I just have to react to your message posted on the Microscopy Listserver as a reply to Doug Keene's question. I fully agree with you that limited or lack of penetration into the section interior may very well be the cause for negative results. I had a message from Doug where he indicates that he will be looking into this, using FITC labels for LM first and ultra small colloidal gold labels with silver enhancement for EM if the first approach proves to be successful. In your answer you say that "you have to use a nanogold reagent, not colloidal gold". As the principal inventors of ultra small colloidal gold particles we demonstrated already back in 1988 that conjugates based on ultra small colloidal particles are very well able to penetrate under circumstances where 5 or 10 nm particle based conjugates will not do so. If you like I will be happy to send you the documentation. The reagents you refer to didn't even exist at that time, although undecagold was known and succesfully used for high resolution immunoelectron microscopy without silver enhancement. The colloidal ultra small labels (sold by several companies with a reputation in gold labels) as well as gold clusters may solve the problem.
Jan
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Managing Director Costerweg 5, 6702 AA Wageningen The Netherlands
AutoMatch is a public domain macro written for the public domain software NIH Image. It is described in an article by Swidbert R. Ott, Microscopy Research and Technique 38:335-339(1997). The montage is automated. The article indicates that the software is available via FTP from zippy.nimh.nih.gov/pub/nih-image/contrib/
Mark, try the imtools package from the SDSC group. Specifically, it contains a program alled imstoryboard that allows you to make a montage of many images into a single file ready for printing. You have control over spacing between images, between images and the edges, the background color, etc. Another way would be to use the netpbm package. Using pnmcat one can concatenate multiple images into a single image. As this one works with standard in and output, quite possibly one could simply redirect the output of it straight to the printer, as in:
pnmcat pnmfile1 pnmfile2 ... | lp (or lpr)
Hope this helps.
Jaap
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Tue, 30 Jun 1998 mark_munro-at-bio-rad.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } does anyone know of a graphics package that can take several separate } digital image files and montage them on a single page ready for printing? } } Thanks, } } } Mark Munro }
We have the same software and might be able to help with some questions, at least on the image processing side.
Remember, the imaging part is Noesis Vision's Visilog product rolled into the Oxford suite of products.
If anyone would be interested in discussing either Visilog, or ImQuant in particular, I would be willing to set up a mailing list for discussion. Please contact me directly.
At 03:25 PM 6/30/98 +0000, you wrote: } } Has anyone got some tutorials for this Image software. The manual is } too complicated... } } F. (frank.sarrazit)
For calibrating our Mettler FP82HT hot stage, we have been using the melting points of organic compounds supplied with the Kofler Hotbench (Eichsubstanzen fuer Kofler Heizbank). These are now several years old, and someone has suggested that liquid crystals might be more apporopriate.
I would be glad if anyone can update me as to what is state-of-the-art in calibrating hotstages.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Doug Keene wrote, } } Dear Microscopists: } } I am in a mild state of confusion regarding an immuno-EM } experiment where it seems that the secondary antibody (GAM } 5 nm) does not recognize the primary (mouse IgG) once the } primary is bound to its corresponding antigen in tissue. } We expect that the antigen is present with periodicity on } fibrils in the connective tissue matrix. After the tissue } is emersed in antibody, we see periodic decoration of the } fibrils, which indicates to us that the antibody is bound. } However, secondary antibody does not bind to the tissue. We } are convinced that the secondary conjugate is not } defective, as we use it for other experiments. Has anyone } else experienced a situation where a secondary does not } recognize a primary once the primary is bound to its } corresponding antigen? } } The problem is not that the secondary does not recognize your } primary. It is the fact that colloidal gold reagents can't penetrate } into tissues or cells. The gold particles are too large. These } reagents are used for labeling surfaces of sections. If you want to use } a gold conjugate for whole or thick tissues sections, prior to } embedding, you have to use a nanogold reagent, not colloidal gold. } } What do you mean that, "After the tissue is emersed in } antibody, we see periodic decoration of the fibrils. How do you } visualize this? } } Joseph Goodhouse } Confocal / EM Core Lab Manager } Department of Molecular Biology } Princeton University } jgoodhose-at-molbio.princeton.edu } 609-258-5432 } Info and Images at } http://www.molbio.princeton.edu/confocal/CF-EM-HOME.html
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
We are getting good results with the Polaroid scanner which is really the Microtek scanner in a Polaroid box. It is not as good resolution for 35mm as the Sprintscan, but in some respects it is better in that a whole bunch of frames can be digitized at once. But as for NT compatibility, we have not tried it on any of the NT machines so we don't know.
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://leper1.ca.aecom.yu.edu/aif/ --------------------------------------------
On Fri, 26 Jun 1998, Joiner Cartwright, Jr. wrote: } } we want to buy a scanner which can be used for both 35 mm and 70 mm EM } } film as well as 81x100 mm plates, probably with different adapters (?).
Our imaging database, ElectroImages, can montage images easily and with great flexibility. ElectroImages also allows the user to place text captions under images. Captions can include title, date, source, extensive descriptions, filename, filepath, and up to 6 user-definable fields. You can also easily print a footer and header to give your page a finished look.
Please contact me off-list and I will send you our trial copy of this program.
Matt Irwin ElectroImage, Inc. 277 Northern Blvd. Suite 101 Great Neck, NY 11021
LEO Electron Microscopy is hiring SEM service people in California. If you are a proven SEM service/support person, and would like to practise your art for LEO in the SF or LA areas, please contact me direct (billneill-at-csi.com)
} Experienced Electron Microscopist } } A prominent cardiac muscle physiology laboratory at the } University of Pennsylvania has an opening for an experienced electron } microscopist. The individual must be proficient in all aspects of } conventional transmission EM from preparation of the samples to } publication } quality photography. Knowledge of some standard biochemical } procedures } including electrophoresis would be helpful but not essential. We are } looking for an individual who is willing and can demonstrate the } ability to } learn and execute new techniques. A B.S. degree is highly desirable } but not } absolutely essential. Salary will be commensurate with experience. } Please } contact Dr. Saul Winegrad. } } E-mail address is bsg-at-mail.med.upenn.edu } } } } }
I am looking for an instruction of sufficient detection of apoptotic cells in paraplast embedded tissue. Is anybody able to point me to a reference with an adequate recipe or can mail me one?
-- Mit freundlichen Gruessen Yours sincerely *************************************************************** * T. J. Filler | * * Westfalian Wilhelms-Univ.| phone:*49 251 83 552-26 Fax: -41 * * Institute of Anatomy | e-Mail: filler-at-uni-muenster.de * * Vesaliusweg 2-4 | D-48149 Muenster Germany * ****** http://medweb.uni-muenster.de/institute/anat ***********
Warren wrote: "Offhand, I would be suspicious of the cooling fan/system. I have seen a number of computers get flaky once the processor overheated. My first experience of that type was after installing a 486-50 overdrive processor in a PS/1. (Nobody told me it needed a cooling fan.) Things come to a halt very quickly once the processor overheats, and it doesn't take all that long to warm up. 5-10 minutes sounds like plenty long enough. I have had the same experience with trying to drive a processor a little to hard, like when I was trying to push my 120 MHz up to 133 MHz. It didn't seem like much of a push, but the manufacturers have already well pushed to the edge so that there is no room for a cooling fan to fail"
But the real truth is that CPUs are rated extremely conservatively. Typically, the maximum speed diminishes as they age, and no responsible manufacturer wants to risk having to replace millions of microprocessors on warranty. That is why there can be considerable ability to "hot rod" CPUs, and still keep them running-for a while. Today's CPUs have the clock speed burned in at the factory, and will be reliable for very long periods of time.
-----Message d'origine----- De : Lucille A. Giannuzzi {lag-at-pegasus.cc.ucf.edu} =C0 : microscopy listserver {microscopy-at-sparc5.microscopy.com} Date : mercredi 1 juillet 1998 05:10 Objet : SF6 upgrades for JEOL TEMs
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Lucille
The new norm about CFC's regulation don't take care of the use you'll mak= e of freon. The important thing is to not let freon going to the atmosphere and you cannot be sure of that during emergency on the gun or on the HT tank. This pollution is too bad and freon will not be available soon. I think your provider has to sell his stock which we will not be abble to u= se later.
I received a message from a coworker asking if I had any information to help you.
I am sending you a web site that deals with old scientific instruments and of course includes microscopes. I purchased two book from them, which might help:
1. Turner, G.L'E. COLLECTING MICROSCOPES $25.00 Christie's International Collectors Series New York: Mayflower Books, 1981 8vo, pp 120; cloth, dj; new copies (out-of-print) with 102 instruments illustrated, many in color
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; AFM Technology Leaders for Environmental / In Vitro AFM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
The Adem was ahead of it's time. It has a six axis stage, fully integrated imaging, and X-ray analysis. The column can accept very large samples, and it has a motor for everything. They were coming out with a FE model when they pulled the plug back in the early 90's. This was due to several factors. The 5500 series 2 X-ray system was nearing the end of it's sales cycle, and the other Microscope vendors did not take too kindly to the competition. This caused sales alliances with the OEM's to collapse. Throw in the fact that Noran was bought and sold twice in that time frame. So the new management felt they had to save the core business. There were still some working units out there when I left Noran 2 years ago.I worked on them briefly and I found them to be difficult to work on. The concept of a fully integrated SEM was sound, however, they made it way too complicated. If Noran could have held on for a while longer they would probably have a good market share. I found it interesting that the chief designer Fred Schamber went to R J Lee and designed the personal SEM. The same concept only on a much simpler scale.
Thu, 2 Jul 1998 09:25:13 +0100 Received: from localhost by spsscsc2.rdg.ac.uk (8.8.5/8.8.5) with SMTP id JAA24200; Thu, 2 Jul 1998 09:25:00 +0100 (BST)
On Tue, 30 Jun 1998 mark_munro-at-bio-rad.com wrote:
} Dear all, } does anyone know of a graphics package that can take several separate } digital image files and montage them on a single page ready for printing?
For a GENERALLY USEFUL package which can do things in a nice way, try downloading a trial version of Corel Xara! 2.0 from:
http://xaraxone.i-us.com/
I have only recently acquired this, so I haven't tried out all its functions, but I have started converting some of my black-and-white TEM images to duotone using this package.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
If you want to impress the kiddies, go ahead and use those liquid crystals. After all, it sounds much fancier that way.
But I certainly wouldn't bother. There is nothing at all wrong with regular melting point standard substances. And viewing in cross polarizers, the melting temperature is always quite clear. I certainly wouldn't buy anything new if I were sure that the existing materials were OK. (If they're not, the melting point is no longer sharp. You can test this in a DSC.)
Was the person suggesting other standards in any chance a salesman wanting to sell them to you?
Cynthia Bennett Hoechst Diafoil
The opinions expressed here are solely my own. Don't blame my employer.
------------------------------------------------------------------------ - Robert Olley wrote:
For calibrating our Mettler FP82HT hot stage, we have been using the melting points of organic compounds supplied with the Kofler Hotbench (Eichsubstanzen fuer Kofler Heizbank). These are now several years old, and someone has suggested that liquid crystals might be more apporopriate.
I would be glad if anyone can update me as to what is state-of-the-art in calibrating hotstages.
+----------------------------------------------------------------------- -+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley |
There will be a meeting of the MSA Standards Committee and ASTM committees E42-96 (US TAG for ISO TC202 Microbeam Analysis) and E42-15 (Electron Probe Microanalysis/Electron Microscopy) on Thursday July 16th at 12:30 PM in Rm. 275-W of the Georgia World Congress Center. For additional information contact john.small-at-nist.gov.
Hi! Does anybody know if there is any problems (eg precipitate within the tissue, etc) if tissue is fixed in 2% osmium tetroxide within a stainless steel container? Thanks Dorota
Everyone was so helpful when I was looking for a large SEM chamber. Can you help again?
I had a call from a science museum in Virginia. They are setting up a display and would like an illustration of the crystallographic structure of aluminum and if possible, a 7079 alloy.
I have at my ready grasp a drawing of graphite, which is close. But museums tend to be very particular and they should have aluminum. They would like the drawing for reference in constructing a model.
If you have such a drawing, please contact Mr. Brian Alfano at balfano-at-smv.mus.va.us
We have funding to buy a new SEM and have at least one application for an = environmental(variable pressure) microscope. Those of you who have = experience with environmental scopes, especially the Hitachi 3500N, please = let me know the good and bad points about your scope. Most of our samples are animal tissue and some of my reading has = suggested that to prevent tissue drying, when working at low vacuum, it is = necessary to use a cold stage. How do you rate the importance of a cold = stage for this type of specimen? =20 Finally, those of you who have a cold substage from Fullam... have you = been satisfied with its performance? =20 Any comments are welcomed. Thanks.
Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il 62794-1220 217-782-0898
Here's a general question that should stir up some comment...
My department has sent out a standard questionaire for us to estimate any problems our lab computers, software, or equipment might have with the year 2000 date problem. Perhaps some of you have already gone through this. I'm not a computer wiz, and naively thought that everything I use would be fine, since I don't do any date calculations like folks in the banking, insurance, etc. businesses do. However, the more I hear about this problem the more confused I get. Is it true that some older CPU's or BIOS programs will not handle the "00" date? So some of our older equipment might not work even though the software we are aware of has little reliance on dates?? Any microscope or software vendors want to comment??
Hope this isn't stirring up needless fears or urban legends, but I have definitely heard some alarmist opinions and would like to consult a more technically aware and level-headed source (i.e. yourselves!).
Thanks, Karen
-- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
I wish to inject ferritin into the hemolymph (blood) of crabs to determine which spaces are blood spaces. (Crab blood has copper-based hemocyanin, rather than hemoglobin).
Sigma carries several varieties of Ferritin, including Apoferritin. Any suggestions as to which one I should use?
Does anyone have a protocol to recommend?
What final concentration should I shoot for in the blood stream?
Thanks to all respondents.
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
A recent breakthrough in environmental microscopy technology is MAC Mode-AFM. This will allow you to do in vitro, time lapse imaging at a nanometer scale resolution.
You can use a 37 C temperature control with flow through liquid cell to change your biological buffers and observe conformational change at the molecular level.
Also you can characterize attractive and repulsive forces between antibody - antigen or other bio agents. A great paper, "Antibody-Antigen Recognition Detected By Ultra sensitive Force Microscopy" will be presented at Protein 98 san Diego by Peter Hinterdofer of the Schindler group from Linz Austria.
George
At 05:55 PM 7/2/98 -0600, Donna Wagahoff wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have experience with e.g. marine samples in ESEM? Or tissue in buffers? I am curious to know whether they are feasible. With samples, such as small planktonic organisms which are living in what is essentially 3% sodium chloride solution, can they be dried off without getting a coating of salt crystals? I know from cryoSEM that once the water is removed, you get the covering of salts. I imagine that some items might withstand a quick rinse in fresh water, but not everything.
Karen, Most old PCs (but not Macs or most Unix) will not transfer the date correctly to the year 2000 although many will handle dates after Jan 1, 2000 if told explicitly (i.e. with 4 figure years). You can get free test software for PCs off the www. I found one from National Software Testing Laboratories (NSTL) that seemed pretty useful. If your old PC will not do the transition automatically you can either roll the date forward manually (if it can do this), use a false date (such as 1990) but obviously this could be misleading in some cases, or scrap the PC and buy a new one.
For the most of our PCs that control lab equipment in our group I know that they will not roll the date over automatically, so I am likely to keep all the data backed up, wait till the end of 1999 and see what happens. If one dies for some reason then we can always replace it with another cheap PC.
Hope this helps
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
This is a multi-part message in MIME format. --------------99D16DAA7889B787EDE9016F Content-Type: text/plain; charset=big5 Content-Transfer-Encoding: 7bit
A postdoctoral position in scanning tunneling microscope investigation on the interfacial reactions of metal thin films on silicon is available at the
IC Thin Film Lab. Department of Materials Science and Engineering National Tsing Hua University Hsinchu, Taiwan, Republic of China
The position is for one year and renewable for two more years. Interested persons should have experiences in UHV-STM. Please contact Professor Lih J. Chen directly either by e-mail or by fax at 886-3-5718328.
--------------99D16DAA7889B787EDE9016F Content-Type: text/x-vcard; charset=big5; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Lih J. Chen Content-Disposition: attachment; filename="vcard.vcf"
begin: vcard fn: Lih J. Chen n: Chen;Lih J. org: Department of Materials Science and Engineering, National Tsing Hua University adr: Department of Materials Science, National Tsing Hua University;;;Hsinchu;Taiwan;300;Republic of China email;internet: ljchen-at-mse.nthu.edu.tw title: Professor tel;work: 886-3-5731166 tel;fax: 886-3-5718328 x-mozilla-cpt: ;0 x-mozilla-html: FALSE version: 2.1 end: vcard
The question has been posed whether cells grown in culture or cells from sea water can be imaged using ESEM. As a general principle I have found it possible to image cells in culture medium - crystallisation upon drying. It is possible to image cells from 0.9% saline and in such cases the NaCl crystals are widely dispersed and mostly do not interfere with imaging (sod's law not withstanding!) I can try 3% saline to see what happens and will let you know.
Another question needs to be addressed however. What are you trying to see? I have much success by viewing glutaraldehyde fixed samples washed in water and imaged with ESEM. The cells remain hydrated are much more robust (resistant to accidental drying, withstanding beam damage). On most occasions I have not seen structural differences between fresh hydrated and fixed hydrated. It is the drying which causes changes not fixation.
As with all SEM low kV should be used (hard work below 5KV unless you have Brendan Griffin detector modification). Check the pressure temperature curve for water vapour pressure but 3 - 4 degrees C with a pressure of 5 - 6 torr should keep specimens hydrated.
Good luck Let me know if I can help further.
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
If you are attending the MSA Meeting in Atlanta, Mike Boykin and I would like to invite you to a gay social on Monday, July 13th at 8:00pm. The party will be held at Mike's home in midtown Atlanta. Directions to Mike's house will be available via email or at the Information and Hospitality Booth. Also, copies of Southern Voice (a gay weekly newspaper) will be available. If you are planning on attending the social please RSVP to me. See y'all there, Beth Richardson
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
For those interested, I thought I would summarize the comments received regarding my last querry to the listserver, which was:
Dear Microscopists,
I am in a mild state of confusion regarding an immuno-EM experiment where it seems that the secondary antibody (GAM 5 nm) does not recognize the primary (mouse IgG) once the primary is bound to its corresponding antigen in tissue. We expect that the antigen is present with periodicity on fibrils in the connective tissue matrix. After the tissue is emersed in antibody, we see periodic decoration of the fibrils, which indicates to us that the antibody is bound.
However, secondary antibody does not bind to the tissue. We are convinced that the secondary conjugate is not defective, as we use it for other experiments. Has anyone else experienced a situation where a secondary does not recognize a primary once the primary is bound to its corresponding antigen?
Responses:
Several responses pointed out that secondary antibody conjugates are sub-type specific, meaning that if my primary was really an IgM and not an IgG, that a GAM IgG would not recogize the IgM. In my case, I know that the primary is an IgG.
Several responses suggested that I might not reasonably expect primary and secondaries to penetrate into the connective tissue matrix, that it might be to dense. Therefore I should consider a surface labeling procedure during which the antigen is exposed via sectioning. Unfortunately, this antibody will not work in surface labeling procedures, even though I have tried many different fixation/embedding techniques. Also, we have had years of success diffusing antibody into the CT space, including antibodies to other components of this same microfibril. However, as pointed out by another response, the gold particule we are using (5 nm) may be to big, and steric hindrance might be an issue. The suggestion is to try LM first, with a smaller FITC conjugate, then if positive use a smaller (1 nm) secondary for EM. This is the procedure we are about to try.
Many people still call this meeting MSA, but I do believe that the meeting is sponsored by the Microbeam Analysis Society and the Microscopy Society of America and that is why it is known as Microcopy and Microanalysis 98. There seems to be a general lack of understanding that the meeting is a joint affair of the two socities.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This sounds more probable with Environmental SPM imaging in the 3% sodium chloride solution / buffer. There is not need to dry it of and you can image in vitro.
