfm-at-engin.umich.edu } URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html } Location: Lat. 42° 16' 48" Long. 83° 43' 48" } }
Does anyone know a recipe for chemically etching GaN and/or InN? The material is already thinned and ready for TEM, we just want a final surface etching to enhance dislocation imaging.
Thanks,
Mick
------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Angela Klaus } Sent: Tuesday, November 6, 2001 2:00 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Info needed: JEOL 100 CX TEMSCAN } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all... } } Does anyone know what type of microscope the JEOL 100 CX TEMSCAN is/was? } I've come across a reference to it in a 1982 paper by Eddy and Koehler and } need to describe the SEM imaging technology for something I'm writing. } } Thanks in advance for any help. } } Best, } } Angela } } Angela V. Klaus } } Director, Core Imaging Facility } American Museum of Natural History } Central Park West at 79th Street } New York, NY 10024 USA } } Email: avklaus-at-amnh.org } Tel: 212-769-5977 } } }
} -----Original Message----- } From: Sy Griffey } Sent: Wednesday, November 07, 2001 11:48 AM } To: Jacqueline D. Garfield } Subject: Processing of tendon samples } } } } If anyone has some experience processing human tendon samples for EM } and/or histology we would appreciate your help. } } Problem: We have begun processing human tendon samples for both EM and } standard histology but are having trouble getting good sections and tissue } quality. The samples for histology can be as thick as 5-8 mm. } } For the EM samples we have only done a freeze-substitution protocol } to date but the tissue was extremely hard before embedding and even thick } sectioning was a challenge. We anticipate similar problems when doing } standard EM processing based on experience with histology processing thus } far........The tissue in the histology sections appears cracked, } exhibiting separation within the fibrous bundles of the tendon when viewed } in X-sxn. The samples were actually somewhat flexible just prior to } embedding. All processing steps (graded alcohol, clearing solution and } paraffin) have been done at ambient temperature and pressure. The } incubation times, in general, have been in the 45-60 min. range. We are } thinking about increasing the incubation times and perhaps doing some } vacuum infiltration but would welcome any suggestions. } } Thank you! } } Jackie Garfield and Sy Griffey } } Lifecell Corporation } One Millennium Way } Branchburg, NJ 08876 USA } (908) 947-1182 (Jackie) } (908) 947-1085 (Fax) } jgarfield-at-lifecell.com
Thank you for the many replies regarding the problem of firmly-attached carbon.
It appears that in removing the metal coating left on the glass surface in the bell jar's previous life in a physics lab, the aqua regia that I ended up using activated the glass surface, perhaps by producing a micro-roughness. So the first few uses produced a firmly-attached carbon coating.
I was able to remove the carbon with a mildly-abrasive ("without harsh scratching") household cleaner, plus lots of rubbing.
I then wiped the surface with a rag wetted with kerosine, then with a clean dry rag. This left sufficient kerosine to prevent a good vacuum being obtained, so I sprayed it with a household general-purpose multi-solvent cleaner ("Ajax Spray 'n' Wipe), and wiped that off with a clean dry rag.
This has left the surface sufficiently deactivated so that I can remove new carbon simply by spraying with Spray 'n' Wipe and wiping.
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
A graduate student in microbiology has just finished her first course in electron microscopy, and she informs her advisor that she wants to study mitochondrial structure and function in several related strains of yeast, for one of which she fortuitously has electron micrographs of mitochondrial profiles that are, she has been told, quite publishable.
Her advisor suggests that she start by comparing 3-4 strains with respect to number of mitochondria per cell and total mitochondrial cristal area under identical conditions of culture.
The student happily responds that she already has micrographs for regularly sampled sections from serial sections she learned to make of the strain she used in her EM course. She returns the next week with estimates of cristal area and numbers of mitochondria per cell based on morphometric analysis of those regularly sampled sections.
If the cell is 12-25um in length (haploid and diploid) and always between 3-5um in diameter, and if the mitochondrial profiles are in the range of 1-2um (circular and ovoid, and sometimes or often branched), what should the sampling criteria be for cells that are serially sectioned with sections approximately 90nm in thickness?
How could one derive the same, but apparently much more statistically satisfactory, data from the 40+ cell profiles per section of cells randomly oriented and not necessarily (or importantly) sectioned or analyzed completely.
All who take on this task should know there is an extremely important, and possibly significant, hint that can be observed at: http://www.msg.ucsf.edu/sedat/marsh/mito.html. I sometimes think it is well to know of such hints (or at least to be aware of their possibility) before attempting a task. Assumptions are almost always incorrect unless rigorously defined and applied, and very often difficult to prove as such or to isolate in generating an experimental design without flaws.
Knowing that reality often gets in the way of statistics, and vice versa, I am
Yours most respectfully, because I wrestle with this problem every once in a while and I haven't yet determined a satisfactory answer.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
I just looked in Yahoo and found the website. It's www.powersof10.com.
Edward Haller University of South Florida Pathology Department Tampa, Florida
Caroline Schooley wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear All, } } } } I think someone had once posted a URL to a website with an HTML } } presentation that moved the observer from the sub-atomic viewing level to } } one outside of our solar system using metric increments. Could someone } } please send me that URL again? } } } } Thanks, } } } } Louis - } } That would be "Powers of 10", by Eames. I know it as a video, book, and } flip book (see the MICRO bibliography, URL below), but not as a website. } Will you please post the URL on the list when you get it? } } Caroline Schooley } Project MICRO Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Can anyone in the DFW area perform these types of analysis? Respond to me with details and contact information.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
-----Original Message----- } From: Rajan Vempati [mailto:rvempati-at-mail.smu.edu] Sent: Wednesday, November 07, 2001 10:16 PM To: rbeavers-at-mail.smu.edu
I am looking for a simple, reliable method for obtaining thin sections for EM from thick (1-2 microns) epoxy resin sections that have been cut, mounted on slides, and stained for preliminary examination with the LM .
Hi All, After Caroline's message I did a web search for "powers of 10" and found this web site which seems to fit what was requested. http://www.wordwizz.com Steve
Steve Buckingham Dir. of Process Development Excellatron Solid State LLC 1640 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 617 812 5920
} -----Original Message----- } From: Boche, Jo Ellen } Sent: Wednesday, November 07, 2001 4:33 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: TEM Request for method for cutting thin sections from } thick. } } } } I am looking for a simple, reliable method for obtaining thin sections } for EM from thick (1-2 microns) epoxy resin sections that have been } cut, mounted on slides, and stained for preliminary examination with } the LM. } } bochej-at-boystown.org } } Jo Ellen Boche } Neuroanatomy Laboratory } Boys Town National Research Hospital } Omaha, NE 68131 } (402) 498-6531 } } .
You can "just" re-embed the sections directly on the slide (invert the slide-bound sections on BEEM capsules filled with resin or put the capsule on the slide), repolymerize, and cut. I always manage to make a serious mess with the resin, but it does work.
You may also be able to glue the thick section to a polymerized blank block and then pop it off the slide with a heat/cold regimen - I haven't tried this but it is supposed to work. I think it may be more useful/less disastrous with a REALLY thick section than a normal thick section.
Good luck!
Tamara
On Wed, 7 Nov 2001, Boche, Jo Ellen wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } -----Original Message----- } } From: Boche, Jo Ellen } } Sent: Wednesday, November 07, 2001 4:33 PM } } To: 'Microscopy-at-MSA.Microscopy.Com' } } Subject: TEM Request for method for cutting thin sections from } } thick. } } } } } } } } I am looking for a simple, reliable method for obtaining thin sections } } for EM from thick (1-2 microns) epoxy resin sections that have been } } cut, mounted on slides, and stained for preliminary examination with } } the LM. } } } } bochej-at-boystown.org } } } } Jo Ellen Boche } } Neuroanatomy Laboratory } } Boys Town National Research Hospital } } Omaha, NE 68131 } } (402) 498-6531 } } } } . } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
For this reason it's more correctly to call it: "mitochondrion" (M), instead mitochondria. In my point of view, on EM level only 3D reconstruction from the serial sections may give you truthful info about this structure.
By the way, mitochondria/mitochondrion is known as an extremely sensitive organelle: to oxidative stress etc&etc. Its structure may be changed by very slight variation of condition/procedure. Simple example: you are looking how some particular mutation affected M structure. It's very possible, that your mutation will affect the membrane permeability nor M at all. On EM level this may cause the different conditions during sample fixation/processing. Finally you will detect M changes, because the processing conditions were different for control and mutant. Another example: mutants are growing slowly at many instances, it's very difficult to have the culture's "age" similar. People usually grow cells to the some particular optical density (at 450 nm?). So the control will grow up to 1 OD, let say 5 h, but the mutant - 18+ h. Are the "age" of the cells the same? I guess, not. The structure of the M in the cells of different "age" may (and could) be different. So, we have to be careful with M as, from another hand, all EM stuff we have deal with. Sergey
At 12:53 PM 11/7/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (george_greg-at-msn.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, November 7, 2001 at 19:50:43 ---------------------------------------------------------------------------
Email: george_greg-at-msn.com Name: Greg George
Education: 9-12th Grade High School
Location: Pearl, Mississippi USA
Question: Dear Sir, I recently received, as a gift, a used lab grade microscope without a user manual. Three of the four (oil immersion) objectives are spring retractable. Does this feature mean that I can touch the slide surface without fear of damaging the lens? Thank you for your kind reply, Greg George
Powers of 10 is also available as a Java applet at http://micro.magnet.fsu.edu/primer/java/scienceopticsu/powersof10/ At this moment, the website is inaccessible to me so I cannot verify for sure. Best Wishes.
-----Original Message----- } From: Caroline Schooley [SMTP:schooley-at-mcn.org] Sent: Thursday, November 08, 2001 9:25 AM To: Louis Somlai Cc: Microscopy-at-sparc5.microscopy.com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
That would be "Powers of 10", by Eames. I know it as a video, book, and flip book (see the MICRO bibliography, URL below), but not as a website. Will you please post the URL on the list when you get it?
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
The first site mentions Eames and would be the wanted site. The second site too is very good and worth a long look. Its part of the molecular expressions site at Florida State Uni. The third site gives a power of ten perspective on computer data sizes. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, November 08, 2001 1:46 AM, Caroline Schooley [SMTP:schooley-at-mcn.org] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear All, } } } } I think someone had once posted a URL to a website with an HTML } } presentation that moved the observer from the sub-atomic viewing level to } } one outside of our solar system using metric increments. Could someone } } please send me that URL again? } } } } Thanks, } } } } Louis - } } That would be "Powers of 10", by Eames. I know it as a video, book, and } flip book (see the MICRO bibliography, URL below), but not as a website. } Will you please post the URL on the list when you get it? } } } Caroline Schooley } Project MICRO Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html }
on 11/7/01 10:46 AM, Yan Xin at xin-at-magnet.fsu.edu wrote: } We are interested in doing EELS work with a LaB6 filament TEM. Can anyone } tell us how best is the energy resolution with this type of filament? } Dear Yan Xin, As I remember it, the energy spread in a LaB6 filament is about 1-2 eV. The energy resolution obtainable in EELS is affected by factors other than the filament, however, and I do not know the best that can be obtained with your instrument. Yours, Bill Tivol
We are trying to section a tampon in to about five micrometers but any normal sample prep involves EtOH or some other fluid that will be absorbed. i was wondering if anyone knew a method to keep it in at its normal size but in a form that can be cut very thin thanks john
} ===== Original Message From "Monson, Frederick C." {fmonson-at-wcupa.edu} ===== } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
With effect from 1st November 2001 the Polaron Range of EM preparation instruments will change ownership from Thermo VG Scientific to Quorum Technologies Ltd.
Quorum Technologies Ltd. is the company formed by Bob Kenhard, Mike Wombwell, David Cheshire and the existing staff within Thermo VG Scientific involved in the management, development, sales and manufacturing of the Polaron Range.
We will continue to support the many Polaron instruments out in the field (including EM preparation instruments branded Polaron Equipment Ltd, Emscope, Bio-Rad Microscience, Fisons Instruments and VG). Additional details are available on the new Quorum WWW site ( http://www.quorumtech.com).
The new contact address (effective Nov. 12) will be
Quorum Technologies Unit 15A Euro Business Park New Road Newhaven East Sussex U.K. BN9 0DQ
Contacts: Mike.wombwell - sales (mike.wombwell-at-vgscientific.com - after 12th November - mike.wombwell-at-quorumtech.com) David Cheshire - technical support (david.cheshire-at-vgscientific.com - after 12th November - dave.cheshire-at-quorumtech.com) Bob Kenhard - sales and other enquiries (bob.kenhard-at-vgscientific.com - after 12th November - bob.kenhard-at-quorumtech.com) WWW: http://www.quorumtech.com US Contact: Energy Beam Sciences (telephone: 413 786 9322 E-mail - sales-at-ebsciences.com)
Mike Wombwell Polaron Range Sales Director Quorum Technologies
At 14:32 2001-11-06 -0600, Louis Somlai wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
Louis Somlai wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All, } } I think someone had once posted a URL to a website with an HTML } presentation that moved the observer from the sub-atomic viewing level to } one outside of our solar system using metric increments. Could someone } please send me that URL again? } } Thanks, } } Louis } Louis.Somlai-at-usm.edu
Would anyone know how to go about getting official certification for histo in the KS and MO region.
Thanks Tim Quinn University of Kansas Research Assistant Natural History Museum and Biodiversity Research Center Dyche Hall Room 414 Lawrence, KS 6604-2454 785-864-4556 tquinn-at-ku.edu
With effect from 1st November 2001 the Polaron Range of EM preparation instruments will change ownership from Thermo VG Scientific to Quorum Technologies Ltd.
Quorum Technologies Ltd. is the company formed by Bob Kenhard, Mike Wombwell, David Cheshire and the existing staff within Thermo VG Scientific involved in the management, development, sales and manufacturing of the Polaron Range.
We will continue to support the many Polaron instruments out in the field (including EM preparation instruments branded Polaron Equipment Ltd, Emscope, Bio-Rad Microscience, Fisons Instruments and VG). Additional details are available on the new Quorum WWW site ( http://www.quorumtech.com).
The new contact address (effective Nov. 12) will be
Quorum Technologies Unit 15A Euro Business Park New Road Newhaven East Sussex U.K. BN9 0DQ
Contacts: Mike.wombwell - sales (mike.wombwell-at-vgscientific.com - after 12th November - mike.wombwell-at-quorumtech.com) David Cheshire - technical support (david.cheshire-at-vgscientific.com - after 12th November - dave.cheshire-at-quorumtech.com) Bob Kenhard - sales and other enquiries (bob.kenhard-at-vgscientific.com - after 12th November - bob.kenhard-at-quorumtech.com) WWW: http://www.quorumtech.com US Contact: Energy Beam Sciences (telephone: 413 786 9322 E-mail - sales-at-ebsciences.com)
Mike Wombwell Polaron Range Sales Director Quorum Technologies
At the risk of looking very stupid, I will take your challenge.
Am I correct in understanding that your question has two parts? 1. What is the mean number of mitochondria per yeast cell in a population. 2. What is the mean area of cristae membranes.
If this is so, then there are actually three challenges here because to get an absolute number for the cristae surface area and the number of mitochondria per cell we also need to have an estimate for a reference space in actual numbers. A good reference space to estimate is cell volume.
Obtaining mean number of mitochondria per cell is actually very easy. I would first throw away the images already obtained because they were subjectively sampled.
We must then remove bias that may be a result of centrifugation - heavier (more dense) cells will pellet to the bottom, with the possibility that a gradient is formed in the pellet. To avoid this, embed the cells first in fast-setting gelatin or agarose, being careful to make sure the cells have been thoroughly mixed. We could of course do it properly and apply the "orintator" to make sure our sampling is isotropic (Mattfeldt et al 1989 Acta Stereol. 8:671-676).
Fix the block and cut it into slabs, rods and then cubes. Using a sequential random sampling protocol, take every "nth" block (decided using random number generator) and embed them separately in resin (see Lucucq 1993 Trends Cell Biol 3:345-358 for details).
Section for counting. A good way to sample the cells to count particles (here is where the catch comes in) is to only count the tops of the particles. With the yeast mitochondria, the 2-D profile does not always represent an individual particle. A profile may represent one single mitochondrion, or may represent a multiply branched, single mitochondrion, depending on the level of branching that is occurring. By counting the tops, connected profiles are eliminated from the sampled population. (for details on a possible method for estimating the amount of branching that is present, look for "Star Volume" in Gundersen et al 1986 J. Microsc. 143:3-45)
How to sample? Use the "Disector" (in this instance, the word means "2 sectors"). The reference is D.C. Stereo (a pseudonym for HJG Gundersen) 1984. J. Microsc. 134:127-136.
To apply, cut two sequential sections and take similar fields of view from both. Use one field to examine for mitochondrial profiles and the other to check if the profile is also present there ( the look-up frame). If a profile is present in one field but not present in the other, it is counted as a "top". (It is a variation of the 2-D counting rule, the associated tangent rule).
Knowing the area of the field of view and the thickness separating the two sequential sections, we can work out the volume being sampled. This will give us an estimate of the number of mitochondria per unit volume. This is not the number of mitochondria per cell. We still need to estimate the cell volume.
To estimate surface area, we can use simple cross lattice overlays and count the number of times a membrane profile crosses a test line. From this we can estimate length of membrane, and by knowing section thickness, can obtain an estimate of membrane area. (see H.J.G. Gundersen et al 1986 J. Microsc. 143:3-45. 1988 APMIS 96:379-394 & 857-881, T.M. Mayhew 1991 Exp. Physiol. 76:639-663, J. Lucocq for details on how to do this) We still need to estimate the cell volume.
How am I doing so far, Fred?
Now, a question for you. I can provide estimates of relative values for number and surface area. However, I do need a reference space to obtain absolute values. How would I obtain a reliable estimation of the mean cell volume of the yeast population? Presumably they are a rapidly dividing population.
Regards
Paul Webster.
on 11/7/01 12:53 PM, Monson, Frederick C. at fmonson-at-wcupa.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hypothetical! } } A graduate student in microbiology has just finished her first course in } electron microscopy, and she informs her advisor that she wants to study } mitochondrial structure and function in several related strains of yeast, } for one of which she fortuitously has electron micrographs of mitochondrial } profiles that are, she has been told, quite publishable. } } Her advisor suggests that she start by comparing 3-4 strains with respect to } number of mitochondria per cell and total mitochondrial cristal area under } identical conditions of culture. } } The student happily responds that she already has micrographs for regularly } sampled sections from serial sections she learned to make of the strain she } used in her EM course. She returns the next week with estimates of cristal } area and numbers of mitochondria per cell based on morphometric analysis of } those regularly sampled sections. } } If the cell is 12-25um in length (haploid and diploid) and always between } 3-5um in diameter, and if the mitochondrial profiles are in the range of } 1-2um (circular and ovoid, and sometimes or often branched), what should } the sampling criteria be for cells that are serially sectioned with sections } approximately 90nm in thickness? } } How could one derive the same, but apparently much more statistically } satisfactory, data from the 40+ cell profiles per section of cells randomly } oriented and not necessarily (or importantly) sectioned or analyzed } completely. } } All who take on this task should know there is an extremely important, and } possibly significant, hint that can be observed at: } http://www.msg.ucsf.edu/sedat/marsh/mito.html. I sometimes think it is well } to know of such hints (or at least to be aware of their possibility) before } attempting a task. Assumptions are almost always incorrect unless } rigorously defined and applied, and very often difficult to prove as such or } to isolate in generating an experimental design without flaws. } } Knowing that reality often gets in the way of statistics, and vice versa, I } am } } Yours most respectfully, because I wrestle with this problem every once in a } while and I haven't yet determined a satisfactory answer. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } CASI Home Page: http://darwin.wcupa.edu/casi/ } South Church Street } West Chester, PA, 19383 } Office: SS024 } Phone: 610-738-0437 } FAX: 610-436-3036 } eMail: fmonson-at-wcupa.edu } Please call before visiting } }
I am looking for a transformer to go into the Ion getter pump powersupply for a Philips 400T. The power supply transformer in this microscope is fried and needs to be replaced. I'm not sure who to contact since the microscope division of Philips is now owned by FEI. Any information would be appreciated.
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We have a Biorad MRC-1000 (1994) with the original video card, and do not have a service contract. The video signal goes into an Indec Systems box with an output RGB + horiz. + vert. via BNC connectors to the monitor. Lately we have been having sporadic severe problems where both the frame and image do not appear on the monitor. Instead a series of dots and/or lines moving across the monitor screen appear. This problem may persist for days and then disappear and the normal frame and image reappear. Sometimes the problem appears momentarily and then corrects. Jiggling the cables during the malfunction changes the rate of dots and lines, but does not bring the frame or image back. Most experts have said this is a monitor problem, but some suggest the video card. The monitor itself is specialized in that ordinary BNC monitors will not work satisfactorily (we have tried). A new monitor from Biorad is several thousand dollars. We would hate to buy a new monitor only to discover it really is the card. Any suggestions as to what additional troubleshooting can be done, or other insights, would be appreciated. Bruce Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Laboratory University of Kansas
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id TAA24931 for dist-Microscopy; Thu, 8 Nov 2001 19:25:10 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id TAA24924 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 8 Nov 2001 19:24:39 -0600 (CST) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id TAA24916 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 8 Nov 2001 19:24:28 -0600 (CST) Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {p05100303b810df6617d6-at-[206.69.208.21]}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (reimar_gaertner-at-wsib.on.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, November 8, 2001 at 12:19:54 ---------------------------------------------------------------------------
Question: My 8-year-old daughter and I have developed an interest in microscopy as a hobby. We have a Meade 9400 student microscope with built-in light (no field diaphram) and a fixed condenser (with a rotating aperture wheel). I am able to "focus" the condenser by inserting blocks under the slide (and lowering the stage to focus). This has allowed me to get closer to the critical focusing position (the surface of the light source now focuses about 1 cm above the stage. I need this for an effective dark-field effect at 100x magnification (10x10). I have heard that in order to best see thick specimens (especially at 400x), we need Kohler illumination. Do you think this will really make a difference? If so, what modifications would I need to make to achieve this? Many thanks. Reimar
Hi Listers, I am looking for independent providers of service contracts for a JSM-5310 SEM with Digital Image Grabber in Southern Arizona. I would appreciate it if anyone who could provide this service, or anyone who has experiences, good or bad, would email me off list.
on 11/8/01 3:48 PM, Paul Webster at pwebster-at-hei.org wrote:
} } At the risk of looking very stupid, I will take your challenge. } } Am I correct in understanding that your question has two parts? 1. What is } the mean number of mitochondria per yeast cell in a population. 2. What is } the mean area of cristae membranes. } } If this is so, then there are actually three challenges here because to get } an absolute number for the cristae surface area and the number of } mitochondria per cell we also need to have an estimate for a reference space } in actual numbers. A good reference space to estimate is cell volume. [skip] } } Regards
} Paul Webster.
Dear Paul, The risk of your looking stupid is very minimal. I'd use a mito-specific dye and LM to count mitos/cell (and get the distribution of cell sizes, info on whether [mitos] is greater for smaller/larger cells or is constant, etc.). I also have a bias for using HVEM tomography on thick sections to determine cristal-area/mito. Otherwise, yours seems like a very good plan. Yours, Bill Tivol
on 11/8/01 1:03 PM, Dusevich, Vladimir at DusevichV-at-umkc.edu wrote:
} } I am looking for publications of cell cultures observation in a ESEM. So } far not a great success. Any help is greatly appreciated. } Dear Vladimir, I don't know about ESEM, but I did look at cultured PTK1 cells in an environmental chamber in the HVEM (a TEM). I presented a few images at a Biophysical Society meeting about 15 years ago, so I do not remember many details. I'd be happy to discuss this with you off line if you want. Yours, Bill Tivol
Following is an announcement of a position opening at Washington University in St. Louis. I would appreciate your bringing this to the attention of anyone who might be interested. The workhorses of our facilities are a JEOL733 electron microprobe with Advanced Microbeam automation and image analysis, and a Rigaku Geigerflex X-ray diffractometer. Please respond to {blj-at-levee.wustl.edu} if you have any questions and not to the listserver.
Thanks!
Brad Jolliff _________
Research Specialist/Electron Probe Microanalysis
Washington University in St. Louis is seeking a research scientist who will operate and enhance the Electron Microprobe and X-ray Diffractometer facilities and conduct independent and collaborative research in petrology and geochemistry in the Department of Earth and Planetary Sciences. This is a Research Specialist position and is not tenure-track. Duties will include day-to-day management and operation of the EMP and XRD laboratories as well as enhancement of systems and analytical capabilities. Ph.D. preferred. For his or her research, the Specialist would have access to the wide array of analytical facilities within the Department of Earth and Planetary Sciences and in the McDonnell Center for the Space Sciences. These facilities include microbeam techniques (SIMS, laser-ablation ICP-MS, Raman spectroscopy), major- and trace-element chemical analysis (INAA, XRF, ICP-MS), mass spectrometers for stable and radiogenic isotope analysis, and a variety of other spectrometers including visible, IR, and Moessbauer.
To apply, submit a resume, statement of research interests, and names and contact information of three professional references to:
Bradley L. Jolliff Department of Earth and Planetary Sciences Washington University Campus Box 1169 One Brookings Drive St. Louis, MO 63130
Applications should be submitted by Dec 21, 2001. Women and underrepresented minorities are encouraged to apply. Washington University is an equal opportunity/affirmative action employer. Employment eligibility verification is required upon employment. __________
department web site: {http://epsc.wustl.edu/} __________
Bradley L. Jolliff, Ph.D Research Associate Professor Dept. of Earth & Planetary Sciences Campus Box 1169 Washington University One Brookings Drive St. Louis, MO 63130
I know you are looking for an independent service provider. However, in the past, I had been very pleased with JEOL service. Currently, I am working on a Hitachi EM, but for 12 years prior I worked on JEOL scopes (both TEM and SEM). The lab that I worked in was on the east coast in New Jersey. I'm not certain how the service is on the west coast, but the number one reason why I purchased new JEOL EMs was for their service history. We had an old JSM-35 and wanted to replaced it as well as purchase a new TEM. Based on the service we had received in the past, we immediately considered JEOL's line of EMs. Granted, this was about 7 years ago since the purchase and about 2 years since I've been out of that particular lab; however I was very pleased with the service from the north east division of JEOL. Good Luck with your search.
I intended on writing you right away, but I got too busy in the lab. I have tried this procedure a few times in the past with some success. I have two articles which I can fax to you if you would like. Sorry, I do not have an electronic copy. The references are:
Gonzalez-Angulo A, Ruiz De Chavez I, Castaneda M: A Reliable Method for Electron Microscopic Examination of Specific Areas from Paraffin-embedded Tissue Mounted on Glass Slides. American Society of Clinical Pathologists (1978) 697-700.
Bretschneider A, Burns W, Morrison A: "Pop-off" Technic. The Ultrastucture of Parafin-embedded Sections. American Journal of Clinical Pathology. Vol. 76, No. 4, October 1981. 450-453.
Jackie Garfield Reseach & Development Lifecell Corporation One Millennium Way Branchburg, NJ 08876 (908) 947-1182 jgarfield-at-lifecell.com
Pardon the off-topic post, but this is the only list to which I subscribe which has a significant number of members in Australia and East Asia, where the meteor shower has potential to be a once or twice in a lifetime experience this year:
For North America, best watching period is 4AM to 6AM EST on Nov. 18. For Australia and East Asia, predicted maximum is in the hours between midnight and about 4AM, depending on location, with possible maximum ZHR (zenithal hourly rates) approaching 10,000 per hour from dark-sky sites.