George
At 08:16 AM 7/3/98 +0100, Keith Ryan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Donald, } } I wish to inject ferritin into the hemolymph (blood) of crabs to determine } which spaces are blood spaces. (Crab blood has copper-based hemocyanin, } rather than hemoglobin). } } Sigma carries several varieties of Ferritin, including Apoferritin. Any } suggestions as to which one I should use? } I think the apoferritin is the protein without the iron; if so, it would be a very poor tracer.
} Does anyone have a protocol to recommend? } If you can map the iron, you can see where it appears and where it is co-localized with copper. If you do not have element-mapping capability, the problem is much more difficult. In the latter case, some gold-labelled anti-ferritin (or forgetting the ferritin altogether, gold-labelled anti- hemocyanin) could work.
} What final concentration should I shoot for in the blood stream? } For EDS you would need a reasonable fraction of 1% iron; for WDS you need much less--I'll let the WDS experts tackle this one. Good luck. Yours, Bill Tivol
MSA's middle school microscpoy manual has JUST been published! Here's a brief description (taken from the Project MICRO bibliography):
Brady, S. and Willard, C. 1998 Microscopic Explorations 158pp, paperback, 8.5x11". ISBN 0-924886-00-5 Lawrence Hall of Science, University of California, Berkeley, CA 94720-5200; 510-642-7771 or 7262, gems-at-uclink.berkeley.edu. A collaboration between the Microscopy Society of America and the LHS has produced an outstanding Great Explorations in Math and Science (GEMS) guide. It's written in "festival" format, with a dozen explorations that can be presented simultaneously to circulating groups of students. There is a rich assortment of supplemental information on microscopes and how to buy them, curriculum extensions, further reading, and sources of help. The units are more classic than unique; subjects include crystals, color printing, fingerprints, pond water, brine shrimp, etc. Its uniqueness lies in the carefully written "inquiry science" presentation of those topics and the thorough classroom testing of content that a GEMS guide receives. It will work well in any classroom; teachers aren't expected to have special skills. Information on other GEMS guides (there are over 50!) is available at www.lhs.berkeley.edu/GEMS. Grades 4-8.
The retail price is $21.00, plus $4.00 shipping (in the U.S.); MSA members may request a 15% discount. A limited number of copies will be available at the Project MICRO booth in Atlanta. See y'all there!
Caroline Schooley Educational Outreach 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/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I've put some of the material from our LASER diffraction workshop on the web*, so as to share the fun. This began as a 2.5 hour workshop for gifted high school students participating in a six week program (the "Engleman Institute") here at UM-StL. A bit of thought, however, has suggested that even some of our diffraction experts might enjoy both the subjects, and the competition.
I would encourage microscopists to check out there computers with regard to the Y2K problem. However, I expect that the impact will be minimal, if not trivial, in practically all cases. We don't have the same sort of issues to deal with as the financial and insurance people.
I just checked out our computers using the test program from NSTL described by Ian below (the software is available from http://www.nstl.com/html/y2k_faq.html). It seems to do a good job of finding the problems, if any.
Our new computers (Pentiums and newer) checked out okay.
Our two remaining 486 machines reported minimal problems. The date would not correctly rollover on 1-1-2000. If the computer was powered off and on after 1-1-2000, the system would come up with a date of 1980. However, if the DOS DATE command was given on or after 1-1-2000 and the system restarted, it would come up with a correct date. There is a "century bit" in the BIOS info somewhere that will stay set once set, but does not appear to roll over correctly on these old machines.
Therefore, we can simply use the DOS DATE command after 1-1-2000 and reboot. Or I found that a utility form RightTime (www.RightTime.com) called YEAR2000.COM will fix the BIOS so that the computer passes the NSTL test. I do not know if it needs to be kept loaded after 1-1-2000. I would definitely want to check before I removed it.
The Year2000.txt file that comes with the RightTime utility does a very good job describing what the technical issues are for those who care to look deeper.
BIOS fixes are available for many computers to fix the problem. However, they may not be woth the hassle unless they are needed to fix another problem.
As far as application problems, I cannot imagine much happening in microscopy programs. If programs use only a two character year code, the code should still get transferred correctly. The problems will probably crop up when sorting or searching according to date. I have noticed the following behavior with some of our programs.
Access 7.0 (for 95) - Entering a value of 1/1/00 will be interpreted as 1900. You can and must enter 2000 (four digits) as the year for it to register as the next century. Otherwise 00 and 01 will precede 98 and 99 when sorted in ascending order.
Excel 95 - Entering 2-digit dates 00 through 19 is treated as 2001 to 2019. (You can check by formating cells to show the full 4-digit date.) 2-digit dates from 20 through 99 are treated as 1920 through 1999. In other words, their two digit century runs from 1920 through 2019.
Paradox database (as used by Quartz PCI, and others) - A search for dates after 1/1/30 return no files, while a search for files before 12/31/29 returnes all files. Their two digit century apparently is defined to cover the years 1930 through 2029.
To Summarize: A - Nothing broke in a real bad way. Even if something did, I suppose that 1999 could be extended artifically by backdating the computer until a suitable fix was found.
B - Many 486 and earlier computers will probably need some attention, but can probably get by with manual use of the DATE command after 1/1/2000. BIOS fixes are available for many computers if someone wants to go that route.
C - The data may have the correct two digit year code, but you may have to be wary how you search or sort the data. As long as manufacturers have written their code so that the two digit century encompasses the full range of our data collection, we should be all right. And since I don't have any data files collected between 1920 and 1929, I think I can safely assume that two digit codes representing the years 1930 through 2019 for my files will be treated correctly.
Hoping this helps, Warren Straszheim
At 10:26 AM 7/3/98 +0100, Ian Mc Laren wrote: } Karen, } Most old PCs (but not Macs or most Unix) will not transfer the date } correctly to the year 2000 although many will handle dates after Jan 1, } 2000 if told explicitly (i.e. with 4 figure years). You can get free test } software for PCs off the www. I found one from National Software Testing } Laboratories (NSTL) that seemed pretty useful. If your old PC will not do } the transition automatically you can either roll the date forward manually } (if it can do this), use a false date (such as 1990) but obviously this } could be misleading in some cases, or scrap the PC and buy a new one. } } For the most of our PCs that control lab equipment in our group I know that } they will not roll the date over automatically, so I am likely to keep all } the data backed up, wait till the end of 1999 and see what happens. If one } dies for some reason then we can always replace it with another cheap PC.
I am looking for PTFE tubing to fit a Hexland cryostage. Does anyone know who carries parts for this unit?
Thanks, Carl
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
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Hi Donna, Hope all goes well with you. Haven't talked to you ikn a long time. ESEM There is actually only one true environmental scanning EM and that is the = Philips Electroscan. The advantage is that samples are kept humid and do = not dry out. Any other "Vairiable pressure" SEM dries out the sample = with only short times to look at it. SO, as everything else, it depends = on what you want to do with the scope. If you are really looking at wet = samples, then the true environmental SEM seems appropriate. If that = isn't of interest then the others are cheaper. My two cents, Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
We have funding to buy a new SEM and have at least one application for an = environmental(variable pressure) microscope. Those of you who have = experience with environmental scopes, especially the Hitachi 3500N, please= let me know the good and bad points about your scope. Most of our samples are animal tissue and some of my reading has = suggested that to prevent tissue drying, when working at low vacuum, it = is necessary to use a cold stage. How do you rate the importance of a = cold stage for this type of specimen? Finally, those of you who have a cold substage from Fullam... have you = been satisfied with its performance? Any comments are welcomed. Thanks.
Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il 62794-1220 217-782-0898
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Dear List,
We recently purchased a microwave system for tissue fixation/embedding. Unfortunately, the instruction book that we ordered on it's use did not come and we now find out that it is no longer available (through Ted Pella). Does anyone have any information on procotols or know of any good reference books on microwave fixation and embedding? Any information that will get us going would be greatly appreciated.
William R.McManus Supervisor Electron Microscope Facility Department of Biology Logan UT 84322-5305 billEMac-at-cc.usu.edu
Looking for a new/used copy of: THE STUDY OF PLANT STRUCTURE: PRICIPLES AND SELECTED METHODS. 1981. O'Brien, T.P. and McCully, M.E. Termarcarphi Pty. Ltd. Melbourne, Australia.
Also, looking for steel (non-disposible, and glass blades for a Spencer 820 American Optical Rotary Microtome.
Thank you in advance Please reply via email: crow-at-aloha.net
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear List, } } We recently purchased a microwave system for tissue fixation/embedding. } Unfortunately, the instruction book that we ordered on it's use did not } come and we now find out that it is no longer available (through Ted } Pella). Does anyone have any information on procotols or know of any good } reference books on microwave fixation and embedding? Any information that } will get us going would be greatly appreciated. } } William R.McManus } Supervisor } Electron Microscope Facility } Department of Biology } Logan UT 84322-5305 } billEMac-at-cc.usu.edu } } } Bill, Here is a reference which may be of some help. Ted Pella equipment was used throughout: J Vet Diagn Invest 9:61-67 (1997) "Four-hour processing of clinical/diagnostic specimens for electron microscopy using microwave technique."
The Microwave Tool Book--A practical guide for microscopists by Gary Login and Ann Dvorak ISBN 0-9642675-0-0
Microwave Cookbook for Microscopists--Art and Science of Visualization by L. P. Kok and M. E. Boon ISBN 90-71421-20-1
Rick Giberson at Ted Pella should be able to give you some starting point protocols.
Patty Jansma Tel:520-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
On Mon, 6 Jul 1998 billemac-at-cc.usu.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear List, } } We recently purchased a microwave system for tissue fixation/embedding. } Unfortunately, the instruction book that we ordered on it's use did not } come and we now find out that it is no longer available (through Ted } Pella). Does anyone have any information on procotols or know of any good } reference books on microwave fixation and embedding? Any information that } will get us going would be greatly appreciated. } } William R.McManus } Supervisor } Electron Microscope Facility } Department of Biology } Logan UT 84322-5305 } billEMac-at-cc.usu.edu } }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Greetings, } } I am looking for PTFE tubing to fit a Hexland cryostage. Does anyone know } who carries parts for this unit? } } Thanks, } Carl
Hi Carl, Hexland are now part of Oxford Instruments, Research Instruments, Tubney Woods, Abingdon, Oxford, OX13 5QS. UK. +44 1865 393200. Or try your local agent. If you do not know a local contact e-mail Judith Brock on judith.brock-at-oxinst.co.uk.
Regards, Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
We are looking for flat molds for embedding in acrylic resins which could be hermitically closed. Can one out of you send me the name of the companies which sells them.
Thanks a lot
Marc
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
We are currently trying to establish capability in our lab to look at die cross-sections. In previous work we mounted them in epoxy or acrylic and coated the samples, but I'd like to be able to do this without mounting and coating. I've been told that there are simple polishing fixtures one can by to do this, but I can't seem to find where to buy them. Perhaps someone out there could suggest a source? Thanks,
Janet Rice MCC Senior Member Technical Staaff rice-at-mcc.com 512-338-3266
} We are currently trying to establish capability in our lab to look at di= e } cross-sections. In previous work we mounted them in epoxy or acrylic an= d } coated the samples, but I'd like to be able to do this without mounting and } coating. I've been told that there are simple polishing fixtures one ca= n } by to do this, but I can't seem to find where to buy them. Perhaps someone } out there could suggest a source? Thanks
--------- BUEHLER supplies the MICRO-PRECISE(TM) line of IC cross-sectioning equipment including the Tripoint Polisher for SEM/TEM sectioning. We als= o supply a semi-automated system that might be appropriate for your application as well. For more information, please contact me directly.
Scott D. Holt BUEHLER LTD. PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-6500 http://www.buehlerltd.com
The technique you are describing is very well suited to our Tripod Polish= er and is most likely the tool you have heard about. The Tripod Polisher is=
ideal for preparing a cross section of a specific device and can be used = to prepare both SEM and TEM cross sections. The SEM cross section is actual= ly the "easy" part of the process and is referred to as our "first side polish" when preparing a TEM cross section. The Tripod Polisher is easy = to use and we have developed a large database of technical reports on its us= e. The Tripod Polishing technique and the tool were actually developed at I= BM East Fishkill and have been used in cross sectioning for many years.
Another alternative is our BiPod Polisher which is a simpler version of t= he Tripod Polisher and is designed primarily for SEM cross sectioning. The BiPod polisher can be mounted on our polishing machines for semi-automati= c cross-sectioning. I'll be pleased to send you additional information on any of these products if they sound of interest to you. You can also get=
some of the information off of our web site at http://www.southbaytech.co= m.
If you plan to be at MSA next week in Atlanta, you can get a thorough demonstration of the technique at our booth (#433). =
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Janet Rice } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
We are currently trying to establish capability in our lab to look at die=
cross-sections. In previous work we mounted them in epoxy or acrylic and=
coated the samples, but I'd like to be able to do this without mounting a= nd coating. I've been told that there are simple polishing fixtures one can=
by to do this, but I can't seem to find where to buy them. Perhaps someo= ne out there could suggest a source? Thanks,
} We are looking for flat molds for embedding in acrylic resins which could } be hermitically closed. Can one out of you send me the name of the } companies which sells them.
} Marc -
The Ted Pella company has one. It's teflon, and should be sealed for polymerization with Thermanox cover slips. I know it works, because it was designed by Doug Davis, a tech in my (pre-retirement) lab at U.C.Berkeley.
Caroline Schooley Educational Outreach 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/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Is anyone currently doing the Kleinschmidt procedure for visualization of DNA with TEM? Dave Long at Montana State University needs to cooperate with a microscopy lab to have some DNA samples shadowed at low angle with the Kleinschmidt procedure. Either contact him by phone (406-994-5239) or me by phone (801-378-2451) or e-mail. Thank you. W. M. Hess.
Hi, I was asked to post this information from Cynthia Goldsmith for all M&M '98 attendees.
Taking MARTA (Metro Atlanta Rapid Transport Authority) from Hartsfield International airport to the downtown hotels.
} The MARTA rail can be found at the end of the baggage claim areas. Take } the MARTA Northbound train to the Peachtree Center station (N1), then } follow the signs to either the Marriott Marquis (through the Peachtree } Center Food Court and across a ramp) or the Westin Peachtree Plaza } (across Peachtree Street). } } You could also mention (as Sandy has in the information brochure) that } the rail lines run North-South and East-West, with the transfer point at } the Five Points Station. The GWCC (Georgia World Congress Center) is on the } } E-W Line, at the W1 station (Omni/Dome/GWCC).
see y'all soon, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
We have a couple of older Kevex Delta (III, IV) EDS units based on DEC PDP-1173's running RT-11 ver. 4.05. The units will not rollover to from Dec 31, 1999 to Jan 1, 2000, instead giving a date error. The RT-11 ver. 4.05 operating system is not year 2000 compliant. Once the Kevex software is up and running it will accept the year 2000 dates and run fine, however if you reboot the system the Kevex software takes its date from RT-11 which has to be pre-2000 in order for it to boot. What this means is that if we want to have the correct date on our printouts we'll have to change the dates both in RT-11 and the Kevex software every time we use the system. Currently this is the only date related problem we've noticed with the systems. Of course, we could upgrade to a PC based system or the RT-11operating system and DEC PDP-1173's could be upgraded. Both are now owned and supported by a company called Mentec (http://www.mentec.com/) and the new version of RT-11 (ver. 4.7) is y2000 compliant. An upgrade to ver. 4.7 would cost in the neighborhood of $2k. Mentec has indicated to me that although the new version is y2000 compliant, there are other changes and without input from the developers of the overlying Kevex software they can't guarantee that the Kevex software would run correctly. The PDP-1173 could be upgraded (bournoulli drives are no longer available) with SCSI cards and compatable peripherals and in fact would have to be since Mentec no longer supplys software upgrades on bournoulli's. For now, our strategy will probably be to change dates and hope for an upgrade to PC based systems.
kszaruba-at-MMM.COM wrote:
} Here's a general question that should stir up some comment... } } My department has sent out a standard questionaire for us to estimate } any problems our lab computers, software, or equipment might have with } the year 2000 date problem. Perhaps some of you have already gone } through this. I'm not a computer wiz, and naively thought that } everything I use would be fine, since I don't do any date calculations } like folks in the banking, insurance, etc. businesses do. However, the } more I hear about this problem the more confused I get. Is it true that } some older CPU's or BIOS programs will not handle the "00" date? So } some of our older equipment might not work even though the software we } are aware of has little reliance on dates?? Any microscope or software } vendors want to comment?? } } Hope this isn't stirring up needless fears or urban legends, but I have } definitely heard some alarmist opinions and would like to consult a more } technically aware and level-headed source (i.e. yourselves!). } } Thanks, } Karen } } -- } Karen Zaruba, kszaruba-at-mmm.com } BioMaterials Technology Center } 3M Center Bldg. 270-1S-01 } St. Paul, MN 55144 } } *The opinions above are my own, not necessarily my employer's*
-- ================================================================== 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 ==================================================================
Our Hitachi 3200H SEM came with a Magneto-Optic drive for storing images. When we transfer these to our PC however, the images show up without any of the labels (Mag. KeV, etc.). Has any one experienced this and does any one know how to correct this ?
Thanks to all who replied to this question. I should have said that the specimens, in the first instance, would be barnacle larvae. These are quite tough compared to a lot of other specimens that we have handled over the years. The study is to do with the question of settlement onto substrates.
The ideal would be to look at specimens with ESEM and then to allow them to grow on - do you think this is possible or would ESEM/radiation dose curtail normal development after exposure to the beam?
In this age of digital imaging, is a film recorder still useful? The last presentation I went to was all PowerPoint 'slides' but shown directly from a computer to a video projector, no film to be seen.
I am trying to decide if we should get one, to transfer digital images back to film and/or to make slides for presentations etc. If you have some opinions or experience with these things, could you let me know.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu **Area code changing to 831 as of 7/11/98**
Jordi writes ... } } Hi ! } } Our Hitachi 3200H SEM came with a Magneto-Optic drive for storing } images. When we transfer these to our PC however, the } images show up without any of the labels (Mag. KeV, etc.). } Has any one experienced this ...
Yes ... on a different system ... but at least my JEOL system allows an "export" option which "burns" the info you want into the TIF (... assuming it is TIF files you are archiving ...). I respond because we actually prefer that JEOL doesn't do this ... as we prefer to annotate with better software (e.g., Photoshop). If we've lost track of our notes and the associated mag with each image ... we've realized we can load the image into Windows' Wordpad and search for the mag in the TIF header ... we can then pick an appropriate micron bar dimension from a table associated with micron/pixel for each mag ...
Also ... you might ask Photoshop to show you the TIF header info ... you might be able to find the info you're after if your software adhered to TIF's standard comment fields whereas JEOL didn't.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
In response to Donald Lovett who wanted to know about using ferritin as a tracer for the crab. I did a study several years ago using ferritin mixed with india ink as a tracer to study circulation in the squid Loligo. The india ink helped us see when the solution was in the circulatory system. Mix 5 mls of 2X artificial sea water with 5 mls concentrated ferritin (Calbiochem) and 2 drops of india ink (sonicate the ink for 15 mins to break up the chunks). You need to spin down the ferritin for 30 mins -at- 30K to concentrate the ferritin. Then perfuse the animal-it might be hard in a crab-for 30 mins -at- room temperature. Then submerge into a high osmolarity fix (ie 2.5% glut in 0.1M cacodylate buffer with 0.8M sucrose- 1500 mosm). It worked really well. We also tried lanthanum and HRP. The combo of ink and ferritn was the best and easy to see in the TEM. Good luck! JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94022
Jon, Film recorders, as you are aware, are quite expensive ($10K for a good one). For the same amount of money you could buy the computer and projector to show them with. It's tempting, for sure. I am dithering with this same question myself. The only difficulty with going digital is the complete reliance on the technology: what happens if your computer crashes, the digital projector breaks, or both are incompatible. In getting ready for the MSA meeting (right now, in fact), I opted to do the textual slides in PP but to produce conventional slides for the non-textual material. Regular slides are always going to be superior (as is the photographic medium) and everyone has a conventional projector. Now, if you are pressed for time and have access to the technology, then digital is wonderful. If you have to make the purchase now, then go digital unless you really need the resolution.
} In this age of digital imaging, is a film recorder still useful? The last } presentation I went to was all PowerPoint 'slides' but shown directly from } a computer to a video projector, no film to be seen. } } I am trying to decide if we should get one, to transfer digital images back } to film and/or to make slides for presentations etc. If you have some } opinions or experience with these things, could you let me know.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 ####################################################################
Jon, we use and will continue to use a film recorder. In our case a Lasergraphics machine. Our lab is entirely digital and to be honest rapid computer based Powerpoint presentations are easily produced and often used. However, when it counts (ie: in public, to your peers) nothing is as good as an excellent high resolution 35mm slide. Whether it is to show illusive immunogold particles on cryosections "I am not sure whether you can see this at the back" or the minutae of confocal or decon image stacks there is NO substitute for a decent film recorder used to make images from high resolution digital images.
Simon C. Watkins Ph.D. MRC Path Associate Professor, Director CBI University of Pittsburgh
Allied High Tech specializes in Materiallographic, SEM and TEM sample preparation. We have the necessary tools, techniques, and supplies to ensure the cross-section you achieve is excellent. Allied also has a new polishing system with a micro-positioning head and polisher for doing precision cross sections, the Multi-Prep and TechPrep 8. If a micro-precision head is not what you want, we also have a easy to use hand tool. The procedures and techniques we have developed work very well. We also have a new Diamond Lapping film brochure containing a pictorial guide illustrating the results that can be achieved with Allied's Diamond Lapping Film.
If you are attending the Microscopy show in Atlanta our booth number is 548.
Gary Liechty is your representative in Texas, but any of the staff at Allied would be happy to assist you with any specific question you may have. Allied's goal is total customer satisfaction and the entire staff is committed to it.
For more information you may contact us at 800-675-1118 and visit our web site at: http://www.alliedhightech.com
We look forward to serving you.
Sincerely,
Edward Hirsch
At 09:06 AM 7/7/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com *************************************************
Is there an independent technican and/or dose any one on this list know an independent technican they could recommend to dissamble a Philips 300 TEM, located in Montreal, Canada?
} In this age of digital imaging, is a film recorder still useful? The last } presentation I went to was all PowerPoint 'slides' but shown directly from } a computer to a video projector, no film to be seen.
Joe: A very good question. I have been on both sides of this issue. Below are deciding factors in terms of which format I use. 1) How solid is the technology at the point where the lecture is to be given? The concerns here: I have Mac, they always use PC's, the right cable is not there. I made my presentation on Office 98 under Mac OS8.1 and they have System 7.1 loaded with PowerPoint 3. The mismatches can be endless and much harder to resolve when compared to changing a bulb in a slide projector. 2) Is the PowerPoint presentation better than using the slides? Is animation a key feature of the presentation? Or sound? Are the flying bullets better than the sharpness of the slide? How important is screen lumens? Bright slides work better than dim PowerPoint. It is hard to beat 2000 plus lines of slide resolution when compared to 640 by 480 pixels. They have a video projector with a bulb 500 hours on the otherside of dead. 3) How important is it to fix the presentation on the fly? You sized your slides to work in a 30 foot long room and you find yourself in a 100 foot hall. You can not resize 35 mm slides two hours before the lecture. Of course you can extensively rearrange a lecture in an airplane seat; but you sure can't do that with some 35 mm slides. 4) Can your presentation work best with twin screens: two images at a time? This is usually a snap with 35 mm slides but it is still pretty rough with PowerPoint. 5) One must always keep in mind the audience. Recently I saw a really slick computer controlled presentation. Some in the audience were put off by the excellent presentation because it was too slick. An analysis might help here: it was a research lecture being used to strut one's wares for a tenure-track teaching position. The students in the audience thought it was great. Older faculty who did not use computer media thought it was needlessly glitzy. Media proficient faculty were more focused on how the presentation was assembled rather than the message. Predictable but it still caught me off guard.
Finally back to your question. I use both and probably will continue to do so for the next several years. For really important lectures on the road, I have 35 mm slides for backup. The bottom line is, however, which format will the audience respond to best.
Blystone in Texas
Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu} Professor of Biology Trinity University San Antonio, Texas 78212 210.736-7243 210.736-7229 FAX
I am trying to respond to a message from Reggie Beijer of the Netherlands Forensic Sciences Laboratory. Unfortunately, I have no contact information. Can anyone provide me with an e-mail or FAX address? Please do not reply to the ListServer. Thank you for your assistance (TIA).
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
The Laboratoire d=92Analyse des Materiaux (LAM) of the Centre de Recherche Public-Centre Universitaire of Luxembourg has immediate opening, on a fixed term basis, for a maintenance engineer
The LAM facility includes 1 SEM, 1 TEM, 2 D-SIMS (CAMECA IMS-4f, CAMECA IMS-LAM), 1 S-SIMS (CAMECA/Ion Tof SIMS III) and one machine under development optimized for MCs+ clusters. The machines in operation are used to perform analysis for industries or by students preparing their PhDs.
The maintenance engineer will take care of all instruments existing at LAM (trouble shooting on electronics, maintenance of the physics).
Candidates should have a degree in electronics (baccalaureat plus two to three years), a first experience in maintenance of UHV equipment and be fluent in English. Interviews will take place at LAM, no funding will be provided for overseas trips.
Please send a resume and a list of references to:=20 CRP-CU 162a, avenue de la Fa=EFencerie L-1511 Luxembourg
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu
We just got a film recorder here and I definitely think that they are worth it: (1) good 35mm film recorders record 4,000 lines of resolution (resolution up to a maximum of 4096 horizontal by 2732 vertical lines, or 4096 x 2732) , which generally excedes the resolution of most 35mm films (looking under a LM I can't see the scan lines on 100ASA/ISA film - NO pixelation), (2) the HIGHEST resolution computer projection systems I have seen are 1024 x 860 - for text these are "o.k." but we work so hard to get excellent microscopic images and do we want to loose it all? (Besides a good film recorder costs the same or less than the best projection system, and you don't have to drag the thing with you to insure you have the best images you can) (3) many MANY places don't have good quality projector systems, but almost every where has a slide projector, (4) images take a LOT of disk storage space, so the image presentations don't fit on a diskette - you really need to burn a CD to take along to the meeting (can you guareentee they'll have a Zip drive? Maybe?), (5) Big image files are slow on all but the best computers systems, (6) which system to design the program for? Mac? PC? (7) want to use your notebook computer? Is it upto the task? do you want to play driver games the hour before your talk? (8) For B&W images the best quality I've seen are reproductions on true B&W media (as many have said this holds ture for printers as well), what I do is record the B&W images in negative to standard B&W film (i.e. T-max 100), develop in my darkroom (20-30mins) dry and mount - beautiful positive B&W slides! (Warning: look carefully, apparently negative film is much more senesitive than slide films so you have to get a good film recorder for this.
The one thing I haven't tried yet is taking true digital images back through film, back to a photographic enlargment, and comparing this to a dye-sub print.
O.k., so that's my two cents.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
Look for the "Microwave Cookbook of Pathology. The art of microscopic visualization". by Mathilde Boon and L.P. Kok. Coulomb Press Leyden, Leiden, The Netherlands. ISBN 90-71421-10-4 bound. I have the second revised edition from 1988, but probably there is a newer one available now.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } On Mon, 6 Jul 1998 billemac-at-cc.usu.edu wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear List, } } } } We recently purchased a microwave system for tissue fixation/embedding. } } Unfortunately, the instruction book that we ordered on it's use did not } } come and we now find out that it is no longer available (through Ted } } Pella). Does anyone have any information on procotols or know of any good } } reference books on microwave fixation and embedding? Any information that } } will get us going would be greatly appreciated. } } } } William R.McManus } } Supervisor } } Electron Microscope Facility } } Department of Biology } } Logan UT 84322-5305 } } billEMac-at-cc.usu.edu } } } } } } }
Bill, }
Here is a reference which may be of some help. Ted Pella } equipment was used throughout: J Vet Diagn Invest 9:61-67 (1997) } "Four-hour processing of clinical/diagnostic specimens for electron } microscopy using microwave technique." } } Drs. Marcel Paques Wageningen Centre for Food Sciences p.a. NIZO Food Research Kernhemseweg 2, 6718 ZB Ede P.O.Box 20, 6710 BA Ede The Netherlands Phone: (31) 318-659690 Fax: (31) 318-650400 e-mail: paques-at-nizo.nl
Jon, "Yes" everyone is going to PowerPoint slides but" no" not everyone has a powerbook that they can take to meetings, courses, etc. Having made countless slides using PowerPoint and a digital slide maker, I don't know how we would manage without both at this point.
Example, I recently returned from a 3 week trip to China. Both my husband and I gave talks at 2 major Universityies on what was otherwise a vacation trip. There was no way we were going to drag a powerbook along. Even if we had, the University lecture halls were not equipped with digital projectors or LCD panels for projection of the PowerPoint slides directly from the computer program.
We have been in countless other situations where we wanted to make informal presentations in rooms which were not equipped with digital projectors such as when our extension educators go out to give talks to local groups. It is easy to come up with a 35mm slide projector, set it up and in no time be able to informally present data in the form of slides but more difficult to find suitable digital projectors, etc. for multiple people from one department to use at the same time in different parts of the state.
It is also very easy to tailor a presentation in just a few minutes by having an assortment of slides and picking and chosing for the audience at hand....much faster and easier that going through multiple PowerPoint files to find the slide desired and then copying and rearranging into a new presentation.
Debby Sherman +++++++
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
Dear List:
In this age of digital imaging, is a film recorder still useful? The last presentation I went to was all PowerPoint 'slides' but shown directly from a computer to a video projector, no film to be seen.
I am trying to decide if we should get one, to transfer digital images back to film and/or to make slides for presentations etc. If you have some opinions or experience with these things, could you let me know.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu **Area code changing to 831 as of 7/11/98**
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Would you please recommend a short course for Light Element X-Ray Analysis
I have missed Lehigh's class that took place in June of '98.
Thank you in advance.
Darlene Peterson Harvey Eltech Research Center Fairport Harbor, Ohio
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We will be setting up interviews for candidates for the Fall teaching = position at San Joaquin Delta College for electron microscopy. Course = details are below. If you are interested in teaching all 4 courses or 2 = of them, please contact me in Atlanta at the Microscopy and Microanalysis = Meeting July 12-16, 1998. I will be staying at the Marriott Marquis = Hotel. You can leave a message there or on the meeting bulletin board. = A message left at the hotel might be faster. I will be coming into = Atlanta Saturday July 11 and leaving Thursday evening, July 16, 1998.
Please bring your active resume with you. I will have official SJDC = applications which you can fill out. You will of course need names, = addresses and dates of past employers and names, addresses, and phone = numbers for recommendations and typical application type information. We = can fax them to our Human Resources Dept, and then, if appropriate, set = up a conference interview with the search committee from Atlanta. Please = contact me as early as possible in the week. We are up against an = extremely tight deadline.
Thanks
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
URGENT: ADJUNCT FACULTY NEEDED FALL Semester 1998 (Aug. 12 - Dec 18, 1998)
A San Joaquin Delta College Faculty member of the Microscopy Technology = Center is planning a Sabbatical Leave for the Fall '98 semester. San = Joaquin Delta Community College is currently searching for an EM = Instructor Adjunct / Temporary One Semester Contract for Fall 98 (Aug. 12 = - Dec 18, 1998).
MINIMUM QUALIFICATIONS: Bachelor's Degree plus two years of directly related experience OR an = Associate Degree plus six years of directly related experience OR a valid = credential.
DESIRABLE QUALIFICATIONS: Master's or PhD Degree in a Biological = Science; Experience in teaching / practice of Electron Microscopy
COURSES TO BE TAUGHT:
Introductory Techniques for Transmission Electron Microscopy (EM21) This is a lecture/lab course which includes beginning Transmission = Electron Microscopy dealing with the alignment and operation of the TEM, = vacuum techniques, photographic techniques, as well as the preparation of = particles and replicas for viewing in the TEM. Includes individual = training in the use of the TEM, preparation techniques, and written and = oral reports. (Lec - 2 hrs; Lab - 3 hrs/wk)
Biological Ultrastructure (EM28) Course contents include specific information about the fine structure and = function of cells and tissue at the ultrastructure level. Videos, slides = and micrograph examination will be correlated with the lectures so that = students will learn to recognize the fine structure of cells and tissues = in relationship to their function. (Lec-2 hrs/wk)
Advanced Techniques in Biological Electron Microscopy (EM37) Course contents include lecture and laboratory which covers advanced = techniques for biological specimen preparation in TEM including an = advanced research project.( Lec - 1 hr; Lab - 6 hr/wks)
** Current Microscopies: Optics, Theory and Application (EM30) Course contents include information related to the physical laws and = applications of the various types of current microscopies e.g. = TEM,SEM,FIB, AFM, and confocal microscopy, as well as other current = topics e.g. asbestos analysis, lab design, etc. (Lec - 2 hrs; Lab - 3 = hrs/wk) ** This course is negotiable. A portion of this assignment would be = paid as an additional overload.
The above teaching load would not require development of new courses as = these courses have been fully developed. Course materials are available = to aid in the instruction. TERMS OF EMPLOYMENT: Full semester, Non-tenured track position with Full = Benefits (Medical, Dental, & Vision). SALARY for Fall 98 semester: BA/BS w/ 2 yrs experience or AA w/ 6 yrs experience$17,498 - 25,333 for Fall 98 sem
MA/MS$19, 219 - 28,137 for Fall 98 semester
PhD$ 22,528 - 31,739 for Fall 98 semester
Stockton, CA is 90 miles Northeast of San Francisco and 50 miles South of = Sacramento. It is in lush, green, San Joaquin Valley. We do not suffer = from the high rent districts of San Francisco and Silicon Valley, both of = which are our neighbors.
Additional SJDC Microscopy Technology Program Information available at = http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html/
If you do not contact me in Atlanta, you can contact Human Resources at = the below number however we are up against an extremely tight deadline = and a search committee that may only be available during the Atlanta = meeting. I will have official applications at the meeting and will fax = them to Human Resources.
APPLICATION: Contact Human Resources at 209/954-5056. Human Resources San Joaquin Delta College Admin. Bldg. Rm 202 5151 Pacific Ave Stockton, CA 95207
by gcsin1.gecm.com (8.9.0/8.9.0) with SMTP id SAA16375; Wed, 8 Jul 1998 18:04:29 +0100 (BST) Received: from gcschm.geccs.gecm.com by gcsin3.geccs.gecm.com; (5.65/1.1.8.2/13Mar95-1139AM) id AA18376; Wed, 8 Jul 1998 18:03:57 +0100 Disclose-Recipients: prohibited
Janet, you can either spend lots of dosh on a tripod polisher and polishing system, or do the following;
1) Get some glycothalate (?spelling) thermoplastic wax, such as 'crystalbond' (lots of electron microscopy suppliers sell it).
2) Get some epoxy - anything will do, 5 minute (Devcon) saves a lot of time
3) Glue up a 'sandwich' of three pieces of silicon using the epoxy, with the die you want in the middle. Do it one at a time, using as little glue as possible. Make sure everything is really clean, and squidge the samples about from side to side to squeejee the excess glue away. You can speed up curing on a hotplate. Make sure that all the pieces line up on one edge so that it stands on edge for when you section it.
4) Mount the sandwich, edge on, in the wax (melts at about 120C, nice and runny at 180C) in the centre of a standard polishing platen. Put three blocks of silicon round the edge.
5) Grind till flat on 1200 grit, polish for a few seconds on 2400 grit, fine polish on 1um diamond and a short nap felt pad (slow rotation) for 10 mins.
This is essentially tripod polishing without spending any money. If you want to hit a specific area, you can use a piece of glass instead of the top silicon piece, or even a small piece of silicon so you can see the uncovered part of the die and work out where you are from a (previously taken!) photo.
Let me know if you want any more details,
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
I think that it is absolutely worthwhile to get a film graphics recorder. While computers with video projectors are probably the wave of the future, at most presentations that I attend, there is the old Kodak Carousel and the 35 mm slides being projected. While a lot of commercial enterprises have the new computerized slide shows, I still believe that 35 mm slides are going to be around for a long time to come, especially in academic institutions. Not every institution nor department can afford the notebook and video projector just to be state of the art in presentation technology. Nor can many investigators for that matter.
I get called on a lot to either shoot 35 mm slides on my film graphics recorder for presentations and often at short notice.
The other thing that you should do as regards the need in your own facility is to ask your customers (the investigators at UCSC) if they use 35 mm slides in their presentations in order to gauge the need for a film graphics recorder. I will bet that the majority of them do. Their Powerpoint presentations as well as any digital microscopic images (confocal or otherwise) can easily be shot on 35 mm slide film with a film recorder.
Matthew J. Schibler Ph.D. Imaging Facilities Core Coordinator UCLA Brain Research Institute 73-384 CHS 951761 Los Angeles, CA 90095-1761
(310) 825-9783 FAX (310) 206-5855 E-mail: mschibler-at-bri.medsch.ucla.edu http:/btrip.mednet.ucla.edu/bri/homepage.htm } ---------- } From: jmkrupp-at-cats.ucsc.edu[SMTP:jmkrupp-at-cats.ucsc.edu] } Sent: Tuesday, July 07, 1998 3:40 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: LM/EM Film recorders? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Dear List: } } In this age of digital imaging, is a film recorder still useful? The } last } presentation I went to was all PowerPoint 'slides' but shown directly } from } a computer to a video projector, no film to be seen. } } I am trying to decide if we should get one, to transfer digital images } back } to film and/or to make slides for presentations etc. If you have some } opinions or experience with these things, could you let me know. } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } **Area code changing to } 831 as of 7/11/98** } }
One feature of film that has not been mentioned is its archival qualities. It is quite likely that digital presentations will not be very accessible in five years due to hardware and software changes. Properly processed film should last more than 50 years--probably more than 100 years. A slide can be viewed without any eletricity--hold it up to the sky! Try that with a Zip disk.
Has anyone used Image Space Software for looking at dual channel "single section" images. Specifically using the dual channel overlays with two colours. I may be missing something but I cannot get the dual or stereo applications to work with single images. Probably this is not possible with this software it seems to be looking for depth information.
Also which antifade agent do people recommend for fluorescent antibody probes for use with confocal.
I've been following the thread and I have two cents to add about something that I don't think has been touched upon.
I gave an internal presentation that had TEM images that were acquired from a Gatan 1024 x 1024 size CCD camera. The presentation was put together in powerpoint and printed out on a sub-dye printer on overhead transparencies. My manager asked me why I didn't do the presentation with the computer/projection system like several other people have recently done very elegantly. I told them that they didn't have images in their presentations and could get away with it. He said "what does that have to do with anything?"
So I told him the following things: 1) the best projection systems today have about 1024x780 pixel resolution. 2) my images are 1024 x1024 in size. If I wanted to show all the available details in the image, I would have to set the zoom on the powerpoint window so that only the image was visible and none of the title info or other stuff that I have on the slide would be visible. 3) on the sub-dye printer which has 300 dpi resolution, the same 1024 x1024 image would be 3.4 inches in size and I could put several images on the same slide which is then projected with all the details of all of the images in the slide.
Now I would prefer to use 35 mm slides when the absolute best quality in the images are required. However, in the last several years, I have converted to using overhead slides because of the ease of producing good quality overheads in a very short period of time and because slide projectors are becoming less and less available in some locations, particularly in the government labs. When I was at the Materials Lab at Wright Patterson Air Force Base, it was an ordeal to find a working slide projector when someone was coming through the lab that had slides. It is about the same here.
Ok, can I have the change left over from my two cents?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
In this age of digital imaging, is a film recorder still useful? The last presentation I went to was all PowerPoint 'slides' but shown directly from a computer to a video projector, no film to be seen.
I am trying to decide if we should get one, to transfer digital images back to film and/or to make slides for presentations etc. If you have some opinions or experience with these things, could you let me know.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu **Area code changing to 831 as of 7/11/98**
It has become my task to convince our University here of the strategic importance in maintaining sophisticated microscopy, ie Confocal Microscopy, Electron Microscopy etc as a service to research and teaching.