Astronomy is just like microscopy except we look at bigger things with bigger instruments, right?
thurston e herricksthurst0n-at-u.washington.edu11/8/01 3:59 PM
Thurston,
FEI is located in Hillsboro, OR.
Contact them at: 1-503-640-7500.
Good luck!
Gib
} Hello, } } I am looking for a transformer to go into the Ion getter pump powersupply } for a Philips 400T. The power supply transformer in this microscope is } fried and needs to be replaced. I'm not sure who to contact since the } microscope division of Philips is now owned by FEI. Any information would be } appreciated. } } Take care } } Thurston Herricks -- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
I have been asked to prepare some samples of brown adipose tissue for TEM, to look at mitochondrial structure and density. Has anyone got any suggestions for fixative and processing schedule for this type of material.
thanks
Kevin Electron Microscope unit Department of Zoology University of Aberdeen Aberdeen AB24 2TZ
Tel 01224-272847 Fax 01224-272396 ------------ Kevin Mackenzie k.s.mackenzie-at-abdn.ac.uk
} Question: My 8-year-old daughter and I have developed an interest in } microscopy as a hobby. } We have a Meade 9400 student microscope with built-in light (no field } diaphram) and a fixed condenser (with a rotating aperture wheel). } I am able to "focus" the condenser by inserting blocks under the } slide (and lowering the stage to focus). } This has allowed me to get closer to the critical focusing position } (the surface of the light source now focuses about 1 cm above the } stage. } I need this for an effective dark-field effect at 100x magnification } (10x10). I have heard that in order to best see thick specimens } (especially at 400x), we need Kohler illumination. } Do you think this will really make a difference? If so, what } modifications would I need to make to achieve this? } Many thanks. } --------------------------------------------------------------------------- Reimar -
You'll find several suggestions for modifying your scope in Stevens' "The Microscope on a Budget"; here's the description from the MICRO bibliography (URL below):
Stevens, M.B. 1993 The Microscope on a Budget 247pp, 5.5x8.5", paperback, $13.00. ISBN 0-9638839-1-7 Logical Image Research, P.O.Box 1523, Buda, TX 78610. Also available from Nasco, 4825 Stoddard Rd., Modesto, CA 95356-9318 as #SB23799M; 800-558-9595. Subtitled "a complete guide to the low cost light microscope for the laboratory, photographers, and hobbyists", this book delivers solid information; it's a good buy. Homemade equipment (including exciting but seldom-discussed things like darkfield and Rhineberg illumination) is described in detail for the low-budget teacher or (adult-supervised) student. Both basic biological microtechnique and microcrystal preparation are described in detail. There are photography, supply source, reference, glossary, and index sections. High school - adult. RECOMMENDED
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Just wanted to send out a reminder about the ballot and M&M Journal subscription deadlines.
Ballots MUST BE postmarked by December 1, 2001 to be counted. M&M Journal subscription forms MUST BE returned by December 15, 2001 for the 2002 subscriptions.
Both the ballots and the subscription forms can be returned with your MAS renewal application.
If you have any questions, please contact me at rosslm-at-missouri.edu or (573) 882-4777 or at the the MAS email address or phone number listed below.
Thanks, Lou Ross MAS Membership Services masmembership-at-excite.com 1-800-4MASMEM (1-800-462-7636) www.microanalysis.org
on 11/9/01 11:01 AM, Rick Mott at rickmott-at-alumni.princeton.edu wrote: } } Astronomy is just like microscopy except we look at } bigger things with bigger instruments, right? } Dear Rick, That, and the fact that no one has yet invented electron astronomy (although study of the solar wind could be called proton astronomy)--let alone scanning probe astronomy (scanning black-hole force astronomy anyone?). :-) Yours, Bill Tivol
Counting the number of simple convex 3-D particles from some 3-D reference space (i.e. cell) using 2-D sections is not a trivial task. The sectioning process reduces the dimension of the particles to 2-D in the section. This 2-D sample of a particle is usually referred to as a profile. Many people have counted the number of profiles in a section and used this number to estimate the number of particles in the 3-D reference space. This of course is usually not a very good estimate of the TRUTH since the number of profiles in the 2-D sample is related not only to the number of particles but also the volume of the particles (more correctly the length of the particle perpendicular to the section plane) in the 3-D reference space. As Paul Webster pointed out in his earlier submission, Hans Gundersen solved the problem of counting particles in sections in his DISECTOR paper (J Microscopy 134:127-136, 1984).
Determining the number of complex particles, such as branching mitochondria adds complexity to the counting task. Luckily the hypothetical graduate student in Fred Monson's example did not take her EM class last year since Gundersen only solved the problem of easily counting complex particles using 2-D sections in September 2001 (J Microscopy 203:314-320). In this paper he introduces the ConnEulor principle for counting complex particles. Coincidentally the example he uses to demonstrate the ConnEulor principle is to determine the number of mitochondria in a cell. This technique like the original disector technique gives a density measure (number/volume) and you must still multiply by the average cell volume to obtain your estimate of particle number per cell.
A practical grad student might wonder if number of mitochondria is necessary and decide to measure the total volume of mitochondria within the cell (a much easier parameter to measure).
John
} } Hi Fred, } } At the risk of looking very stupid, I will take your challenge. } } Am I correct in understanding that your question has two parts? 1. What is } the mean number of mitochondria per yeast cell in a population. 2. What is } the mean area of cristae membranes. } } If this is so, then there are actually three challenges here because to get } an absolute number for the cristae surface area and the number of } mitochondria per cell we also need to have an estimate for a reference space } in actual numbers. A good reference space to estimate is cell volume. } } Obtaining mean number of mitochondria per cell is actually very easy. I } would first throw away the images already obtained because they were } subjectively sampled. } } We must then remove bias that may be a result of centrifugation - heavier } (more dense) cells will pellet to the bottom, with the possibility that a } gradient is formed in the pellet. To avoid this, embed the cells first in } fast-setting gelatin or agarose, being careful to make sure the cells have } been thoroughly mixed. We could of course do it properly and apply the } "orintator" to make sure our sampling is isotropic (Mattfeldt et al 1989 } Acta Stereol. 8:671-676). } } Fix the block and cut it into slabs, rods and then cubes. Using a } sequential random sampling protocol, take every "nth" block (decided using } random number generator) and embed them separately in resin (see Lucucq 1993 } Trends Cell Biol 3:345-358 for details). } } Section for counting. A good way to sample the cells to count particles } (here is where the catch comes in) is to only count the tops of the } particles. With the yeast mitochondria, the 2-D profile does not always } represent an individual particle. A profile may represent one single } mitochondrion, or may represent a multiply branched, single mitochondrion, } depending on the level of branching that is occurring. By counting the } tops, connected profiles are eliminated from the sampled population. (for } details on a possible method for estimating the amount of branching that is } present, look for "Star Volume" in Gundersen et al 1986 J. Microsc. } 143:3-45) } } How to sample? Use the "Disector" (in this instance, the word means "2 } sectors"). The reference is D.C. Stereo (a pseudonym for HJG Gundersen) } 1984. J. Microsc. 134:127-136. } } To apply, cut two sequential sections and take similar fields of view from } both. Use one field to examine for mitochondrial profiles and the other to } check if the profile is also present there ( the look-up frame). If a } profile is present in one field but not present in the other, it is counted } as a "top". (It is a variation of the 2-D counting rule, the associated } tangent rule). } } Knowing the area of the field of view and the thickness separating the two } sequential sections, we can work out the volume being sampled. This will } give us an estimate of the number of mitochondria per unit volume. This is } not the number of mitochondria per cell. We still need to estimate the cell } volume. } } To estimate surface area, we can use simple cross lattice overlays and count } the number of times a membrane profile crosses a test line. From this we } can estimate length of membrane, and by knowing section thickness, can } obtain an estimate of membrane area. (see H.J.G. Gundersen et al 1986 J. } Microsc. 143:3-45. 1988 APMIS 96:379-394 & 857-881, T.M. Mayhew 1991 Exp. } Physiol. 76:639-663, J. Lucocq for details on how to do this) We still need } to estimate the cell volume. } } How am I doing so far, Fred? } } Now, a question for you. I can provide estimates of relative values for } number and surface area. However, I do need a reference space to obtain } absolute values. How would I obtain a reliable estimation of the mean cell } volume of the yeast population? Presumably they are a rapidly dividing } population. } } Regards } } Paul Webster. } } } } } } } on 11/7/01 12:53 PM, Monson, Frederick C. at fmonson-at-wcupa.edu wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hypothetical! } } } } A graduate student in microbiology has just finished her first course in } } electron microscopy, and she informs her advisor that she wants to study } } mitochondrial structure and function in several related strains of yeast, } } for one of which she fortuitously has electron micrographs of mitochondrial } } profiles that are, she has been told, quite publishable. } } } } Her advisor suggests that she start by comparing 3-4 strains with respect } } to } } number of mitochondria per cell and total mitochondrial cristal area under } } identical conditions of culture. } } } } The student happily responds that she already has micrographs for regularly } } sampled sections from serial sections she learned to make of the strain she } } used in her EM course. She returns the next week with estimates of cristal } } area and numbers of mitochondria per cell based on morphometric analysis of } } those regularly sampled sections. } } } } If the cell is 12-25um in length (haploid and diploid) and always between } } 3-5um in diameter, and if the mitochondrial profiles are in the range of } } 1-2um (circular and ovoid, and sometimes or often branched), what should } } the sampling criteria be for cells that are serially sectioned with } } sections } } approximately 90nm in thickness? } } } } How could one derive the same, but apparently much more statistically } } satisfactory, data from the 40+ cell profiles per section of cells randomly } } oriented and not necessarily (or importantly) sectioned or analyzed } } completely. } } } } All who take on this task should know there is an extremely important, and } } possibly significant, hint that can be observed at: } } http://www.msg.ucsf.edu/sedat/marsh/mito.html. I sometimes think it is } } well } } to know of such hints (or at least to be aware of their possibility) before } } attempting a task. Assumptions are almost always incorrect unless } } rigorously defined and applied, and very often difficult to prove as such } } or } } to isolate in generating an experimental design without flaws. } } } } Knowing that reality often gets in the way of statistics, and vice versa, I } } am } } } } Yours most respectfully, because I wrestle with this problem every once in } } a } } while and I haven't yet determined a satisfactory answer. } } } } Fred Monson } } } } Frederick C. Monson, PhD } } Center for Advanced Scientific Imaging(CASI) } } West Chester University of Pennsylvania } } Schmucker Science Center II } } CASI Home Page: http://darwin.wcupa.edu/casi/ } } South Church Street } } West Chester, PA, 19383 } } Office: SS024 } } Phone: 610-738-0437 } } FAX: 610-436-3036 } } eMail: fmonson-at-wcupa.edu } } Please call before visiting } } } } } } } } .
John M. Basgen Department of Pediatrics University of Minnesota Mayo Mail Code 491 420 Delaware Street SE Minneapolis, MN 55455 USA Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-umn.edu
the much anticipated Leonid meteor show next Monday morning (Australian time) promises to be a spectacular event if the various predictions are even half correct. Unfortunately there are no guarantees (unlike eclipse predictions) so those of us making travel plans to observe the Leonids risk being in the wrong place at the wrong time. Personally I think the risk is worth taking considering the spectacular sight of thousands of meteors per hour if things work out.
In 1999 I drove for 5 hrs west of Sydney with a friend and my son in an attempt to observe a predicted reasonable Leonid shower. As it turned out we were treated to a much closer light show. The most ferocious thunder storm for twenty years devastate the local orchards in the area we chose to observe from and chased us all the way back to Sydney. We did manage to see a few dozen bright meteors through patchy clouds early in the night so it wasn't a total loss astronomically speaking.
This year I will be observing from central Australia, just a few kilometers from Ayres Rock. I figure that even if the sky show is clouded out or the predictions don't hold up, at least I can get a few good bush walks done during the days.
I'll give a brief report next week. Cheers,
Mark Blackford Materials Division, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} ---------- } From: Rick Mott } Sent: Saturday, November 10, 2001 3:01 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: OT: possible spectacular Leonid meteors, Nov 18/19 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Pardon the off-topic post, but this is the only } list to which I subscribe which has a significant } number of members in Australia and East Asia, } where the meteor shower has potential to be } a once or twice in a lifetime experience this } year: } } } } } http://science.nasa.gov/headlines/y2001/ast08nov_1.htm?list534615 } } } } For North America, best watching period is 4AM } to 6AM EST on Nov. 18. For Australia and East } Asia, predicted maximum is in the hours between } midnight and about 4AM, depending on location, } with possible maximum ZHR (zenithal hourly rates) } approaching 10,000 per hour from dark-sky sites. } } Astronomy is just like microscopy except we look at } bigger things with bigger instruments, right? } } Rick Mott } }
-----Original Message----- } From: reimar_gaertner-at-wsib.on.ca [mailto:reimar_gaertner-at-wsib.on.ca] Sent: Thursday, November 08, 2001 8:19 PM Hi Reimar, Here's a cheap substitute for a Kohler type illumination I learned early in my microscopy career. Put a piece of translucent tape (magic mending type) on the rear of the slide. While this is not Kohler it comes close as it mimics the effect of light emitted close to the sample. The thinner the slide the better e.g. a cover slip. Good luck and have fun. Russ Gillmeister Xerox ~~~~~~~~~~~~~
Question: My 8-year-old daughter and I have developed an interest in microscopy as a hobby. We have a Meade 9400 student microscope with built-in light (no field diaphram) and a fixed condenser (with a rotating aperture wheel). I am able to "focus" the condenser by inserting blocks under the slide (and lowering the stage to focus). This has allowed me to get closer to the critical focusing position (the surface of the light source now focuses about 1 cm above the stage. I need this for an effective dark-field effect at 100x magnification (10x10). I have heard that in order to best see thick specimens (especially at 400x), we need Kohler illumination. Do you think this will really make a difference? If so, what modifications would I need to make to achieve this? Many thanks. Reimar
Unfortunately, we used our old JSM-35 as a trade-in. The purchase was about 7 years ago. The scope was in good shape, so I am sure it got a good home. However, you can check with JEOL for any used scopes. With a little luck, they just might have something. Also, you may want to check with Dick Daniel from Radco, Inc. (201) 891-8647. He services quite a bit of SEM equipment (mostly Denton products), and has many contacts throughout the United States. Perhaps, he knows of a good used SEM. It is worth a call. Good Luck,
Nanotubes and Peapods - Synthesis, Processing, Structure, Properties
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The experimental program involves electron microscopy including in-situ experiments, chemical synthesis and processing, and physical property measurements. The successful candidate should work well in a group, but be interested in developing and driving an aggressive, independent research effort.
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Be aware, however, that most tape is anisotropic, subject to strain polarization, and will have a slight polarizing effect on transmitted light, perhaps preventing one from obtaining complete extinction between crossed polars or otherwise modifying expected optical effects.
John Twilley Conservation Scientist
Gillmeister, Russ wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } -----Original Message----- } } } From: reimar_gaertner-at-wsib.on.ca [mailto:reimar_gaertner-at-wsib.on.ca] } } Sent: Thursday, November 08, 2001 8:19 PM } Hi Reimar, Here's a cheap substitute for a Kohler type illumination I } learned early in my microscopy career. Put a piece of translucent tape } (magic mending type) on the rear of the slide. While this is not Kohler it } comes close as it mimics the effect of light emitted close to the sample. } The thinner the slide the better e.g. a cover slip. Good luck and have fun. } Russ Gillmeister } Xerox } ~~~~~~~~~~~~~ } } } Email: reimar_gaertner-at-wsib.on.ca } Name: Reimar Gaertner } } Organization: WSIB } } Education: Graduate College } } Location: Toronto, Ontario, Canada } } Question: My 8-year-old daughter and I have developed an interest in } microscopy as a hobby. } We have a Meade 9400 student microscope with built-in light (no field } diaphram) and a fixed condenser (with a rotating aperture wheel). } I am able to "focus" the condenser by inserting blocks under the } slide (and lowering the stage to focus). } This has allowed me to get closer to the critical focusing position } (the surface of the light source now focuses about 1 cm above the } stage. } I need this for an effective dark-field effect at 100x magnification } (10x10). I have heard that in order to best see thick specimens } (especially at 400x), we need Kohler illumination. } Do you think this will really make a difference? If so, what } modifications would I need to make to achieve this? } Many thanks. } Reimar } } --------------------------------------------------------------------------- } } }
In its trinocular head there sits a CF PL 5X Projection lens, above that is a Microflex HFX-IIA Photomicrographic Attachment, and onto that is a "35mm camera adapter A", then what is called in the HFX-IIA manual a "Motorized dark box FX-35WA". The latter looks more like a camera body to me.
We want to be able to mount our Nikon D 1 body onto the microscope, with or without the HFX-IIA, whatever it takes.
Is there anyone out there who can tell, from the above, what I need to do to acheive this?
I would have thought that all we need to do would be to mount the D 1 body directly onto the trinocular vertical tube, but then I know almost nothing about light microscopy.
tia
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Does anyone know who services or can tell me about how the mechanism for the LKB knifebreaker works? The knob that you turn to put pressure on the glass to break is very difficult to turn. Is there an easy way to clean or free it up? Will some solvent work or WD40? Or is there something else that I can do?
Thanks in advance.
Mei Lie -- Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
Dr Sims, first of all you need to get a C-Mount for the microscope, at the bottom of the trinocular head there are 3 hex-screw use them to remove the tube and insert the c-mount with your D1 camera, connect all the wires on the D1, and enjoy.
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My name is Kelly Stanard and I am a lab tech in the Image Analysis Laboratory at National Cancer Institute at Frederick, Maryland. I have been left with a Balzers freeze-fracture unit and some basic instructions but I have no idea what I am doing. Although a few of my attempts have produced decent replicas, I am very uncomfortable with the unit. What I am in need of are a few guidelines as to what all the buttons are for and when would be a good time to push them!! What I believe I have sitting here is a High Vacuum Coating Unit BAF 300 purchased in May, 1976. Quartz Crystal Thin Film Monitor QSG 201 Freeze Etching Unit Control BMS 101 Commutator Unit BCM 101 Pumping Unit Control DPA 101 Control Unit EVM 052A Microtom Movement Control BMB 101 Chamber IKR 010
If you can help me in anyway it would be greatly appreciated.
Thank you,
Kelly Stanard Image Analysis Laboratory NCI- Frederick P.O. Box B Frederick, MD 21702 301-846-1528 kstanard-at-ncifcrf.gov
Dr Sims, first of all you need to get a C-Mount for the microscope, at the bottom of the trinocular head there are 3 hex-screw use them to remove the tube and insert the c-mount with your D1 camera, connect all the wires on the D1, and enjoy.
Alfredo Hernandez
"Ritchie Sims" {r.sims-at-auckland.ac.nz} wrote:
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You can try calling Leica for service, however you may have to send the knife-maker out. I use a company called Dolbey-Jamison, located in PA. They do in-house service. Good Luck,
CRYOMICROSCOPY GROUP ANNUAL MEETING EM Centre, University of Birmingham, Birmingham, UK
Wednesday 21st November 2001
PROGRAMME :
9.30 - 10.30 Registration, Coffee and Trades Exhibition 10.30 Chairman's Welcome and Introduction 10.35 -11.15 Charge elimination for the X-ray microanalysis of frozen hydrated specimens Patrick Echlin, Cambridge Analytical Microscopy, Cambridge 11.15 -12.00 Freeze-substitution and 3D reconstruction of muscle Pradeep Luther, Imperial College, London 12.00 -12.45 Silver enhancement for immunolabelling Jan Leunissen, Aurion, Wageningen, The Netherlands 12.45 - 14.00 Lunch , Trades Exhibition & Posters 14.00 -14.15 Annual General Meeting 14.15 - 15.00 RMS Beginners Competition 15.00 -15.45 Ultrastructural localisation of Prion protein in the brain using cryo-immunogold electron microscopy Peter Peters, Netherlands Cancer Institute, Amsterdam 15.45 - 16.00 Problems with preparing articular cartilage for microscopic study Iolo ap Gwynn, S C Wade and R G Richards. University of Wales, Aberystwyth 16.00 Closing Remarks and Tea
FURTHER DETAILS : http://www.cryomicroscopygroup.org.uk/index.htm
Dr Laurence Tetley Division of Infection & Immunity, IBLS, Integrated Microscopy Facility Joseph Black Building University of Glasgow Glasgow G12 8QQ
l.tetley-at-bio.gla.ac.uk tel 0141 330 4431 FAX 0141 330 3516
Use acetonitrile instead of ethanol or acetone for dehydration. Also, use imidazole buffered osmium to postfix. The mitochondria come out beautifully. I have done this and it works. The references are:
Edwards, H. H. et al. 1992. Microscopy Research and Technique Volume 21, pp 39-50.
Angermuller, S. and H. D. Fahimi. 1982. Histochemical Journal Volume 14, pp. 823-835.
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Dear Listers, I am expecting the following samples for paraffin embedding sometime soon -- "wolf ovaries have been preserved in tequila for the past 6 months or so". I first assumed that this was a joke but apparently they are serious--this is a reproductive physiology project from an ecology lab. Any comments are welcome--levity included. I can hear your laughter already! My questions: --has anyone used tequila as a fixative? -- is the embedding worth pursuing? Rosemary Walsh
We service and sell reconditioned LKB Knife Makers. Please contact me off line. We are closed from Nov. 15 to 26. Regards, Microscopy Labs P.O.Box 338 61 West Street Red Bank, NJ 07701 732 727 6228 fax 732 758 9142
----- Original Message ----- } From: "Mei Lie Wong" {wong-at-msg.ucsf.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, November 12, 2001 8:43 PM
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No experience and can't think of any jokes but .....
I guess tequila is about 40% ethanol or is this home-brewed stuff?? Standardisation in the future could be a problem - will they be using the same brand next time?
Limited lipid extraction in that case - wash in PBS and cryosection?
You could rehydrate to PBS and refix in buffered formaldehyde - could help to protect against processing.
I guess that wolf ovaries are resonably large so you could try various combinations?
I would be interested to hear how it goes. Go for it and educate us all!
At 19:46 2001-11-13 -0500, Rosemary Walsh wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
I haven't worked in biological EM for some time. I do, however, have some experience with the stated topic. One evening some years ago I partook a bit too much tequila and had a howling good time. Unfortunately, the next morning I found that I was very well fixed indeed. Needed the hair of the dog (or wolf as the case may be) to survive the day. That memory has been well embedded in my memory as I wax nostalgic.
I have no other comments aside from this nonsense. Good luck.
Gary M. Brown
Rosemary Walsh {rw9-at-psu.edu} To: Microscopy-at-sparc5.microscopy.com cc: Subject: a sample prep queri 11/13/01 06:46 PM
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Dear Listers, I am expecting the following samples for paraffin embedding sometime soon -- "wolf ovaries have been preserved in tequila for the past 6 months or so". I first assumed that this was a joke but apparently they are serious--this is a reproductive physiology project from an ecology lab. Any comments are welcome--levity included. I can hear your laughter already! My questions: --has anyone used tequila as a fixative? -- is the embedding worth pursuing? Rosemary Walsh
Dear listers, Some time ago one of you discussed a combination of resins that snapped off culture dishes fairly easily. Could you post that again as I need the information and can't seem to locate it in the archives. I've tried several combinations of resins from different sources on different brands of plates and the results are not great. There also appear to be differences in the "snapability" between the multiple well plates and the individual dishes from the same companies, which adds to the problem. I'd really appreciate knowing the magic combination of resin components if you could readdress the issue. Thanks, Mary Gail Engle
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
} --has anyone used tequila as a fixative? } -- is the embedding worth pursuing? } Rosemary Walsh ****************** Hi Rosemary, that's a new one.
My first question is, what proof was the tequila? If it was potent stuff, the ethanol level may have been high enough to act as a coagulative fixative. Many years ago I was asked to do EM of the gills of clams that had been fixed in 70% ethanol. I rehydrated them to buffer and then processed for EM as usual (glut, osmium, etc). The ultrastructure, while clearly compromised was far better than I had expected. You can try this with the ovary samples. The fact that they are destined for paraffin means that you won't have the very fine structure to worry about. You might want to use a higher than usual concentration of pfa, or even throw in a little glut to lock in whatever is left.
Good luck, Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I would like to contract out cryosectioning of some carbon black - filled rubber material. We desire sections on the order of 1 micron thick amenable to light microscope/image analysis measurements that we will perform. There is plenty of sample material and we don't need serial sectioning. We are hoping to have this done within weeks.
If you are interested or know of someone who may be, and/or have questions, please contact me off line at tgruber-at-phelpsd.com.
Not too dumb on the part of the enviro's. After all, Tequila does the worm! Tequila, as I quickly discovered, can have between 35-55% ethanol.
My recommendation would be to proceed with your own fixation and not worry. They used their heads when they realized they went into the field unprepared, or they had an accident, or they couldn't take the fixative with them and couldn't find any there. You treat the tissue as if it just got to you. The nitrogens are likely still waiting for the formaldehyde. Or, you could transfer them to Clarke's 3:1(Absolute ethanol:Glacial Acetic Acid), and have a really granular texture in the sections. Or Bouin's, which would also complete the job on the worm!
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Rosemary Walsh } Sent: Tuesday, November 13, 2001 7:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: a sample prep queri } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } I am expecting the following samples for paraffin embedding sometime } } soon -- "wolf ovaries have been preserved in tequila for the past 6 months } } or so". I first assumed that this was a joke but apparently they are } serious--this is a reproductive physiology project from an ecology } lab. Any comments are welcome--levity included. I can hear your laughter } } already! My questions: } --has anyone used tequila as a fixative? } -- is the embedding worth pursuing? } Rosemary Walsh } } }
As I recall the lower range is around that for normal cells and the upper range is something like 15-40um (BIG!!!). Just hope that J. Kiernan sees your email so that you get a more refined result.
Fred Monson
} ---------- } From: Cheri Owen } Sent: Tuesday, November 13, 2001 11:09 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Range of size of neuronal nuclei } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Does anyone know off the top of their head what the range of size of } neuronal nuclei is? } } Cheri Owen } Dept. Emergency Medicine/Physiology } Wayne State University } } }
Position Open Microscopy and Microanalysis Research Assistant
The NCSU Analytical Instrumentation Facility (AIF) is seeking qualified applicants for a SEM Microscopist position opening December 11, 2001.
Duties and responsibilities include: Operation and maintenance of SEM/EDS instrumentation and sample preparation and analysis equipment; scheduling of access to and oversight of the above instrumentation; user training and assistance; assistance with the teaching of electron microscopy laboratory classes, and analysis of a wide variety of samples. Qualifications must include a minimum of a BS with 5 years hands on SEM/EDS experience, MS with 2 years hands on SEM/EDS experience or a Ph.D. with 1 year hands on SEM/EDS experience. Experience must be in a non biological materials related discipline. Required skills include: extensive hands on experience with SEM and related techniques and accessories (e.g. EDS, specimen preparation and associated analytical tools). Preferred qualifications include: teaching and user training; familiarity with modern electronics; and experience with vacuum systems.
Please send resume and three letters of reference to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 318A EGRC; 1010 Main Campus Drive; Raleigh, NC 27695-7531 or email PRUSSELL-at-NCSU.EDU.
North Carolina State University is an Equal Opportunity and Affirmative Action Employer. ADA Accommodations: Phil Russell, prussell-at-ncsu.edu, 919-515-7501. In its commitment to diversity and equity, North Carolina State University seeks applications from women, minorities, and persons with disabilities
It's possible, that the ovaries are fixed OK, provided they were cut open when placed into the tequila. Normally, 70% ethanol would have been better, maybe they should have used for alcohol fixation. If they have not been cut open prior to fixation, then the odds are that the morphology of the internal areas of the ovary will not be very good. However, try one sample, trim to desired sized and post-fix in 10% formalin overnight, process for paraffin embedding.
Good Luck!
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642
-----Original Message----- } From: Rosemary Walsh [mailto:rw9-at-psu.edu] Sent: Tuesday, November 13, 2001 7:46 PM To: Microscopy-at-sparc5.microscopy.com
Dear Listers, I am expecting the following samples for paraffin embedding sometime
soon -- "wolf ovaries have been preserved in tequila for the past 6 months or so". I first assumed that this was a joke but apparently they are serious--this is a reproductive physiology project from an ecology lab. Any comments are welcome--levity included. I can hear your laughter already! My questions: --has anyone used tequila as a fixative? -- is the embedding worth pursuing? Rosemary Walsh
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
My name is Kelly Stanard and I am a lab tech in the Image Analysis Laboratory at National Cancer Institute at Frederick, Maryland. I have been left with a Balzers freeze-fracture unit and some basic instructions but I have no idea what I am doing. Although a few of my attempts have produced decent replicas, I am very uncomfortable with the unit. What I am in need of are a few guidelines as to what all the buttons are for and when would be a good time to push them!! What I believe I have sitting here is a High Vacuum Coating Unit BAF 300 purchased in May, 1976. Quartz Crystal Thin Film Monitor QSG 201 Freeze Etching Unit Control BMS 101 Commutator Unit BCM 101 Pumping Unit Control DPA 101 Control Unit EVM 052A Microtom Movement Control BMB 101 Chamber IKR 010
If you can help me in anyway it would be greatly appreciated.
Thank you,
Kelly Stanard Image Analysis Laboratory NCI- Frederick P.O. Box B Frederick, MD 21702 301-846-1528 kstanard-at-ncifcrf.gov
The CCD on the D1 is 23.7mm x 15.6mm, so a projection lens of 1.5x should be sufficient. Do you have some kind of C-mount to Nikon mount converter? The D1 uses the same Nikon "F" mount that Nikon SLR bodies have used for years, so without a converter it will not mount directly to a C-mount.
George
George Laing National Graphic Supply
The only question now is which C-mount. Whereas the CCD for Coolpix mounts requires 1X, the CCD for the D1 is larger ... 1.5X, 2X???
C-Mount is just an adapter and wont requiere any additional lens, You'll get a direct image. You can use different c-mounts with zoom in them in order to get a larger field of view. The lens you require will depend on the chip size of your camera (DN100 has 1/2" chip size) so if you want a larger field of view you probably want to buy a C mount + TV adapter 0.45X The DXM1200 has a 2/3" chip size therefore the best combination will be C mount + TV adapter 0.6X
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Hi folks, Thanks for the replies. Apparently this was serious inquiry. Samples were taken in a remote area in Alaska. The interest is in determining the number of ovulations compared with the number of pregnancies in dominant wolves versus pseudo pregnancies in other pack females. We'll probably rehydrate to PBS and fix in 4% paraformaldehyde, process and embedd in paraffin, stain with H&E for LM to see what we have. Another option is to go directly to 10% formalin. If I can obtain some sample I intend to process for TEM "just for the record". This should take us awhile but I promise to keep you posted. Thanks again to all of you wonderful folks who responded. Not yet destined for Margaritaville! Rosemary
Here are the replies No experience and can't think of any jokes but .....
I guess tequila is about 40% ethanol or is this home-brewed stuff?? Standardisation in the future could be a problem - will they be using the same brand next time?Limited lipid extraction in that case - wash in PBS and cryosection? You could rehydrate to PBS and refix in buffered formaldehyde - could help to protect against processing. I guess that wolf ovaries are resonably large so you could try various combinations? Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
Sheesh! What an ecology lab. Were they getting the wolves drunk and then ripping out their ovaries and plunking them in tequila? I've never tried preserving anything in tequila, but it probably works for livers. Good luck (and you should charge these idiots A LOT). Mary McKee MGH Renal Unit Charlestown, MA 02129 (617)726-3696
Hi Rosemary, After 6 months or so, do you suppose that the ovaries are "Wasting Away in Margaritaville"? JME Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA phone: 506-364-2519 fax: 506-364-2505 email: jehrman-at-mta.ca
In "The collection and preservation of Animal Parasites" by G.O.W. Kruse e M.H. Pritchard (Tech. Bull. no 1, The Harold W. Manter Laboratory- University of Nebraska Press, 1982) the use of any available local brew to fix and preserve parasites is reported, if no fixative is available.Transfer of parasites to 70% EtOH as soon as the latter is available, after carefully pouring out the spirit.Bruno Dore
I have been present at the AFIP when they got specimens similar to your wolf ovaries. Some of us wanted to go do field work with the guys who came up with this as they said they use tequila alot, for many things. Our imaginations went active. You might try contacting the Histology laboratory there and find out what they do exactly. I can give you some advise a fellow histologist and I will forward yourmessage to HistoNet as someone there may be able to assist you also. The first issue is the fixation, at this point both fixation and dehydration are complete. Do you plan to dissect the specimens into blocks, if so what sizes? I have a protocol using butanol to clear the specimen prior to infiltration with paraffin. It is very gentle and would hopefully allow the perimeters to stay softer and easier to section. Let me know if I can assist you or you need the protocol. I am generally in from 9AM to 5PM. Today will be the exception we have meetings until 1PM.
Pamela A. Marcum Histology/Microscopy Product Development Manager 400 Valley Road Warrington, PA 18976 Phone: 800-523-2575 Ext 167 215-343-6484 Ext 167 Fax: 215-343-0214 E-mail: pmarcum-at-polysciences.com
My first question is, what proof was the tequila? If it was potent stuff, the ethanol level may have been high enough to act as a coagulative fixative. Many years ago I was asked to do EM of the gills of clams that had been fixed in 70% ethanol. I rehydrated them to buffer and then processed for EM as usual (glut, osmium, etc). The ultrastructure, while clearly compromised was far better than I had expected. You can try this with the ovary samples. The fact that they are destined for paraffin means that you won't have the very fine structure to worry about. You might want to use a higher than usual concentration of pfa, or even throw in a little glut to lock in whatever is left. Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
For TEM I have looked at skin and underlying connective tissue from all kinds of strange scenarios, eg. collected from a dead emu under freezing weather conditions for a number of days. Expect loss of fine detail and for certain organelles to be much more poorly preserved than others. You might take some samples and run them down to buffer then refix in buffered formaldehyde. Embed some other samples straight from the tequila. I suspect the tequila is between 40-60% ethanol. Don't waste the tequila, maybe you can start a fad, a pleasent way to increase fertility, or some other patent medicine application. Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Laboratory University of Kansas
Not too dumb on the part of the enviro's. After all, Tequila does the worm! Tequila, as I quickly discovered, can have between 35-55% ethanol. My recommendation would be to proceed with your own fixation and not worry. They used their heads when they realized they went into the field unprepared, or they had an accident, or they couldn't take the fixative with them and couldn't find any there. You treat the tissue as if it just got to you. The nitrogens are likely still waiting for the formaldehyde. Or, you could transfer them to Clarke's 3:1(Absolute ethanol:Glacial Acetic Acid), and have a really granular texture in the sections. Or Bouin's, which would also complete the job on the worm.Regards,Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu
I do a lot of ovaries here - A LOT, but they are usually rodent and are fixed in 10% formalin, Bouins or 70% ETOH not tequila (what a waste of good tequila - assuming that it was of the good variety). I hope that the ovaries were small or trimmed before they were plunked into the tequila. I'd process them mostly out of curiosity to see how they might turn out. The ultrastructure wouldn't be much to speak of but there might be some histology to see. One reply on the microscopy listserver stated that tequila is about 40% alcohol. If so I'd trim the samples if necessary and process as I usually do by starting in 70% ETOH through 80%, 95% x2, 100% x3, Xylene x 2, paraffin x 2. The replier on microscopy also said to rehydrate in PBS and refix in formalin. You could do that. In fact this might be a good idea it might help to preserve what hasn't already been destroyed. I hope Rosemary processes the wolf ovaries and gives a report on the outcome.
Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : Sr. Research Specialist University of Arizona : (office: AHSC 4212) P.O. Box 245044 : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : (FAX: 520-626-2097) (email: algranth-at-u.arizona.edu) :
I think the active ingredient would be the ethanol content of tequila. No, I am not laughing, when Lord Byron died somewhere in Mediterrean area or whereever, his body was preserved in a wine or brandy cask and shipped home (to England?) for burial.
She might want to know what the percentage of alcohol is, what proof, and work on processing from that point. I would love to know the outcome of this, I used Chinese vodka as a means to clean a scratch in skin while in that country and prevent a skin infection, it worked but I had a quart of the stuff, gave it to some person who loved the stuff in coca cola! Skin healed well, his liver suffered badly!
As for humor, it could be said the tissues were well "pickled", same as people are when they ingest too much of this stuff!! Gayle Callis MT,HT,HTL(ASCP) Histopathology Supervisor Veterinary Molecular Biology - Marsh Lab Montana State University - Bozeman 19th and Lincoln St Bozeman MT 59717-3610
406 994-6367 406 994-4303 (FAX)
It's possible, that the ovaries are fixed OK, provided they were cut open when placed into the tequila. Normally, 70% ethanol would have been better, maybe they should have used for alcohol fixation. If they have not been cut open prior to fixation, then the odds are that the morphology of the internal areas of the ovary will not be very good. However, try one sample, trim to desired sized and post-fix in 10% formalin overnight, process for paraffin embedding. Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642
The C-mount interface suggestions below should work and will result in an image being available to your D1 sensor. However, if your solution requires that you invest more money into the process, you might consider additional alternatives geared more towards 35mm SLR photography to optimize your results. With the D1 being an SLR camera body, and given the pixel resolution of the sensor, for our D1 customers, we have, instead, used one of two options which, we believe, result in optimal sensor fill and end results.
1.) Diagnostic Instruments has a photographic adapter series for coupling 35mm SLR cameras to microscopes. Within that series, the PA1-12A with the NIKC-T2 adapter usually fits the bill for most Nikon scopes. Diagnostic Instruments recommends a Nikon 1.6x photoeyepiece for image projection.
2.) Alternatively, I believe Nikon now has a direct Nikon option for the D1 series of cameras. It does pretty much the same thing as Option 1, providing the 1.6x projection, except it all comes from Nikon, and might be slightly more "refined".
Diagnostic Instruments products are available through a network of dealers, most of which are also microscope dealers. Nikon products are (obviously) available from your local Nikon dealer. You might consult your Nikon dealer in regard to either solution. They can probably offer both.
PLEASE NOTE: This e-mail is in no way meant to pass judgment on the suggestions of other list members. I only provide the above info based on opinion and the approach we have taken in this situation.
-----Original Message----- } From: "tartenon-at-netscape.net"-at-sparc5.microscopy.com [mailto:"tartenon-at-netscape.net"-at-sparc5.microscopy.com] Sent: Wednesday, November 14, 2001 4:08 PM To: Microscopy-at-sparc5.microscopy.com
C-Mount is just an adapter and wont requiere any additional lens, You'll get a direct image. You can use different c-mounts with zoom in them in order to get a larger field of view. The lens you require will depend on the chip size of your camera (DN100 has 1/2" chip size) so if you want a larger field of view you probably want to buy a C mount + TV adapter 0.45X The DXM1200 has a 2/3" chip size therefore the best combination will be C mount + TV adapter 0.6X
} } Dear listers, } Some time ago one of you discussed a combination of resins that } snapped off culture dishes fairly easily. Could you post that again } as I need the information and can't seem to locate it in the } archives. I've tried several combinations of resins from different } sources on different brands of plates and the results are not great. } There also appear to be differences in the "snapability" between the } multiple well plates and the individual dishes from the same } companies, which adds to the problem. I'd really appreciate knowing } the magic combination of resin components if you could readdress the } issue. } Thanks, } Mary Gail Engle } **************** HI Mary Gail,
I was the one who posted my "magic mixture". It is this: LX112 (Ladd Industries), DDSA (Electron Microscopy Sciences), NMA (also EMS) and DMP-30 (Ladd). I use a "standard" Epon recipe, nothing secret there. After the last 100% ethanol, I pour a shallow layer of resin into the dish and insert (tubes made by cutting the ends off BEEM capsules) into the resin over areas of interest. Be sure that the molded end of the tube is facing down so that you have a perfectly flat edge. Polymerize overnight, then fill just the tubes with more resin. Return to the oven for 24-36 hr. when you remove then dish from the oven, just grasp the embedding tube with a pair of needle-nosed pliers and snap. If anything, a small amount of the dish may come off with your block, but it shoul.d not leave any cells behind. I haven't seen a difference between single and multi-well plates, although the small diameter wells often don't work well....I think its tough to get a good exchange of the last alcohol and the blocks tend to be gummy.
If you have any more questions, contact me off-list and will will elaborate.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Hi listers, With the agave shortage a couple of years back, tequila is getting expensive. What about gin? Or whiskey? Then we could give the fixations cute names like 'The Limey' or 'Kentucky's Finest' (respectively). The ethanol content would be about the same, and there would be less impurities (unless those impurities are what makes it a good fixative). Just a thought.
Gareth Morgan wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } No experience and can't think of any jokes but ..... } } I guess tequila is about 40% ethanol or is this home-brewed stuff?? } Standardisation in the future could be a problem - will they be using the } same brand next time? } } Limited lipid extraction in that case - wash in PBS and cryosection? } } You could rehydrate to PBS and refix in buffered formaldehyde - could help } to protect against processing. } } I guess that wolf ovaries are resonably large so you could try various } combinations? } } I would be interested to hear how it goes. Go for it and educate us all! } } At 19:46 2001-11-13 -0500, Rosemary Walsh wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Listers, } } I am expecting the following samples for paraffin embedding sometime } } soon -- "wolf ovaries have been preserved in tequila for the past 6 months } } or so". I first assumed that this was a joke but apparently they are } } serious--this is a reproductive physiology project from an ecology } } lab. Any comments are welcome--levity included. I can hear your laughter } } already! My questions: } } --has anyone used tequila as a fixative? } } -- is the embedding worth pursuing? } } Rosemary Walsh } } } } } } Med vänliga hälsningar/With best wishes } } Gareth } } "Close your eyes and look. What you saw at first is there no more; and what } you will see next has not yet come to life" Leonardo da Vinci (1452-1519). } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-impi.ki.se } } Tel +46 8 728 3734 } Fax +46 8 728 3688 } } Gareth Morgan MPhil MSc FIBMS, } Institutionen för Mikrobiologi, } Patologi och Immunologi(IMPI), H5, } Karolinska Institutet, } Dept of Biomedical Laboratory Science, } Lindhagensgatan 92, Box 12773, } S 112 96, Stockholm } Sweden } } OBS! Besöksadress: Lindhagensgatan 92 } NB! Visiting address:Lindhagensgatan 92
-- Russell McConnell Confocal Imaging Facility Technician Department of Neuroscience Tufts University School of Medicine M&V Building room #137 136 Harrison Ave. Boston, MA 02111 Tel. (617) 636-3795
Hi It all depends on which measurements you have in mind. In the simplest case, you can use topographic imaging to trace the changes in topography during oxidation. From these measurements, you can detect the formation of oxide if it is accompanied by significant morphological changes, e.g. instead of flat surface with atomic steps you see large (10-100 nm) oxide crystallites. Of course, if oxide is uniform, you wouldn't see much. If you want to get oxide thickness, you can etch it locally and measure the step heigh between etched and unetched parts. Electrostatic force microscopy and surface potential microscopies are completely different story. In principle, on the potential map you can see charged grain boundaries, trapped charges and other electroactive defects. You can also measure CPD variations between different phases. Some useful leads can be found in papers by J.W.P. Hsu, D.A. Bonnell et al and Ludeke and Cartier, among others. However, interpretation of the SSPM images can be tricky. The usual caveat is topographical artifacts, etc. You might also consider asking the same question at DI forum at spm-at-di.com Hope this helps Sergei
} } Hi, I have a question about the GaSb. } } How to use AFM/EFM to characterize the oxidation layer which is above the } GaSb surface. } } Thank you for your help! } } Xianglin Li } } University of Massachusetts } 978-934-3411
-- Sergei V. Kalinin Dept. Mat. Sci. Eng. University of Pennsylvania, 3231 Walnut St, Philadelphia PA 19104 Tel: (215) 898-3446 Fax: (215) 573-2128 E-mail: sergei2-at-seas.upenn.edu URL: http://www.seas.upenn.edu/~sergei2
Bacardi 151 proof rum made a good fixative for marine organisms ...
Phil
} Hi listers, } With the agave shortage a couple of years back, tequila is getting } expensive. What about gin? Or whiskey? Then we could give the } fixations cute names like 'The Limey' or 'Kentucky's Finest' } (respectively). The ethanol content would be about the same, and there } would be less impurities (unless those impurities are what makes it a } good fixative). Just a thought. } } Gareth Morgan wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } No experience and can't think of any jokes but ..... } } } } I guess tequila is about 40% ethanol or is this home-brewed stuff?? } } Standardisation in the future could be a problem - will they be using the } } same brand next time? } } } } Limited lipid extraction in that case - wash in PBS and cryosection? } } } } You could rehydrate to PBS and refix in buffered formaldehyde - could help } } to protect against processing. } } } } I guess that wolf ovaries are resonably large so you could try various } } combinations? } } } } I would be interested to hear how it goes. Go for it and educate us all! } } } } At 19:46 2001-11-13 -0500, Rosemary Walsh wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear Listers, } } } I am expecting the following samples for paraffin embedding sometime } } } soon -- "wolf ovaries have been preserved in tequila for the past 6 months } } } or so". I first assumed that this was a joke but apparently they are } } } serious--this is a reproductive physiology project from an ecology } } } lab. Any comments are welcome--levity included. I can hear your laughter } } } already! My questions: } } } --has anyone used tequila as a fixative? } } } -- is the embedding worth pursuing? } } } Rosemary Walsh } } } } } } } } } } Med vänliga hälsningar/With best wishes } } } } Gareth } } } } "Close your eyes and look. What you saw at first is there no more; and what } } you will see next has not yet come to life" Leonardo da Vinci (1452-1519). } } } } http://www.ki.se/biomedlab } } e-mail Gareth.Morgan-at-impi.ki.se } } } } Tel +46 8 728 3734 } } Fax +46 8 728 3688 } } } } Gareth Morgan MPhil MSc FIBMS, } } Institutionen för Mikrobiologi, } } Patologi och Immunologi(IMPI), H5, } } Karolinska Institutet, } } Dept of Biomedical Laboratory Science, } } Lindhagensgatan 92, Box 12773, } } S 112 96, Stockholm } } Sweden } } } } OBS! Besöksadress: Lindhagensgatan 92 } } NB! Visiting address:Lindhagensgatan 92 } } -- } Russell McConnell } Confocal Imaging Facility Technician } Department of Neuroscience } Tufts University School of Medicine } M&V Building room #137 } 136 Harrison Ave. } Boston, MA 02111 } Tel. (617) 636-3795
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Microscopy technician vacancy in Albany, California (East San Francisco Bay Area), near Berkeley, California.
An individual is needed to provide assistance and expertise in scanning electron and light microscopy on various projects in food and non-food applied research. In addition, the person will provide expertise in photography, imaging, and preparation of graphics for presentation and publication using digital cameras and Windows-based computers. Applicants will be considered at the GS-5 through GS-7 levels.
Those interested, please see the Federal website for further information about the position and how to apply.
I am looking for a supplier for Barium permanganate (for use as a TEM Stain), and I can't find anyone who carries it (Fisher, Sigma, EMS, Pella, SPI, Ladd, etc.).
Any suggestions?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
I have just seen an Imax film "Cosmic Journey" which expands on the plot of "Powers of Ten" adds some graphics and presents a (speeded up!) depiction of the early development of the Universe and all that. Its worth looking out for.
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
Hello, I have been loaned a microtome (Leitz 1512) by a colleague which according to the instruction manual requires regular oiling. The microtome did not come with any oil and the price I have been quoted by a local supplier for microtome oil was about 75$US (including VAT) for 50ml (SAKURA 1447). I found the price a bit steep and am left wondering if it is absolutely necessary to buy "microtome" oil or if I can use an off the shelf oil (e.g 3 in 1 or something like that). I do not want to compromise the performance of the microtome as it is not mine and would appreciate any advice from those with experience. Thanks for any help. Sincerely, Jonathan
Jonathan Wilson (PhD) CIIMAR Rua do Campo Alegre 823 4150-180 Porto, Portugal tel: 351 22 608 0470 / 71 fax: 351 22 606 0423 e-mail: wilson_jm-at-cimar.org web: www.cimar.org
Hello, I have been loaned a microtome (Leitz 1512) by a colleague which according to the instruction manual requires regular oiling. The microtome did not come with any oil and the price I have been quoted by a local supplier for microtome oil was about 75$US (including VAT) for 50ml (SAKURA 1447). I found the price a bit steep and am left wondering if it is absolutely necessary to buy "microtome" oil or if I can use an off the shelf oil (e.g 3 in 1 or something like that). I do not want to compromise the performance of the microtome as it is not mine and would appreciate any advice from those with experience. Thanks for any help. Sincerely, Jonathan
Jonathan Wilson (PhD) CIIMAR Rua do Campo Alegre 823 4150-180 Porto, Portugal tel: 351 22 608 0470 / 71 fax: 351 22 606 0423 e-mail: wilson_jm-at-cimar.org web: www.cimar.org
We are trying to microscopy a microorganism, spiroplasma using TEM. Every time we had problem to see clearly the membrane around the organism. We tried to increase the time of Osmium (4%) fixation, 3-5 hours at RT after glutaldehyde fixation. But the membrane preservation is still terrible. I would very much appreciate any of suggestions for that.
The Integrated Microscopy Core, Department of Molecular and Cellular Biology, Baylor College of Medicine has an immediate full-time opening for an electron microscopy technician. The Integrated Microscopy Core is a busy, state-of-the-art facility with a new Hitachi H7500 equipped with Gatan’s new 2Kx2K CCD camera, deconvolution, laser scanning confocal, and 2 CCD-based upright and inverted epifluorescence microscopes.
The applicant should have at least one year of experience in various aspects of sample preparation for biological TEM. This should include fixation, embedding, ultrathin sectioning staining and darkroom procedures, TEM operation and knowledge of computers. Other duties include preparation of solutions, embedding media and excellent organizational skills.
The position offers opportunities for training in advanced microscopy techniques, including digital TEM, fluorescence, deconvolution and laser scanning confocal microscopy. Excellent communication skills and a Bachelor's degree are required. A competitive salary and benefit package commensurate with experience is offered. Please email a cover letter expressing your research/technical interests and resume to:
Michael A. Mancini, Ph.D. Assistant Professor Director, Integrated Microscopy Department of Molecular and Cellular Biology Baylor College of Medicine Houston, TX 77030 mancini-at-bcm.tmc.edu http://microscopy.bcm.tmc.edu 713 798 8952
Hi Y'All, I'm interested to know what exactly is left behind after Ag (or C) paint (used as an electrical contact on a sample stub) has dried. I have used this method of attaching samples to holders in ultra high vacuum systems and after the paint is thoroughly dried there is no outgassing in uhv to 10-10 torr. However, I wondered if anyone has any idea what the remaining constituents or composition of the paint would be after drying.
Thanks in advance,
Steve
Steve Buckingham Dir. of Process Development Excellatron Solid State LLC 1640 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 617 812 5920
Michelle, I have good results staining respiratory mucosal tissue embedded in Araldite 502 with Alcian blue (8GS?) with or without PAS and in conjugation with Weigert's hematoxlyn. The plastic is first etched 1 to 3 minutes with sodium ethoxide, one:one, with toluene. After that I followed standard histological protocols. However, the staining times need to be increased and/or heated to compensate for the reduced number of available staining sites in semithins. The sections were spectacular I don't mind saying.
Hank Adams Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77030
Hello fellow microscopists-
Does anyone have any experience with staining thick (semi-thin) sections with Luxol Fast Blue?
I believe it's the nature of the Spiroplasma genus membrane structure rather than the poor preservation from your side. The usual glut/OsO4 fixation should be just fine, the problem is in visualization - lack of contrast. There is a good site on http://www.oardc.ohio-state.edu/spiroplasma/what.htm explaining evolutionary degeneration of the cell walls of the whole class Mollicutes. You may want to try some other contrast enhancing methods in addition to traditional heavy metal post-staining. Feel free to contact me off-line.
Alice Dohnalkova Environmental Microbiology Battelle, Pacific Northwest National Lab Richland, WA 99352 (509)372-0692
-----Original Message----- } From: Haixin Xu To: Mel Dickson Cc: microscopy-at-sparc5.microscopy.com Sent: 11/16/2001 9:23 AM
Hi Listers,
We are trying to microscopy a microorganism, spiroplasma using TEM. Every time we had problem to see clearly the membrane around the organism. We tried to increase the time of Osmium (4%) fixation, 3-5 hours at RT after glutaldehyde fixation. But the membrane preservation is still terrible. I would very much appreciate any of suggestions for that.
Steve Buckingham wrote: ========================================================== } I'm interested to know what exactly is left behind after Ag (or C) } paint (used as an electrical contact on a sample stub) has dried. I have used } this method of attaching samples to holders in ultra high vacuum systems and } after the paint is thoroughly dried there is no outgassing in uhv to 10-10 } torr. However, I wondered if anyone has any idea what the remaining } constituents or composition of the paint would be after drying. ========================================================== Our firm has had hands-on invovlement in the formulation of both silver and carbon "paints" for SEM (including UHV) applications going back to the early 1970's. I think I can comment on this topic.
The silver and carbon systems have to be discussed separately.
In the case of the silver system, of the leading brands used in microscopy, there is some considerable difference in the size and shape of the silver colloid, the suspending organic carrier and the composition and loading of polymer(s) that are present to impart to the paint different characteristics . We have learned over the years that some polymer is important in order to give the final paint layer the kind of adhesive properites users desire. More can sometimes be "too much" and some polymers tend to be more effective as an adhesive at low levels than other polymers. The shape of the colloid (e.g. flat platelet vs. round sphere) is important as well, since some sizes/shapes of the silver colloid lead to a higher likelihood of a skin forming during drying, which then acts as a transport barrier so that there is a wet region underneath. When such a skin-over-wet sample is inserted into the vacuum system, we all know the consequences.
Finally, we have found that not all silver paints found in the microscopy market have the same high purity, although we would be the first to recognize that for most SEM users, the presence of such impurities (such as contaminant SiO2 particles, otherwise known as glass frit) at low levels would not make any difference. There is a difference in the % silver loadings, something many just don't appreciate, and sometimes what appears to be a "better price" deal in fact is a product that is "cheaper" because of lower silver solids.......full disclosure on the percent silver solids at least for the SPI Supplies products are given on our website. Further evidence of the differences between products are the differences in flash points as disclosed in MSDS information.
So to now answer the question, with regard to silver, after the liquid carrier evaporates completely, with no wetness under a skin, there should be only the silver colloid, polymer at a low level, and otheriwise unintended impurities. Most people report that when the dried SPI Supplies silver paint is inserted into a UHV sustem, they find no indication of a diminuition of the quality of the vacuum.