Here in New Zealand the bulk of scientific research funding comes from the government via a small number of government funding agencies. The research climate is rapidly changing. It would seem the government is moving towards a situation where only a specific number of 'identifiable and targeted' research outcomes will be contested for research funding. If your research activity does not fall into one of these 'targeted' areas and you can not get funding from some other source then you are essentially out of the 'research game'.
With regard to major scientific equipment purchases, such as electron microscopes, NMR's etc, this means that new equipment initiatives (and presumably replacement of existing equipment) will become 'end user driven' rather than, as in the past, 'provider push'.
Traditionally Universities here have obtained research funding on the understanding that the infrastructure to do that research was already in place within the university and, if major equipment purchases were required, then funding was based on current and anticipated usage from a number of researchers or groups, with funds coming from a number of different funding sources (most of which have gone now) and including a contribution from the university itself based on a teaching component. This model is now almost gone.
In the new model it would seem that equipment will only be purchased if it is integral to the efforts of a particular targeted outcome. The 'funding for teaching' aspect is yet to be resolved however it would seem that of the income received by universities from the government to teach students, a 'research component' will also be contestable. This makes capital equipment, such as electron microscopes, NMR's etc, quite vulnerable unless the University can be convinced that it is in it's strategic interest to maintain the availability of such technology and techniques (funding of university teaching activity is also probably going to change soon with the possible introduction of a 'student voucher' system).
As I said above it has become my task to convince our University of the strategic importance in maintaining sophisticated microscopy, ie Confocal Microscopy, Electron Microscopy etc as a service to research and teaching. If such technology and techniques are considered of strategic importance then there is a better chance that the university will continue to maintain such facilities. Obviously I have a vested interest in the long term survival of our Microscopy Unit but I also believe that microscopy as a technique and technology continues to have vital role to play in many scientific disciplines and industrial processes therefore must continue to be taught to our science students.
1. Has anybody out there in 'microscopy land' had to convince their institution of the strategic importance in maintaining sophisticated microscopy within that institution ?
2. If so, I would appreciate some feed back on what arguments you thought worked and what arguments failed.
3. Did you succeed ?
All feedback would be appreciated.
Allan Mitchell
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
Microscopy/Microscopy Education offers a full range of customized, on-site courses. For more information, see our web-site at {http://www.MME-Microscopy.com/education} or call our offices.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site: {http://www.MME-Microscopy.com/education} ****************************************************** MME: America's first national consortium dedicated to customized on-site training in all areas of microscopy, sample preparation, and image analysis. Our goal: immediate growth in your productivity!
At 09:24 AM 7/8/98 -0500, Darlene Harvey wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
OK, I'll use propane instead of isopentane. Can someone tell me what purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane available. Airgas also has what they call "Natural grade (Grade 1.6)", 96%. (I assume there's a reason not to use propane containing mercaptans, oil, and who knows what else, from the cylinder of my propane torch or camping equipment. Right?) Can you recommend any US suppliers for 2 to 20 pound bottles? (2 pounds costs almost the same as 20 pounds. I don't need 100 pounds of propane.)
Thanks! Richard
} } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane. Its easy to store in a lab a small gas cylinder with a blunt needle on a bit of tubing as the outlet. With little gas flow rub the needle over the small metal cup that is cooled by liq N2. Soon you will have a couple of ml of liquid propane. Do this in a fumehood, which is a good idea when using solvents too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
I have a few extra rooms reserved at the Atlanta Marriott Marquis that I will not be using. I know the city is crowded next week so if anyone is looking for a room, please let me know ASAP as I plan to cancel these ext= ra rooms on Friday morning.
Use camping gas and I doubt you will find Any difference. None of the plunge cooling organic cryogens are as good as high pressure freezing (BUT that option although the best is expensive)
Patrick Echlion Multi-Imaging Centre University of Cambridge
On Wed, 8 Jul 1998, Richard Thrift wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, I'll use propane instead of isopentane. Can someone tell me what } purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane } available. Airgas also has what they call "Natural grade (Grade 1.6)", } 96%. (I assume there's a reason not to use propane containing } mercaptans, oil, and who knows what else, from the cylinder of my } propane torch or camping equipment. Right?) Can you recommend any } US suppliers for 2 to 20 pound bottles? (2 pounds costs almost the same } as 20 pounds. I don't need 100 pounds of propane.) } } Thanks! } Richard } } } } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } } Quite right, but now the discussion goes to: Which is the better cryoagent } and that was a topic here a few months ago. } Propane gas liquefied by cooling is a much, much better cryo-agent than } is isopentane. Its easy to store in a lab a small gas cylinder with a blunt } needle on a bit of tubing as the outlet. With little gas flow rub the needle } over the small metal cup that is cooled by liq N2. Soon you will have a } couple of ml of liquid propane. Do this in a fumehood, which is a good } idea when using solvents too. } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } **************************** www.proscitech.com.au ***** } }
We run courses "in house", in your own laboratory on your own equipment, on all aspects of electron microscopy. Please take a look at our web site for more information, we are able to tune a course to any requirement.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
For a quick review of the new technologies on exhibit at next week's Microscopy & Microanalysis meeting, see the July issue of American Lab: "Focus on Microscopy: What's New at M&M '98", pp. 41-45. (Hot off the presses!)
Extra copies will be available at the meeting at the Microscopy/Microscopy Education Booth (#510).
Richard, you should not have visited that propane variety store, its too confusing. I have used the propane supplied for camping and home stoves. Very likely this contains minor contaminants, but it worked well. Contaminants would change the freezing point in a minor way but I think it very unlikely that they would penetrate and affect cell structure. However, there is an "interesting" research project for some doubter. Please let me know after such "propane purity" research has been accepted for publication. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes service-at-proscitech.com.au *** www.proscitech.com.au
-----Original Message-----
OK, I'll use propane instead of isopentane. Can someone tell me what purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane available. Airgas also has what they call "Natural grade (Grade 1.6)", 96%. (I assume there's a reason not to use propane containing mercaptans, oil, and who knows what else, from the cylinder of my propane torch or camping equipment. Right?) Can you recommend any US suppliers for 2 to 20 pound bottles? (2 pounds costs almost the same as 20 pounds. I don't need 100 pounds of propane.)
Thanks! Richard
} } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane. Its easy to store in a lab a small gas cylinder with a blunt needle on a bit of tubing as the outlet. With little gas flow rub the needle over the small metal cup that is cooled by liq N2. Soon you will have a couple of ml of liquid propane. Do this in a fumehood, which is a good idea when using solvents too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
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OK, I'll use propane instead of isopentane. Can someone tell me what purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane available. Airgas also has what they call "Natural grade (Grade 1.6)", 96%. (I assume there's a reason not to use propane containing mercaptans, oil, and who knows what else, from the cylinder of my propane torch or camping equipment. Right?) Can you recommend any US suppliers for 2 to 20 pound bottles? (2 pounds costs almost the same as 20 pounds. I don't need 100 pounds of propane.)
Thanks! Richard
} } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane. Its easy to store in a lab a small gas cylinder with a blunt needle on a bit of tubing as the outlet. With little gas flow rub the needle over the small metal cup that is cooled by liq N2. Soon you will have a couple of ml of liquid propane. Do this in a fumehood, which is a good idea when using solvents too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
We are using LR White for post-embedding staining with colloidal gold. It is absolutely imperative that we get the best possible fixation for synapses. Synapses have a large lipid component. Has anyone tried tannic acid in combination with LR White? If so, how? p-phenylenediamine is known to protect against lipid loss during alcohol dehydration. Has anyone used it with LR White? If so, how? Anyone have any other ideas? We have at our disposal UV light and Progressive Lowering of Temperature techniques, as well as standard oven polymerization. We would so much appreciate any help we can get. We absolutely have to have good fixation of synapses this summer in order to get our next grant (and keep our jobs!) Our antigens tend to be very difficult to locate with Au in material fixed with osmium and embedded in epoxides, which, of course, would give us our best fixation of synapses.
"Can someone tell me what purity of propane is required? I see 99%, 99.5% & 99.99 (!) % propane available. Airgas also has what they call "Natural grade (Grade 1.6)", 96%. (I assume there's a reason not to use propane containing mercaptans, oil, and who knows what else, from the cylinder of my propane torch or camping equipment. Right?) "
I have just constructed an apparatus for freeze spraying in propane, with the help of instructions from Scott Russell, and have been told barbecue gas propane is just fine....in fact the impurities help to reduce the freezing point (I think that's right!) Good job, because the pure stuff costs a bomb!
-- Amanda Wilson awilson-at-sghms.ac.uk Assistant Manager, Electron Microscope Unit, St George's Hospital Medical School, Tooting, London, UK. http://sghms.ac.uk/em tel:? 0181 725 5220 fax:? 0181 725 3326
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{HTML} {BODY BGCOLOR=3D"#FFFFFF"} Richard wrote:
{P} "Can someone tell me what purity of propane is required?=A0 I see 99%, 99.5% & 99.99 (!) % propane available.=A0 Airgas also has what they c= all "Natural grade (Grade 1.6)", {BR} 96%.=A0=A0 (I assume there's a reason not to use propane containing {BR} mercaptans, oil, and who knows what else, from the cylinder of my {BR} propane torch or camping equipment.=A0 Right?) " {BR} =A0
{P} I have just constructed an apparatus for freeze spraying in propane, with the help of instructions from Scott Russell, and have been told barb= ecue gas propane is just fine....in fact the impurities help to reduce the fre= ezing point (I think that's right!)=A0 Good job, because the pure stuff costs a bomb! {BR} =A0
} OK, I'll use propane instead of isopentane. Can someone tell me what } purity of propane is required?...
In my experience, commercial grade propane, such as is sold for use in your barbecue, propane torch and campstove, works perfectly well for plunge freezeing or propane jet freezing. The major differences that I found between chemically pure propane (99%) and commercial grade propane were of course due to the impurites in the latter. For example: (a) the viscosity of commercial grade propane is higher, so I had to enlarge the bore of the propane jets to obtain the same jet velocity (and thus freezing quality), and (b) the freezing point of commerical grade propane is depressed several degrees, and the higher viscosity may slow nucleation, so that I had more time to prepare my specimen before the propane froze. I have also saved a bundle of money and a lot of trouble (since it's so readly available) with the commercial grade stuff. FYI, I have been using 20 lb containers for jet freezing and campstove containers for plunge freezing.
Bill McManus asked: } We recently purchased a microwave system for tissue fixation/embedding. } Unfortunately, the instruction book that we ordered on it's use did not } come and we now find out that it is no longer available (through Ted } Pella). Does anyone have any information on procotols or know of any good } reference books on microwave fixation and embedding? Any information that } will get us going would be greatly appreciated.
I'm not certain which specific book Bill meant, but we sell the Kok & Boon _Microwave Cookbook for Microscopy_ and Gary Login's _Microwave Toolbook_. We also have several microwave processing and microwave fixation procedures at our web site (address in my .sig, below).
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Just a quick note to say that my experience with propane freezing has been very similar to that of John Gilkey's.
---------------------
" In my experience, commercial grade propane, such as is sold for use in your barbecue, propane torch and campstove, works perfectly well for plunge freezeing or propane jet freezing. ......................" J. Gilkey
"Breakthroughs in biology are preceded by breakthroughs in microscopy."
I cordially invite you to come see Molecular Imaging's technology breakthrough in microscopy, which may dramatically change and speed up the processes in drug discovery.
This "Environmental / In Vitro Atomic Force Microscopy" (AFM) can deliver nanometer resolution time lapse imaging and physical property characterization.
Please come to scientific talks at The Protein Society, 12th Symposium in San Diego, July 25-29, 1998, on this technology and its revolutionary applications: - Prof. Stuart Lindsay, ASU; Oscillating Probe AFM Study of Titin Unfolding, Mon. July 27, 1998, 10:15AM - Dr. Peter Hinterdorfer, University of Linz; Austria (Molecular Recognition Force Microscopy) Antibody-Antigen Recognition Detected By Ultrasensitive Force Microscopy Sunday July 26, 1998, Molecular Recognition, Poster Session, 3:15 - 5:30PM
Or come for a live demonstration by Molecular Imaging at exhibition booth # 613.
If you can not join us in San Diego, please visit us at http://molec.com/BIO-AFM/protein
Respectfully
George Sibbald
PS: If you can not get to Protein come to the Microscopy Society of America Show in Atlanta, July 13-16, 1998 ____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; "Innovating Probe Microscopy" 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Dear All: We are seeking to purchase a TEM simulation software named Mc Tempas runed on Mcintosh computer. We appreciate it if any one can provid the information about where we can buy it, or suggest a better one that can perform interface structure simulation with supercell generation option. Thank you.
---------------------------------------- Dr. Dai Jiyan IMRE National Univ. of Singapore 10 Kent Ridge Crescent Singapore 119260 ----------------------------------------
I am interested in touring the CDC in Atlanta while attending the M&M98. Are there any official tours set up for this? It would be very interesting if we could have a tour geared towards microscopists. Is any one interested?
Sally Burns
Center for Electron Optics burnssal-at-pilot.msu.edu (517) 355-5004
Attention Poor and Desparate EM facilities: -I have three DENTON 515-DV available, used, operational, crated and ready for surplus to any Govt agency who wants it. Just let me know and get your GBL ready. If there are no Gov't takers, then Universities, then Schools are eligible. -I also have two LN2 tanks, on wheels, probably 25L size. Ready to go. -Misc LKB Ultramicrotome repair parts, like fuses, washers, ETC, freebies. -Eight LAB6 filaments for a JEOL 35, never used. You will need the ultra-vacuum pump for these. We never did buy one. -Small Sorvall Histo-knifemaker. That's it for now. I will ship small items today, or after the EMSA meeting. If you are there, I will bring them for you. BIG items will be shipped when the shipping paper work is completed. Please use E-Mail, so I will have a record of the time and date that you responded. Most equipment has service record and instructions. Thank you, Thomas A Baginski, USUHS, Bethesda, MD 20814 Email: tombg-at-bictom.usuf1.usuhs.mil or Voice phone for specific questions 301 295 5691
HILDEGARD CROWLEY wrote: } } Hi, } } We are using LR White for post-embedding staining with colloidal } gold. It is absolutely imperative that we get the best possible fixation for synapses. Synapses have a large lipid component. Has anyone tried tannic acid in combination with LR White? If so, how? } p-phenylenediamine is known to protect against lipid loss during } alcohol dehydration } snip {
Hildy:
I'm pretty sure p-phenyleaminediamine only protects against lipid loss when used after osmium. Sorry I don't have more to offer.
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 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I would like to look at the cross section of silicon nitride right next to the fracture surface. There is not enough room for dimpling . I was considering tripod polishing, creating a wedge on the side face perpendicular to the fracture surface and hopefully I will get a thin region that is close enough to the fracture. I would appreciate comments or suggestions from people who have tried this.
Hildy, You did not indicate whether it is possible to perfuse fix your material. If so, you might want to check the reference "Liposits, ZS etal (1986). A combined light and electron microscopic immunocytochemical method for the simultaneous localization of multiple tissue antigens. Histochemistry 85:95-106"
I worked with Zsolt on this publication which involved perfusion of rat brains with high and low pH paraformaldehyde, vibratoming the brains, immunocytochemical reaction using the PAP-DAB method, OsO4 fixation and finally embedding in epoxy. It gave very good localization and excellent ultrastructure since membranes were fixed with osmium after ICC. We worked out a simple method of flat embedding the desired areas of the sections between plastic coverslips and then serial sectioning them so we could trace the synapses.
I'm sure that Zsolt has worked out a method to use colloidal gold since that time....probably using nanogold coupled with silver intensification. You should be able to do a literature search and find more recent publications.
Although I do not presently work with neuronal tissue, I would be happy to dig back into my notes after returning from MSA. Do feel free to contact me if you want additional information.
Debby Sherman =================================================== Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Hi,
We are using LR White for post-embedding staining with colloidal gold. It is absolutely imperative that we get the best possible fixation for synapses. Synapses have a large lipid component. Has anyone tried tannic acid in combination with LR White? If so, how? p-phenylenediamine is known to protect against lipid loss during alcohol dehydration. Has anyone used it with LR White? If so, how? Anyone have any other ideas? We have at our disposal UV light and Progressive Lowering of Temperature techniques, as well as standard oven polymerization. We would so much appreciate any help we can get. We absolutely have to have good fixation of synapses this summer in order to get our next grant (and keep our jobs!) Our antigens tend to be very difficult to locate with Au in material fixed with osmium and embedded in epoxides, which, of course, would give us our best fixation of synapses.
So long, Hildy
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Fellow microscopists:
I am looking for suggestions/references on doing immuno on lens from human eyes. For some reason I am under the impression that the texture of the lens may require special fixation and penetration of Ab's may be difficult. Does it become brittle under certain conditions, i.e. would paraffin or cryo- sectioning be better - that, of course, may depend partly on the Ab's.
Also, does lens contain any endogenous autofluorescence?
Thank you for any assistance.
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
Hi Folks: I want to make some resin casts for SEM of murine skin. two questions, 1: which is the least viscous resin that works well (I need to get at the capillary microstructure) 2: which is the best perfusion method for this type of perfusion (we normally perfuse intracardially) Of course I would welcome any other tips from the wise on this method Thanks a lot Simon
----------------------------------------------------------------------- Simon C. Watkins Ph.D. M.R.C.Path Associate Professor Director, Center for Biologic Imaging University of Pittsburgh Pittburgh PA 15261 tel:412-648-3051 fax:412-648-8330 URL: http://sbic6.sbic.pitt.edu
Sorry if I am sending this message a second time, but I suspect the original message was not ssent out.
Fellow microscopists:
I am looking for suggestions/references on doing immuno on lens from human eyes. For some reason I am under the impression that the texture of the lens may require special fixation and penetration of Ab's may be difficult. Does it become brittle under certain conditions, i.e. would paraffin or cryo- sectioning be better - that, of course, may depend partly on the Ab's.
Also, does lens contain any endogenous autofluorescence?
Thank you for any assistance.
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
Listers-I thought most of you would appreciate the moral of this story. I know it happens quite frequently.
Paula :-)
} } } } This is a weird but true story (with a moral) } } } } } } } } A complaint was received by the Pontiac Division of General Motors: } } } } } } } } "This is the second time I have written you, and I don't blame you } } for } } } } not answering me, because I kind of sounded crazy, but it is a fact } } } that } } } } we have a tradition in our family of ice cream for dessert after } } dinner } } } } each night. But the kind of ice cream varies so, every night, after } } } } we've eaten, the whole family votes on which kind of ice cream we } } } should } } } } have and I drive down to the store to get it. } } } } } } } } It's also a fact that I recently purchased a new Pontiac and since } } then } } } } my trips to the store have created a problem. You see, every time I } } } } } buy } } } } vanilla ice cream, when I start back from the store my car won't } } start. } } } } If I get any other kind of ice cream, the car starts just fine. I } } want } } } } you to know I'm serious about this question, no matter how silly it } } } } sounds: } } } } } } } } 'What is there about a Pontiac that makes it not start when I get } } } } vanilla ice cream, and easy to start whenever I get any other } } kind?'" } } } } } } } } The Pontiac President was understandably skeptical about the letter, } } } } } but } } } } sent an engineer to check it out anyway. The latter was surprised } } to } } } be } } } } greeted by a successful, obviously well educated man in a fine } } } } neighborhood. He had arranged to meet the man just after dinner } } time, } } } } so the two hopped into the car and drove to the ice cream store. } } } } } } } } It was vanilla ice cream that night and, sure enough, after they } } came } } } } back to the car, it wouldn't start. } } } } } } } } The engineer returned for three more nights. The first night, the } } man } } } } got chocolate. The car started. The second night, he got } } strawberry. } } } } } } } } The car started. The third night he ordered vanilla. The car } } failed } } } to } } } } start. } } } } } } } } Now the engineer, being a logical man, refused to believe that this } } } } man's car was allergic to vanilla ice cream. He arranged to } } continue } } } } his visits for as long as it took to solve the problem. } } } } } } } } And toward this end he began to take notes: he jotted down all sorts } } } } } of } } } } data, time of day, type of gas used, time to drive back and forth, } } etc. } } } } } } } } In a short time, he had a clue: the man took less time to buy } } vanilla } } } } than any other flavor. Why? The answer was in the layout of the } } } store. } } } } Vanilla, being the most popular flavor, was in a separate case at } } the } } } } front of the store for quick pickup. All the other flavors were kept } } in } } } } the back of the store at a different counter where it took } } considerably } } } } longer to find the flavor and get checked out. } } } } } } } } Now the question for the engineer was why the car wouldn't start } } when } } } it } } } } took less time. Once time became the problem - - not the vanilla } } ice } } } } cream - the engineer quickly came up with the answer: vapor lock. } } } } } } } } It was happening every night, but the extra time taken to get the } } other } } } } flavors allowed the engine to cool down sufficiently to start. } } } } } } } } When the man got vanilla, the engine was still too hot for the vapor } } } } lock to dissipate. } } } } } } } } Moral of the story: even insane looking problems are sometimes real. } } } } } } } } A better moral: chocolate ice cream cures vapor lock! } } } } } } } } } } } } }
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207
Does anyone know of a list server or organization similar to MSA for on-line exchange of information to people in the Molecular Biology field? Thanks in advance for any suggestions.