For the carbon paint, the system is a bit easier to describe, the colloid is present much as is the silver colloid, a bit of a polymer to give the carbon paint greater adhesion properties, and a low level of impurities. Again, the carbon paints are not "all the same". There are differences in the carbon colloid size, difference in the polymer, and differences in the impurity elements that might be found. When the carbon paint dries, there is also a tendency for a skin to form, and since there are differences in the formulations, there are differences in the tendency for skin formation. But when fully dry, and there is no organic carrier underneath a skin, there should be only the carbon colloid, the polymer, and unintended impurities (which vary from product to product). We have had similar reports with regard to the SPI Carbon Paint product, that when inserted into a UHV system , there is no indication of a diminuiution of the quality of the vacuum.
Disclaimer: For nearly thirty years, SPI Supplies has formulated silver and carbon paints for SEM/EDS and other vacuum applications, and we have been able to do extensive testing on our own in-house equipment of various formulations. Now despite our obvious eagerness to sell more, I would urge consideration of the dry adhesives, mainly the double sided conductive carbon sheets and the analogous double sided silver sheet products. They too seem to be equally vacuum compatible and one eliminates the uncertainly of any kind of "wet" region underneath a skin on the surface. One also eliminates the possibility of inhalation of organic vapors. Such double sided (dry) adhesives are available from SPI Supplies as well as from the main supply sources for consumables for SEM and TEM. But like with the silver and carbon paint products, they don't all "come from the same place" and your experiences with one might not necessarily be the same as with the others. You might want to try more than one kind and then draw your own conclusions.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Erinasilk-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, November 17, 2001 at 15:31:20 ---------------------------------------------------------------------------
Email: Erinasilk-at-aol.com Name: Erin Silk
Organization: Summit Middle Schoo
Education: 6-8th Grade Middle School
Location: Boulder,Colorado,USA
Question: What power microscope do I need to see bacteria grow on samples of meat. I have two microscopes one that is up to 43x and another at up to 900x. I want to see and measure if and how much bacteria is present in my slide samples.
I am looking for a strange device, but I suppose already someone see something like hat.
A friend of mine is an optician and got a slit lamp.
He is now picking up pictures to be stored in a computer with a TV camera.
Obviously, after the frame grabber, he get an ugly quality of the image.
The slit lamp is equipped with a camera attachment and he got a camera too, but the procedure of taking a picture with a film, develop it, scan and store it is too long.
I remember of a converter device that fit in any 35 mm camera with a CCD sensor able to convert a normal film camera in a digital one.
BIMS will soon be profiled by some major analysts and newsletters along with the release of significant news regarding explosive sales for the Company. There will be huge volume and a strong increase in price for several days. The same groups that featured TIWI will begin coverage on BIMS. TIWI exploded from $ .44 to $2.41 in 4 days!! We know for certain that the same groups are going to feature BIMS and even better returns are expected.
We are very proud that we can share this information with you so that you can make a profit out of it. It is highly advisable to take a position in BIMS as soon as possible, today before the market closes or tomorrow.
The stock could easily reach $5.00 in less than a month on the strength of their upcoming contract announcements and Strong Analyst Buy Recomendations.
At 11:09 PM 11/17/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Name: Erin Silk } } Organization: Summit Middle School } } Education: 6-8th Grade Middle School } } Location: Boulder,Colorado,USA } } Question: What power microscope do I need to see bacteria grow on } samples of meat. I have two microscopes one that is up to 43x and } another at up to 900x. I want to see and measure if and how much } bacteria is present in my slide samples. } Erin -
The "900x" may do the job for you. But since anything above 400x SHOULD require what a microscopist calls an "oil immersion objective", I fear that your lenses will not give you a clear image. The best way available to you for actually seeing the bacteria will be to use a sterile swab to take a sample to smear on a slide; you'll probably need to stain them to make them visible. Getting accurate quantitative data probably won't be possible with the methods available to you. Look in catalogs like Flinn Scientific, Carolina Biological, or Edmund Scientific for kits with stains and instructions. If you can't see them, consider trying to grow the bacteria on culture medium in petrie dishes. You can find supplies in the same catalogs.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
This depends on whose paint you use. Ted Pella, SPI, EMS, etc. all use different solvents (such as methylethylketone, acetone, and others), and they use some sort of polymer (acrylic?) as a binder. The best thing is to get the MSD sheet for the particular brand you use, and call the folks from whom you bought the paint.
Phi
} Hi Y'All, } I'm interested to know what exactly is left behind after Ag (or C) } paint (used as an electrical contact on a sample stub) has dried. I have } used this method of attaching samples to holders in ultra high vacuum } systems and after the paint is thoroughly dried there is no outgassing in } uhv to 10-10 torr. However, I wondered if anyone has any idea what the } remaining constituents or composition of the paint would be after drying. } } Thanks in advance, } } Steve } } Steve Buckingham } Dir. of Process Development } Excellatron Solid State LLC } 1640 Roswell Street, Suite J } Smyrna, GA 30080 } phone 770 438 2201 } fax 617 812 5920
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Unless I'm dreadfully mistaken a microtome does not produce extreme heat (like an engine) and would not be used at -100C. The oil is required for lubrication and should cause little drag due to high viscosity. Sounds like sewing machine oil to me. If it was my microtome, I'd use some Molyslip (molybdenum disulfide), which lubricates better than oil, does not lose its properties, sticks better then grease and does not increase drag like grease. Until better reasons are provided, it appears somebody is selling snake-oil. Many years ago a top engineer from Ford Motors compared fancy oil additives with quality engine oil thus: They are mouse milk and very precious, but don't do anything that Cows-milk won't do.
On second thoughts Jonathan: I'll provide twice the volume and include freight cost. Maybe we can corner the world market in "microtome oils". Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, November 17, 2001 7:52 AM, Jonathan Wilson [SMTP:wilson_jm-at-cimar.org] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I have been loaned a microtome (Leitz 1512) by a colleague which } according to the instruction manual requires regular oiling. The microtome } did not come with any oil and the price I have been quoted by a local } supplier for microtome oil was about 75$US (including VAT) for 50ml (SAKURA } 1447). I found the price a bit steep and am left wondering if it is } absolutely necessary to buy "microtome" oil or if I can use an off the shelf } oil (e.g 3 in 1 or something like that). I do not want to compromise the } performance of the microtome as it is not mine and would appreciate any } advice from those with experience. } Thanks for any help. } Sincerely, } Jonathan } } Jonathan Wilson (PhD) } CIIMAR } Rua do Campo Alegre 823 } 4150-180 Porto, Portugal } tel: 351 22 608 0470 / 71 } fax: 351 22 606 0423 } e-mail: wilson_jm-at-cimar.org } web: www.cimar.org
Hi, I am trying to build fluorescence confocal microscope using a laser,a dichroic mirror, an objective(numerical aperture=0.7;working distance = 6 mm; focal length = 2 mm; focus depth 0.6 micron)which focuses into a cryostat,the emmited light is focused into the monochromator with a mirror in, and then to the CCD.
However, I did not achieve wide illumination, I've checked the spot size with the CCD but it had hardly changed even though i added a short focus lens in the exciting beam path and a diffuser. I also tried to put a longer focusing lens in the exciting beam path so that the focal point will be inside the objective but the image didn't widened.
Does anyone has an idea how to solve the problem and what lens or optics i should use in the path of the exciting laser so that i would get wide illumination. and a wide image as well.
hello everyone, I very rarely send out a blanket message - please bear with me, but you might like to consider this virus warning. best wishes, Jeremy
--- "Sanderson, Yvonne (REPP)" {Yvonne.Sanderson-at-repp.co.uk} wrote: } From: "Sanderson, Yvonne (REPP)" } {Yvonne.Sanderson-at-repp.co.uk} } To: "'jb_sanderson-at-yahoo.com'" } {jb_sanderson-at-yahoo.com} } Subject: FW: New Virus - Important } Date: Mon, 19 Nov 2001 14:19:21 -0000
} Importance: High } } Subject: New Virus - Important } Someone is sending out a very cute screensaver of the Budweiser Frogs. } } } } } } } } } } If you download it, you will lose everything! Your hard } drive will crash } } } } } } } } } } and someone from the Internet will } get your screen name and } } } } } } password! } } } } } } } DO } } } } } } } } } } NOT DOWNLOAD IT UNDER ANY } CIRCUMSTANCES! } } } } } } } } } } It just went into circulation } yesterday. Please distribute } } this } } } } } } } } } } message.This is a new, very } malicious virus and not many } } people } } } } } } know } } } } } } } } about } } } } } } } } } } it. This information was announced } yesterday morning from } } } } } } Microsoft. } } } } } } } } } } Please share it with everyone that } might access the } Internet. } } } } } } } } *************************************************************************************** } The information in this internet eMail is } confidential and is intended } solely for the addressee(s). Access, copying, } dissemination or re-use } of information in it by anyone else is unauthorised. } Any views or opinions } presented are solely those of the author and do not } necessarily represent } those of Reed Educational & Professional Publishing } or any of its affiliates. } If you are not the intended recipient please contact } } Reed Educational & Professional Publishing, Oxford, } +44 1865 311366. } *************************************************************************************** }
__________________________________________________ Do You Yahoo!? Everything you'll ever need on one web page from News and Sport to Email and Music Charts http://uk.my.yahoo.com
You may try using low molecular weight tannic acid after osmium fixation.
Reference: Nicolae Simionescu and Maia Simionescu 1976. Galloylglycoses of low molecular weight as mordant in electron microscopy. The Journal of Cell Biology. Vol 70, 608-621
Annfook Yang
} } } Haixin Xu {xu-at-gl.umbc.edu} 11/16/01 12:23PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Listers,
We are trying to microscopy a microorganism, spiroplasma using TEM. Every time we had problem to see clearly the membrane around the organism. We tried to increase the time of Osmium (4%) fixation, 3-5 hours at RT after glutaldehyde fixation. But the membrane preservation is still terrible. I would very much appreciate any of suggestions for that.
Microscopy Society of America Undergraduate Research Scholarship Program
With this year's call for applications the MSA Undergraduate Research Scholarship Program begins its 14th year providing funding for undergraduate research. To date over 71 projects covering a wide range of topics in the physical and biological sciences have received support through this program. Over the years nearly all the scholarship recipients have maintained a strong interest in imaging sciences and have gone on to graduate school, professional school, teaching, or industry positions. The program, which is funded by MSA and by matching funds from MSA Sustaining Members, is able to support approximately 30% to 40% of applicants. This past year the MSA Executive Council approved an increased in the amount provided by MSA. This, plus matching funds, will permit funding of additional scholarships for 2002. The maximal award for the Undergraduate Scholarships is $3000 and helps to provide student stipends, supply costs, and limited travel expenses associated with the research. Additional support in the form of instrument use time, equipment purchases, etc. is generally provided by the student's supervisor and/or through the sponsoring institution. Abstracts reporting the research results, are prepared by scholarship awardees, and published in "Microscopy and Microanalysis." The program actively seeks external sources of matching funds in order to maintain the favorable levels of support both in terms of the number of projects supported and the level of support for each. We are extremely grateful for the matching support provided by MSA sustaining members and individuals. Their support over the years has enabled the program to increase both the number of awards and the maximum amount of each award.
The MSA Undergraduate Research Scholarship Program is currently solicting applications from students interested in conducting a research project which involves the use of any microscopy technique. Students should be sponsored by a member of MSA. The maximal award is $3000. The application deadline is Dec 31, 2001. Applications can be obtained from the MSA Business Office, businessoffice-at-msa.microscopy.com or call toll free at (800) 538-3672. If you have any questions or require additional information regarding the program please contact either:
Dr. Ralph Albrecht, University of Wisconsin 1675 Observatory Drive, Madison, WI 53706 (608) 263-3952 or 263-4162; (608) 262-5157 FAX; albrecht-at-ahabs.wisc.edu
Dr. Richard Ornberg, Monsanto Company Analytical Sciences Center, 800 N. Lindbergh Blvd., St. Louis, MO 63167 (314) 694-1184; (314) 694-6727 FAX; rlornb-at-ccmail.monsanto.com
This Year's MSA Undergraduate Research Scholarship Awardees and Alternates were:
Awardees: JORGE GARCIA "Frictional and Mechanical Interactions between Nanotubes and Self-Assembled Monolayers: An Atomic Force Microscope Study" University of Texas-El Paso/Univ. Wisconsin, Madison Advisor: Robert Carpick
JAYAN RAMMOHAN "Imaging Aggregation Proteins InVitro Using Atomic Force Microscopy" Case Western Reserve University Advisor: Steven Eppell
PETER KRSKO "Electron Beam Lithography for Micro-Patterning of Bioactive Surfaces" Stevens Institute of Technology Advisor: Matthew Libera
DANIEL J. MAZEAU "Three-Dimensional Structure of the Human TFIIH-IIE Complex" University of California-Berkeley Advisor: Eva Nogales
DANIEL L. PECHKIS "The Impact of Processing on the Interface between Thin Film Dielectrics and their Silicon Substrates" Southern Connecticut State University Advisor: Christine Broadbridge
EUGENE KUNG "HR-TEM Studies of Polymer-Carbon Nanotube Composites" Princeton University Advisor: Nan Yao
Alternates RENEE LOPEZ SMITH "Comparisons Between Pre-Released and Swimming Sperm Cells of the Fern Lygodium: A Correlated SEM, TEM, and Light Microscope Study" Southern Illinois University Advisor: Karen Renzaglia
ERIN GRUND "Quantitation of Glomerular Cell Number in Diabetic Patients and Normal Controls" University of Minnesota Advisor: John Basgen
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Thanks to all who replied to my posting last week about camera-to-microscope interfacing, and for the suggestions, many of which suggested contacting my local Nikon rep.
I would be perfectly happy to use Nikon parts, after all, I'm trying to arrange a marriage between a Nikon camera and a Nikon microscope, so it shouldn't be THAT difficult.
Unfortunately, in my country there is no direct Nikon presence, only local representatives, but one company handles cameras, another handles microscopes, so the question of their interfacing kind of falls into a small but deep void.
If any expert from Nikon is reading this list, I would be grateful if they would reply to me directly, so that we could engage in a constructive dialogue.
thanx
rtch
} } Hi } } We have a Nikon Labophot-POL microscope. } } In its trinocular head there sits a CF PL 5X Projection lens, above } that is a Microflex HFX-IIA Photomicrographic Attachment, and onto } that is a "35mm camera adapter A", then what is called in the } HFX-IIA manual a "Motorized dark box FX-35WA". The latter looks more } like a camera body to me. } } We want to be able to mount our Nikon D 1 body onto the microscope, } with or without the HFX-IIA, whatever it takes. } } Is there anyone out there who can tell, from the above, what I need } to do to acheive this? } } I would have thought that all we need to do would be to mount the D } 1 body directly onto the trinocular vertical tube, but then I know } almost nothing about light microscopy. }
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I just read Gib Ahistrand's short in "Microscopy Today" about cleaning rolls in a laser printer. When I have problems with rubber rolls, either ink jet printers or laser printers, I rub the rubber with a lintless cloth moistened with isopropanol. It removes the "shine" that Gib is talking about. It definitely fixes the problem on inkjet printers where the paper will not feed from the stack. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Maybe try fast freezing plus freeze substitution? Either high pressure freezing or slam freezing? This might better capture the membrane in its native state. Then a freeze sub into osmium tetroxide in acetone should give good contrast to the membrane.
David H. Hall Center for C. elegans Anatomy Department of Neuroscience 1410 Pelham Parkway Albert Einstein College of Medicine Bronx, NY 10461
I am organizing a Leica confocal user/administrator listserver in order to facilitate and encourage dialogue regarding the problems, idiosyncrasies and use of the Leica SP platform for confocal and multiphoton microscopy. I am hoping that such a forum would be regarded as a logical extension of the existing microscopy listserver for Leica confocal users to voice Leica-specific questions/concerns.
A Leica LCSM/MP Listserver will allow us to archive questions and answers, and may offset some of the perceived lack of documentation regarding the software. As I have witnessed recently on the general confocal listserver, Leica is paying attention to users' sharing their dilemmas, concerns and successes. It would be advantageous for system administrators, end users and Leica representatives to continue to build momentum in this direction.
LeicaSP_LCS_MP_Users is a subscriber access list-server devoted to questions and issues unique and/or specific to the Leica SP Series Spectral Confocal and Multiphoton System.
If you would like to subscribe, please send an e-mail message to:
listserv-at-listserv.uiuc.edu
with the body of the message consisting of:
subscribe LEICASP_LCS_MP_USERS {full name}
I will be delighted to manage/maintain a Leica confocal/mp listserver resource. Let me know if you think if you have any thoughts, suggestions or concerns regarding this user forum at your convenience. Thank you.
Best Regards, Karl G.
_______________________________________________ Karl Garsha Specialist in Light Microscopy Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana, IL 61801 Room B650J Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
I work for a company in Lafayette, Louisiana and we have an interest in obtaining some SEM and TEM images of chitin isolated from crab shell waste. Particularly, I am interested if anyone knows of institutions that provide such a service, be it private or public, the cost of such services and contact information. The chitin samples we have are in a flake form about 5 - 7mm square. None of the samples are ready for imaging, so they would need to be prepared for the respective imaging processes. We would prefer the images to be in a digital format.
Thank you in advance for any information received.
André N. Blanchard, Ph.D. Manager-Technical and Business Development The Venture Group, Inc PO Box 53631 Lafayette, LA 70505-3631 U.S.A.
A Leica listserv already exists and the reasons for the establishment of that list several years ago were similar to those being touched upon here. Traffic on that list has been essentially nil for the past two years. This tells me one of three things: a) that nobody has any Leica-specific problems that they feel can be solved by list interaction, b) that the interaction received through the list was too limited to make the list a useful resource, or c) that the list consists of "power users" who fix their own problems. I suspect the answer is a mix of a and c. Certainly a lot of Leica systems have been installed since the list was first put together, and I am the first to admit that it was never well publicized, partly for reasons that may become apparent below.
If you desire to move forward with establishment of this list (and I am not discouraging you from doing so), then I would ask you to consider a few points regarding how listservs work and who stands to benefit from the list. 1) is the list set up as "reply to all" or "reply to sender"? 2) can anyone subscribe (it would appear so) and post, or can only those people with legitimate interests in Leica systems subscribe? 3) will Leica monitor the list? will they respond to posts to the list? These two items are important. With the earlier (still existing) list, the answers are "yes" and "no". I hope you can achieve a "yes" for both questions, but you must at least achieve a "yes" for the first question, otherwise it can be a needlessly unpleasant experience for the company itself - very much a blindside tackle. If people simply want to talk about how terrible tech support or service can be, then the list is pretty useless (witness current BioRad postings) - everyone knows this is a sore point with virtually every fancy piece of equipment sold from microscopes to FACS machines. We don't need "venting" lists, particularly if the people who should be "vented upon" are not listening (if they were, we wouldn't be venting now would we??).
We are about to have a new SP2-MP-UV system installed here. My prediction is that the learning curve on this system will be no steeper than that on the TCS-NT system - fairly shallow but with a few specific headaches (really just finding out which buttons to push when). What would really be useful is a list for people who still have older instruments (e.g., TCS4D) on which the learning curve is not only very steep but also for which the repository of that knowledge is a handful of people worldwide. The final point I would like to bring up is that the hardware/software configurations of the various Leica machines sold over the years (even just SP/SP2 series) can be very different, so it can be dangerous to generalize.
Rob Palmer Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
Hi all, We have been using our Gatan duo mill for some time now with a copper exhaust line on the rotary pump. It has worked fine, but now we have someone who would like to use the iodine guns and I am wondering if we should use a copper exhaust line with a corrosive gas like iodine. Does anyone know if copper is an appropriate material to use for an exhaust line when milling with iodine? Any additional pointers for milling with iodine? TIA. Greg
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
It is difficult to answer the question Maxine Daws raised about the operation of the gun and comumn valves in her electron microscope without having a diagram of the vacuum system; however, operating procedures for a variety of different vacuum systems (with a variety of different kinds of pumps) are described in some detail in Chapter 9 of my book 'Vacuum Methods in Electron Microscopy' (see http://www.2spi.com/catalog/books/book48.html and http://pup.princeton.edu/titles/6484.html, for a description). If you were to read this chapter, I believe that you could figure out what the proper operating sequence might be. Otherwise, Maxine, if you will send me a diagram of your system I will see if I can answer your question.
Good luck & Happy Thanksgiving, W. C. Bigelow -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
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For the heck of it I did a quick & dirty EDS (thin window detector) on some old carbon and Ag paint spots, no idea of who distributed this paint. The carbon had significant peaks at C, O & P. The Ag had significant peaks at Ag, C & S. The only surprise to me was the P peak with the carbon paint. The S on the Ag spot probably came from Ag tarnishing in air. For what its worth..... Bruce Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Lab University of Kansas
} } Hi Y'All, } } I'm interested to know what exactly is left behind after Ag (or C) } } paint (used as an electrical contact on a sample stub) has dried. I have } } used this method of attaching samples to holders in ultra high vacuum } } systems and after the paint is thoroughly dried there is no outgassing in } } uhv to 10-10 torr. However, I wondered if anyone has any idea what the } } remaining constituents or composition of the paint would be after drying. } } } } Thanks in advance, } } } } Steve } } } } Steve Buckingham } } Dir. of Process Development } } Excellatron Solid State LLC } } 1640 Roswell Street, Suite J } } Smyrna, GA 30080 } } phone 770 438 2201 } } fax 617 812 5920
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id QAA18046 for dist-Microscopy; Mon, 19 Nov 2001 16:59:09 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id QAA18039 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 19 Nov 2001 16:58:39 -0600 (CST) Received: from postal1.lbl.gov (postal1.lbl.gov [128.3.7.82]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id QAA18032 for {microscopy-at-sparc5.microscopy.com} ; Mon, 19 Nov 2001 16:58:26 -0600 (CST) Received: from SpamWall.lbl.gov (localhost [127.0.0.1]) by postal1.lbl.gov (8.11.2/8.11.2) with ESMTP id fAJMsqD26340 for {microscopy-at-sparc5.microscopy.com} ; Mon, 19 Nov 2001 14:54:52 -0800 (PST) Received: from lbl.gov (maokeefe.wins.lbl.gov [131.243.3.225]) by SpamWall.lbl.gov (8.11.2/8.11.2) with ESMTP id fAJMsq526332; Mon, 19 Nov 2001 14:54:52 -0800 (PST) Message-ID: {3BF98EA8.C7832787-at-lbl.gov}
I believe that this is a hoax. See: http://www.f-secure.com/hoaxes/budfrogs.shtml -Mike
Jeremy Sanderson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } hello everyone, } I very rarely send out a blanket message - please bear } with me, but you might like to consider this virus } warning. } best wishes, Jeremy } } --- "Sanderson, Yvonne (REPP)" } {Yvonne.Sanderson-at-repp.co.uk} wrote: } From: } "Sanderson, Yvonne (REPP)" } } {Yvonne.Sanderson-at-repp.co.uk} } } To: "'jb_sanderson-at-yahoo.com'" } } {jb_sanderson-at-yahoo.com} } } Subject: FW: New Virus - Important } } Date: Mon, 19 Nov 2001 14:19:21 -0000 } } } Importance: High } } } } Subject: New Virus - Important } } } Someone is sending out a very cute screensaver of the } Budweiser Frogs. } } } } } } } } } } } } If you download it, you will lose everything! Your } hard } } drive will crash } } } } } } } } } } } and someone from the Internet will } } get your screen name and } } } } } } } password! } } } } } } } } DO } } } } } } } } } } } NOT DOWNLOAD IT UNDER ANY } } CIRCUMSTANCES! } } } } } } } } } } } It just went into circulation } } yesterday. Please distribute } } } this } } } } } } } } } } } message.This is a new, very } } malicious virus and not many } } } people } } } } } } } know } } } } } } } } } about } } } } } } } } } } } it. This information was announced } } yesterday morning from } } } } } } } Microsoft. } } } } } } } } } } } Please share it with everyone that } } might access the } } Internet. } } } } } } } } } } } } } } *************************************************************************************** } } The information in this internet eMail is } } confidential and is intended } } solely for the addressee(s). Access, copying, } } dissemination or re-use } } of information in it by anyone else is unauthorised. } } Any views or opinions } } presented are solely those of the author and do not } } necessarily represent } } those of Reed Educational & Professional Publishing } } or any of its affiliates. } } If you are not the intended recipient please contact } } } } Reed Educational & Professional Publishing, Oxford, } } +44 1865 311366. } } } *************************************************************************************** } } } } __________________________________________________ } Do You Yahoo!? } Everything you'll ever need on one web page from News and Sport to Email and Music Charts } http://uk.my.yahoo.com
You need an activated carbon trap between your mechanical pump and your turbo. When I was at the University of Alabama at Birmingham, students on two occasions left the iodine gun valves open. They destroyed the mechanical pump springs and rendered it useless. I found out that we were not the only ones that this happened to. Gatan fixed a bunch of mechanical pumps under warranty and came up with a fix -a carbon trap with a sight glass on the mechanical pump side. (Our second pump failure was not covered.) Inside the tube where you could see, there was a silver strip that would change to black when the trap got saturated. You regenerated the trap by replacing the charge because otherwise you would be getting iodine into the mechanical pump. I would worry that copper might react similarly to iodine as the silver. The iodine does come out the exhaust of the mechanical pump.
I thought that the fix was rather expensive at the time, but it did work. You could probably get a zeolite trap and replace the zeolite with activated carbon from a pet store that sells aquarium supplies. Ten years ago, Gatan wanted $90 for a charge of activated carbon (about a quart jar). You could get twice that amount at a pet store for less than $10.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
-----Original Message----- } From: Greg Strout [mailto:stro8031-at-msmailhub.oulan.ou.edu] Sent: Monday, November 19, 2001 5:17 PM To: Microscopy-at-sparc5.microscopy.com
Hi all, We have been using our Gatan duo mill for some time now with a copper exhaust line on the rotary pump. It has worked fine, but now we have someone who would like to use the iodine guns and I am wondering if we should use a copper exhaust line with a corrosive gas like iodine. Does anyone know if copper is an appropriate material to use for an exhaust line when milling with iodine? Any additional pointers for milling with iodine? TIA. Greg
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
I think you need to check your supply of paints. I never had anything behind C and O on carbon paint, do not remember S on Ag either.
Vladimir
} } For the heck of it I did a quick & dirty EDS (thin window } detector) on some old } carbon and Ag paint spots, no idea of who distributed this } paint. The carbon had } significant peaks at C, O & P. The Ag had significant peaks } at Ag, C & S. The } only surprise to me was the P peak with the carbon paint. The } S on the Ag spot } probably came from Ag tarnishing in air. } For what its worth..... } Bruce } Bruce Cutler, Ph.D. } Director, Microscopy & Electronic Imaging Lab } University of Kansas } } } } } Hi Y'All, } } } I'm interested to know what exactly is left behind } after Ag (or C) } } } paint (used as an electrical contact on a sample stub) has } dried. I have } } } used this method of attaching samples to holders in ultra } high vacuum } } } systems and after the paint is thoroughly dried there is } no outgassing in } } } uhv to 10-10 torr. However, I wondered if anyone has any } idea what the } } } remaining constituents or composition of the paint would } be after drying. } } } } } } Thanks in advance, } } } } } } Steve } } } } } } Steve Buckingham } } } Dir. of Process Development } } } Excellatron Solid State LLC } } } 1640 Roswell Street, Suite J } } } Smyrna, GA 30080 } } } phone 770 438 2201 } } } fax 617 812 5920 } } } } } }
I suggest that you try Structural Research Inc. in Mobil, Alabama, (205) 649-9740. They come recommended although I have no personal experience with them.
Best regards,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Michael O'Keefe" To: listserver a {MAOKeefe-at-lbl.g {microscopy-at-sparc5.microscopy.com} ov} cc: Subject: Re: Fwd: FW: New Virus - Important
11/19/01 04:58 PM
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I believe that this is a hoax. See: http://www.f-secure.com/hoaxes/budfrogs.shtml -Mike
Jeremy Sanderson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } hello everyone, } I very rarely send out a blanket message - please bear } with me, but you might like to consider this virus } warning. } best wishes, Jeremy } } --- "Sanderson, Yvonne (REPP)" } {Yvonne.Sanderson-at-repp.co.uk} wrote: } From: } "Sanderson, Yvonne (REPP)" } } {Yvonne.Sanderson-at-repp.co.uk} } } To: "'jb_sanderson-at-yahoo.com'" } } {jb_sanderson-at-yahoo.com} } } Subject: FW: New Virus - Important } } Date: Mon, 19 Nov 2001 14:19:21 -0000 } } } Importance: High } } } } Subject: New Virus - Important } } } Someone is sending out a very cute screensaver of the } Budweiser Frogs. } } } } } } } } } } } } If you download it, you will lose everything! Your } hard } } drive will crash } } } } } } } } } } } and someone from the Internet will } } get your screen name and } } } } } } } password! } } } } } } } } DO } } } } } } } } } } } NOT DOWNLOAD IT UNDER ANY } } CIRCUMSTANCES! } } } } } } } } } } } It just went into circulation } } yesterday. Please distribute } } } this } } } } } } } } } } } message.This is a new, very } } malicious virus and not many } } } people } } } } } } } know } } } } } } } } } about } } } } } } } } } } } it. This information was announced } } yesterday morning from } } } } } } } Microsoft. } } } } } } } } } } } Please share it with everyone that } } might access the } } Internet. } } } } } } } } } } } } } } ***************************************************************************************
} } The information in this internet eMail is } } confidential and is intended } } solely for the addressee(s). Access, copying, } } dissemination or re-use } } of information in it by anyone else is unauthorised. } } Any views or opinions } } presented are solely those of the author and do not } } necessarily represent } } those of Reed Educational & Professional Publishing } } or any of its affiliates. } } If you are not the intended recipient please contact } } } } Reed Educational & Professional Publishing, Oxford, } } +44 1865 311366. } } } ***************************************************************************************
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Isn't it true some roughing pumps that are intended for this type of application are teflon coated internally for a barrier from corrosive gases?
Scott: By the way, the springs in that pump could have been replaced...
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282
Walck, Scott D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } You need an activated carbon trap between your mechanical pump and your turbo. When I was at the University of Alabama at Birmingham, students on two occasions left the iodine gun valves open. They destroyed the mechanical pump springs and rendered it useless. I found out that we were not the only ones that this happened to. Gatan fixed a bunch of mechanical pumps under warranty and came up with a fix -a carbon trap with a sight glass on the mechanical pump side. (Our second pump failure was not covered.) Inside the tube where you could see, there was a silver strip that would change to black when the trap got saturated. You regenerated the trap by replacing the charge because otherwise you would be getting iodine into the mechanical pump. I would worry that copper might react similarly to iodine as the silver. The iodine does come out the exhaust of the mechanical pump. } } I thought that the fix was rather expensive at the time, but it did work. You could probably get a zeolite trap and replace the zeolite with activated carbon from a pet store that sells aquarium supplies. Ten years ago, Gatan wanted $90 for a charge of activated carbon (about a quart jar). You could get twice that amount at a pet store for less than $10. } } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } P. O. Box 11472 (letters) } Guys Run Rd. (packages) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8515 (fax) } } "The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." } } } } -----Original Message----- } } } From: Greg Strout [mailto:stro8031-at-msmailhub.oulan.ou.edu] } } } Sent: Monday, November 19, 2001 5:17 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Exhaust line for ion milling } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } We have been using our Gatan duo mill for some time now with a copper } exhaust line on the rotary pump. It has worked fine, but now we have } someone who would like to use the iodine guns and I am wondering if we } should use a copper exhaust line with a corrosive gas like iodine. Does } anyone know if copper is an appropriate material to use for an exhaust } line when milling with iodine? Any additional pointers for milling with } iodine? TIA. } Greg } } -- } ================================================================== } Greg Strout } Electron Microscopist, University of Oklahoma } WWW Virtual Library for Microscopy: } http://www.ou.edu/research/electron/www-vl/ } e-mail: gstrout-at-ou.edu } Opinions expressed herein are mine and not necessarily those of } the University of Oklahoma } ================================================================== } } } } }
I was analyzing a situation where some electroless (Cu bath that plates w/o electricity) copper plate was peeling away from a copper substrate.
I have to clues. Oxidation and small blisters.
When I do a tape test I get a continuous sheet of electroless Cu on the back of the tape and also a fresh surface of the Cu substrate. The first clue is that with the naked eye you can see oxidation on both surfaces. To me this means there must have been trapped Oxidation during the plating process. Using SEM/EDS at low and medium kV, I did not observe any contamination on either surface, just very thin oxidation.
The next clue was that when I took a photomicrograph with the polarity of the Everhart and Thornley detector inverted I see a two types of topographies. I see the expected micro-etch of the Cu substrate, but also smooth bumps at 5-10 microns, as if the electroless plated some droplets. I think these droplets are really blisters that occurred at very beginning of the plating process. I feel that the blister cause the poor adhesion. I am not sure what caused the blisters, maybe internal stress in the plated deposit together with oxidized or very thin organic contamination of the substrate. Or possibly gas evolution. The substrate has a very even etch topography across the surface, no bumps. I would gladly e-mail these images to anyone interested in seeing this unexpected use of a replicated surface. I would also appreciate any comments.
We are looking into buying a Nikon coolpix 995 digital camera to be used to take mostly brightfield images in a Zeiss Axioplan microscope. So I would like to know how good this camera is for photomicrography. I also found out Nikon has a new coolpix 5000 in the market. Does anyone have good experience with this new one for our kind of application ?
Thanks a lot,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
I would like to receive any information about review papers on the basic principles of histochemical techniques. I am quite interested to receive references of papers on the basic characteristics of a histochemical method (reproducibility, specificity etc).
Many thanks.
Fabio Grizzi Scientific Direction Istituto Clinico HUMANITAS Milan, Italy
I would like to get in touch with Tina to ask her a question. I cannot seem to find her email address. Can anyone help?
Better yet, Tina, if you're listening, could you please reply?
Thanks.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
There is no such thing... Recently we have more and more spams and hoaxes, unfortunately. This virus warning (and the others) is a big fat hoax (http://www.symantec.com/avcenter/venc/data/buddylst.zip.html), only just for clogging the net, generating chain-mails... If you receive any "Important-Virus warning" in the future, please check the Symantec page for hoaxes first (http://www.symantec.com/avcenter/hoax.html). Don't forward these messages automatically, especially not for lists or discussion forums... Regards,
Imre
Imre Kovacs M.D., Ph.D. Research Fellow Genetics and Aging Unit Harvard Medical School Massachusetts General Hospital East Building 114, 16th Street Charlestown, MA, 02129-4404 Phone: 617-724-3253 Fax: 617-724-1823
On Tue, 20 Nov 2001 gary.m.brown-at-exxonmobil.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Michael, } } I suggest that you try Structural Research Inc. in Mobil, Alabama, (205) } 649-9740. They come recommended although I have no personal experience with } them. } } Best regards, } } Gary M. Brown } ExxonMobil Chemical Company } Baytown Polymers Center } 5200 Bayway Drive } Baytown, Texas 77520-2101 } phone: (281) 834-2387 } fax: (281) 834-2395 } e-mail: Gary.M.Brown-at-ExxonMobil.com
"Histological and Histochemical Methods" by John Kiernan
Most (all?) editions of "Cell and Tissue Biology" by Leon Weiss (editor) have a nice chapter on histochemistry by Helen Padykula.
Grizzi Fabio ICH wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All } } I would like to receive any information about review papers on the basic } principles of histochemical techniques. I am quite interested to receive } references of papers on the basic characteristics of a histochemical method } (reproducibility, specificity etc). } } Many thanks. } } Fabio Grizzi } Scientific Direction } Istituto Clinico HUMANITAS } Milan, Italy
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (reimar_gaertner-at-wsib.on.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, November 19, 2001 at 14:48:41 ---------------------------------------------------------------------------
Question: My daughter and I have come across a most curious pond-water creature that we can't identify in any of our books. It is large - about 400 um in diameter and round. It has movable internal organs that move food around - especially two pincer-like claws that appear to be macerating captured food. It therefore must be an animal - no cilia are apparent. It has a unique method of feeding: it unfolds a clear satalite-dish-like catchers mit with which it directs passing prey into it's mouth area. It can quickly retract the antena array when touched or jolted. What is this marvelous thing?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (elir-at-chem.ch.huji.ac.il) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, November 18, 2001 at 13:55:49 ---------------------------------------------------------------------------
Email: elir-at-chem.ch.huji.ac.il Name: Eli rothenberg
Organization: Hebrew University
Education: Graduate College
Location: Jerusalem, Israel
Question: I would like to know what is the correct setup for a wide illumination in a confocal microscopy arrangement, that is, what optics should i use in the path of the laser, if i want to get the maximum illuminated area through my objective, the setting is as follows: laser-collimated beam- dichroic mirror-objective-illuminated surface. (p.s I thought of using a short focusing lens that will focus half way through the objective so maybe that will give me a wide illumination on the surface, and/or maybe a diffuser to go along with it?)
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (malone1527-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, November 18, 2001 at 19:17:02 ---------------------------------------------------------------------------
Email: malone1527-at-aol.com Name: Gene Cimino
Organization: None-Microscopy is a hobby for me. I am 58 years old
Education: Undergraduate College
Location: Hydes,MD,USA
Question: Is Balsam still considered a good mounting medium? If so, can it be kept fluid enough to apply by adding Xylol to it? I am curious about this as I want to make some mounts of insect structures. I know about Permount, but I wanted to try the older Balsam method.By the way,I got my BS in Biology in 1967 from Towson University.
It sounds to me very much like a rotifer: these are very common, and can survive considerable desiccation, and so are often found in roof gutters (where I remember finding them). I would guess that the "satellite dish" is the ciliated "wheel" from which the phylum gets its name - I think that the cilia are either moving too fast or simply too small to be seen at whatever magnification you are using. This generates a current, and I remember as a boy watching green spherical algae being sucked in and macerated.
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
----- Original Message ----- } From: {reimar_gaertner-at-wsib.on.ca} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, November 21, 2001 3:34 AM
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I am trying to find out what has been recently published in the field of ink-jet materials (dyes, pigments) using transmission electron microscopy. If you know of any reviews or research articles on this specific subject I would appreciate your feedback!
Thank you very much,
Pedro Miguel Rodrigues de Almeida, Dr. Sc. Agfa-Gevaert N.V. RDM/PA-Mo - Electron Microscopy Laboratory Septestraat 27 BE-2640 Mortsel
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Dear expert microscopist, I'm a very beginner with SEM imaging and particularly with EDS Microanalysis. I'm trying to work with my own standard (Oxford ME-....) and the standard-loading job is not quite simple for me. I've got I few questions tha you might find stupid but I would appreciate any help!!! 1) The SEM is a LEO 4400 2) I work with geological sample 3) The EDS software package is ISIS 300 4) What are the best conditions for loasing the standard? i.e. 15 or 20 Kv EHT? 600 ma I-Probe or more? 50 or 100 sec. Preset Time? I have mostly pure metals and compounds plus Natural Albite, Ortoclase and Wollastonite. 5) Once I've performed the calibration using Cobalt, must I fix the counts per second to the same value I obtained for the Cobalt for any further standard acquisition? 6) Before preparing the element profiles, what's the best way to strip peaks away from the spectrum: basic strip or advanced strip? This is particularly important for silicates where Si, Al and Na peaks are close (i.e. whithin the -600/+600 ev energy windows). 7) How can I perform deconvolution of peaks (Si and Al) before saving the profiles? Is it possible to "erase" unwanted peaks from the standard spectrum without altering the background? Thank You for any answer or suggestion. I will appreciate. Melina Meli
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id XAA06086 for dist-Microscopy; Wed, 21 Nov 2001 23:35:31 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id XAA06083 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 21 Nov 2001 23:35:00 -0600 (CST) Received: from bloodwood.adelaide.edu.au (bloodwood.adelaide.edu.au [129.127.43.1]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id XAA06076 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 21 Nov 2001 23:34:48 -0600 (CST) Received: from adelaide.edu.au ([129.127.101.59]) by bloodwood.adelaide.edu.au (Netscape Messaging Server 4.15) with ESMTP id GN6SMM00.90Z for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 22 Nov 2001 16:00:22 +1030 Message-ID: {3BFC8D51.E5B293F-at-adelaide.edu.au}
Hi all,
I have been asked to remove a specimen from a sem stub turn it over and photograph the other side. The specimen is an organically preserved plant fossil and very fragile, it is attached to the sem stub using double sided black tape called STR tape made by Shinto Chemitron Co. Ltd. I have tried soaking a piece of the tape in acetone and also in ethanol to see if this would dissolve the tape. Neither did anything and the tape is still sticky. Any suggestions would be greatly appreciated
Jo Shrapnel Environmental Biology Adelaide University
Dear Listers, As my wife and I drove and hiked around the Northeast one last time to see the fall colors before driving across from New York to California, I thought of the differences among what could be seen while walking, driving, and flying. It occurred to me that these modes of transportation differed in rates by roughly factors of ten: a brisk walk is about 2 m/s; a leisurely drive is about 20 m/s; an airliner travels at about 200 m/s. As I thought further, I realized that a weather satellite at a height of about 1000 km would have an orbital speed of about 2 km/s, and scanning through a TEM grid looking for suitably crystallized outer mitochondrial membranes would happen at a speed of about 2 um/s. (I can examine the ~10 cm screen at 40 kx in about 1 s.). Also, a trip across the solar system at 0.01 c would take about a day. Other possibilities for filling in this series could be the scan speed of AFM on one end and the rate that the Hubble telescope can record a field for the deep field survey at the other. (Someone knowledgeable in astronomy could calculate the apparent speed by figuring how much distance is represented by the angular width of the Hubble field of view at a depth of 10^10 light-years and dividing by the exposure time.) Can we put together a complete list of powers-of-ten speeds, along with how these can be realized and what typically can be seen? I'll start.
?? AFM individual atoms 2 um/s TEM organelles 2 m/s walk individual rocks, trees, streams, etc. 20 m/s drive mountains, lakes, forests, etc. 200 m/s fly mountain ranges, storms, deserts, etc 2 km/s orbit weather patterns, continents 200 Mm/s ion drive? solar system ?? Hubble scan quasar field I hope you will have as much fun with this as I have. Yours, Bill Tivol
I'm not familiar with that particular product, but adhesives that remain sticky (rather than dry out) usually respond very well to naptha. Lighter fluid for zippo type cigarette lighters contains naptha and is an inexpensive and readily available source.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
-----Original Message----- } From: Joanne Colmer [SMTP:joanne.colmer-at-adelaide.edu.au] Sent: Wednesday, November 21, 2001 9:30 PM To: Microscopy-at-sparc5.microscopy.com
Hi all,
I have been asked to remove a specimen from a sem stub turn it over and photograph the other side. The specimen is an organically preserved plant fossil and very fragile, it is attached to the sem stub using double sided black tape called STR tape made by Shinto Chemitron Co. Ltd. I have tried soaking a piece of the tape in acetone and also in ethanol to see if this would dissolve the tape. Neither did anything and the tape is still sticky. Any suggestions would be greatly appreciated
Jo Shrapnel Environmental Biology Adelaide University
Dear all Most photographers are only too aware that flash intensity diminishes according to the square of the distance to the subject, making it difficult to use flash for distance photogrpahy with a lens or telescope. Some years ago I considered the possibility of illuminating a subject viewed with a telescope or telephoto lens by using the same optics to focus an electronic flash on the subject. A problem was how short would the flash duration have to be for the all illuminating light to have left the optics before image-forming light started to come back. I concluded that for an object distance of 50m, i.e. a total light-path of 100m, would be about a third of a microsecond, and gave up at that point, having no way of generating flashes that short.
An orders of magnitude table (in 3-orders of magnitude steps) for distance travelled by a light flash looks like this: 1 second 300,000 km 1 millisecond 300 km (this is typical electronic flash duration) 1 microsecond 300 m 1 nanosecond 0.3 m 1 picosecond 0.3 mm 1 femtosecond 0.3 µm (30nm)
so a light "beam" of 1 femtosecond duration, such as can be produced by lasers, is in fact a light disc!
Not a lot of people know that :-) Best wishes Chris
Date sent: Thu, 22 Nov 2001 03:10:18 -0500
Joanne
Chloroform appears to soften our sticky tape and its adhesive within a few minutes - if you have any spare STR tape you could try it first. It should be easy to separate the sample then. However I don't know whether chloroform will damage your sample, you will probably need to rinse the surface of the sample with fresh solvent and remember to use the chloroform in the fume hood.
Malcolm Haswell e.m. unit, School of Sciences, University of Sunderland, UK (+44)0191 515 2872
Joanne Colmer wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all, } } I have been asked to remove a specimen from a sem stub turn it over and } photograph the other side. } The specimen is an organically preserved plant fossil and very fragile, } it is attached to the sem stub using double sided black tape called STR } tape made by Shinto Chemitron Co. Ltd. } I have tried soaking a piece of the tape in acetone and also in ethanol } to see if this would dissolve the tape. } Neither did anything and the tape is still sticky. } Any suggestions would be greatly appreciated } } Jo Shrapnel } Environmental Biology } Adelaide University
Dear Colleagues, A colleague of mine, working on a thesis in pathology is looking for review papers on the application of confocal microscopy in cancer research, diagnosis and prognosis. Can anyone cite useful recent references for him, or possesses or has authored a favored reference? many thanks, Jeremy Sanderson
__________________________________________________ Do You Yahoo!? Everything you'll ever need on one web page from News and Sport to Email and Music Charts http://uk.my.yahoo.com
First posting to the list. I originally posted this on Sci.techniques.microscopy but was advised that better response may be obtained here. I'm a post doc reseacher at the Uni of B'Ham, UK.
I was wondering whether anybody had any experince of using colloidal gold in ESEM. I wish to visualise fungal extracellular matrices released onto glass or plastic surfaces in "wet" mode. If any body has any advice on this, or has used colloidal gold in ESEM and can pass on a few tips it would be most appreciated.
Regards
Steve Thomas
********************************* Dr Steve Thomas School Of Biosciences The University of Birmingham Edgbaston Birmingham B15 2TT 0121 414 5573 Email: S.Thomas-at-bham.ac.uk
Jo Shrapnel wrote: ======================================================= I have been asked to remove a specimen from a sem stub turn it over and photograph the other side. The specimen is an organically preserved plant fossil and very fragile, it is attached to the sem stub using double sided black tape called STR tape made by Shinto Chemitron Co. Ltd. I have tried soaking a piece of the tape in acetone and also in ethanol to see if this would dissolve the tape. Neither did anything and the tape is still sticky. Any suggestions would be greatly appreciated ======================================================== The adhesive system will not really "dissolve" in any common solvents. It can be swollen and loosened, such as in chloroform, which can aid in its removal, but it will not actually be dissolved away in the traditional sense . For a flat surface often times such solvent treatment is enough but for your kind of sample, I would trust it would not be enough.
But in a sense you are fortuante in that your sample of interest is not organic but is in fact inorganic in nature. You should be able to put it into a small glass petri dish, without cover, contaminating adhesive side "up", and then using an oxygen plasma, such as would be generated in the SPI Supplies® Plasma Prep™ II plasma etcher, the organics should be etched away and the substrate (relatively) untouched. I say "relatively" because eventually, in the limit, the oxygen will cause changes in the substrate, but the etch rate for the organic adhesive is many times faster than for the inorganic substrate. So you want to be careful to etch only to the extend you need to etch and to try not etching beyond that point.
Special note and disclaimer: SPI Supplies manufactures the Plasma Prep II plasma etcher so we would naturally have a vested interest in expanding its use. However, the same physics found in the SPI Plasma Prep II unit also exist in several other table-top systems being manufactured such as by Denton Vacuum and Polaron (Now Quorum Technologies, formerly, VG Microtech). More about the Plasma Prep II plasma etcher can be found at URL http://www.2spi.com/catalog/instruments/etchers1.html
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We are trying to locate a journal in the medical or cellular fields with an online version that accepts videoclips. Thank you in advance for your help.
as there was a little interest in the Leonid meteors on this list server last week I thought would report my observations from Ayers Rock, Australia.
Central Australia was expected to be reasonably placed to observe the peak activity early on Monday morning, Nov 19th. The sky was clear and dark from sunset to sunrise and relatively warm (about 16°C). From the sand dunes I spent a very pleasant 5 or 6 hours under the stars and counted approximately 300 meteors in the first 90 minutes. After that the rate was too great to keep count. I estimate seeing approximately 2000 to 3000 meteors, with many others that I missed because I was looking the wrong way at the time. I can well imagine whiplash injuries being commonplace amongst those of us fortunate enough to be in the right place at the right time with the right weather conditions.
By comparison, the next morning I saw only 8 meteors in 90 minutes, and only 2 of these were Leonids.
Nestor, I managed to capture a few meteors in photos (one 15 minute exposure has over 20 meteors visible on the print). I'll bring some of the better ones to Adelaide. Cheers,
Mark Blackford Materials Division, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} From: Maike Andersson [m.andersson-at-t-online.de] Sent: Mittwoch, 14. November 2001 21:27 To: Microscopy
I am trying to characterize the water pathway in plant tissue by the water soluble, apoplastic tracer sulforhodamin. In order to prevent secondary movement of the tracer, I want to freeze substitute under anhydrous conditions prior to embeddment in Spurr. What substitution media do you recommend? Does polar acetone lead to loss or secondary movement of the tracer? Is apolar diethylether a better choice and how can I overcome the very slow rewarming protocols of it? Thanks in advance
Maike Andersson Institut for Applied Botany Hamburg / Germany
Listers; The University of Calgary has a terrific job opening in the Microscopy and Imaging Facility.
Please check it out. Calgary is one of the most prime cities to live in. We are 70 miles from Banff and the best mountain activities in the world. Our province has no sales tax!
Because of government regulations we must first look at qualified Canadian citizens or Landed Immigrants and then folks from other nations. We of course are an equal opportunity employer.
Jon McGovern Manager, Microscopy and Imaging Facility University of Calgary Calgary, Alberta, Canada.
Position Available
The Microscopy and Imaging Facility of the University of Calgary requires an experienced EM technologist. The Microscopy and Imaging facility houses three transmission and three scanning electron microscopes. The flagship of these being a newly acquired FEI/Philips Tecnai F20 equipped with EDX, STEM, EELS, and Cryo imaging. This instrument is the only one of its kind in Canada. The successful candidate should have a very strong background in TEM, preferably with cryo microtomy and cryo observation. Experience in operational instruction to a wide group of users is a necessity. Overseas training on the Tecnai F20 will be provided. It is expected that the successful candidate will assist with the SEM side of the facility as required. Although there are no strict educational requirements, candidates should possess the appropriate qualifications and training for the position. Experience in operating EM’s, routine maitenance, management of the day-to-day operation of the instruments, the ability to instruct new users, and work in a multi-user environment would be an asset. This position will appeal to persons who thrive on challenges and are willing to take on demanding tasks. Applicants are invited to forward their resumes, three letters of reference and if possible examples of their expertise to:
Dr. John Reynolds (reynolds-at-ucalgary.ca) Head, Department of Cell Biology and Anatomy Faculty of Medicine University of Calgary 3330 Hospital Dr NW Calgary, Alberta Canada T2N 1N4
I've never looked at colloidal gold in an ESEM before, although I've imaged it in a FESEM, but my immediate reaction is that you would have two things working against you. One is that to image a wet or uncoated sample, you would have to introduce sufficient gas into the chamber to seriously degrade your resolution. The second is that you need to use the backscatter detector to image the gold particles, as you know, which again is significantly lower resolution than the secondary imaging mode. With these two resolution-losers combined, I seriously doubt that you could image gold particles down into the nanometer range.
But, that said, I always tell people that only way to really know for sure is to try. But, is there some compelling reason why the sample must be viewed in "wet" mode, or for that mattter, in an SEM at all? Is there a reason not label and embed your samples for TEM?
Good luck, Randy Tindall EM Core University of Missouri Columbia, MO
-----Original Message----- } From: Steve Thomas To: Microscopy-at-sparc5.microscopy.com Sent: 11/22/01 10:28 AM
Hello,
First posting to the list. I originally posted this on Sci.techniques.microscopy but was advised that better response may be obtained here. I'm a post doc reseacher at the Uni of B'Ham, UK.
I was wondering whether anybody had any experince of using colloidal gold in ESEM. I wish to visualise fungal extracellular matrices released onto glass or plastic surfaces in "wet" mode. If any body has any advice on this, or has used colloidal gold in ESEM and can pass on a few tips it would be most appreciated.
Regards
Steve Thomas
********************************* Dr Steve Thomas School Of Biosciences The University of Birmingham Edgbaston Birmingham B15 2TT 0121 414 5573 Email: S.Thomas-at-bham.ac.uk
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bob-at-rockisland.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, November 24, 2001 at 15:06:36 ---------------------------------------------------------------------------
Email: bob-at-rockisland.com Name: Bob Carter
Organization: independant researcher
Education: Graduate College
Location: Lopez Island WA 98261 USA
Question: Hi. I recently purchased a Vickers M171237 (Patholux???). It is a compound "research " type similar to the Leitz Orthomat. It has a quick change 5 objective nose, digital readout, substage and epi lighting, plus a box under the binocular with a beamsplitter, pol, and other optics I am unfamiliar with. I am restoring my Vickers for daily usage. I am looking for technical information, and user or repair guides. I completely disassembled, cleaned, reassembled, and adjusted an AO MicroStar, but this microscope is far more complicated. If you know of any resources, it would be greatly apreceated. Thanks, Bob
Bob Carter 2000 Bayshore Road Lopez Island, WA USA 98261
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Is there any direct way to quantitatively determine and measure the actual spot size of a SEM beam?
Given various KV, WD, final aperture sizes, condenser lens settings, etc., is there a way to truly find out what the real spot size is on the specimen?
In TEM we measure the actual contamination spot "burned" into the carbon film by the beam. I use a grid with calibration spheres for reference. With the appropriate organic film, a similar technique could work with SEM in spot mode?? I'm not sure how much accuracy you're looking for, but it might give you an idea.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Sunday, November 25, 2001 8:59 PM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } Hi all: } } Is there any direct way to quantitatively determine } and measure the actual spot size of a SEM beam? } } Given various KV, WD, final aperture sizes, condenser } lens } settings, etc., is there a way to truly find out what } the real spot size is on the specimen? } } Any ideas? } } gary g.
Gary, One technique I've seen used is to mount an aperture over a hole (on carbon is best), or better yet, a thinned foil for a sharper edge, and use a line-profile mode to scan across after focussing at a very high mag. Absorbed current can be used instead of secondary for a signal, although the latter works well, especially on a thinned foil. The curve you get will resemble a rise-time curve and can be treated in a similar manner. Mark the 10% and 90% rise points on the curve, then measure the horizontal distance between those two points in units appropriate for the mag. This is generally considered to be the beam diameter given the Gaussian distribution of electrons across the beam.