Patrice Abell-Aleff Electron Microscopy Core Facility Mayo Clinic 200 1st Street SW Rochester, Mn. 55905 phone: 507-284-3148 fax: 507-284-9349 e-mail: abellaleff.patrice-at-mayo.edu
There is an opening for a TEM technician at Seagate in the Minneapolis area. Interested individuals please reply directly to "Deb", at address shown at the bottom of the position description by fax or by US mail. Please do not reply by email. Thank you.
Augusto Morrone Seagate Technology Augusto_A_Morrone-at-notes.seagate.com (612)844-5838
Position Open at Seagate Technology: TEM Technician. A TEM technician position is open at Seagate Technology, RHO, in Bloomington, MN. Applicants should have at least a 2-year technical college degree, or equivalent experience, in TEM basic operation and sample preparation techniques in the physical sciences. The main duties involve TEM sample preparation using standard grinding, polishing, dimpling and ion milling techniques, operation and maintenance of sample preparation and darkroom equipment, and may include operation of the TEM, and handling TEM data in the form of negatives and digital images. The TEM facility is installing a Philips CM200 and is projecting the purchase of a FEG TEM within a year. Seagate offers an attractive benefits package and salary commensurate with experience. Please send your resume to:
Seagate Technology Staffing Department 7801 Computer Ave South Bloomington, MN 55435 Fax: (612) 844-7008 Attn: Deb
I am going to purchase a digital camera for general lab usage. I need a digital camera that has a capacity of producing high-quality outputs from both color and B/W photos (the resolution has to be 1024 x 800). Your kind suggestions and advice on the right make and good deal in the market will be appreciated.
Thank you very much for your help.
H. You ********************************************************************* Hong You, Ph.D Dept. of Cell Biology College of Medicine University of Cincinnati Cincinnati, OH 45267-0521 Voice: (513) 558 3709 Fax: (513) 558 4454 email: youhg-at-email.uc.edu *********************************************************************
Dear colleagues, I am trying to find a relatively fast method to count synapses on rat brain sections. I have done immunohistochemistry, but no matter how thin I cut, the synapses are so numerous that it is impossible to count. In old papers they used to do densitometry, but I don't think that is accurate way of counting. I have tried on fresh- frozen or perfused brains embedded in paraffin. I also tried to plastic embed and attempted to count under LM using oil immersion objective. Also tried to do immuno on LR White embedded thin plastic sections, which didn't work very well. Everything to avoid using the EM (too much time consuming, considering that I have six groups of different animals to compare). I would very much appreciate receiving your suggestions. Thank you, Lilith
G'day all from Hot and Muggy Atlanta (at least from a Chicagoan's perspective).
Just a single announcement that this is the Microscopy &Microanalysis 98 Meeting week. I'm in Atlanta now and hopefully all will run smoothly on the Listserver.
Live video and Video Conferences are being broadcast/held from the Meeting Site, in addition a daily Newsletter of events is being posted all to the MSA WWW site.
You may login and virtually join the conference at the MSA URL of:
Dear all, As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
On 9 Jul 98 at 14:21, The slowly moving finger of Mandayam V. Parthasarathy wrote:
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } ---. } } Just a quick note to say that my experience with propane freezing } has been very similar to that of John Gilkey's. } } --------------------- } } " In my experience, commercial grade propane, such as is sold for } use in } your barbecue, propane torch and campstove, works perfectly well for } plunge freezeing or propane jet freezing. ......................" J. } Gilkey } } ******************************************************************* } } M.V. Parthasarathy } Prof. of Plant Biology, Adjunct Prof. of Anatomy (Vet), &
On a completely different note. Has any work been done to characterise the Freon replacement gases ?? I can't find any ready reference to this and would like to find a safe replacement to Freon 12 and 22. Thanks John John Findlay Science Faculty EM Facility. Edinburgh University. Daniel Rutherford Bldg. Kings Buildings. Edinburgh EH9 3JH. tel. 0131-650-5344 fax. 0131-650-6563 John.Findlay-at-ed.ac.uk
Try Muller T, Moser S, Vogt M, Daugherty C & Parasarathy MV (1993). Optimisation and application of jet freezing. Scanning Microscopy 7, 1295-1310. Using HCFC 124 (SUVA 124-CHClFCF3) with thin titanium supports, cooling rates were obtained similar to those from propane and standard copper supports. This was suggestedt (I hesitate to plug this !!) in Ryan KP (1992) Cryofixation of tissues for electron microsocpy: a review of plunge cooling methods. Scanning Microsc. 6, 715-743. See next-to-last page , p. 742 in Discussion with Rerviewers section..
} I suggest that it may be necessary for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. Ian:
Well put, Ian. I agree. The sex post was the pits.
Blystone in Texas
Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu} Professor of Biology Trinity University San Antonio, Texas 78212 210.736-7243 210.736-7229 FAX
Just use the delete key. Blocking postings from non-listers is a real drag. We have had many many many more legit postings from non-list members than from the ocational spammer. And the thing is, the spammers will co-evolve and get by whatever you try and throw at them. A lot of folks don't like to join because of the high volume of messages that are germane in general but not to them in particular. Just use your delete key. My 2 to the tenth electrons, Tobias } Dear all, } As you will have noticed, there have been two recent ads posted to the list } from non members (one for a sex site, and one for an email Marketing } program). Whilst the vendors who subscribe to this group almost always } behave in a responsible manner by sticking to the charter and not sending } overtly commercial mailings and Nestor deals with the occasional abuse, we } have no control about non list members who post onto the list. } } I am aware that some other discussion groups use software that blocks } postings from non list members. I suggest that it may be necessary for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. } } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Ian MacLaren, Tel: (44) (0) 121 414 3447 } IRC in Materials for FAX: (44) (0) 121 414 3441 } High Performance Applications, email: I.MacLaren-at-bham.ac.uk } The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ } Birmingham B15 2TT, } England. } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
I totally agree with you. I belong to several other list serv's none of which can be posted to unless you are a registered member. As an aside ... the sex site that was spammed to the list also got to several universities.......
} } As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. { {
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
I concur with Tobias. As obnoxious as they are, the best thing to do with junk mail is ignore it and throw it out. Remember the golden rule for junk mailers - never respond! Even telling them to leave you alone simply validates your mail address on their list which WILL be sold to someone else.
I think the value of this mail list far outweighs the junk mails I get because of it!
I hate when we get bogged-down in all of these "administrative" discussions, but since this could affect the list, here's my opinion:
Spamming is the e-mail version of junk-mail, and I haven't heard of any completely proven manner of removing junk mail without risking the reliability of the "real" mail. When you go through your mail at home, I'm assuming that you don't open everything mailed to you. You can guess that if it promises a million dollars, or has the mark of some credit-card company that you have never dealt with, you know its an ad, and can throw it away immediately, and not lose sleep over it.
My philosophy is: If the subject line doesn't have a topic related to my research, THEN I IMMEDIATELY DELETE IT, even if someone forgets to put a subject line. The subject line is your "filter" that only requires a couple of your own brain cells to activate. Let's face it: I don't know any e-mail software that doesn't show the subject line before opening messages, and if you're worried about spamming, I don't think I've seen any "spam" that didn't have a suspicious sender address (No professional that I know of would send and e-mail from "Lisa" with no last name!), or a suspicious or missing subject line.
This listserver is a forum for discussion, so I have responded to this topic, but let's please not get away from the SCIENTIFIC DISCUSSIONS!
Thank you for listening.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA Note: This message is my opinion and has nothing to do with my employer. } -----Original Message----- } From: Ian MacLaren [SMTP:I.MacLaren-at-BHAM.AC.UK] } Sent: Monday, July 13, 1998 5:57 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ads from non list members } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } As you will have noticed, there have been two recent ads posted to the } list } from non members (one for a sex site, and one for an email Marketing } program). Whilst the vendors who subscribe to this group almost } always } behave in a responsible manner by sticking to the charter and not } sending } overtly commercial mailings and Nestor deals with the occasional } abuse, we } have no control about non list members who post onto the list. } } I am aware that some other discussion groups use software that blocks } postings from non list members. I suggest that it may be necessary } for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. } } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Ian MacLaren, Tel: (44) (0) 121 414 3447 } IRC in Materials for FAX: (44) (0) 121 414 3441 } High Performance Applications, email: I.MacLaren-at-bham.ac.uk } The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ } Birmingham B15 2TT, } England. } ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ }
Another method that does require some practice but is faster than the jigs and glass wheels is a very simple 2 step method that can be done when time is crucial. Obtain:
1) Pencil size XACTO knife holder. 2) 600 grit SiC paper. 3) MICROCLOTH - polishing cloth from Buehler Ltd. 4) 0.05 micron deagglomerated alumina or 0.02 micron colloidal silica.
The procedure is as follows:
1) Insert the die of interest into the end of the XACTO. 2) With a polish wheel rotating ( in the CCW direction with 600 grit paper, hold the die and holder assembly at 45 degree angle and begin to slowly grind (on the right hand side of the wheel) to the area you desire to cross section. You must frequently monitor the progress with an optical microscope. 3) Once you are 10 - 15 microns away (or so) from the area of interest, stop grinding. Rinse well and dry. 4) Mix up a slurry of 0.05 micron powder and apply to a MICROCLOTH. 5) Begin polishing the die on the left hand side of the wheel at a 90 degree angle to the plane of the wheel. Again, frequent inspection of the progress is needed. The colloidal silica can be used if a very fine artifact free finish is needed, but note that the silica will NOT polish tungsten plugs. When using the colloidal silica, try to find a polish pad that is made of spongy material.....i.e. neoprene or some such, the silica works with MICROCLOTH but a sponge-like material works better. The CMP supplier RODEL has some nice 8" CMP pads that fit the bill. 6) Rinse, clean and dry the specimen and then delineate layers with the appropriate acids
Hope this helps out. We use both methods in our lab but when time is crucial and the volume is large this technique saves the day.
John Staman Consulting FA Engineer Analytical Services Lab Symbios Inc. Colorado Springs, CO. 719-573-3282 ---------- -----------------------------------------------------------------------.
We are currently trying to establish capability in our lab to look at die cross-sections. In previous work we mounted them in epoxy or acrylic and coated the samples, but I'd like to be able to do this without mounting and coating. I've been told that there are simple polishing fixtures one can by to do this, but I can't seem to find where to buy them. Perhaps someone out there could suggest a source? Thanks,
Janet Rice MCC Senior Member Technical Staaff rice-at-mcc.com 512-338-3266
I agree with Tobias that at the moment, there are not sufficient violations of the rules to prevent legitimate postings from non-members. However, this could change. The net is still in its infancy. Ciao for now, Ken
I strongly disaggree with those of you who would simply roll-over and live with junk e-mail posted to a private e-mail list. Will you still be hitting the delete key when half the postings were never invited? ... or will you be unsubscribing?? I belong to several lists and monitor one myself and can attest to this list being especially vunerable (... for some reason ...). This list's monitor really should look into different list server software.
[to Nestor: I really do understand you have a real job ... and do sympathize with some of this not being under your control ... that is, I don't get to choose the list software this university uses ... but please pass these concerns on to your lists-meister ...]
... my $0.02 :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I agree that these Spam postings are an annoyance. It would be nice if one could reply to them and overload their server. I know in fact that this is a bad idea, as it gobbles bandwidth, but it would just feel good to give them a dose of their own clutter. I know there is a web site to deal with this issue, I will find it and if there is any useful information I will post to the server. For now the delete key sounds best and fastest, if not the most emotionally satisfying.
-- Respectfully, Bob ( Robert G. ) Lawrence Failure Analyst Motorola Phoenix Corporate Research Lab 2100 E. Elliot Rd. MD EL-703 Tempe, AZ 85284-1806 Phone: 602-413-5848 Fax: 602-413-4952 Pager: 1-800-759-7243 PIN 834-2458
} [snip] } } ...I suggest that it may be necessary for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. }
It is easy to set most software to not allow non-members to post to a list. However, turning off automatic subscription and then vetting would-be subscribers is a tremendous amount of work. Since there is no vetting for subscription to this list, there is no protection from folk automatically subscribing to post an ad. I'll even bet that's how those ads got on here.
The degree of security one imposes on a list is directly related to the amount of work necessary to administer and maintain the list.
While I subscribe to a couple of lists where subscription is not automatic, they all cater to rather small groups. It's a bit much to ask Nestor to perform the vetting function. Until such time as Nestor gets extra pay to maintain the list and until such time as we decide to pay for his infrastructure, I suggest that Nestor can do whatever the hell he wants.
Dear all, As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Nestor gave a report on this very problem yesterday to MSA Council, sponsors of this List. He is well aware of the problem but the solutions are difficult. Nestor's a little busy right now here at M&M'98 in Atlanta (check out the WWW site), but will probably reply himself when he has a chance.
Ernie Hall MSA Secretary
---------- From: Ian MacLaren[SMTP:I.MacLaren-at-BHAM.AC.UK] Sent: Monday, July 13, 1998 5:57 AM To: Microscopy-at-sparc5.microscopy.com Subject: Ads from non list members
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society 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, As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
One camera that might be suitable for your requirements is the svMicro digital camera. This new camera will be available September 1st; it is a c-mount, resolution of 800x1000, 3 shot color camera, live black and white video preview & operates as a Photoshop plugin. On a MAC the svMicro is a SCSI device; on a PC, it is a parallel device. The CMOS chip is sensitive enough to image bright fluorescent subjects. The price is $2,200 as a c-mount and $2,600 with a package that includes lenses & extension tubes. For more information, please feel free to contact me at telephone 516-773-4305.
Matt Irwin ElectroImage, Inc. 277 Northern Blvd Suite 101 Great Neck, NY 11021
I am in the process of creating a web page for irusers-l, an internet listserv for scientists and conservators around the world who use infrared spectroscopy to study historic and artistic works and other cultural property.
The page will include links to commercial suppliers of FT-IR instrumentation and materials.
If lurking company reps would like to e-mail me company names, contact information, and URLs I'll try to include them in the page.
I'm realizing my PhD and I research about an halophyte, Atriplex nummularia Lindl. I work with plant anatomy and I'm looking for a stain that marks the vesicular hairs that are in this specie. Would you help me? Thanks.
i am working on the localization of antigens being involved in the adhesion/invasion process of pathogenic bacteria. Pre-embedding labelled samples are embedded in a resin which is suitable for perfoming post-embedding labelling afterwards like Lowicryls etc. I would like to know if someone has got any experience in using silver-enhancement kits for enlarging the gold-particles of the pre-embedding labelling with subsequent post-embedding labelling of the same ultrathin section.
- does the used resin has any effect on enhancing the gold-particles of the pre-embedding label
- does the silver-enhancement solution penetrates the entire ultrathin section and developes not only those gold-particles being exposed on the surface of the ultrathin section
- does silver-enhancement allows to perform postembedding labelling afterwards.
In the past, I've seen discussions on the list about the effects of vibrations from roadways, "floods" (both water and liquid nitrogen), ambient dust etc. on electron microscopes, but none have addressed lightning.
It may sound funny, but this is serious business.
It wasn't a direct hit, but it certainly "fried" a good deal of the electronics in our SEM. 2 1/2 weeks ago lightning struck the nearby power plant that supplies us with electricity, wreaking havoc all about in our industrial park. All the local production facilities managed to get back running within hours or a day of the disaster. Our SEM is *still* down, despite repeated visits of the SEM manufacturer's technical service people, who have been having trouble finding all the problems. This does not reflect well on the manufacturer...nor on us, for choosing this manufacturer.
But I'm interested in preventing similar problems in the future. Here in Germany, lightning is not all that frequent and at home I have *never* experienced a power outage due to thunderstorm action (20 years). But I think our power plant in the industrial park may be "lightning prone", since this is the second hit it's taken in the past 5-6 years. (The first time, we had no SEM.)
I'd be interested in suggestions about protecting our SEM from such hits to the power line in the future. Surely there are SEMs in more lightning-prone areas (Natal, South Africa? Southeast US?), where such protection must be routine. Can anybody give me some ideas?
Cynthia Bennett Hoechst Diafoil Germany
************** The opinions expressed here are solely my own and not the fault of my employer. **************
I think that very general "what do you people recommend" inquiries are best sent to the inquirer only and that person could then broadcast a summary. Another camera was touted here, so I wish to advise that Pixera has: 1260x940 pixel, comes complete with lens and other accessories, has new, superior software, offers most pixel/$ (even before the recent substantial price-reduction). The Pixera patent also gives better depths-of-field for it's pixel size than any other camera. A lot of technical info is provided in our online. Disclaimer: Yes, ProSciTech supplies Pixera and I would not have made this rather commercial posting had not another supplier preceded. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
From: H.You {youhg-at-email.uc.edu} To: microscopy-at-Sparc5.Microscopy.Com {microscopy-at-Sparc5.Microscopy.Com}
Lightning strikes and the resulting spikes are a problem, especially for us on the lightning-prone 'Highveld' of South Africa. Ordinary lightning protection diode and voltage dependent resistors are not necessarily fast enough to protect your EM. We use a UPS to protect EM's. You do not need a bigger capacity than one that will carry the microscope for a few minutes, so they are not too expensive. You do, however, need one that fully isolates the microscope from the mains supply. Take care to get one that outputs a clean sine waveform, even under full load - some of the UPS systems produce a pretty nasty square wave when working hard.
Jan Coetzee
} } Hi Listers, } } In the past, I've seen discussions on the list about the effects of } vibrations from roadways, "floods" (both water and liquid nitrogen), } ambient dust etc. on electron microscopes, but none have addressed } lightning. } } It may sound funny, but this is serious business. } } It wasn't a direct hit, but it certainly "fried" a good deal of the } electronics in our SEM. 2 1/2 weeks ago lightning struck the nearby } power plant that supplies us with electricity, wreaking havoc all } about in our industrial park. All the local production facilities } managed to get back running within hours or a day of the disaster. } Our SEM is *still* down, despite repeated visits of the SEM } manufacturer's technical service people, who have been having } trouble finding all the problems. This does not reflect well on the } manufacturer...nor on us, for choosing this manufacturer. } } But I'm interested in preventing similar problems in the future. } Here in Germany, lightning is not all that frequent and at home I } have *never* experienced a power outage due to thunderstorm action } (20 years). But I think our power plant in the industrial park may } be "lightning prone", since this is the second hit it's taken in the } past 5-6 years. (The first time, we had no SEM.) } } I'd be interested in suggestions about protecting our SEM from such } hits to the power line in the future. Surely there are SEMs in more } lightning-prone areas (Natal, South Africa? Southeast US?), where } such protection must be routine. Can anybody give me some ideas? } } Cynthia Bennett } Hoechst Diafoil } Germany }
Prof Jan Coetzee Head: Lab for Microscopy and Microanalysis Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-362-5150 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa http://www.up.ac.za/science/electron/emunit1.htm
Cynthia Bennett wrote ================================== In the past, I've seen discussions on the list about the effects of vibrations from roadways, "floods" (both water and liquid nitrogen), ambient dust etc. on electron microscopes, but none have addressed lightning.
It may sound funny, but this is serious business.
It wasn't a direct hit, but it certainly "fried" a good deal of the electronics in our SEM. 2 1/2 weeks ago lightning struck the nearby power plant that supplies us with electricity, wreaking havoc all about in our industrial park. All the local production facilities managed to get back running within hours or a day of the disaster. Our SEM is *still* down, despite repeated visits of the SEM manufacturer's technical service people, who have been having trouble finding all the problems. This does not reflect well on the manufacturer...nor on us, for choosing this manufacturer.
But I'm interested in preventing similar problems in the future. Here in Germany, lightning is not all that frequent and at home I have *never* experienced a power outage due to thunderstorm action (20 years). But I think our power plant in the industrial park may be "lightning prone", since this is the second hit it's taken in the past 5-6 years. (The first time, we had no SEM.)
I'd be interested in suggestions about protecting our SEM from such hits to the power line in the future. Surely there are SEMs in more lightning-prone areas (Natal, South Africa? Southeast US?), where such protection must be routine. Can anybody give me some ideas? ===============================================
I think the reason is in temporary increase of mains voltage. Methods of protection: - autonomous generator - most reliable method, - motor-generator with additional protection of the motor, - high-power BACK UPS (with constant converting of mains voltage) with additional protection of input circuits - most modern method. Regards.
Victor Sidorenko, ANTRON Co. Ltd., Moscow, Russia. antron-at-space.ru
For what it's worth, I agree with Gregg. I'm not brain dead yet (despite the long hours) and I can figure out which e-mail I want to read and which I want to delete. "For your eyes only", as a subject line, was a dead giveaway!!!
I'm sure that Nestor has enough to do without having to protect us all from a few pranksters.
} Hello all, } } Does anyone know of a list server or organization similar to MSA for } on-line exchange of information to people in the Molecular Biology field? } Thanks in advance for any suggestions. } } Patrice Abell-Aleff } Electron Microscopy Core Facility } Mayo Clinic } 200 1st Street SW } Rochester, Mn. 55905 } phone: 507-284-3148 } fax: 507-284-9349 } e-mail: abellaleff.patrice-at-mayo.edu
Nestor needn't go to the trouble to monitor subscriptions to the list or block mail from certain domains, but as the 'list owner' he should probably do his part to eliminate the spam at its source (see below), if he isn't already. Unsolicited bulk commerical email is a huge drain on the network bandwidth, disk space and other resources of network providers and SMTP relay sites. Furthermore, unlike bulk postal mail, for whch the sender pays at least part of the cost, the cost of bulk email is paid by us consumers in increased ISP charges, slow netowrks, etc. I started to receive several spams a day in my personal email about two years ago. Aside from the tangible and intangible costs, I consider spamming to be in intrusive and irresponsible use of network bandwidth, so I started sending messages to the postmaster and administrative contact (obtained from a WHOIS lookup, http://rs.internic.net/cgi-bin/whois) of the originating sites (watch for forged entries, though), their ISP, if any, and the postmaster of all relay sites listed in the message header (who can put pressure on the originating site to take action - again, watch for forged entries). By last spring, I had received notification of the termination of the accounts of several dozen of these spammers, and the rate of spamming had dropped to a trickle (1-2 per week). Some might think that by sending these messages, I was only adding to the noise, but while a single bulk mailing consists of thousands of messages, only a handful of people complain, and this is sufficient to alert postmasters to the problem and eliminate it at the source. I have in fact found that postmasters appreciate someone informing them of what is generally unwanted network traffic (for the relay sites) or a violation of the conditions of use (of a provider's services), so that they can deal with it effectively, since it is such a huge drain on their resources. Those spammers who have persisted and been brought to trial have generally been convicted on "theft of services" charges for this innapropriate use of other's equipment and abuse of priveleges. There is some remedial legislation in Congress, but unfortunately it does not go so far as to place the same sanctions on unsolicited commercial email as were placed on unsolicited commercial faxes ($500/message or cost to recipient, whichever is greater); see:
We are selling a JEOL 9000 freeze fracture unit. It needs a little work, but we never use it so we never had it repaired. We will accept the best offer we receive. You will have to pay for the shipping. If you're interested in this lovely putty & orange colored machine, please contact me. I promise that we will clean all of our junk off of it [we've been using it as a workbench ;)].
Looking forward to seeing the floor underneath it,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207
In Tampa Bay, (FL, USA), touted as the lightening capital of the world, = lightening protection is a must. The local TV stations enjoy giving out = frivolous statistics such as the number of surface-to-cloud strikes per = hour. For example, night before last, there were more than 9000 strikes = in a one hour period. Hence, the potential of lightening damage is taken = seriously and has spawned an entire industry of consultants, manufacturers = and installers. The first line of defense seems to be the protection of = the building (every corner, projection etc. has a 12-18" lightening rod = and these are all connected), and there is massive protection on the main = power lines and communication lines. In fact, our computer network lines = are fiber even between floors of the main Hospital here. Most of this = falls under the responsibility of our main Hospital engineer and costs = many tens of thousands of dollars to install and maintain.
Although this is adequate for many applications, sensitive equipment = requires much more and that is even more expensive. I teamed up with the = rest of the Laboratory, and had installed a very large battery powered, = online UPS (takes up a whole room). From this we can run all essential = equipment for about ten minutes in the event of an outage, time enough for = the generators to kick-in. This device also smooths out generator noise, = and brown outs, (low voltage). In extremely simplistic terms, the = incoming current is disassembled on the incoming side, and reconstituted = on the outgoing side. The batteries not only act as a reserve and buffer, = but also a filter. All grounds are on this side of the UPS to prevent = ground loops, et cetera. At the time (about five years ago), this was the = most cost effective way to cover a multitude of bases for a large clinical = laboratory. =20
Intermediate-sized equipment, not in proximity to the large UPS, is on = individual battery powered online UPS. There is a wide range of sizes = available from vendors (for example, APC) for a wide range of dollars. = This is what I would expect is best for a single instrument such as a SEM. = Pay attention to wave shape and ratio for computerized equipment (vendors = can advise). If you cannot locate any local outlets, your network gurus = may be able to point you to their sources. Most network servers have such = equipment installed, albeit smaller than you probably require. The same = vendors usually sell the larger models as well (5 to 35 KW).
Our desktop computers are NOT on these devices. Instead, they are on = individual surge protectors only (for example, Tripplite surge protectors) = with no battery backups. One aspect about computers is that the majority = of strikes come across phone lines so a surge protector with phone line = protection is a must! That is, unless your institution has a switchboard, = which is already protected. At home, I experience at least one or two = such strikes a year that fries the modem protection on my computers. =20
A note of caution, do not place the inexpensive types of surge protectors = in serial, as it is possible to create destructive harmonics between the = devices.
Of course, like insurance, you must gauge the investment and risk with the = cost of protection.
Regards,
Peter O. Steele, Ph.D., PMIAC Dir., Special Anatomic Pathology Unit Pathology & Laboratory Medicine All Children's Hospital Saint Petersburg, Florida 33731-8920 v-mail: 813/892-4465 e-mail: steelep-at-allkids.org
We are thinking of putting uninteruptable power supplies (UPS) on our electron microscopes (Philips & Zeiss TEM's).
Any suggestions or comments?
Thanks - Dennis
------------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: winkler-at-calvin.niams.nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
I am trying to use a quantitative analysis program to analyze EDS spectra collected with TEM. I have 2 questions: (1) I have oxide inclusions in a diamond matrix . I think there are absorption problems (Carbon absorbing oxygen). Is there anyway of getting around it? I am using the "thin-section analysis" as an action. Should I use " bulk sample analysis"?
(2) I am using Nickel grid. So, the spectrum has Ni as an element in it. How do I calculate the formula of the mineral? Do I perform the quantitative analysis without the Ni? Or do I normalize the other elements without taking into account that the Ni is present? Is there anyway of determining if I do have Ni in my sample (or finding out proportions of Ni coming from the grid and from the sample?)
} We are thinking of putting uninteruptable power supplies (UPS) on our
} electron microscopes (Philips & Zeiss TEM's).
}
} Any suggestions or comments?
}
Dennis,
We have the Ferrups unit from Best Power on several instruments. There are two things that I would keep in mind:
1. make sure that you have the capability to handle the power surges that come from motors switching on and off (ie, pumps and compressors). For Best Power, this is the DVR option.
2. make sure that your physical plant people RTFM when installing the unit. For example, Best requires grounding cable that is *at least* the same gauge as the hot and common wiring. This requirement is more stringent than the typical electrical code.
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 7 - October 15, 1998
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2050 (Includes room and board)
Application Deadline: August 4, 1998
Admission application and information: Carol Harnel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratio-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual research problems with the faculty.
} We are thinking of putting uninteruptable power supplies (UPS) on our } electron microscopes (Philips & Zeiss TEM's). } } Any suggestions or comments?
Here's how it works in our lab:
The SEM and TEM are both running on a 220 V AC mains separated from the "ordinary". The separation is done by a 220 V AC generator driven by a 220 V AC electric motor. It seems to be a dull solution but this way we completely eliminate all electric surge and unwanted noise from the mains and besides the inertia of the two rotating iron cores is enough to overcome about 1/10 sec dropouts of the "ordinary" mains supply. We really do not need a UPS (taking into consideration the high power of TEMs and SEMs the UPS should be dimensioned quite big and expensive). During the past more than twenty years we had about two or three dropuouts long enough to shut down the microscopes. And the generator/motor system runs without problems, almost no maintenance, and cheap, even if you take into account the efficiency loss of the system. We even operate some sensitive computer servers and other instruments on this same local and very "clean" mains.
Kris
PS: The above solution also provides extremely effective protection against lightnings, see posting of Peter Steele ---------------------------------------------------------------------------- --------- Kristof Kovacs, Ph.D. Head, Central Laboratory University of Veszprem, P.O.Box 158, Veszprem H-8201 Hungary Phone: +36-(88)-421-684 Fax: +36-(88)-328-643
Hello, Do any of you know where I might obtain a copy of the book "Cell and Tissue Ultastructure- A Functional Perspective" by Patricia C. Cross and K. Lynne Mercer. W.H. Freeman and Co. 1993? It is out of print and I have checked the obvious resources. This is an excellent resource which I would very much like to add to my library.
I just joined the list. I love the infinite complex world. I really want a microscope and am interested in building one if it's cheaper. I've been researching home made telescopes too but cant find any information on making microscopes. Could someone help me? A powerful microscope would be great, like 1000x or larger, if possible (smaller's ok, I'd like at least 100x), but I'm assuming its similar to telescopes in that you can make pretty much any size given your time, money, and experience level. Perhaps someone knows of urls or books relevant?
thanks,
danny
ps- has anyone heard of Ernst Haeckel? He was a biologist who drew images of small complex life forms, here are a few examples of his art... I would love to be able to see these under a microscope
{A HREF="http://www.comptime.com/hoti/Haeckelview.html"} Haeckelview.html at www.comptime.com {/A}
In a message dated 7/15/98 12:28:50 AM US Eastern Standard Time, CALYK-at-aol.com writes:
} Microscopy-at-sparc5.microscopy.com
Hi there, when I was a younger person I used a book called wonders through the microscope to build a projection microscope. Gee, they even had plans in there to build your own arc lamp to use with it........a wonder we didn't burn the house down....
Anyway Check your local library there still may be a copy of this old tome there.
If all else fails maybe we even have an extra copy of it in the library here that we could trade off. The authors were the editors of popular science monthly. Edward Sharpe, Archivist SMECC
An NIH Director's Wednesday Afternoon Lecture Series Event Sponsor: NIH Inter-Institute Mitochondria Interest Group (MIG)
A Day-long Minisymposium=20 Mitochondria: Genetics, Health, and Disease 2 December 1998 Featuring The Wednesday Afternoon Lecture by Dr. Eric A. Schon (Columbia) Molecular Genetics of Human Mitochondrial Disease
Jack Masur Auditorium, Clinical Center, NIH For special accommodation needs call 301-594-5595 CME credit awarded=20 MITOCHONDRIA: GENETICS, HEALTH, AND DISEASE MINISYMPOSIUM 2 DECEMBER 1998
Lectures Venue: Masur Auditorium, Clinical Center, NIH Poster/Exhibitor Venue: Visitor Information Center, Clinical Center, NIH
* 0745 Registration, Poster Set-up, Continental Breakfast in Exhibit = Area * 0830 Dr. David A. Clayton (HHMI): Mitochondrial DNA Control Features * 0905 Dr. William C. Copeland (NIEHS): Avoidance of Mitochondrial DNA = Mutations by DNA Polymerase Gamma * 0940 Dr. Vilhelm A. Bohr (NIA): Oxidative DNA Damage Repair in = Mammalian Mitochondria * 1015 Poster Session/Coffee Break in Product Exhibit Area * 1045 Dr. Tracey Rouault (NICHD): Abnormalities of Mitochondrial Iron = Metabolism and Human Disease * 1120 Dr. Steven J. Zullo (NIMH): In situ Localization of the common = Human 4977bp Mitochondrial DNA Deletion Mutation * 1200 Lunch Break * 1330 Dr. Mariana Gerschenson (NCI): Mitochondrial Genotoxic and = Functional Consequences of Chemotherapeutic Drugs * 1400 Poster Session/Coffee Break in Product Exhibit Area * 1500 Wednesday Afternoon Lecture: Dr. Eric A. Schon = (Columbia):Molecular Genetics of Human Mitochondrial Disease * 1600 Reception/Poster Session in Product Exhibit Area * 1700 Poster Session/Product Exhibition Closes
Continental Breakfast, Coffee Breaks, Lunch Break, and Reception = sponsored by the Technical Sales Association Attendance/Poster Registration Form Mitochondria: Genetics, Health, and Disease Wednesday Afternoon Lecture Series Minisymposium 2 December 1998 Masur Auditorium and Visitor Information Center Clinical Center (Building 10), NIH Bethesda, MD
Note: There is no registration fee to attend this minisymposium! Name: Title: Affiliation: Address: State: Country: Postal Code: Telephone: Fax: E-mail: Web Page URL: I will attend and present a poster I will attend but not = present a poster________ Poster Title: Abstract (Abstract Booklet available at the meeting):
Do you need special accommodations while at NIH?______________ For a map of NIH and area, please check the following Web Site: = http://www.nih.gov/welcome/maps.html WARNING: Parking is limited on = campus, plan to use METRO! A block of rooms at a special meeting rate has been reserved at The = Bethesda Ramada. Call 800-272-6232, or 301-654-2703, before 9 November = 1998. Mention NIH Minisymposium, group # 6210. For other = accommodations in the area, please check the following Web Site: = http://www.patsys.com/ftd/city.cgi?pcityid=3D2521&pcity=3DBethesda Submit advanced registration via Minisymposium web site at = http://www-lecb.ncifcrf.gov/~zullo/migDB/symposium.html or via e-mail to: zullo-at-helix.nih.gov deadline by e-mail is 8 November = 1998=20 or via regular mail to: Steven J. Zullo, PhD postmark deadline is 2 = November 1998 Building 10, Room 2D54 NIH Bethesda, MD 20892
Steven J. Zullo, PhD Laboratory of Biochemical Genetics NIMH-NIH; Bldg. 10, Rm. 2D56; 9000 Rockville Pike Bethesda, MD 20892 301-435-3576; FAX 301-480-9862 zullo-at-helix.nih.gov Mitochondria Interest Group Web Page: = http://www-lecb.ncifcrf.gov/~zullo/migDB/
I was wondering if anyone has experience with the new Leica CPC unit. We are using this for freezing thin films to get virtified ice on holey carbon grids. We recentlly purchased one of these units and I would be extremely grateful for any tips and advice from someone with experience of using the unit for this application. How do you avoid ice contamination?
Many thanks in advance,
Dr Timothy F. Booth Canadian Food Inspection Agency National Centre For Foreign Animal Disease Suite T2300 1015 Arlington St. Winnipeg Manitoba R3E 3M4 CANADA http://www.hc-sc.gc.ca/main/lcdc/web/bmb/fedlab_e.html#toc email tbooth-at-em.agr.ca Tel 204 789 2022 Fax 204 789 2038
There are a number of sources of plans for home made microscopes; however, I really don't think you will save any money over the very low cost microscopes currently imported from China.
Here is the design I consider the most realistic for construction using only hand tools: Curry, Alan; Grayson, Robin F.; and Hosey, Geoffrey R. "Under the Microscope"; Van Nostrand Reinhold, New York, 1982; pp 31-40. (This design is/was featured on the Boston Museum of Science Web page)
I have a few ideas on how to simplify this even further, contact me if you are interested.
What is the current, generally accepted way of disposing of osmium tetroxide?
I use 1% osmium tetroxide in sodium cacodylate buffer. After removing this solution from my tissue, I pour it into a hazardous waste container that is 2/3 filled with corn oil to bind the osmium solution. We've done this for years, but our new hazardous waste/disposal monitor needs to have an official source with exact quantities of oil to osmium tetroxide ratios.
Can you help me? If there is safer, more acceptable method of disposal, your suggestions would be greatly appreciated.
Thank you in advance for you help and guidance.
Lucy Stribling Air Force Research Lab Brooks AFB Texas
I am considering the purchase of an evaporative carbon coater for use on coal clinker specimens that have a history of bad charging under the SEM. The evaporative carbon coaters (carbon rod source) come in two types: high vacuum (10^-5 torr) and low vacuum (10^-2 torr). The high vacuum coaters (with a turbo pump) are significantly more expensive. Are they worth the extra cost?
I'm told that www.abebooks.com is an excellent web site for out-of-print and hard-to-find books in the sciences and other areas. Haven't yet tried it personally, but a friend has had good success.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
We maintain a JEOL 2010 FEG-TEM for which a UPS is regarded by the manufacturer as essential. We use a Phase One Series 700 12.5 KVA unit that was suggested to us by JEOL. It works great and has plenty of capacity (right now it is running the scope and some accessories and only outputing 50% of its full capacity). It and the microscope have been in use for 5 months and with all the power outages due to thunderstorms here in Houston I don't know how we could live without it. If I could I would get one for our other microscope and every other piece of major equipment in our facility. So my thoughts these days are: UPS, Gotta' have it!
Disclaimer: I have no financial interest in any of the abovementioned companies
TTFN
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Roy Christoffersen Materials Science Research and Engineering Center University of Houston 3201 Cullen Houston, TX 77204-5932 roy-at-bayou.uh.edu (713) 743-8273 FAX: (713) 743-2787
I'm told that www.abebooks.com is an excellent web site for out-of-print and hard-to-find books in the sciences and other areas. Haven't yet tried it personally, but a friend has had good success.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I'm told that www.abebooks.com is an excellent web site for out-of-print and hard-to-find books in the sciences and other areas. Haven't yet tried it personally, but a friend has had good success.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Dear Barna, The science of EDS analysis on a TEM is a less well-characterized than on an SEM. I think that the results are semi-quantitative at best and it is difficult to know the exact conditions of film thickness and other sample characteristics for good analysis. Good quantitative analysis will require some known standards with a composition close to your unknowns. However, in answer to your questions: (1) Carbon will not appreciably absorb oxygen in your oxides. X-rays are absorbed by elements heavier than the emitting element. The absorbtion correction for oxygen in a carbon matrix, particularly in a thin film, will be very small. Use the "thin-film" analysis mode. (2) When you calculate the formula of your mineral, just leave the element of your grid out of the element list. The only way I know to tell if you have Ni in your sample is to use another grid. Copper, nylon, berylium, molybdenum, etc. are all available. Or, if your diamond matrix is strong enough, use a large-mesh grid, slot or single-hole grid so you can do measurements a long way from the grid material and see if your Ni response drops down to a minimal signal. You wrote: } } } Hi everyone, } } I am trying to use a quantitative analysis program to analyze EDS spectra } collected with TEM. } I have 2 questions: } (1) I have oxide inclusions in a diamond matrix . I think there are } absorption problems (Carbon absorbing oxygen). Is there anyway of getting } around it? I am using the "thin-section analysis" as an action. Should I } use " bulk sample analysis"? } } (2) I am using Nickel grid. So, the spectrum has Ni as an element in it. } How do I calculate the formula of the mineral? Do I perform the } quantitative analysis without the Ni? Or do I normalize the other elements } without taking into account that the Ni is present? } Is there anyway of determining if I do have Ni in my sample (or finding } out proportions of Ni coming from the grid and from the sample?) } } Thanks very much for your time and help, } } Please reply to: barna-at-geo.princeton.edu } } } } Barna } Best of luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
... i am not quite sure, but I think that there has been a discussion about different dye-sub printers on this list-server... sorry for coming back to the same point, but here is my problem: we are planning to buy a high-end "photorealistic" printer. For certain reasons we tend to a Codonics ... Could anybody explain to me, what the advantage of the additionally implemented "direct thermal" technology (-} Codonics NP1660) is? The main task for our printer would B&W prints. The maximum percentage of colour prints is estimated to be in the region of 10 to 20%; pages/year appr.: 750..1000 ). It would be also very interesting to to have a comparison about the costs per page (Codonics NP1600 versus NP1660) including paper and ribbons, but excluding the investing costs for the printer itself.