Ken Converse owner Quality Images third party SEM service Delta, PA
Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all: } } Is there any direct way to quantitatively determine } and measure the actual spot size of a SEM beam? } } Given various KV, WD, final aperture sizes, condenser lens } settings, etc., is there a way to truly find out what } the real spot size is on the specimen? } } Any ideas? } } gary g. } } } }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (twiinie-at-aolcom) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, November 25, 2001 at 22:36:35 ---------------------------------------------------------------------------
Email: twiinie-at-aolcom Name: Brett Chamberlin
Education: 6-8th Grade Middle School
Location: Gresham, Oregon USA
Question: We recently purchased a fairly old Bushnell Microscope. It is all metal construction with a service sticker. The service sticker reads a last serviced date of 1986.
Where might I find a manual and/or some instruction on the features of this microscope. There are some attachments that I do not understand the function of.
It says "Magna" on the front of it.
Thank you very much for any help that you can offer.
Any calculation would require factors specific for any one make of instrument. Anyway, doing it experimentally is more fun. Microanalysts need to know the actual location of the spot on the specimen and for that observe the beam spot on a daily basis. For WDS an SEM needs among other accessories a light microscope. With that and with the beam on a fluorescent mineral the beam spot is readily visible.
Usually in SEM the spot is busy scanning, so spot mode is required and for that the SEM scan coils are switched off. (Also, turn down fluorescent screen/ monitors, to avoid burn-marks. With a graticule in the eyepiece and micron- step stage movements the beam can be measured, but . . . For small spots, suitable for SEM the microscope on a microprobe will not have enough power.
The alternative is to burn spots onto a smooth specimen in a location which could be found under a high power light microscope. A grided coverslip with a light carbon film may be suitable. By parking the beam spot near line intersections the burn mark could later be found and measured. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Monday, November 26, 2001 11:59 AM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all: } } Is there any direct way to quantitatively determine } and measure the actual spot size of a SEM beam? } } Given various KV, WD, final aperture sizes, condenser lens } settings, etc., is there a way to truly find out what } the real spot size is on the specimen? } } Any ideas? } } gary g.
This method has promise....it is not too difficult to do! Sounds sort of like a Faraday cup affair. I could try this with 35u, 70u and 100u apertures mounted on a Carbon stub.
In general terms, what IS a typical spot size? Is it Angstroms, microns, or what? I never really thought about the true spot size--only relative, based on CL setting. Higher setting, smaller spot size. If a spot is used with the SEM in spot mode, the issue is to find out how big the spot actually is. This affects spillover of data when doing x-ray probing of very thin layers in microchips.
thanks to all, gary g.
At 05:53 AM 11/26/2001, you wrote: } Gary, } One technique I've seen used is to mount an aperture over a hole (on } carbon is best), or better yet, a thinned foil for a sharper edge, and use } a line-profile mode to scan across after focussing at a very high } mag. Absorbed current can be used instead of secondary for a signal, } although the latter works well, especially on a thinned foil. The curve } you get will resemble a rise-time curve and can be treated in a similar } manner. Mark the 10% and 90% rise points on the curve, then measure the } horizontal distance between those two points in units appropriate for the } mag. This is generally considered to be the beam diameter given the } Gaussian distribution of electrons across the beam. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } Gary Gaugler wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi: I am looking to replace my old Epson flat bed scanner and want a high quality scanner. This instrument is used primarily for scanning TEM negatives, 35 mm slides, some OCR and needs to be function in a multi-user environment. I have been considering the UMAX Powerlook 3000 and the Heidelberg Linoscan 2650 due to their reported 3.6 dynamic range and 42 bit pixel depth. Does anyone have experience with these machines? Is there something better?
Thanks!
Ken --
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323
Dear Gary, The classic method is to take the highest magnification photo that will show you the resolution limit, defined as the closest two points can be and still be determined as two points instead of one. I just take a hi-mag photo of a highly detailed surface and measure the smallest detail I can see and divide by the magnification. This will be the resolution at the conditions currently set. I was always told that the resolution of the SEM was equal to the diameter of the electron beam. For a sample I use an evaporated gold film on a graphite substate, to reduce the contribution of back-scattered electrons. At 05:59 PM 11/25/01 -0800, you wrote: } } Hi all: } } Is there any direct way to quantitatively determine } and measure the actual spot size of a SEM beam? } } Given various KV, WD, final aperture sizes, condenser lens } settings, etc., is there a way to truly find out what } the real spot size is on the specimen? } } Any ideas? } } gary g. } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
I am using a stereo microscope equipped for epiflourescent GFP excitation and emission. I am looking at live GFP transgenic mice and while the technique works quite well, there is a problem with autoflourescence of the skin surface.
These are mouse pups with little or no hair, and the skin has dander or flakes that seems to autofluor. I was wondering if anyone might know of a solution I could quench this external autofluor with? Something mild would be in order since these are delicate little critters.
Thanks, Mike
--
________________________________________________________ / Michael J. Herron, U of MN, Dept. of Pediatrics/BMT / / herro001-at-umn.edu / / 612-626-4321 Mpls MN 55455 / /_______________________________________________________/
I have _usually_ assumed for purposes of x-ray analysis that the SEM spot size is zero. I know it is not, but considering the interaction volume inside the specimen, the beam size is almost negligible. The various Monte Carlo programs tell me that I must consider an interaction volume on the order of a micron or more at 15 kV depending on my substrate. I am pretty sure that my beam is not half a micron in diameter, so how much will it affect the interaction volume?
I suppose the experiment to try would be to use an x-ray signal along with the secondary or absorbed current signal to check the effective spot size. And I suppose I should check the backscattered signal too while I am at it.
"Fortunately", much of my work has not involved the small structures where we are striving to keep the beam within the very thin layers. But those problems are coming along with increasing frequency. One of these days I will probably need to delve into the deconvolutions needed to determine the compositions of phases smaller than the excitation volume. I recall seeing an example of principle component analysis at MSA (2000?) where spectrum imaging allowed the discernment of a thin phase at an interface that was ordinarily undiscernible. Of course I do not have the program or equipment that those folks at Oak Ridge and NORAN had, so it will be a while before I could repeat the experiment. I wonder if anyone has developed a good, public-domain version of the routine.
Warren
At 06:20 AM 11/26/01 -0800, Gary wrote:
} Ken: } } This method has promise....it is not too difficult to do! } Sounds sort of like a Faraday cup affair. I could try } this with 35u, 70u and 100u apertures mounted on } a Carbon stub. } } In general terms, what IS a typical spot size? Is it } Angstroms, microns, or what? I never really thought } about the true spot size--only relative, based on CL setting. } Higher setting, smaller spot size. If a spot is used with } the SEM in spot mode, the issue is to find out how big } the spot actually is. This affects spillover of data when } doing x-ray probing of very thin layers in microchips. } } thanks to all, } gary g. } } } At 05:53 AM 11/26/2001, you wrote: } } Gary, } } One technique I've seen used is to mount an aperture over a hole (on } } carbon is best), or better yet, a thinned foil for a sharper edge, and } } use a line-profile mode to scan across after focussing at a very high } } mag. Absorbed current can be used instead of secondary for a signal, } } although the latter works well, especially on a thinned foil. The curve } } you get will resemble a rise-time curve and can be treated in a similar } } manner. Mark the 10% and 90% rise points on the curve, then measure the } } horizontal distance between those two points in units appropriate for the } } mag. This is generally considered to be the beam diameter given the } } Gaussian distribution of electrons across the beam. } } } } Ken Converse } } owner } } Quality Images } } third party SEM service } } Delta, PA } } } } Gary Gaugler wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for a hydrophobic heavy metal stain to stain the hydrophobic core of micelles suspended in water. I need to view the micelles with TEM. Non-water soluable hydrophobic drug molecules partition rapidly into the core; and I suspect a hydrophobic heavy metal such as a stable organo metallic would also, such as lead ethylhexanoate. Any suggestions or experience you can share with me?
We would like to dimple silver for TEM sample prep. When we used our standard methods, it dimpled exceedingly slowly (perhaps because it is soft).
Does anyone have experience in dimpling silver successfully? -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Is this a silver film on a substrate of some sort or is it a lump of Ag? If it is a piece of silver why don't you electroplish it? Far fewer artifacts.
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com NOTE CHANGES: 1-845-892-2225 Office; -6899 FAX; 1-800-352-4732, pin 8604 Pager; 1-914-453-2917 Personal Cell Phone
Alwyn Eades {jae5-at-lehigh.edu} on 11/26/2001 01:15:31 PM
To: EMNET {microscopy-at-sparc5.microscopy.com} cc:
We would like to dimple silver for TEM sample prep. When we used our standard methods, it dimpled exceedingly slowly (perhaps because it is soft).
Does anyone have experience in dimpling silver successfully? -- ......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
This is a good point; the interaction volume overwhelms the actual "spot size" under most circumstances. But since Gary asked for some actual numbers: In TEM (W filament) I generally find the spot ~200nm or less by measuring the contamination spot. As a matter of interest, I believe for NVLAP asbestos analysis a spot {250nm is required. That is, the *minimum* obtainable spot must be {250nm. Since, as Ken mentioned, the distribution is presumed Gaussian there is a bit of subjectivity as to where you pick the "edge" and how long you burn....not to mention time-dependent drift.
I have a table of "theoretical" spot sizes for my ESEM-FEG: they range from {1nm to about 40nm depending on kV and spot size selected.
Matt
On Monday, November 26, 2001 11:20 AM, Warren E Straszheim [SMTP:wesaia-at-iastate.edu] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } I have _usually_ assumed for purposes of x-ray analysis } that the SEM spot } size is zero. I know it is not, but considering the } interaction volume } inside the specimen, the beam size is almost negligible. } The various Monte } Carlo programs tell me that I must consider an interaction } volume on the } order of a micron or more at 15 kV depending on my } substrate. I am pretty } sure that my beam is not half a micron in diameter, so how } much will it } affect the interaction volume? } } I suppose the experiment to try would be to use an x-ray } signal along with } the secondary or absorbed current signal to check the } effective spot size. } And I suppose I should check the backscattered signal too } while I am at it. } } "Fortunately", much of my work has not involved the small } structures where } we are striving to keep the beam within the very thin } layers. But those } problems are coming along with increasing frequency. One } of these days I } will probably need to delve into the deconvolutions needed } to determine the } compositions of phases smaller than the excitation volume. } I recall seeing } an example of principle component analysis at MSA (2000?) } where spectrum } imaging allowed the discernment of a thin phase at an } interface that was } ordinarily undiscernible. Of course I do not have the } program or equipment } that those folks at Oak Ridge and NORAN had, so it will be } a while before I } could repeat the experiment. I wonder if anyone has } developed a good, } public-domain version of the routine. } } Warren } } At 06:20 AM 11/26/01 -0800, Gary wrote: } } } Ken: } } } } This method has promise....it is not too difficult to do! } } } } Sounds sort of like a Faraday cup affair. I could try } } this with 35u, 70u and 100u apertures mounted on } } a Carbon stub. } } } } In general terms, what IS a typical spot size? Is it } } Angstroms, microns, or what? I never really thought } } about the true spot size--only relative, based on CL } } setting. } } Higher setting, smaller spot size. If a spot is used } } with } } the SEM in spot mode, the issue is to find out how big } } the spot actually is. This affects spillover of data } } when } } doing x-ray probing of very thin layers in microchips. } } } } thanks to all, } } gary g. } } } } } } At 05:53 AM 11/26/2001, you wrote: } } } Gary, } } } One technique I've seen used is to mount an aperture } } } over a hole (on } } } carbon is best), or better yet, a thinned foil for a } } } sharper edge, and } } } use a line-profile mode to scan across after focussing } } } at a very high } } } mag. Absorbed current can be used instead of secondary } } } for a signal, } } } although the latter works well, especially on a thinned } } } foil. The curve } } } you get will resemble a rise-time curve and can be } } } treated in a similar } } } manner. Mark the 10% and 90% rise points on the curve, } } } then measure the } } } horizontal distance between those two points in units } } } appropriate for the } } } mag. This is generally considered to be the beam } } } diameter given the } } } Gaussian distribution of electrons across the beam. } } } } } } Ken Converse } } } owner } } } Quality Images } } } third party SEM service } } } Delta, PA } } } } } } Gary Gaugler wrote: } } } } } } } ------------------------------------------------------- } } } } ----------------- } } } } The Microscopy ListServer -- Sponsor: The Microscopy } } } } Society of America } } } } To Subscribe/Unsubscribe -- Send Email to } } } } ListServer-at-MSA.Microscopy.Com } } } } On-Line Help } } } } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FA Q.htm } } } } l } } } } ------------------------------------------------------- } } } } ----------------. } } } } } } } } Hi all: } } } } Is there any direct way to quantitatively determine } } } } and measure the actual spot size of a SEM beam? } } } } Given various KV, WD, final aperture sizes, condenser } } } } lens } } } } settings, etc., is there a way to truly find out what } } } } the real spot size is on the specimen? } } } } Any ideas? } } } } gary g. } }
I am trying to find information on remote control and remote viewing on the SEM. I am currently trying to use Microsoft Net Meeting. Does anybody have a better way to do these things?
Is there a reliable method of detecting Anthrax spores within a very short time, through the use of Electron Microscopy? Do you know anyone working with this method of detection of Anthrax spores? The system I am looking into involves using sticky pads to capture spores in work areas to include heating and cooling vents. The material collected is then taken to an area where a microscope is used to determine the presence or no-presence of the spores. Within a few hours, 2-4 hours, there is a determination made if anthrax spores are in fact present.
which also presents some other solutions. This is a pdf copy of the slides I presented during a tutorial at the M&M2000 meeting. That tutorial was video taped so if you are really keen you can order a copy of the tape from the MSA Education Committee.
} Email: twiinie-at-aolcom } Name: Brett Chamberlin } } Education: 6-8th Grade Middle School } } Location: Gresham, Oregon USA } } Question: We recently purchased a fairly old Bushnell Microscope. It } is all metal construction with a service sticker. The service sticker } reads a last serviced date of 1986. } } Where might I find a manual and/or some instruction on the features } of this microscope. There are some attachments that I do not } understand the function of. } } It says "Magna" on the front of it. } } Thank you very much for any help that you can offer. } ---------------------------------------------------------------------------
Brett -
If you can't find a manual, the general advice in this book will be very helpful:
Nachtigall, W. 1995 Exploring With the Microscope 160 pp. hardbound. 6.5x9.5". $19.95. ISBN 0-8069-0866-1 Sterling Publishing Co., NY. Although this book is intended for adult amateur microscopists, it is well written and will provide teachers and classroom volunteers with much useful information on "serious" light microscopy; it's unequalled as a basic reference for beginners. Almost half of the book is devoted to simple preparation methods for biological specimens and descriptions (with good illustrations) of commonly encountered organisms. Adult. RECOMMENDED
The description above was taken from the MICRO bibliography; you'll find other helpful books there. URL below.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
In a message dated 11/26/2001 10:23:39 AM US Mountain Standard Time, herro001-at-umn.edu writes:
{ { I am using a stereo microscope equipped for epiflourescent GFP excitation and emission. I am looking at live GFP transgenic mice and while the technique works quite well, there is a problem with autoflourescence of the skin surface.
These are mouse pups with little or no hair, and the skin has dander or flakes that seems to autofluor. I was wondering if anyone might know of a solution I could quench this external autofluor with? Something mild would be in order since these are delicate little critters. } }
Mike,
Is the autofluorescence yellowish? Rather than try and scrub a mouse pup, I would first try filtering out the offending autofluor color with the scope, assuming it probably isn't at exactly the same wavelength as the GFP signal.
It depends on your scope and what kind of filters you are using, but I would contact either the manufacturer of the scope or the fluorescence rig and inquire about narrow bandpass filters for GFP (both excitation and emission). Some filter sets have longpass filters in them that pass the required signals plus everything on either side for quite a distance out.
If you can get narrow bandpass filters, this should help a lot. If you already have narrow bandpass filter in your scope...never mind the above advice! I wonder how baby shampoo works on newborn mice...??
FYI, we have had good success using the advice of the folks at Chroma Technology Corp., especially Paul Millman. You can contact Paul at Choma by calling (800) 824-7662 in the U.S. They're experts at everything that's fluorescence-related.
Good Luck!
Bob Chiovetti GTI Microsystems Tucson, AZ Leica Exclusive Regional Dealers (AZ, NM, West TX)
Would appreciate some help with preparation of gram -ve bacteria for SEM.
I plan to do a standard gold coating but was wondering if anyone has some practical experience on fixation before proceeding with the rest of the protocol.
Many thanks in advance.
Lachlan
________________________ Lachlan McDonald EnGeneIc Pty. Ltd. 105 Delhi Road Riverside Corporate Park North Ryde NSW 2113 AUSTRALIA
} Hi, } } We are trying to salvage our 8 year old Cambridge Instruments Quantimet 570 } image analysis workstation. However there is no sign of any manuals. Does } anybody have a manual that they would be willing to photocopy for us, } donate, orcan you put me in contact with someone who might have one?? } } Any help appreciated, } } Jim } } } Jim Buckman } Department of Petroleum Engineering } Heriot-Watt University } Riccarton } Edinburgh } EH14 4AS } Scotland } } jim.buckman-at-pet.hw.ac.uk
I don't have experience with either of the two scanners you mentioned. For our customers, we usually recommend Microtek. We have had good success with the (previously discussed here) ArtixScan 2500 which is 36-bit with a 3.4 dynamic range. Resolution is 2500 dpi (optical) and the list price is $4,499. (Usually available for less.) (10,000 element CCD)
Microtek also offers (but I have yet to use) the ArtixScan 1100 which is closer to the ones you mentioned. Optical resolution is 2000 x 1000, 42-bit, and a higher reported dynamic range of 3.9. This one has a list price of $1,499. (8,000 element CCD)
Both of these use dual plate technology including Microtek's patented Emulsion Direct Imaging Technology for incredibly sharp transparency scanning. With E.D.I.T., there is no glass between the scanning lens, film, or light source, eliminating distortion and Newton rings that occur with traditional transparency adapters.
I hope this helps. If I can be of any additional assistance, please let me know.
*Kind Regards, *Dave Hall *Resolution Technology, Inc - (614) 921-0045
-----Original Message----- } From: Ken Bart [mailto:kbart-at-hamilton.edu] Sent: Monday, November 26, 2001 10:09 AM To: microscopy-at-sparc5.microscopy.com
Hi: I am looking to replace my old Epson flat bed scanner and want a high quality scanner. This instrument is used primarily for scanning TEM negatives, 35 mm slides, some OCR and needs to be function in a multi-user environment. I have been considering the UMAX Powerlook 3000 and the Heidelberg Linoscan 2650 due to their reported 3.6 dynamic range and 42 bit pixel depth. Does anyone have experience with these machines? Is there something better?
Thanks!
Ken --
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323
Mike, Why don't you try something like magic mending tape and do a few strips on the areas of interest. The tape is quit good at cleaning surfaces and it should be gentle enough and leave no residue. Good Luck, Russ Gillmeister Xerox ~~~~~~~~~~~~~
-----Original Message----- } From: Michael Herron [mailto:herro001-at-umn.edu] Sent: Monday, November 26, 2001 12:15 PM To: microscopy-at-sparc5.microscopy.com
All,
I am using a stereo microscope equipped for epiflourescent GFP excitation and emission. I am looking at live GFP transgenic mice and while the technique works quite well, there is a problem with autoflourescence of the skin surface.
These are mouse pups with little or no hair, and the skin has dander or flakes that seems to autofluor. I was wondering if anyone might know of a solution I could quench this external autofluor with? Something mild would be in order since these are delicate little critters.
Thanks, Mike
--
________________________________________________________ / Michael J. Herron, U of MN, Dept. of Pediatrics/BMT / / herro001-at-umn.edu / / 612-626-4321 Mpls MN 55455 / /_______________________________________________________/
The answer is complex. Probably, the bottom line for practicality is "no", not easily or rapidly. It could be done with the right equipment and the right reagents.
Bacillis anthrasis spores morphologically resemble spores of other Bacillus species (e.g., B. cereus). Some of these other bacilli are ubiquitous (e.g., on food). Thus, it would be impossible to differentiate, based solely on appearance, what species the spores were.
One possibility would be to react an antibody against B. anthrasis with them and then tag that first antibody with a secondary antibody that had a marker recognizable in the electron microscope. This could be done either by scanning electron microscopy (SEM) of the surface of spores, or by transmission electron microscopy (TEM) of ultrathin sections or of negatively stained bacteria--although negative staining would not be ideal because of the large size of the organism. (Negative staining is usually used for small things like viruses).
This immunolabeling technique would not be particularly fast (a matter of hours, rather than the days it would take to grow and speciate the bacteria, but not minutes). It would also be rather cumbersome in requiring immunological reagents and operators familiar with immunolabeling and thin sectioning. It would also require that the bacteria be handled as a biohazard and be killed before examination to prevent contamination.
Another possibility for identification, or for at least preliminary indication that malice was intended, would be to examine the preparation with Xray microanalysis. Most times, weaponized B. anthrasis spores are mixed with a chemical that causes them not to form large clumps. (The smaller particle size allows them to get further into the small airways of the lung.) This powder could be detected with microprobe analysis. However, one could question whether a smart crook might use other kinds of powder with different components to disguise the dispersant. Also, one would need to kill the spores while working on them to prevent infection.
I hope this helps. While I¹m in the business of using the electron to diagnose infectious diseases (we examine almost a thousand samples a year for viruses), and welcome any new use, I want to acknowledge the limitations.
Sara Miller
(P. S. The genus and species should be either italicized or underlined; my primitive email program won¹t do this.)
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
On Mon, 26 Nov 2001, Norton, Buddy wrote:
} Date: Mon, 26 Nov 2001 19:35:31 -0600 } From: Norton, Buddy {hnorton-at-richmond.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: detecting Anthrax spores using EM? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is there a reliable method of detecting Anthrax spores within a very short } time, through the use of Electron } Microscopy? Do you know anyone working with this method of detection of } Anthrax spores? The system I am looking into } involves using sticky pads to capture spores in work areas to include } heating and cooling vents. The material collected is then } taken to an area where a microscope is used to determine the presence or } no-presence of the spores. Within a few hours, 2-4 } hours, there is a determination made if anthrax spores are in fact present. } } } $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ } } "Buddy" } } Howard B. Norton } Captain } Deputy Chief of Police - Operations } } Office: 804-289-8723 } Pager: 804-346-6263 } Fax: 804-289-8720 } } hnorton-at-richmond.edu } } 31 UR Drive } Special Programs Building } University of Richmond Police Department } University of Richmond, Virginia 23173 } } For Sale - 1985 Prowler Camper Trailer 32' } } Those who row the boat seldom have time to rock it. } } $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ } } }
Our lab. had a failure of a thermal imaging system. The system is used for thermal mapping integrated circuits. We are interested in using a contract service to do measurements for us while our hardware is being repaired. Does anyone know of a contract lab., preferably on the East Coast, that has this capability for hire? I may consider a thermal AFM if available. The system we have now is optical and low resolution compared to an AFM. Resolution is ~ 10 uM.
Regards, Peter Tomic Failure Analysis & Analytical Services Group Anadigics, Inc. Warren, New Jersey
The New England Society for Microscopy (NESM) will be holding it's 35th Annual Fall Symposium at UMASS-Lowell in Lowell, Massachusetts on Friday, Dec. 7, 2001.
The meeting will begin at 12 Noon with Registration in the Mil Conference Center-Wannalancit Building (North Campus).
There will be 3 Sessions: A special student session beginning at 1:05 pm with 4 presentations, followed by a Poster session and coffee break.
Session II (Biological) begins at 2:20 pm and has as it's speaker, Ken Moore (University of Iowa), the National MSA Speaker. His talk will be: "Application of Microscopy Techniques to the Study of Genetic Therapy Research".
Another Poster Session and Afternoon Coffee Break will follow at 3:20 pm.
Session III (Materials Science) will begin at 3:40 pm and have 2 speakers: Elen Humphreys from M.I.T. and Koichi Nishikida from Thermo Spectra-Tech.
The Annual Business Meeting will convene at 5:00 pm and will include the election of NESM officers for 2002.
A Dinner and after-dinner speaker will follow at 6 pm.
Advance registration is due by Friday, November 30th. Registration after November 30th will NOT include dinner. The registration for NESM members is $10.00 and $25.00 for non-members. Dinner is $15.00 extra.
For more detailed information, please contact Mary McCann, NESM Treasurer at mccanns-at-tiac.net or NESM's website: The New England Society for Microscopy (NESM) will be holding it's 35th Annual Fall Symposium at UMASS-Lowell in Lowell, Massachusetts on Friday, Dec. 7, 2001.
The meeting will begin at 12 Noon with Registration in the Mil Conference Center-Wannalancit Building (North Campus).
There will be 3 Sessions: A special student session beginning at 1:05 pm with 4 presentations, followed by a Poster session and coffee break.
Session II (Biological) begins at 2:20 pm and has as it's speaker, Ken Moore (University of Iowa), the National MSA Speaker. His talk will be: "Application of Microscopy Techniques to the Study of Genetic Therapy Research".
Another Poster Session and Afternoon Coffee Break will follow at 3:20 pm.
Session III (Materials Science) will begin at 3:40 pm and have 2 speakers: Elen Humphreys from M.I.T. and Koichi Nishikida from Thermo Spectra-Tech.
The Annual Business Meeting will convene at 5:00 pm and will include the election of NESM officers for 2002.
A Dinner and after-dinner speaker will follow at 6 pm.
Advance registration is due by Friday, November 30th. Registration after November 30th will NOT include dinner. The registration for NESM members is $10.00 and $25.00 for non-members. Dinner is $15.00 extra.
For more detailed information, please contact Mary McCann, NESM Treasurer at mccanns-at-tiac.net or NESM's website: http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
There is a SEM/EDS software "gunshot inspection" for particle inspection automatically. I use this system for particle analysis on hard drive disk, a 95 mm diameter disk with 0.5 um resolution in whole disk scaning. The particle size range can be set so that the software ignores the larger one or the smaller one (dirt), and locates the particles and does EDS analysis automatically. I do not think you need to do EDS, but the image capture and classification functions may be good enough for you.
Contact Oxford Instrument (I have this system) or Noran Instrument (they make the similar system) for more information. Your sample should be no problem (particles on sticky pad).
Hope this helps,
Best regards,
Zhiyu Wang Sr. Engineer Maxtor Corp. Milpitas, CA (408)432-4421
-----Original Message----- } From: zhiyu wang [mailto:zhiyuw-at-home.com] Sent: Monday, November 26, 2001 2:39 PM To: zhiyu_wang-at-maxtor.com
The problem with this method is that most Bacillus species spores will look alike by SEM . . . and Bacillus species are very common contaminants. Most Bacillus species are not pathogenic. Some of the species spores, such as Bacillus stearothemophilus are used on purpose, like in Spore Strips, which are put into autoclaves to make sure the proper kill time and temp are reached. Unless there are some very distinctive characteristics only found on Bacillus anthracis spores, SEM or any other strictly visual method will not work to prove the presence of anthrax-causing spores.
Dr.Tina S. Schwach, President Microscopy Consulting Services Inc. 651-681-0112
----- Original Message ----- } From: "Norton, Buddy" {hnorton-at-richmond.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, November 26, 2001 7:35 PM
As discussed by Sara Miller and Tina Schwach, the genus Bacillus can not be distinguished on the basis of morphology alone. One needs specific tests: (1) immunological tests such as fluorescent antibody staining and light microscopy (LM) or even antibody-labeled latex spheres that would attach to the spores (LM and EM), (2) molecular amplification techniques where unique sequences are amplified and identified. Either of these tests take hours rather than days. The gold standard is still growing the organisms from the spores since even a single spore is detectable by this method. The downside is that culturing can take many days.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
With this instrumentation, one might be able to detect particles the size of bacillus spores, but would not be able to tell the difference between spores of the different bacillus species.
Sara Miller
On Tue, 27 Nov 2001, Wang, Zhiyu wrote:
} Date: Tue, 27 Nov 2001 09:45:59 -0700 } From: Wang, Zhiyu {Zhiyu_Wang-at-Maxtor.com} } To: "'Norton, Buddy'" {hnorton-at-richmond.edu} } Cc: "'microscopy-at-MSA.Microscopy.com'" {microscopy-at-sparc5.microscopy.com} } Subject: RE: detecting Anthrax spores using EM? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, Norton: } } There is a SEM/EDS software "gunshot inspection" for particle inspection } automatically. I use this system for particle analysis on hard drive disk, } a 95 mm diameter disk with 0.5 um resolution in whole disk scaning. The } particle size range can be set so that the software ignores the larger one } or the smaller one (dirt), and locates the particles and does EDS analysis } automatically. I do not think you need to do EDS, but the image capture and } classification functions may be good enough for you. } } Contact Oxford Instrument (I have this system) or Noran Instrument (they } make the similar system) for more information. Your sample should be no } problem (particles on sticky pad). } } Hope this helps, } } Best regards, } } Zhiyu Wang } Sr. Engineer } Maxtor Corp. } Milpitas, CA } (408)432-4421 } } -----Original Message----- } } From: zhiyu wang [mailto:zhiyuw-at-home.com] } Sent: Monday, November 26, 2001 2:39 PM } To: zhiyu_wang-at-maxtor.com } Subject: Fw: detecting Anthrax spores using EM? } } } } ----- Original Message ----- } } From: "Norton, Buddy" {hnorton-at-richmond.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Tuesday, November 27, 2001 1:35 AM } Subject: detecting Anthrax spores using EM? } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Is there a reliable method of detecting Anthrax spores within a very short } } time, through the use of Electron } } Microscopy? Do you know anyone working with this method of detection of } } Anthrax spores? The system I am looking into } } involves using sticky pads to capture spores in work areas to include } } heating and cooling vents. The material collected is then } } taken to an area where a microscope is used to determine the presence or } } no-presence of the spores. Within a few hours, 2-4 } } hours, there is a determination made if anthrax spores are in fact } present. } } } } } } $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ } } } } "Buddy" } } } } Howard B. Norton } } Captain } } Deputy Chief of Police - Operations } } } } Office: 804-289-8723 } } Pager: 804-346-6263 } } Fax: 804-289-8720 } } } } hnorton-at-richmond.edu } } } } 31 UR Drive } } Special Programs Building } } University of Richmond Police Department } } University of Richmond, Virginia 23173 } } } } For Sale - 1985 Prowler Camper Trailer 32' } } } } Those who row the boat seldom have time to rock it. } } } } $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Endospores are produced by many bacteria in the genera Bacillus and Clostridium. Many of these bacteria are quite common and most are harmless. EM could doubtless identify a bacterial endospore as such but could not, by itself, identify the species as Bacillus anthracis. Even so, a positive result should elevate your typical "suspicious white poweder" to the status of a "very credible biohazard threat" and elicit a vigorous response from the FBI, CDC etc.
Phase contrast light microscopy would be just as capable for this purpose, and much more convenient than EM. LM and EM would also both provide some characterization of dispersants mixed with the spores (i.e. bentonite, etc.).
Sample preparation and microscopy should be done with appropriate biosafety containment measures. Anthrax requires Biosafety II containment with special attention to elimination of aerosols.
I have designed sampling and inspection procedures for our campus. If you need further details you can contanct me off list.
At 07:35 PM 11/26/01 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mike Dalbey
Lecturer in Biology University of California, Santa Cruz
Hi I was wondering if any one has a good electropolish recipe, using a double jet-polisher to thin a niobium sample, with out using HF if possible. The sample is needed for high resolution TEM imaging.
Thanks,
Cindy Kleier EPIC Facility Northwestern University j-kleier-at-northwestern.edu
Mike, In my experiments with skin imaging, the dryer the skin (and flakes) the brighter the autofluorescence. How would the pups respond to a little hand lotion? (no experience with this, do lotions autofluoresce?)
As an aside, has anyone else observed that the yellow stratum corneum autofluorescence (in unfixed skin cross sections) actually increases with time exposed to the light beam? Is this just localized drying, or something else (the sections are mounted in 50% glycerol)? Added fluorescent dyes photobleach as expected. Karen -------------------------------------------------------------------------------- Karen S. Zaruba BioMaterials Technology Center 3M Center Bldg. 270-1S-01 Tel: (651) 737-2971 Fax: (651) 737-5645
Michael Herron {herro001-at-umn.edu} 11/26/01 11:15 AM Please respond to herro001
To: microscopy-at-sparc5.microscopy.com cc: (bcc: Karen S. Zaruba/US-Corporate/3M/US) Subject: Whole body fluorescence microscopy
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
All,
I am using a stereo microscope equipped for epiflourescent GFP excitation and emission. I am looking at live GFP transgenic mice and while the technique works quite well, there is a problem with autoflourescence of the skin surface.
These are mouse pups with little or no hair, and the skin has dander or flakes that seems to autofluor. I was wondering if anyone might know of a solution I could quench this external autofluor with? Something mild would be in order since these are delicate little critters.
Thanks, Mike
--
________________________________________________________ / Michael J. Herron, U of MN, Dept. of Pediatrics/BMT / / herro001-at-umn.edu / / 612-626-4321 Mpls MN 55455 / /_______________________________________________________/
I have gotten an old Cooke Vacuum Products CV 301 evaporator to restore but have no manuals. I received an email 'manual' on how to operate the vacuum system valves but not how to connect the transformer connections behind a clear plastic panel to do an evaporation.
There are six wing nuts with connections on a phenolic mounting board behind a plastic panel and two copper bus bars can be connected about seven different ways. I believe you connect the bottom right and bottom middle wing nuts together with one bus bar (the common ground). That connects the two 6 volt center taps together with the two right hand feed throughs in the bell jar. Also the top middle nut probably swings to either of the two bell jar connections in order to do an evaporation. Just guessing.
Volt meter readings could not sort out what the big step down transformer internal wiring was.
Please email me privately if you know how to connect them without me shorting the transformer out. The 2 transformer secondary tap SETS are labeled zero 0, 6 and 12 (volts). I would be glad to call you for your advice.
Any operating manual concerning the proper connection sequence would be nice to get in the mail. The manufacturer says he has none for this old instrument.
Paul Beauregard Senior Research Associate PPG Industries 440 College Park Drive Monroeville, PA 15146
Ron, Right you are! Oops! Actually, the beam could be even wider if the distibution wasn't Gaussian. Guess this requires a more expert source of information.
Ken Converse owner Qualtiy Images third party SEM service Delta, PA
Ronald Anderson wrote:
} Ken, } } You are only seeing the rise part of the beam width distribution curve. } Wouldn't the distance between the points be the beam radius and not the } beam diameter? } } Ron } } Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com } NOTE CHANGES: 1-845-892-2225 Office; -6899 FAX; 1-800-352-4732, pin 8604 } Pager; 1-914-453-2917 Personal Cell Phone } } }
Hi All, In summer it is difficult to get nice Formvar-carbon films for electron microscopy grids. So we prefer to prepare them in autumn or winter, because the humidity is less important in our laboratory. My question is, according your experiences, how many times can we conserve the Formvar-carbon coated grids and at which temperature.
Thank you very much,
Robert Alain Microscopie Electronique Inrs-Institut Armand-Frappier Laval, H7V 1B7 Robert.alain-at-inrs-iaf.uquebec.ca
I have full paper technical data for q570, but it is too long to you, I have this email for q570 "ia-support-at-leica.co.uk", ask people on this adress (Richard Hayden ?), or find Leica www page and call to him with this problem, this q570 is very good image analyser
best regards
Krzysztof Jan Hübner Foundry Reserach Institute, ul. Zakopianska 73 Kraków, Poland
Hi, Niobium is polished with good results in an acid and water free solution (H and O atoms are absorbed by unprotected Nb surfaces) using a solution of 10g Magnesium Perchlorate (Mg[ClO4]2)in 1 liter Methanol (Struers Tenupol: 60 V; 85 mA; temperature -5°C; flow rate 5); ref.: T. Schober and V. Sorajic, Metallography 6 (1973) 183. Good luck, H.K. Schmid
Please, please, please DO NOT take sampling and analysis of suspected bioattack materials into your own hands. Contact the authorities. If you are indeed dealing with an agent you have a "hot zone" and have to be in full protective gear. As someone already mentioned, you have to do the work at the highest BSL the unknown might contain. Small quantities of B.anthracis would be BL2 but what if it turns out to be something worse? You are risking turning your labs into hot zones. Also you are dealing with a crime scene or a potential crime scene. Anything you do could be contaminating evidence and jeopardizing the probability of conviction. I know it is tempting to play detective on your own, but there is an infrastructure in place for dealing with these things. Anthrax is not the only agent "out there" and a course of antibiotics isn't going to do the trick for all of them.
My intention is not to slam Buddy and Mike for their comments, as for all I know they are part of the infrastructure and are working with appropriate authorities in what they do. I just don't want anyone to get it into their heads to go sleuthing into potential incidents with an "it's okay, I'm a scientist" attitude. Even the people here, who design biodetectors and train first responders across the country, call in the authorities if they suspect anything.
Dr. Erica R. Valdes US Army Edgewood Chemical Biological Center EC/B Forensic Analytical Center AMSSB-RRT-CF/Building 3160 Aberdeen Proving Ground, MD 21010
-----Original Message----- } From: Mike Dalbey [mailto:dalbey-at-biology.ucsc.edu] Sent: Tuesday, November 27, 2001 2:27 PM To: Microscopy-at-sparc5.microscopy.com
Endospores are produced by many bacteria in the genera Bacillus and Clostridium. Many of these bacteria are quite common and most are harmless. EM could doubtless identify a bacterial endospore as such but could not, by itself, identify the species as Bacillus anthracis. Even so, a positive result should elevate your typical "suspicious white poweder" to the status of a "very credible biohazard threat" and elicit a vigorous response from the FBI, CDC etc.
Phase contrast light microscopy would be just as capable for this purpose, and much more convenient than EM. LM and EM would also both provide some characterization of dispersants mixed with the spores (i.e. bentonite, etc.).
Sample preparation and microscopy should be done with appropriate biosafety containment measures. Anthrax requires Biosafety II containment with special attention to elimination of aerosols.
I have designed sampling and inspection procedures for our campus. If you need further details you can contanct me off list.
At 07:35 PM 11/26/01 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mike Dalbey
Lecturer in Biology University of California, Santa Cruz
} } } } } } Hi All, } } } } } } I have been given a set of blocks of mouse brain (corpus callosum) to } } } cut. The investigator did her own embedding in Embed-812/Araldite } } } and the blocks are actually quite nice at first blush. The } } } semi-thins are gorgeous and I can cut nice light gold-silver } } } sections, but her boss is interested in the interline structure of } } } the myelin sheaths, and wants silver-grey sections. The block won't } } } cut well that thin. I get alternating silver then gold-purple } } } sections and sections either stick to the knife edge or drag over. I } } } have tried all the standard tricks: a newly re-sharpened diamond } } } knife, smallest possible block face, faster cutting speed. I've } } } played with the meniscus. I even put the blocks back in the oven for } } } another day. We will get whatever we can out of these blocks (the } } } result of a many-week treatment), but want to know how she should } } } vary her formula to get harder blocks next time around. She is using } } } a pretty standard recipe (Mollenhauer's Mixture No. 1, 1964). It is: } } } } } } Embed 812 25 ml } } } Araldite 15 ml } } } DDSA 55 ml } } } DMP-30 1.5% } } } } } } } } } I know that there are other recipes out there, but since I'm a } } } Spurr's fan, I'm not familiar enough with them to know which ones } } } will yield what we need. We probably need just a tad more hardness } } } to enable the really thin sections. } } } } } } Thanks in advance, } } } Lee } } } -- } } } Leona Cohen-Gould, M.S. } } } Sr. Staff Associate } } } Director, Electron Microscopy Core Facility } } } Manager, Optical Microscopy Core Facility } } } Joan & Sanford I. Weill Medical College } } } of Cornell University } } } voice (212)746-6146 } } } fax (212)746-8175
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I looked up two in the Philips Number 5 book that have no HF.
Schrober and Sorajic, 1973, Metallagraphy,6,p.183. 0.05 molar magnesium pechlorate per liter of methanol. 50-70V, -5 deg C.
and another Gidley and Davis, 1967 , J Sci. Instrum. 44, p. 297. 90% methanol 10% perchloric 5.5V, -30C BE CAREFUL WITH Perchloric acid solutions!!!
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: J. Cindy Kleier [mailto:j-kleier-at-northwestern.edu] Sent: Tuesday, November 27, 2001 3:51 PM To: microscopy-at-sparc5.microscopy.com
Hi I was wondering if any one has a good electropolish recipe, using a double jet-polisher to thin a niobium sample, with out using HF if possible. The sample is needed for high resolution TEM imaging.
Thanks,
Cindy Kleier EPIC Facility Northwestern University j-kleier-at-northwestern.edu
We are interested in differentiating between arteries and veins in the adult rat pulmonary vasculature. Are there specific markers or antibodies we could use? A variety of plant lectins exists which label endothelial cells but the literature is confusing about their specificities. They may also preferentially stain only certain organs.
We are perfusing the rats, and cutting thick sections (~100 microns) on a vibratome for confocal microscopy. So staining could be carried out either at the time of perfusion or later, with the sections. Is there enough time for a probe to stick during perfusion? A fluorescent marker is required for the confocal technique.
I look forward to any suggestions. Thank you
Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8 Canada
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} Hi All, } In summer it is difficult to get nice Formvar-carbon films for electron } microscopy grids. So we prefer to prepare them in autumn or winter, } because the humidity is less important in our laboratory. } My question is, according your experiences, how many times can we } conserve the Formvar-carbon coated grids and at which temperature. } } Thank you very much, } } Robert Alain } Microscopie Electronique } Inrs-Institut Armand-Frappier } Laval, H7V 1B7 } Robert.alain-at-inrs-iaf.uquebec.ca
I have kept C coated formvar grids for negative staining for a couple of years in an a/c lab. These usually have a medium to "thickish" C coat. I have prepared these in the summer as well, and eastern KS is fairly humid. Bruce Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Lab University of Kansas
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Hi Judy,
Just a thought: Could you use a fluorescent marker for endothelial cells such as CD-31 (PeCam) and then use the autofluorescence of the elastin sheath that would be around the arteries to distinguish the ateries from the veins?
Bob Underwood U of Washington Seattle
On Wed, 28 Nov 2001, Judy Trogadis wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Fellow Microscopists: } } We are interested in differentiating between arteries and veins in the adult rat pulmonary vasculature. Are there specific markers or antibodies we could use? A variety of plant lectins exists which label endothelial cells but the literature is confusing about their specificities. They may also preferentially stain only certain organs. } } We are perfusing the rats, and cutting thick sections (~100 microns) on a vibratome for confocal microscopy. So staining could be carried out either at the time of perfusion or later, with the sections. Is there enough time for a probe to stick during perfusion? A fluorescent marker is required for the confocal technique. } } I look forward to any suggestions. } Thank you } } Judy } } Judy Trogadis } Bio-Imaging Coordinator } St. Michael's Hospital, 8Q } 30 Bond St. } Toronto, ON M5B 1W8 } Canada } } ph: 416-864-6060 x6337 } fax: 416-864-6043 } trogadisj-at-smh.toronto.on.ca } } } }
Hi Listers, I would appreciate any PRIVATE feedback I can get from USERS of the following:
JEOL JEM 1230 Hitachi 7500 or 7600 Philips Technai 12 LEO 910 Quartz Imaging Intranetworking and Image Database Technology [URL:http://www.quartzimaging.com/index.html]
Thanks for the help to all.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
We have a paper in our archive of technical reports titled "A Jet Electropolishing Solution for Silicon Germanium, Tantalum, Niobium, and Tungsten Rhenium" written by Bernie Kestel. This is for our Model 550 Single Vertical Jet Electropolihser, but it may be useful anyway. I will put a copy in the mail to you.
You can view other papers and application notes on our website at www.southbaytech.com.
I hope this helps.
David
"J. Cindy Kleier" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi I was wondering if any one has a good electropolish recipe, using a } double jet-polisher to thin a niobium sample, with out using HF if } possible. The sample is needed for high resolution TEM imaging. } } Thanks, } } Cindy Kleier } EPIC Facility } Northwestern University } j-kleier-at-northwestern.edu
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
One possibility would be to make the Formvar films whenever the humidity permitted it and then coat them later (weeks, months) as needed. For small particulate particles (e.g., viruses, subcellular organelles) adherence is better with freshly carbon-coated grids. Alternativily, you could glow discharge the stored Formvar and carbon coated grids prior to use to improve spreading and adherence.
Sara Miller
On 28 Nov 2001, Bruce Cutler wrote:
} Date: 28 Nov 2001 10:20:38 -0600 } From: Bruce Cutler {bcutler-at-eureka.idl.ukans.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: TEM Conservation of Formvar-carbon coated grid } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Hi All, } } In summer it is difficult to get nice Formvar-carbon films for electron } } microscopy grids. So we prefer to prepare them in autumn or winter, } } because the humidity is less important in our laboratory. } } My question is, according your experiences, how many times can we } } conserve the Formvar-carbon coated grids and at which temperature. } } } } Thank you very much, } } } } Robert Alain } } Microscopie Electronique } } Inrs-Institut Armand-Frappier } } Laval, H7V 1B7 } } Robert.alain-at-inrs-iaf.uquebec.ca } } } I have kept C coated formvar grids for negative staining for a couple of years } in an a/c lab. These usually have a medium to "thickish" C coat. I have prepared } these in the summer as well, and eastern KS is fairly humid. } Bruce } Bruce Cutler, Ph.D. } Director, Microscopy & Electronic Imaging Lab } University of Kansas } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I agree that sampling and analysis of "suspicous white powders" is not a casual activity and that it must be done with serious consideration of potential risks. Our activity is fully integrated with existing hazmat response procedures followed by the campus police and EH&S. This includes sample collection by EH&S personnel with appropriate personal protection.
Our procedure was created following an incident on campus that the authorities (i.e. FBI) could not effectively respond to because they were overwhelmed by such a large number of similar incidents in the aftermath of the anthrax exposures on the east coast. The FBI agent that did eventually respond had zero knowledge of microbiology and zero personal protection gear. I suspect that waiting for a response by someone prepared to deal with "the highest BSL the unknown might contain" (that would be level IV, I assume) would mean we would be waiting still, and that a major campus building would still be closed. (That material proved to be laundry detergent, by the way.) Based on this experience we concluded that, in fact, there was NOT an infrastructure in place to deal with these situations AT THE LEVEL WE ARE CURRENTLY EXPERIENCING THEM. We hope and expect this will be a very temporary situation.
Mike Dalbey
Lecturer in Biology University of California, Santa Cruz
Dear Listers, I am preparing a report comparing digital cameras for TEMs. We are looking to upgrade our system intending to image negative stains of heterochromatin samples. At this point we do not have cryo capability. If any of you have experience with the digital cameras which can be upgraded for automated tomography, I would appreciate hearing from you. Rosemary Walsh
The incident we had here at Oklahoma State was handled Veterinary medicine and the State Veterinary Diagnostic lab on campus. The Haz Mat guys showed up in moon suits but vet med didn't. They came in rubber boots, mask, coveralls, goggles, gloves and cap the same as they would for a necropsy of a suspicious animal.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger } } } I agree that sampling and analysis of "suspicous white powders" is not a } casual activity and that it must be done with serious consideration of } potential risks. Our activity is fully integrated with existing hazmat } response procedures followed by the campus police and EH&S. This includes } sample collection by EH&S personnel with appropriate personal protection. } } Our procedure was created following an incident on campus that the } authorities (i.e. FBI) could not effectively respond to because they were } overwhelmed by such a large number of similar incidents in the aftermath of } the anthrax exposures on the east coast. The FBI agent that did eventually } respond had zero knowledge of microbiology and zero personal protection } gear. I suspect that waiting for a response by someone prepared to deal } with "the highest BSL the unknown might contain" (that would be level IV, I } assume) would mean we would be waiting still, and that a major campus } building would still be closed. (That material proved to be laundry } detergent, by the way.) Based on this experience we concluded that, in } fact, there was NOT an infrastructure in place to deal with these } situations AT THE LEVEL WE ARE CURRENTLY EXPERIENCING THEM. We hope and } expect this will be a very temporary situation. } } Mike Dalbey } } Lecturer in Biology } University of California, Santa Cruz } } 831-459-3674 } } dalbey-at-biology.ucsc.edu } } }
How many anthrax cases have we had in the United States in the last 50 years?
} From January 1955 to December 1999, there were 236 reported cases of anthrax, most of them cutaneous, in 30 states and the District of Columbia.
When was the last case of inhalational anthrax in the United States?
The last case of inhalational anthrax in the United States, before 2001, was in 1976 in California. A home craftsman, who worked with yarn, died. Bacillus anthracis was isolated from some of the imported yarns used by the patient.
When was the last case of cutaneous anthrax?
The last case of cutaneous anthrax, before 2001, occurred in North Dakota, in 2000
Jim Young
Those who row the boat seldom have time to rock it.
A colleague (Hossler, F, East Tennessee State Med. Sch.) and I have used a method called corrosion casting to study the rich capillary bed underlying the trasitional epithelium of the urinary bladder. We used a modified commercial preparation "MERCOX Blue" for the preparation. The blue dye is, fortuitously, fluorescent. In a report given at the recent Philadelphia meeting of MSA (M&A, Vol6, Suppl 2, pp562-563), Czymmek et al. described the use of confocal microscopy in an analysis of corrosion casts. They used a Zeiss LSM 510 confocal (exciting the dye with the HeNe(Red) laser (633nm)) and detecting via a 660nm longpass emission filter. I have repeated this on an Olympus FV-300 using both the HeNe(R) as above with casts of pig bladder wall prepared by one of the above authors (Hossler). I have a particularly useful z-projection of a large, low power, image stack that I would be willing to send to requestors (69k, jpg) as an example of what can be accomplished with the application of confocal microscopy applied to low-power microanatomic studies of fluorescent objects. I have recommended the use of Evans Blue (C.I. 23860, CAS # 314-13-6) on several occasions in the past but have seen no results. This dye is known to form covalent bonds with albumin stoichimetrically and to be fluorescent in THAT form. It has been used in studies of extravasation around injury sites as well as for vascular volume studies. There is one reported incident of complicity in animal carcinogenesis (bolus, i.p. in rats) mentioned by IPCS INCHEM (http://www.inchem.org/documents/iarc/iarc/iarc121.htm).
Hope this is of some use.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
We are having a sudden rash of test messages. PLEASE review the FAQ for the Microscopy Listserver. Every one of your test messages creates over 3000 email messages and needlessly clogs up the system.
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} } I am developing a research proposal to investigate organic carbon } } colloids as a mechanism for contaminant transport of hydrophobic } } pollutants such as pesticides in surface and ground waters. The } } colloids generally range in size from 0.001 to 1 micron in size. I } } would like to characterize these organic colloids. I know that } } considering the low molecular weight of the carbon that this can be a } } problem for a lot of the microscopes and and complementary analytical } } devices. I was wondering if you had any suggestions for how I might } } characterize these organic colloids? } } } } Thank you for your help, } } Carol Creasey } } creasey-at-cats.ucsc.edu } } --
I have the chance of receiving a surplus PGT EDS detector. It has not been used for two years. My question is would the detector's window still be good? It has been in storage and the detector window has not been kept cold during these past two years. Any advice is appreciated.
Applications are invited for a full-time, non-tenure track position to operate a new electron microprobe/SEM facility in the Department of Geoscience at UNLV. The successful candidate will maintain a user-friendly facility for faculty, students, and visiting users, establish standard operating procedures (some to fulfill DOE quality assurance requirements), and solicit work from outside users. The candidate may conduct research as time permits; however, the primary focus of the position is to maintain the facility and to assist UNLV and outside users. The laboratory currently serves a wide range of research activity in geoscience, biology, chemistry, engineering, and materials science. Minimum requirements include a Master's degree in geology, geochemistry, or an appropriate related field, and electron microprobe and/or SEM experience. Review of applications will begin immediately and will continue until the position is filled. Salary will be commensurate with qualifications and experience. Position is contingent upon funding.
Interested applicants should send a CV, letter of interest detailing your interest in the position as described above, and names, addresses, phone numbers, and email addresses of at least three professional references to Dr. Jean Cline, Department of Geoscience, UNLV, 4505 Maryland Parkway, Box 454010, Las Vegas, NV 89154-4010; phone: 702 895 3262; fax: 702 895 4064, email: jcline-at-nevada.edu.
For information on the Geoscience department at UNLV visit our Web page at http://www.unlv.edu. UNLV is an Equal Opportunity/Affirmative Action employer. Persons are selected on the basis of ability without regard to race, color, sex, age, national origin, sexual orientation, religion, disability or veteran status.
Jean S. Cline Associate Professor, UNLV Department of Geoscience 4505 Maryland Parkway, Box 454010 Las Vegas, NV 89154-4010
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"Histochemistry" by Richard Horobin.
"Histological and Histochemical Methods" by John Kiernan
Most (all?) editions of "Cell and Tissue Biology" by Leon Weiss (editor) have a nice chapter on histochemistry by Helen Padykula.
Grizzi Fabio ICH wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All } } I would like to receive any information about review papers on the basic } principles of histochemical techniques. I am quite interested to receive } references of papers on the basic characteristics of a histochemical method } (reproducibility, specificity etc). } } Many thanks. } } Fabio Grizzi } Scientific Direction } Istituto Clinico HUMANITAS } Milan, Italy
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
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Reimar,
It sounds to me very much like a rotifer: these are very common, and can survive considerable desiccation, and so are often found in roof gutters (where I remember finding them). I would guess that the "satellite dish" is the ciliated "wheel" from which the phylum gets its name - I think that the cilia are either moving too fast or simply too small to be seen at whatever magnification you are using. This generates a current, and I remember as a boy watching green spherical algae being sucked in and macerated.
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
----- Original Message ----- } From: {reimar_gaertner-at-wsib.on.ca} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, November 21, 2001 3:34 AM
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (elir-at-chem.ch.huji.ac.il) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, November 18, 2001 at 13:55:49 ---------------------------------------------------------------------------
Email: elir-at-chem.ch.huji.ac.il Name: Eli rothenberg
Organization: Hebrew University
Education: Graduate College
Location: Jerusalem, Israel
Question: I would like to know what is the correct setup for a wide illumination in a confocal microscopy arrangement, that is, what optics should i use in the path of the laser, if i want to get the maximum illuminated area through my objective, the setting is as follows: laser-collimated beam- dichroic mirror-objective-illuminated surface. (p.s I thought of using a short focusing lens that will focus half way through the objective so maybe that will give me a wide illumination on the surface, and/or maybe a diffuser to go along with it?)