Thank You very much in advance! Dipl.Ing.(FH) Gunnar Glasser, Elektronenmikroskopie
The coater type of choice for your application is the LOW vacuum type. For
some applications, low vacuum coater films are not as "good" as high vacuum types. In your case, however, the lower vacuum means more scattering. With increased carbon scatter, the coating is less "line-of-sight" and will better coat rough, convoluted surfaces.
I would also strongly suggest you investigate low vacuum coaters which use a
carbon yarn filament rather than the carbon rod type.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am considering the purchase of an evaporative carbon coater for use on coal clinker specimens that have a history of bad charging under the SEM. The evaporative carbon coaters (carbon rod source) come in two types: high vacuum (10^-5 torr) and low vacuum (10^-2 torr). The high vacuum coaters (with a turbo pump) are significantly more expensive. Are they worth the extra cost?
by imo24.mx.aol.com (IMOv14_b1.1) id NOCPa22587; Wed, 15 Jul 1998 13:45:13 -0400 (EDT) Message-ID: {7b79c6b5.35aceaab-at-aol.com}
If someone knows a contact at popular science mag. perhaps we can get permission to have a file of the projection microscope that plans that are downloadable on the MSA site?! We have the orig. book here that we could scan but hate to get our hands slapped by a publisher! I will leave this idea in the hands of some able diplomat that would like to contact popular science or who ever owns them now!
Sorry for the several postings about the books---"operator error" is to blame. A newly-learned fact: when a message is returned because of an improper primary address, the "cc:" address still goes out just fine. oops....
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Recently there was a message sent out about the Woods Hole microscopy course for the fall. In error I deleted it and would like to have a copy of it. Anyone have it to forward to me?
Hi, can anyone there help me to put the scale bar onto a confocal image? I saved confocal image with scale bar (with overlay option), but once I open the image with any other imaging program, such as PhotoShop, the scale bar is not on the image any more. I wonder what I did wrong and how can I save the confocal file with scale bar as well as make the scale bar occurs when other imaging softwares other than LSM operating software are used. Your help will be greatly appreciated. Thank you.
I have just acquired two ADEM1 SEMs and am now in a quandary as to what to do with them. It is not economical to get them running and sell them as the upkeep costs for a contract is about $30k per year. I could donate them to a school but I have already donated two Hitachi SEMs in the last two years.
They are worth more in parts than whole but I do not want to dismantle this "work of art". (besides neither the specimen or gun chamber makes an attractive flowerpot).
Both have windowless EDS detectors and a backscatter detector. One has an optical microscope and a WDS system.
Dear Fellow MSA Listserver Members: I noticed the intense debate via email of the recent postings of "junk email" or "spam" on this List. Recently a report from a study sponsored by the Federal Trade Commission (FTC) concludes that there is basically no easy solution to this problem, although the FTC is working to solve it. The Consumer Protection Bureau of the FTC has asked that junk emails be forwarded to them at: uce-at-ftc.gov In the last year, they have prosecuted 5 businesses and warned 1000 more about spamming. This is probably the most effective way to handle the spam problem without one or only a few persons of this list saturating their time to fight against it. See also: http://www.ftc.gov/opa/9807/dozen.htm and http://www.ftc.gov/opa/9807/jbspam.713.htm for more info.
Hi there, are you in Arizona? Heck I would give them a home, besides if you have 2 of something that just means you have spare parts to keep things going with! Of course having electronics background does help.... I have an amr1000 I have yet to get opp. but part of that is my own lack of follow through. Since I have one of them It would be nice to have 2 of them! Therefore, that fact you have 2 is a great asset to the person that wishes to do own maint.
Coaters with only a mechanical pump do not give a satisfactory coating for studying morphology at reasonably high magnifications in SEM and TEM. They are however, quiet satisfactory for EDS or WDS analyses. It's not a matter of money, but most applications simply require a diffusion or turbo pump for carbon coating. Also, sputtering of carbon is unsatisfactory (VERY SLOW) and a carbon sputter head is no solution to carbon coating at all. Rotating the specimen during evaporation is effective for dealing with "difficult to coat" specimens. Disclaimer: ProSciTech distributes EMITECH equipment in SE Australasia only. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
-----Original Message-----
To all you printer users
Why do people prefer the Codonics printer when the Kodak seems to be 3/4 of the price and uses the same engine? I see more comments on the list about the Codonics than the Kodak. Is it just to have a networked printer or is the software better? Our need is primarily TEM (monochrome) but with maybe 10% colour, any comments from users with experience of either (or both) of these would be welcomed.
Thanks Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
I think I know the answer to this one, but does anybody out there have a = need / interest in the manuals described above? I am about to discard = them, but I'm finding hard to let go....
Going, going... gone?
James Wesley-Smith Electron Microscope Unit University of Natal, Durban South Africa
Which scope? If you have a Zeiss 410, write back and I'll pass along the secret :)
Tamara Howard CSHL
On Thu, 16 Jul 1998, Ping Li wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, can anyone there help me to put the scale bar onto a confocal image? } I saved } confocal image with scale bar (with overlay option), but once I open the } image } with any other imaging program, such as PhotoShop, the scale bar is not } on the } image any more. I wonder what I did wrong and how can I save the } confocal file } with scale bar as well as make the scale bar occurs when other imaging } softwares } other than LSM operating software are used. Your help will be greatly } appreciated. Thank you. } } Ping } pli-at-is.dal.ca } } } }
Two ways of doing this without burning the overlay into the image. 1. Perhaps the Zeiss software tells you the size of the field or the size of each pixel. Let's say each pixel is 0.285um and you want a 10um scale bar. Just divide to get the length in pixels to pop in with Photoshop. 2. Take an image of a micrometer slide.
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://www.ca.aecom.yu.edu/aif/ --------------------------------------------
I have been trying to contact RMC at two different # and get a busy signal = for both. Can anyone give me their phone #, or e-mail address or any = explanation of what is going on? Thanks. Donna Wagahoff SIU School of Medicine Springfield,Il. 62794-1220 217-782-0898
Dr. Milos Kalab, a food scientist retired three years ago, has established a web site. Please visit following site if your are interested
http://www.cyberus.ca/~scimat/
He is still working hard in his laboratory.
Ann Fook Yang EM Unit Eastern Cereal and Oilseed Research Centre Agriculture and Agri-Food Canada 960 Carling Ave Central Experimental Farm Ottawa, Ontario Canada K1A 0C6
RMC 3450 So. Broadmont, Suite 100 Tucson, AZ 85713 Tel. (520) 903-9366 Fax (520) 903-0132 http://www.rmc-scientific.com/microtomes
Good luck, hope this helps!
Bob ***************************************** Robert (Bob) Chiovetti rchiovetti-at-aol.com E. Licht Company / 1-800-865-4248 Colorado/Utah/Arizona/Wyoming/ New Mexico/West Texas Representing Leica Since 1967 *****************************************
Two year ago we closed our EM Lab and I guess it is time I unsubscribe from this new group. (If some one would tell me how to.) Before I go I would like to share an essay I wrote just before we sold the EM and closed the lab.
To Whom It May Concern:
It will concern few, almost none, I suppose. Things change, seasons change. What were once considered major advances in the history of the human species, are regarded as important only to the past. I sit in a room which is cool, and dimly lit. There is a constant hum, so natural, that like breathing, it goes unnoticed. Today I hear this sound and try to embed it firmly in my memory. It is the sound of a voice I will not hear again. I focus on the sound, and struggle to imagine the sound of silence. All I can hear is the rhythm of the pump and the buzz of electronic. Most people see it as a huge piece of scientific equipment, just a Transmission Electron Microscope. It is not; it is a place. No No No. It is a magic carpet. It has taken me into the essence of life itself. I have crossed a cell membrane with more ease than sodium ions. I have followed an axon for microns, and watched as its microtubules dance in and out of view. I have observed actin and myosin locked in each other's embrace. I have beheld mitochondria in the thousands and found no two exactly the same. I have watched microvilli wave in rhythm like fields of wheat. I have looked at life (or rather the shadow of life) magnified 50,000 even 100,000 times. People have told me that this technology is obsolete, or they say that it is no longer cost effective for a small research department to support this type facility. Maybe it is their fault. Maybe it is the manufacturers fault, because they made service contracts too high. Maybe it is the government's fault, because it made money so tight. Maybe it is us for we have failed to see the direction science was turning and missed our chance to create the necessary techniques that would make this technology part of the wave of the future. Maybe it is my fault, because I did not show everyone all the things I have seen and places I have been. Astronauts have said that once you walk on the moon you no longer look at it the same way. NASA no longer goes to the moon and although it is still explores space, it seems to only be passing time. Like NASA, science will continue its exploration. Yes, just like NASA. I've not walked on the moon, but I have strolled where few people have tread. For those who have made my journeys possible I send my gratitude. It was definitely an "E" ride.
Thanks to those whom it did concern. To Mike, To Susan, To Ann.
Larry Hawkey, 1996 On the closing of the department's EM Lab.
Larry Hawkey hawkey-at-neuro.duke.edu Department of Neurobiology Duke University Box 3209 DUMC Durham, NC 27710 (919) 681-6425 fax (919)684-4431 http://www.duke.edu/~lah1
It is indeed a sad day when the accountants take over.
Managements general economic philosophy of restructuring, reorganising, reengineering, and retrenching unfortunately has no place in science and technology. Accountants (+snr man.) are reluctant to provide funding to reequip, repair, revitalise or replace.
LEO has a Cambridge S90 SEM for disposal (replaced by a new LEO 435). The SEM is at US Synthetics in Orem Utah. It was in working order when recently decommissioned, and has a BSD. Condition as seen, buyer collects, Any offers to Bill Neill billneill-at-csi.com please.
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
---------- Forwarded message ----------
Dear All,
We would like to purchase a rotatable (360 degree) X-Y stage for our Zeiss Axiovert 35M. Stages that fit the Axiovert 10 also fit this microscope. Even better yet, we would like to purchase just the X-Y part of this stage (Zeiss part number 45 35 60). If anyone has a used stage (or a new one), and would be interested in selling it, please contact me at Schwartz-at-rsbs.anu.edu.au
Thank you very much.
Sincerely,
Owen M. Schwartz
Owen M. Schwartz PhD Research School of Biological Sciences The Australian National University GPO Box 475 Canberra, ACT 2601 Australia
We are in the market for a new knifemaker. Those of you with newer model = knifemakers, please let me know how you rate their performance. Does anyone other that Leica make knifemakers? Thanks.
Donna Wagahoff SIU School of Medicine Springfield, Il. 62794-1220 217-782-0898 fax 217-524-3227
I've got a couple of users who would like to pursue Pre-embedment labling of plant tissue (trials with older knives and glass show excellent results) but are very concerned about potential damage to diamond knife edges. So I'm collecting opinions from experienced pre-embedment microtomists: are biological diamond knife edges basically 'safe' from 10-20 nm gold? Such that future biological sectioning will not suffer from any damage to the knife edge?
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
Try amazon.com on the web. They just found an out-of-print ultrastruct= ure book for me. It took them about 6 months, but they finally came up wit= h a slightly used copy. =
We are putting together a proposal to purchase a inverted microscope system for imaging of live cells. Here are a few general questions that I would love some feed back on:
1. Is Multi-Photon Imaging only available thru BioRad? We don't have the expertise to build one and can't afford the BioRad system.
2. If given the choice between a filter wheel or a monochromator for the excitation wavelengths, what is the better choice? I am concerned about the amount of light you can get through a monochromator. I figure I can always reduce the amoant of light from the filter wheel.
3. Is mercury vapor the best choice for the light source?
4. If you have an environmental chamber, is it also neccessary to have an objective heater or stage heater in order to have versatility in the kind of samples you can image?
5. Have you found that in some circumstances an upright system with a water immersion objective is more useful?
Opinions concerning any of these questions would be greatly appreciated. Thank you.
Bob Underwood Derm Imaging Center U of Washington Box 356524 Seattle WA. 98195-6524
Suggestions for adjustable height chairs (or modifications thereof) that minimize or eliminate fiber shedding and static charge would be appreciated. Thank you.
I am requesting this information for a colleague who is interested in automatic processors for paper and film. Has anyone used the Mohrpro 8 processor and what is their experience with it? You may reply to my email address directly if you wish.
Regarding the Codonics or Kodak printers....They are both built on the Kodak engine and look virtually identical. They also use the Kodak supplies for Dye-Sublimation printing and should give similar output. The difference is indeed in the software. The Codonics has completely rewritten the software and added many unique features which makes the printer much more versatile. I have found that the option to send multiple files to the printer and have them scaled and printed on one page a very desirable feature. That way I can send 4 SEM files at a time and have them printed as 4x5 with the cost of only 1 sheet of paper. That gives quick high quality copies similar to what we get with polaroid but cost is much less with digital capture and dye-sub printing (~$.50 per 4x5). The cost of the 1600 is not very different from the Kodak printer.
The Codonic NP-1660 has the added feature of being able to print on large pieces of sheet film and is very useful for medical imaging. It also will be able to produce high quality thermal prints at a projected cost of about $.50 per 8x10. There have been production problems with the thermal paper but my understanding (having just returned from the MSA meetings where I talked to Codonics representatives) is that they hope to have the media available within 2-3 months. Use of this would drop the cost of 4x5 study prints to a very low $.13/print and is what sold me on this printer. This is a UNIX printer that can be easily networked and image files can be sent postscript, via telnet or via the WWW. There are a huge range of compatable file types.
I have had some techinical problems with our Codonics printer but have had very good service....either another printer has been shipped within 24hrs in the case of hardware problems, or they diagnose and suggest fixes for software problems over the phone. I have been willing to cut them a bit of extra slack over the delay in delivery of the thermal paper because they have been very responsive in many other ways.
Since Kodak may have added features I am not aware of....do carefully check the spec's on both so you can choose the best for your purpose.
Debby Sherman ========================== Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Hi there,
... i am not quite sure, but I think that there has been a discussion about different dye-sub printers on this list-server... sorry for coming back to the same point, but here is my problem: we are planning to buy a high-end "photorealistic" printer. For certain reasons we tend to a Codonics ... Could anybody explain to me, what the advantage of the additionally implemented "direct thermal" technology (-} Codonics NP1660) is? The main task for our printer would B&W prints. The maximum percentage of colour prints is estimated to be in the region of 10 to 20%; pages/year appr.: 750..1000 ). It would be also very interesting to to have a comparison about the costs per page (Codonics NP1600 versus NP1660) including paper and ribbons, but excluding the investing costs for the printer itself.
Thank You very much in advance! Dipl.Ing.(FH) Gunnar Glasser, Elektronenmikroskopie
I don't know about shedding, but I have used a spray can of "Static Guard" or some such product. I think it was made by 3M. It worked very well during our winters when the inside humidity hit bottom. Of course our summers are so humid that we don't have static problems now.
At 11:34 AM 7/17/98 -0400, you wrote: } Suggestions for adjustable height chairs (or modifications thereof) that } minimize or eliminate fiber shedding and static charge would be } appreciated. Thank you. } } James Martin
I am neither a Kodak or Codonics user, but understand the following.=20
The Kodak undoubtedly has some processor built into it to handle the printing functions, and it is apparently adequate for many functions.=20
The Codonics incorporates a Sun Sparcstation as the smarts for their printer. That is apparently what gives it more flexibility, processing (not printing) speed, and network connectivity. Of course it comes at a cost. I do not know what the differences between the two Codonics models are.=20
--------- At 08:28 AM 7/16/98 +0100, you wrote: } Why do people prefer the Codonics printer when the Kodak seems to } be 3/4 of the price and uses the same engine? I see more comments on the=20 } list about the Codonics than the Kodak. Is it just to have a } networked printer or is the software better? Our need is primarily TEM } (monochrome) but with maybe 10% colour, any comments from users with } experience of either (or both) of these would be welcomed. } } Thanks } Ron
---------- and someone else wrote:
Hi there,
... i am not quite sure, but I think that there has been a discussion=20 about different dye-sub printers on this list-server... sorry for=20 coming back to the same point, but here is my problem: we are planning to buy a high-end "photorealistic" printer. For=20 certain reasons we tend to a Codonics ...=20 Could anybody explain to me, what the advantage of the additionally=20 implemented "direct thermal" technology (-} Codonics NP1660) is? The main task for our printer would B&W prints. The maximum=20 percentage of colour prints is estimated to be in the region of 10 to=20 20%; pages/year appr.: 750..1000 ). It would be also very interesting to to have a comparison about the=20 costs per page (Codonics NP1600 versus NP1660) including paper=20 and ribbons, but excluding the investing costs for the printer itself.=20
Thank You very much in advance! Dipl.Ing.(FH) Gunnar Glasser, Elektronenmikroskopie
MME offers customized on-site courses in all areas of microscopy. We have several consultants who work in this area. If we can be of help either visit our website at { {http://MME-Microscopy.com/education} or give me a call here in the office.
FEI Company currently has openings for experienced service engineers on SEM, TEM or FIB systems. Current areas of opportunity include Oregon, Idaho, California, Georgia, Virginia, and Texas. However, we are always looking for experienced engineers in any area.
If you would like to apply or refer an applicant, please contact me directly.
~~~~~~~~~~~~~~~~~~~~~~~~ Sarah A. Boydstun-Brown Sr. Human Resources Generalist FEI Company Phone (503)640-7556 Fax (503)726-2754 www.feic.com ~~~~~~~~~~~~~~~~~~~~~~~~
Dear Barna, } } I am trying to use a quantitative analysis program to analyze EDS spectra } collected with TEM. } I have 2 questions:
Several sessions of the recent MSA-MAS meeting were devoted to just these sorts of questions, and the proceedings are available on-line. In addition to any replies you receive, I suggest you look up those papers which appear relevant to you and contact the authors. This is just the reason why I make every effort to attend the meetings as often as I can; all the experts are within reach.
} (1) I have oxide inclusions in a diamond matrix . I think there are } absorption problems (Carbon absorbing oxygen). Is there anyway of getting } around it? I am using the "thin-section analysis" as an action. Should I } use " bulk sample analysis"? } The only ways "around it" are either to use appropriate standards (which contain a carbon matrix and a known amount of oxygen) or to use a correction to the oxygen intensity which accurately accounts for the absorp- tion. As was repeated several times at the meetings, this correction is usually the one with the greatest uncertainty. If your experimental con- ditions are such that the beam penetrates the specimen with little loss in energy and with little spread, then thin-section analysis is correct, and bulk sample analysis, which includes calculations for the production of x-rays by electrons which have deviated greatly from the incident dir- ection and lost a large fraction of their energy (among other effects), will give the wrong answer. The right procedure to use will always depend on the experimental conditions, and one must understand how the analysis soft- ware relates to those conditions in order to select the appropriate form of analysis.