I apologize if this message has been posted already. I responded with this information several days ago and it did not come back to my Inbox as a message distributed to the List. I wasn't sure if it had made it out or not.
Ken:
I don't have experience with either of the two scanners you mentioned. For our customers, we usually recommend Microtek. We have had good success with the (previously discussed here) ArtixScan 2500 which is 36-bit with a 3.4 dynamic range. Resolution is 2500 dpi (optical) and the list price is $4,499. (Usually available for less.) (10,000 element CCD)
Microtek also offers (but I have yet to use) the ArtixScan 1100 which is closer to the ones you mentioned. Optical resolution is 2000 x 1000, 42-bit, and a higher reported dynamic range of 3.9. This one has a list price of $1,499. (8,000 element CCD)
Both of these use dual plate technology including Microtek's patented Emulsion Direct Imaging Technology for incredibly sharp transparency scanning. With E.D.I.T., there is no glass between the scanning lens, film, or light source, eliminating distortion and Newton rings that occur with traditional transparency adapters.
I hope this helps. If I can be of any additional assistance, please let me know.
*Kind Regards, *Dave Hall *Resolution Technology, Inc - (614) 921-0045
-----Original Message----- } From: Ken Bart [mailto:kbart-at-hamilton.edu] Sent: Monday, November 26, 2001 10:09 AM To: microscopy-at-sparc5.microscopy.com
Hi: I am looking to replace my old Epson flat bed scanner and want a high quality scanner. This instrument is used primarily for scanning TEM negatives, 35 mm slides, some OCR and needs to be function in a multi-user environment. I have been considering the UMAX Powerlook 3000 and the Heidelberg Linoscan 2650 due to their reported 3.6 dynamic range and 42 bit pixel depth. Does anyone have experience with these machines? Is there something better?
Thanks!
Ken --
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323
Has anyone successfully processesed tissue (in our case part of a rat brain) for TEM just after studying brain circulation by fluorescence microscopy? We use rhodamine and fluorescein tagged AB 40 and the experiment runs for about 1 1/2 hours. At that point, I fix the tissue with glutaraldehyde and osmium and embed it in epon-araldite. The problem is that membranes disappear. I'd been warned that would happen and it did. Is there any way to protect the membranes or any other stain or method that might help?
Thanks for any help! Margie Bryant Anatomy Dept. Univ. of South Florida Medical School 12901 Bruce D. Downs Blvd Tampa, Fl. 33612 email address: mbryant-at-com1.med.usf.edu
Dear Listers, Consolidation of our EM facilities has resulted in a surplussing a 1984 Philips 410 LS. It has been factory maintained and remains in excellent condition. The department is asking $30,000, or best offer. For more information or sales inquiry, please contact: Dr. Lesnick Westrum Dept. Neurological Surgery University of Washington (206) 543-5434 westrum-at-u.washington.edu
Regards, Glen -- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax ************************************************************************** C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
I heard on the radio that the economy is in a recession, so I figured I had better get busy and try to save money and recycle some supplies. I also had a big box of used SEM stubs and thought I could start there.
The stubs are left over from various projects, some have silver paint, some with double stick tape, etc. I thought I could loosen things up a bit by soaking them in water and/or an Alconox solution. This fit in very well with my plan for recycling since the plan was to let them soak until the recession is over.
Since there was no way to know when things would return to normal, I put some into an ultrasonic cleaner with Fisher Ultrasonic Cleaning solution. Says it is safe for aluminum and other things.
Now I have a problem. The stubs turned pretty ugly. Big black stains all over the place. Looks like some reaction between the Al and the Alconox. Stubs in plain water are not so bad. Ultrasonic cleaner can't remove the black stuff. I can polish off the black stuff, but that wasn't part of the plan. This was supposed to be simple.
I don't think the black stuff affects the functioning of the stub, it just looks ugly and means I have to explain to some users that they are just as good as the few shiny ones left in the drawer (stubs that is, not users).
So tell me oh mighty metal and materials experts, what is the black stuff, why did it appear, and is there any easy way to get rid of it? Or is this some kind of plan to pull the economy back up to speed by making me buy a bunch of shiny new SEM stubs?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I need practical advice for a colleague on conjugating fluorophores to primary antibodies for immunofluorescence microscopy. She would like to use Alexa and/or Cyanine labels. Are there particular products/vendors that people have used with success? What are the problems and pitfalls to avoid?
Thanks in advance!
Dr. David P. Bazett-Jones Senior Scientist Programme in Cell Biology, Research Institute The Hospital for Sick Children 555 University Avenue Toronto, ON M5G 1X8 Canada
You will want to check if the window is OK, but the fact that the detector has been in storage warm for 2 years does not make it likely that it is broken.
If I had the chance (and needed the detector, of course!), I would accept it. I would pump it (for which you will need a pumping adapter) and see if you get a vacuum (do this *BEFORE* you put liq. N2 in it). You can get by with a rotary pump vacuum, but if you can get a high vacuum (we have a port plumbed into one of our evaporaters, with a valve, so we can pump dtectors), so much the better. If it pumps down, let it pump for a few hours, fill with nitrogen, then plug the pumping port and remove the adapter. Let it cool overnight, then fire it up. If you have access to a Fe55 radioactive source you won't even need to put it on the microscope to check it out, but I guess that you probably don't have one of those!
I have a pumping adapter for very old Kevex detectors with US threads on the plug - more recent detectors, and all those from Link have metric threads. I don't know about PGT, but if yours has US threads, you are welcome to borrow my adapter - I don't think they vary from manufacturer to manufacturer.
Good luck!
Tony Garratt-Reed
At 01:16 PM 11/29/2001 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Our experience is that 0.3 mm is better. 0.5 mm will work, but the width of the hologram for a given fringe spacing will be reduced. It depends on what you mean by "normal holography"...
Larry
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-- Dr. Lawrence F. Allard Distinguished Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
I would ask advise concerning cover glass thickness specs. The objectives on my microscope are specified for .17 mm thick cover slips. I understand this is quite important.
I guess the answer must be obvious, but given that the idea mounting mediun would have the same refractive index as the cover slip, and I think the oil (in case of an immersion situation), how should one deal with this? the objects should be in direct contact with the cover slip?
Obviously this is not possible in all cases. Should one such as myself (unfortunate to have objectives with no correction collar) use even thinner cover slips to compensate for any distance between the object and the bottom side of the cover slip?
If I might pose another perhaps equally obvious question, is the reasoning for use of hanging drop preparations also the need to keep the specimen at this exact distance?
Then what does water do when doing wet mounts?
Sorry for wasting bandwidth on such ridiculous questions,
Alan Davis
-- Alan E. Davis, Science Instructor Marianas High School PMB 30, Box 10006, Saipan, MP 96950 Northern Mariana Islands adavis-at-saipan.com
"An inviscid theory of flow renders the screw useless, but the need for one non-existent." ---Lord Raleigh(aka John William Strutt),or else his son, Jr., who was also a scientist.
We have a collaborator who is having problems using colloidal gold-lectins for the post-embedding labeling of protozoa (lots of non-specific binding, even to Epon and Lowicryl embedding media). Does anyone have any good protocols for the use of these conjugates?
Thanks in advance,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
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I don't know how you could get rid of those ugly stains, but you can probably get your clients to accept the blackened stubs without a murmur by saying that you "antiqued" then after seeing Martha Stewart do it :-)
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, November 29, 2001 8:34 PM
I have seen black "smut" on Al alloys etched with alkalis. Although I never analyzed it, I would guess it was undesolved Al alloying elements (Fe, Cu, etc). A quick dip in a medium strength nitric acid solution, followed by water rinse cleaned it right off.
Woody
} } I heard on the radio that the economy is in a recession, so I } figured I had } better get busy and try to save money and recycle some } supplies. I also had } a big box of used SEM stubs and thought I could start there. } } The stubs are left over from various projects, some have } silver paint, some } with double stick tape, etc. I thought I could loosen things } up a bit by } soaking them in water and/or an Alconox solution. This fit in } very well } with my plan for recycling since the plan was to let them } soak until the } recession is over. } } Since there was no way to know when things would return to } normal, I put } some into an ultrasonic cleaner with Fisher Ultrasonic } Cleaning solution. } Says it is safe for aluminum and other things. } } Now I have a problem. The stubs turned pretty ugly. Big black } stains all } over the place. Looks like some reaction between the Al and } the Alconox. } Stubs in plain water are not so bad. Ultrasonic cleaner can't } remove the } black stuff. I can polish off the black stuff, but that } wasn't part of the } plan. This was supposed to be simple. } } I don't think the black stuff affects the functioning of the } stub, it just } looks ugly and means I have to explain to some users that } they are just as } good as the few shiny ones left in the drawer (stubs that is, } not users). } } So tell me oh mighty metal and materials experts, what is the } black stuff, } why did it appear, and is there any easy way to get rid of } it? Or is this } some kind of plan to pull the economy back up to speed by } making me buy a } bunch of shiny new SEM stubs? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
For those of us with "text-only" email, Larry means 0.3 micrometers.
At 11:16 PM -0500 11/29/01, Larry Allard wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Shu-You: } } Our experience is that 0.3 mm is better. 0.5 mm will work, but the } width of the hologram for a given fringe spacing will be reduced. } It depends on what you mean by "normal holography"... } } Larry } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have used the Alexa kits from Mol Probes to conjugate their visible fluors to protein-G-purified IgG. I have always gotten satisfactory results, although my degree-of-labeling was always less than optimal as reported by Mol Probes. We do almost exclusively laser confocal microscopy, and I can say that the primaries I made are very useful with that approach. The visible Alexa fluors are very bright and very resistant to bleaching. I cannot comment on how they would work in standard epifluorescence, but my guess is that if one has a decent camera (i.e., short exposure times), they should work equally well for this approach. The only practical pitfall in the conjugation process is separation of unbound dye from conjugate. Mol Probes includes a simple gravity-run molecular sizing column in the kit with which to do this. However, if efficiency of labeling is low, it can be difficult to see the band you want and, as with any such separation approach, you will definitely experience some loss of material regardless of how critically you run the column. Also, if you have never set up a column before, this resin is very "gloply" and not trivial to pack. So, I thought, "why not dialyze away the unbound dye?" and Mol Probes says you can do this (they supply the column because you get faster results and they can justify a higher cost for the kits...). As I am never in a huge hurry, I tried this. As far as I could see, there was NO difference in the color intensity of the dialysed material! This suggests that I had 100% labeling (!). The upside on that particular prep was that, as Mol Probes also points out, if your immunofluorescence sample prep is good then you really don't have to worry a lot about unbound dye as it will be washed out any (only the conjugate should stick). This was the case for me with that dialyzed prep.
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Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
Commercially supplied stubs are anodized, which makes them more resistant to oxidizing, but with detergent that barrier breaks down. The dark stuff has to be aluminium oxide, which is much more stable then plain Al. You could argue that those mounts have character and that the appearance does not matter.
I used to recycle a percentage of mounts, but solvents and detergent are just too messy. I prefer a fine flat file (could also use sanding paper, a medium to coarse grade). Peel off any easily removed sticky material and then give the surface of the mount a few strokes on the file. A double layer of paper towel under the file collects most of the rubbish and protects the bench.
Its an ok job, if you don't expect to do that full-time for the rest of your working life. If you are entrepreneurial, leave the set-up with some old mounts on the bench and hide the new mounts. This is doubly effective if some influential people come to contemplate your funding problems while cleaning dirty mounts.
With a bit of tinkering you could have your lab's economy run counter-cyclical to the one Alan Greenspan is worried about. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, November 30, 2001 10:35 AM, Jon Krupp [SMTP:jmkrupp-at-cats.ucsc.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I heard on the radio that the economy is in a recession, so I figured I had } better get busy and try to save money and recycle some supplies. I also had } a big box of used SEM stubs and thought I could start there. } } The stubs are left over from various projects, some have silver paint, some } with double stick tape, etc. I thought I could loosen things up a bit by } soaking them in water and/or an Alconox solution. This fit in very well } with my plan for recycling since the plan was to let them soak until the } recession is over. } } Since there was no way to know when things would return to normal, I put } some into an ultrasonic cleaner with Fisher Ultrasonic Cleaning solution. } Says it is safe for aluminum and other things. } } Now I have a problem. The stubs turned pretty ugly. Big black stains all } over the place. Looks like some reaction between the Al and the Alconox. } Stubs in plain water are not so bad. Ultrasonic cleaner can't remove the } black stuff. I can polish off the black stuff, but that wasn't part of the } plan. This was supposed to be simple. } } I don't think the black stuff affects the functioning of the stub, it just } looks ugly and means I have to explain to some users that they are just as } good as the few shiny ones left in the drawer (stubs that is, not users). } } So tell me oh mighty metal and materials experts, what is the black stuff, } why did it appear, and is there any easy way to get rid of it? Or is this } some kind of plan to pull the economy back up to speed by making me buy a } bunch of shiny new SEM stubs? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu }
I have been asked to prepare some cross sections of hair for inspection with an optical microscope and not having much experience doing this I have gotten some advice locally and tried a few things with an old microtome, but without much success.
I am wondering if someone on this list could offer me some advice on how to prepare the hair for the sectioning. So far working with paraffin has not be successful as the hair seems to not cut well leaving either a hole in the wax strip or a raised and irregular hair.
Alternately I would be interested in talking to a lab near Connecticut that could do this sectioning for me.
Thanks very much for any help you may be able to offer!
Richard Shalvoy Arch Chemicals Cheshire, CT 203-271-4394 rbshalvoy-at-archchemicals.com
} } } I need practical advice for a colleague on conjugating fluorophores to } primary antibodies for immunofluorescence microscopy. She would like to } use Alexa and/or Cyanine labels. Are there particular products/vendors } that people have used with success? What are the problems and pitfalls } to avoid? } } Thanks in advance! } } Dr. David P. Bazett-Jones **************** Dear David, The Alexa family of dyes are sold by Molecular Probes, and if you buy them uncongugated, they come with instructions for congugating them to your Ab. I haven't done it personally, but my coworkers have and tell me its easy.
I have not financial interest in Molecular Probes, I'm just a satisfied customer. Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Jonathan Krupp wrote: ============================================================= I heard on the radio that the economy is in a recession, so I figured I had better get busy and try to save money and recycle some supplies. I also had a big box of used SEM stubs and thought I could start there.
The stubs are left over from various projects, some have silver paint, some with double stick tape, etc. I thought I could loosen things up a bit by soaking them in water and/or an Alconox solution. This fit in very well with my plan for recycling since the plan was to let them soak until the recession is over.
Since there was no way to know when things would return to normal, I put some into an ultrasonic cleaner with Fisher Ultrasonic Cleaning solution. Says it is safe for aluminum and other things.
Now I have a problem. The stubs turned pretty ugly. Big black stains all over the place. Looks like some reaction between the Al and the Alconox. Stubs in plain water are not so bad. Ultrasonic cleaner can't remove the black stuff. I can polish off the black stuff, but that wasn't part of the plan. This was supposed to be simple.
I don't think the black stuff affects the functioning of the stub, it just looks ugly and means I have to explain to some users that they are just as good as the few shiny ones left in the drawer (stubs that is, not users).
So tell me oh mighty metal and materials experts, what is the black stuff, why did it appear, and is there any easy way to get rid of it? Or is this some kind of plan to pull the economy back up to speed by making me buy a bunch of shiny new SEM stubs? =================================================================== In the typical laboratory that "accummulates" no-longer-needed SEM mounts, the "history" of what was originally there on the mounts (e.g. the samples) is often times lost with the passing of the months if not also from the natural turnover of both employees and/or students. We have ourselves considered offering some kind of a "recycling" program for SEM mounts, but the inability to really know what we would be getting would pose to our employees an unacceptable safety risk. Furthermore, from the standpoint of shipping, some might argue that phenomenologially, "waste" mounts are akin in lots of ways to a hazardous waste, which would impose restrictions on how they could be shipped (if indeed they could be shipped easily at all).
Also, some laboratories have a way of accidentally or inadvertently "co- mingling" or mixing up used beryllium mounts with aluminum mounts, and once a part of the kind of "mess" that was described, they look no different from orginary aluminum mounts. Hence you do want to be further cautious when talking about polishing down or grinding, because you absolutely don't want to get into a beryllium dust exposure situation, which can surely occur if the polishing pads or grinding papers dry out.
Disclaimer: SPI Supplies offers new aluminum mounts for all SEMs, and we would like to see more new mounts being used. But we offer the above comment not to encourage the purchase of more new mounts, but only to point out that if you did want to "recycle" used SEM mounts, you do want to be careful not to mix up beryllium mounts with the aluminum and you also want to have some awareness of what kinds of samples, might be on the no-longer- needed mounts.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Molecular probes www.probes.com have outstanding products.
Abizar www.innogenex.com
"David P. Bazett-Jones" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I need practical advice for a colleague on conjugating fluorophores to } primary antibodies for immunofluorescence microscopy. She would like to } use Alexa and/or Cyanine labels. Are there particular products/vendors } that people have used with success? What are the problems and pitfalls } to avoid? } } Thanks in advance! } } Dr. David P. Bazett-Jones } Senior Scientist } Programme in Cell Biology, Research Institute } The Hospital for Sick Children } 555 University Avenue } Toronto, ON M5G 1X8 } Canada } } TEL: 416 813-2181 } FAX: 416 813-2235 } EMail: david.bazett-jones-at-sickkids.ca
Hi Alan, First, the mountant's RI is matched to that of the specimen. That of the immersion oil to that of the glass, although I cheat when I use Cargille oils of non-standard RI to view some difficult coverglassed specimens in order to alter the phase dynamics of the system. Second, the 0.17mm coverglass represents the average thickness of the No 1 coverglass that one normally uses for the purpose of mounting specimens for microscopy. Further, the glass components of the objective have been ground in expectation of that thickness of glass between the objective and the specimen, whether the objective is immersive or not. Even though the use of oil immersion to view a blood smear without a coverglass is common, the oil immersion objective almost always 'says' that it expected a coverglass to be there. Third, with respect to specimen thickness, there is nothing that can change the built-in working parameters of the objectives, thus, you won't be able to use oil immersion to any good effect on a whole mount of any specimen much thicker than 10um. A 40x objective is stressed beyond its limits when one tries to view a whole mounted 24-hr chick embryo. In other words, the working distance, which is not usually given, defines the limit of the distance between the face of the objective and the object one wishes to view. If one places physical impediments between face and specimen that exceed the working distance, then the specimen cannot be viewed by the scientist and is crushed by the freshman biology student who doesn't know about it. On the other side of that is the fact that were the working distance of the oil immersion lens too long, the oil might not be able to bridge the gap between objective and coverglass. Fourth, the information on the barrel of an objective is part of the description of the optical properties of the objective. Its design parameter set, if you will. The 0.17 correction tells much about working distance. The numerical aperture tells a lot about the optical components used inside the objective. Fifth, I always thought that the hanging drop was to provide a method for viewing live material in a confined environmentally friendly volume (cage).
} ---------- } From: Alan Davis } Sent: Thursday, November 29, 2001 5:27 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: cover glass thickness } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would ask advise concerning cover glass thickness specs. The objectives } on my microscope are specified for .17 mm thick cover slips. I understand } this is quite important. } } I guess the answer must be obvious, but given that the idea mounting } mediun would have the same refractive index as the cover slip, and I } think the oil (in case of an immersion situation), how should one deal } with this? the objects should be in direct contact with the cover slip? } } Obviously this is not possible in all cases. Should one such as myself } (unfortunate to have objectives with no correction collar) use even } thinner cover slips to compensate for any distance between the object and } the bottom side of the cover slip? } } If I might pose another perhaps equally obvious question, is the reasoning } for use of hanging drop preparations also the need to keep the specimen at } this exact distance? } } Then what does water do when doing wet mounts? } } Sorry for wasting bandwidth on such ridiculous questions, } } Alan Davis } } -- } Alan E. Davis, Science Instructor } Marianas High School } PMB 30, Box 10006, } Saipan, MP 96950 } Northern Mariana Islands } adavis-at-saipan.com } } } "An inviscid theory of flow renders the screw useless, but the need } for one non-existent." ---Lord Raleigh(aka John William Strutt),or } else } his son, Jr., who was also a scientist. } }
} Commercially supplied stubs are anodized, which makes them } more resistant to } oxidizing
It is my understanding that anodized aluminum has much thicker oxide layer than a natural oxide film, and, as a result, it should lead to the coloring of the metal (which we do not see for the stubs). Im I wrong?
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
The old AO Spencer rotary microtome manual recommends Pike Oil and a Neutral grease. I haven't used Pike oil, but I usually purchase a light machine oil. For use in colder environments such as when used for cryotomy, this manual recommended a lighter Hamilton watch oil. Clock oil will do. Others have suggested Singer sewing machine oil. I have used "Bear Oil" from Norton ("Formerly Pike Oil") which now appears to be Norton Part # 07660 787940 0, 4.5oz "Sharpening Oil" [URL: http://www.nortonconsumer.com/catalog/displaytier.asp?tier_id=D110050050010& display=all].
Hope this helps. Getting the proper lubricating oil is always a problem for these old machines.
P.S.1 I just purchased a new volume of "3 to 1" oil, and was surprised at its viscosity which I consider to be higher than in previous purchases.
P.S.2 A good rule is that if the oil hole contains a wick, the oil should be really light! Whatever, the lubricant should NOT be so light that it leaks out of the bearing.
Regards to all,
Fred Monson
} ---------- } From: Jonathan Wilson } Reply To: Jonathan Wilson } Sent: Friday, November 16, 2001 4:51 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: microtome oil } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I have been loaned a microtome (Leitz 1512) by a colleague which } according to the instruction manual requires regular oiling. The microtome } did not come with any oil and the price I have been quoted by a local } supplier for microtome oil was about 75$US (including VAT) for 50ml } (SAKURA } 1447). I found the price a bit steep and am left wondering if it is } absolutely necessary to buy "microtome" oil or if I can use an off the } shelf } oil (e.g 3 in 1 or something like that). I do not want to compromise the } performance of the microtome as it is not mine and would appreciate any } advice from those with experience. } Thanks for any help. } Sincerely, } Jonathan } } Jonathan Wilson (PhD) } CIIMAR } Rua do Campo Alegre 823 } 4150-180 Porto, Portugal } tel: 351 22 608 0470 / 71 } fax: 351 22 606 0423 } e-mail: wilson_jm-at-cimar.org } web: www.cimar.org } } }
I can't put cute subject headings anymore, so I'm trying not to be too negative ;-).
I'm working in a lab where they have been adding a hypo clearing agent called Orbit Bath into the Kodak rapid fix. Has anybody heard of doing this? I have never heard of this and I can't get the Orbit Bath without promising my first born and pulling a few teeth.
I'd rather go back to the tried and true method of developing the Kodak 4489 in HRP or D-19, fixing in Kodak Rapid Fix (using A & B), dunking in Hypo clear and washing as usual.
If someone can come up with a good reason to keep continuing this practice, or even an explanation as to why they even started it, I'm all ears.
Help clear me of my negative thoughts,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
No. 1 coverslips have a range of 0.13 to 0.17 (for Fisher) or 0.13 to 0.16 (for Corning brand).
No. 1 1/2 range from 0.16 to 0.19 for both Corning and Fisher.
Having used a micrometer on both sizes and brands, I find the most 0.17 coverslips in No. 1 1/2 boxes.
Tom
} } } Hi Alan, } First, the mountant's RI is matched to that of the specimen. That } of the immersion oil to that of the glass, although I cheat when I use } Cargille oils of non-standard RI to view some difficult coverglassed } specimens in order to alter the phase dynamics of the system. } Second, the 0.17mm coverglass represents the average thickness of } the No 1 coverglass that one normally uses for the purpose of mounting } specimens for microscopy. Further, the glass components of the objective } have been ground in expectation of that thickness of glass between the } objective and the specimen, whether the objective is immersive or not. Even } though the use of oil immersion to view a blood smear without a coverglass } is common, the oil immersion objective almost always 'says' that it expected } a coverglass to be there. } Third, with respect to specimen thickness, there is nothing that can } change the built-in working parameters of the objectives, thus, you won't be } able to use oil immersion to any good effect on a whole mount of any } specimen much thicker than 10um. A 40x objective is stressed beyond its } limits when one tries to view a whole mounted 24-hr chick embryo. In other } words, the working distance, which is not usually given, defines the limit } of the distance between the face of the objective and the object one wishes } to view. If one places physical impediments between face and specimen that } exceed the working distance, then the specimen cannot be viewed by the } scientist and is crushed by the freshman biology student who doesn't know } about it. On the other side of that is the fact that were the working } distance of the oil immersion lens too long, the oil might not be able to } bridge the gap between objective and coverglass. } Fourth, the information on the barrel of an objective is part of the } description of the optical properties of the objective. Its design } parameter set, if you will. The 0.17 correction tells much about working } distance. The numerical aperture tells a lot about the optical components } used inside the objective. } Fifth, I always thought that the hanging drop was to provide a } method for viewing live material in a confined environmentally friendly } volume (cage). } } } ---------- } } From: Alan Davis } } Sent: Thursday, November 29, 2001 5:27 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: cover glass thickness } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I would ask advise concerning cover glass thickness specs. The objectives } } on my microscope are specified for .17 mm thick cover slips. I understand } } this is quite important. } } } } I guess the answer must be obvious, but given that the idea mounting } } mediun would have the same refractive index as the cover slip, and I } } think the oil (in case of an immersion situation), how should one deal } } with this? the objects should be in direct contact with the cover slip? } } } } Obviously this is not possible in all cases. Should one such as myself } } (unfortunate to have objectives with no correction collar) use even } } thinner cover slips to compensate for any distance between the object and } } the bottom side of the cover slip? } } } } If I might pose another perhaps equally obvious question, is the reasoning } } for use of hanging drop preparations also the need to keep the specimen at } } this exact distance? } } } } Then what does water do when doing wet mounts? } } } } Sorry for wasting bandwidth on such ridiculous questions, } } } } Alan Davis } } } } -- } } Alan E. Davis, Science Instructor } } Marianas High School } } PMB 30, Box 10006, } } Saipan, MP 96950 } } Northern Mariana Islands } } adavis-at-saipan.com } } } } } } "An inviscid theory of flow renders the screw useless, but the need } } for one non-existent." ---Lord Raleigh(aka John William Strutt),or } } else } } his son, Jr., who was also a scientist. } } } }
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
The thickness of a natural alumina film is in the order of some tens of nanometers. An anodized layer, on the other hand, can reach the micrometer level. However, it is porous. Usually, we can fill those pores simply by immersing parts in boiling or hot water. The film is nearly transparent or translucent. The color we see is actually the color of the dye used during pore filling for decoration purpose. There are many colors we can choose.
Kai Lorcharoensery
Materials Science & Engineering Lehigh University Bethlehem, PA
It is my understanding that anodized aluminum has much thicker oxide layer than a natural oxide film, and, as a result, it should lead to the coloring of the metal (which we do not see for the stubs). Im I wrong?
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
Anodizing of aluminum can be done in many colors, as well as a clear coating. I would not expect SEM stubs to be anodized, as the aluminum oxide coating which forms is an insulator. This would inhibit the removal of the electron charge from the sample. The native oxide film is no where near the thickness of the usual anodized film, and can easily be broken. The thickness of the anodized film can be controlled, but it is still an effective insulator. I am fairly certain that none of the stubs I have ever used were anodized.
Regards, Darrell
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