} (2) I am using Nickel grid. So, the spectrum has Ni as an element in it. } How do I calculate the formula of the mineral? Do I perform the } quantitative analysis without the Ni? Or do I normalize the other elements } without taking into account that the Ni is present? } Is there anyway of determining if I do have Ni in my sample (or finding } out proportions of Ni coming from the grid and from the sample?) } If you were sure that your specimen did not contain nickel, you could subtract the nickel peak along with the continuum background and calculate the formula from what's left. You could put your sample on a different kind of grid and reanalyse it in order to determine whether it contains nickel--if a qualitative analysis shows no nickel, then all the signal is from the grid, and you can safely subtract it. You can also look at an area of the original grid which has the diamond matrix with no inclusions to see if the nickel peak is significantly reduced. In this case, be sure to check the continuum-to-nickel ratio. What you are trying to determine is whether the reduction in nickel signal is due to your moving to an area which contains less (zero) nickel, or whether the reduction is due to fewer electrons being scattered onto the grid bars (or, for that matter, pole pieces or any other nickel-containing part of your instrument) and producing a signal. This can become quite complicated if the average Z of the inclusions is very different from that of the matrix, and can vary depending on how large the inclusions are with respect to beam diameter and specimen thickness (i.e., what fraction of the analysis volume is inclusion). Good luck. Yours, Bill Tivol
I appreciate what Nestor has done and is doing with respect to the listserver and it is a lot of work. Basically, I think that he is aware of the problem and if he wants to tackle the problem, it should be up to him. Until he decides to do something, I am already deleting a lot of stuff that I'm not interested in without reading it. A few extra in a week doesn't bother me. Yes, some of it is offensive, but usually I know from the subject that it is spam and I delete it without reading. Sometimes it gets through. I try not to let it ruin my day as I delete it and forget it.
Essentially, let Nestor decide when enough is enough.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Dear all, As you will have noticed, there have been two recent ads posted to the list from non members (one for a sex site, and one for an email Marketing program). Whilst the vendors who subscribe to this group almost always behave in a responsible manner by sticking to the charter and not sending overtly commercial mailings and Nestor deals with the occasional abuse, we have no control about non list members who post onto the list.
I am aware that some other discussion groups use software that blocks postings from non list members. I suggest that it may be necessary for Nestor to configure the software that is used for this list to prevent posting from non list members.
What do other people think.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
} (1) Carbon will not appreciably absorb oxygen in your oxides. X-rays are } absorbed by elements heavier than the emitting element. The absorbtion } correction for oxygen in a carbon matrix, particularly in a thin film, will } be very small. Use the "thin-film" analysis mode.
X-rays are strongly absorbed when their energy is just above the energy for the transition from a filled state to a vacant state in the absorbing element. This topic is covered in Friedlander, et al. "Nuclear and Radiochemistry", and to quote, "...consider the K-alpha X rays of zinc (Z=30) which have an energy of 8.6 kV. The K absorption edges of Cu and Ni are 9.0 and 8.3 keV respectively. Therefore, nickel is a good absorber for zinc K-alpha X rays, and copper is not..." So your statement that x-rays are absorbed by heavier elements is not correct. The light-ele- ment folks would have memorized the O K-alpha and C K-edge energies (I haven't) and would know whether carbon absorption of O K-alpha is small or not. Yours, Bill Tivol
It depends on your microscope. If you use a coater system that does not have a clean vacuum system such as a turbo pump has, then you can get oil contamination on your sample that will show up in a high beam current density instrument such as a Field emission SEM. However, your coal samples could be a source of contamination even though you went with a more expensive system. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I am considering the purchase of an evaporative carbon coater for use on coal clinker specimens that have a history of bad charging under the SEM. The evaporative carbon coaters (carbon rod source) come in two types: high vacuum (10^-5 torr) and low vacuum (10^-2 torr). The high vacuum coaters (with a turbo pump) are significantly more expensive. Are they worth the extra cost?
Dear Catherine, } } Can anyone please advice on the proper disposal of uranyl acetate? } Thank you. } In New York small amounts of 1% UAc can be poured down the drain. Of course disposal of kg quantities of UAc is a different matter. Also, disposal regulations may vary from state to state, so it is best to check with your safety office. If they don't know, check with your state's environment department (ours is Environmental Conservation, but the name could be different in your state). Yours, Bill Tivol
Dear James, } } Suggestions for adjustable height chairs (or modifications thereof) that } minimize or eliminate fiber shedding and static charge would be } appreciated. Thank you. } Find one made of unuppolstered (sp?) metal, or rip out the uppol- stery and replace it with wood, or cover with aluminum foil (single-use). Yours, Bill Tivol
We've been cutting tissue attached to epoxy coated with 50 nm titanium for years. We keep a special diamond for it, which does need replacing more often than knives used for tissue only. But one can get excellent sections, thick and thin, and the knife does last for a while.
Lesley Weston.
On Fri, 17 Jul 1998 edelmare-at-casmail.muohio.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I've got a couple of users who would like to pursue Pre-embedment labling of plant } tissue (trials with older knives and glass show excellent results) but are very } concerned about potential damage to diamond knife edges. So I'm collecting opinions } from experienced pre-embedment microtomists: are biological diamond knife edges } basically 'safe' from 10-20 nm gold? Such that future biological sectioning will not } suffer from any damage to the knife edge? } } Thanks. } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "WE ARE MICROSOFT. } RESISTANCE IS FUTILE. } YOU WILL BE ASSIMILATED." }
Larry, Your sad commentary could not have reached me at a more pertinent time. Today, at this very hour, a group of faculty sit and discuss the future of our only general use EM facility here at Stanford University. We have tried, as a group of concerned microscopists, to let them know how important it is to keep our facility open. Maybe they need to go on a magic carpet ride to experience the beauty and importance of our "obsolete" technology. Won't there be another mutant that we need to look at soon? JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94022
Our apologies to anyone who is having trouble reaching us. It appears that the area code change to 520 is one culprit, the reassigning of the old number to someone in Phoenix is another.
The fact that US West keeps a seperate directory assistance database from AT&T is a third, moving across town to a new building is a fourth and having a record amount of activity is a fifth.
We have taken all the steps necessary to remedy this including notices to all directory assistance sources we know of and change of address and phone to all internet sources as well.
OUR NEW PHONE IS 520-903-9366, address: 3450 S Broadmont Tucson, AZ 85713
PLEASE NOTE: do not respond to my personal email address on this missive, I am traveling and use this only sporadically. We invite all to visit Tucson to see our new facility and new applications laboratory. We will give a tour of our production facility and even let you use a phone to prove that we have one. Apologies again to anyone that has been inconvenienced.
Regards. Steve Miller Director of Sales, North America
In response to the following posting from Catherine: ================================================================= Wonder if anyone going to the 14th ICEM in Mexico? Any idea of Cancun, such as safety and traveling around? ================================================================= My wife and I are "veterans" of many trips over the years to Cancun, both for holidays and also scientific meetings, and I believe we can speak from the perspective of first hand experience, the most recent trip being about 2 years ago.
The Mexican government designated some years ago certain areas called "zona turisticas" and Cancun was one of the first (if not the first). Basically, these special areas are run from a safety and hygiene standpoint virtually like you were in the USA. The US chain hotels (e.g. Sheraton, Marriott, Hyatt, etc.) seem to be run from a safety, security, and other standpoints as if you really were in the USA. I think the other hotels in the "zona turistica" are also run to that same standard, as are also the local outside-the-hotel restaurants with the zone. We always ask for "agua purificada" and we perhaps take a few other common sense precautions, but in all our many trips to Cancun neither of us has ever suffered any problems. Hotels outside the "zone" would not be necessarily operating to that kind of standard so further precautions should be taken. The one note of caution is this: Just as there is in parts of the US a problem with salmonella in eggs, there is also the problem in Mexico so you would want to make sure that any eggs in your diet were hard boiled or scrambled well done.
So far as "traveling around", we have taken both tours as well as gone off on our own in rented cars. We have never had any problems. Everything is very reasonably priced. We have found the Mexican people to be friendly, hospitable, and genuinely appreciative of those who visit their country, perhaps in part because that part of Mexico is so dependent on tourism. From a purely touristic point of view, there are far more things to see and do than one could possibly have time for doing. The top of my list would include a day trip to Chichen-Itza, and a day trip to the neighboring island of Cozumel. If you are a scuba diver, the "wall" dive on Palancar Reef, just off of Cozumel, is one of the world's greatest dives. However, you might be better off to take a hotel for a few days in Cozumel.
Like anywhere else in the world, common sense must prevail. For example, upon arrival at the airport in Cancun, there are numerous porters to carry your luggage to a waiting taxi. There are no trolleys as there are in most airports for doing this yourself. So if you have more luggage than you can carry on your own, be prepared to take advantage of the local porters. My advice is to convert some money at the currency window right there is in the luggage arrivals area, while you are waiting for the arrival of your luggage. The rate is usually better there than at airports in the US. You should be prepared to tip the porter US $.50 per bag (or the equivalent in pesos). You will need to purchase a ticket for the taxi (really a van holding 8-10 persons) to your hotel. Within the zona turistica, it is all the same taxi rate. The cost is per person and is about US $8-$9. A group of four or more, arriving at the same time and going to the same hotel can get a better deal by reserving an entire taxi. So you should convert enough money to end up with enough pesos in order to be able to purchase your taxi tickets and give the porter a tip and also the driver a tip. If you present for payment for the taxi tickets US dollars, the rate you are given (at least that was the case in the past) is pretty crummy. Therefore the reason to get some pesos. When leaving the airport area, for the taxi ticket stand or anywhere for that matter, I would recommend keeping a close eye on your belongings. I would make this same admonition for someone arriving at JFK in NYC, but this is just common sense.
If you should arrive and one or more of your bags does not arrive, then you want to file the report with the airline before leaving the luggage area and before going through customs. If you are connecting from a different airline to make your final flight to Cancun, make sure when you check in for the Cancun flight you present your bag tags from the previous flight so make sure your bags are in fact transferred onto the Cancun flight. Other wise, with security being what it is, you could end up being separated from your checked luggage.
} From all that I have been able to learn, it really does sound like this ICEM is going to be the best ever. When I attended the MICRO 98 meeting in London a few weeks ago, the "word" was that "everyone" is going. However, there are still luxury hotel rooms available at reasonable rates so if you have not yet made your decision, now is the time to decide! One other note: Don't worry if a hotel is one or more Km from the convention center. A local bus traverses the main road along the hotel strip every few minutes and the cost per ride is roughly US$.25 , so don't let the "distance from the convention center" deter you from a particular hotel (so long as it is along the hotel strip). In any case, riding these local buses is a part of the Cancun experience you would not want to miss anyhow.
Chuck
PS: For prospective exhibitors, I am told there are still some booths (exhibit stands) available.
Disclaimer: These are my own personal opinions and recommendations and might not necessarly reflect the views of the organizers.
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
In response to the following posting from Catherine: ================================================================= Wonder if anyone going to the 14th ICEM in Mexico? Any idea of Cancun, such as safety and traveling around? ================================================================= My wife and I are "veterans" of many trips over the years to Cancun, both for holidays and also scientific meetings, and I believe we can speak from the perspective of first hand experience, the most recent trip being about 2 years ago.
The Mexican government designated some years ago certain areas called "zona turisticas" and Cancun was one of the first (if not the first). Basically, these special areas are run from a safety and hygiene standpoint virtually like you were in the USA. The US chain hotels (e.g. Sheraton, Marriott, Hyatt, etc.) seem to be run from a safety, security, and other standpoints as if you really were in the USA. I think the other hotels in the "zona turistica" are also run to that same standard, as are also the local outside-the-hotel restaurants with the zone. We always ask for "agua purificada" and we perhaps take a few other common sense precautions, but in all our many trips to Cancun neither of us has ever suffered any problems. Hotels outside the "zone" would not be necessarily operating to that kind of standard so further precautions should be taken. The one note of caution is this: Just as there is in parts of the US a problem with salmonella in eggs, there is also the problem in Mexico so you would want to make sure that any eggs in your diet were hard boiled or scrambled well done.
So far as "traveling around", we have taken both tours as well as gone off on our own in rented cars. We have never had any problems. Everything is very reasonably priced. We have found the Mexican people to be friendly, hospitable, and genuinely appreciative of those who visit their country,
perhaps in part because that part of Mexico is so dependent on tourism. } From a purely touristic point of view, there are far more things to see and do than one could possibly have time for doing. The top of my list would include a day trip to Chichen-Itza, and a day trip to the neighboring island of Cozumel. If you are a scuba diver, the "wall" dive on Palancar Reef, just off of Cozumel, is one of the world's greatest dives. However, you might be better off to take a hotel for a few days in Cozumel.
Like anywhere else in the world, common sense must prevail. For example, upon arrival at the airport in Cancun, there are numerous porters to carry your luggage to a waiting taxi. There are no trolleys as there are in most airports for doing this yourself. So if you have more luggage than you can carry on your own, be prepared to take advantage of the local porters. My advice is to convert some money at the currency window right there is in the luggage arrivals area, while you are waiting for the arrival of your luggage. The rate is usually better there than at airports in the US. You should be prepared to tip the porter US $.50 per bag (or the equivalent in pesos). You will need to purchase a ticket for the taxi (really a van holding 8-10 persons) to your hotel. Within the zona turistica, it is all the same taxi rate. The cost is per person and is about US $8-$9. A group of four or more, arriving at the same time and going to the same hotel can get a better deal by reserving an entire taxi. So you should convert enough money to end up with enough pesos in order to be able to purchase your taxi tickets and give the porter a tip and also the driver a tip. If you present for payment for the taxi tickets US dollars, the rate you are given (at least that was the case in the past) is pretty crummy. Therefore the reason to get some pesos. When leaving the airport area, for the taxi ticket stand or anywhere for that matter, I would recommend keeping a close eye on your belongings. I would make this same admonition for someone arriving at JFK in NYC, but this is just common sense.
If you should arrive and one or more of your bags does not arrive, then you want to file the report with the airline before leaving the luggage area and before going through customs. If you are connecting from a different airline to make your final flight to Cancun, make sure when you check in for the Cancun flight you present your bag tags from the previous flight so make sure your bags are in fact transferred onto the Cancun flight. Other wise, with security being what it is, you could end up being separated from your checked luggage.
} From all that I have been able to learn, it really does sound like this ICEM is going to be the best ever. When I attended the MICRO 98 meeting in London a few weeks ago, the "word" was that "everyone" is going. However, there are still luxury hotel rooms available at reasonable rates so if you have not yet made your decision, now is the time to decide! One other note: Don't worry if a hotel is one or more Km from the convention center. A local bus traverses the main road along the hotel strip every few minutes and the cost per ride is roughly US$.25 , so don't let the "distance from the convention center" deter you from a particular hotel (so long as it is along the hotel strip). In any case, riding these local buses is a part of the Cancun experience you would not want to miss anyhow.
Chuck
PS: For prospective exhibitors, I am told there are still some booths (exhibit stands) available.
Disclaimer: These are my own personal opinions and recommendations and might not necessarly reflect the views of the organizers.
============================================
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 ######################## ============================================
FALL 1998 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-V2)
NASSAU COMMUNITY COLLEGE
A fifteen week, fall 1998 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered ONE EVENING PER WEEK, Thursdays, starting at 5:30pm. Classes will begin on Sept. 3 and end on Dec. 10, 1998.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 per credit.
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
For those without www access, the catalog description is specified below. If you have further questions, you should e-mail me directly at the address below.
Interested individuals should register early (by Aug. 10) since the course is limited to a total enrollment of ten (10) students.
Questions regarding the actual registration process can be directed to our registrar at (516) 572-7355. ________________________________________________________________________________
CATALOG DESCRIPTION BIO 221: Transmission Electron Microscopy -- 4 credits Prerequisites: BIO 109-110 or equivalent, CHE 151-152 or equivalent. An introduction to the basic principles of transmission electron microscopy including tissue preparation, microscope (TEM) operation, black & white photography, and micrograph interpretation. The entire laboratory is devoted to the development of skills and preparative techniques involved with the operation of an actual transmission electron microscope. (3 lecture, 3 laboratory hours). Laboratory fee applies. ________________________________________________________________________________
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
FALL 1998 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-V2)
NASSAU COMMUNITY COLLEGE
A fifteen week, fall 1998 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered ONE EVENING PER WEEK, Thursdays, starting at 5:30pm. Classes will begin on Sept. 3 and end on Dec. 10, 1998.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 per credit.
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
For those without www access, the catalog description is specified below. If you have further questions, you should e-mail me directly at the address below.
Interested individuals should register early (by Aug. 10) since the course is limited to a total enrollment of ten (10) students.
Questions regarding the actual registration process can be directed to our registrar at (516) 572-7355. ________________________________________________________________________________
CATALOG DESCRIPTION BIO 221: Transmission Electron Microscopy -- 4 credits Prerequisites: BIO 109-110 or equivalent, CHE 151-152 or equivalent. An introduction to the basic principles of transmission electron microscopy including tissue preparation, microscope (TEM) operation, black & white photography, and micrograph interpretation. The entire laboratory is devoted to the development of skills and preparative techniques involved with the operation of an actual transmission electron microscope. (3 lecture, 3 laboratory hours). Laboratory fee applies. ________________________________________________________________________________
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
I am writing to request some help please. I am a Chiropractic Physician who has taken two training courses in Enderleinian Darkfield Microscopy. I am searching for a used microscope to use for that purpose. I would like your opinion comparing the quality of the Zeiss Universal to a Leitz Orthoplan. Also, is IC optics a significant help with darkfield? Is the image from an Axioskop any better than that obtained from a Universal or ORthoplan? Finally, which type of objectives - plan Achromat. plan-Neoflour, or Plan apochromat would suffice/be recommended? Thank you very much.
Numerous micrographs of bacteria and other specimens can be found in Ruska's book: Ernst Ruska. 1980. The early development of electron lenses and electron microscopy. Translated by Thomas Mulvey. S. Hirzel Verlag (Stuttgart). ISBN 3-7776-0364-3.
A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
-------------------------------------
On Mon, 20 Jul 1998, frank booy wrote:
} I remember seeing an image of a bacterium imaged in/by Ruska's } first microscope. } Can anyone tell me where this is to be found please? } Thanks! } } Frank Booy }
I am currently looking into a new Field Emissions Auger for our lab. I know the general differences between the Cylindrical Mirror and the Hemispherical analysers but has anyone performed a recent comparison. I run a multipurpose system doing powders, fracture surfaces, polished surfaces and anything in between. If anyone has data that they can share with me I would really appreciate it.
Thank you Tamara E. Bloomer Assistant Scientist Ames Laboratory 137 Wilhelm Hall Ames, IA 50011 (515) 294-2564
Donna Wagahoff asked: } We are in the market for a new knifemaker. Those of you with newer model } knifemakers, please let me know how you rate their performance. } Does anyone other that Leica make knifemakers? Thanks.
We (Energy Beam Sciences) manufacture both a triangular and a "Ralph" glass knifemaker. The triangular knifemaker is the descendent of the old DuPont-Sorvall knifemaker, and is designed to cut 10mm and 12mm knives (as opposed to the Leica, which is designed for 6.4mm glass). RMC also make a knifemaker, and there is another one, made in the UK, which is distributed through Agar there.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
First: regarding the choice between Zeiss and Leitz
The best suggestion for your application is that you try your specimen with the microscope you intend to purchase, to see which does the best job for you. I am not familiar with the term "Enderleinian" with respect to Darkfield microscopy. If you can provide a description, I may be able to comment more completely on which optics would perform best.
Secondly: regarding "IC" optics
This nomenclature refers to a different contrast technique, differential interference contrast (DIC), often called "Nomarski" (Dr. Georges Nomarski developed a method for cutting the necessary prisms which enabled the technique to become commercially viable). Since DIC is based on polarized light, it requires strain free optics; the engraving IC indicates that the optics are suitable for this technique. Making the assumption that you are working in transmitted light, a complete DIC system would require a set of objectives fitted with polarizers and DIC prisms (some systems have a single, common prism while others have a separate prism/polarizer set for each objective) as well as a matching condenser (often a turret, if you are going to do DIC with more than one objective), also fitted with polarizers and prisms.=20
These two techniques work on very different aspects of the sample:
In Darkfield, the light approaching the sample comes in at such a high angle that it cannot be collected by the objective. As a result, the background is dark. However, any feature in the sample (little grains, strings, pits, scrathes, dust) which can scatter light will do so. The scattered light is collected by the objective and used to form the image. The result: bright objects against a dark background. Since scatter is highly wavelength dependent, darkfield can also cause is extraneous color.
DIC is a bit too complex to explain completely here but basically, it detects gradients (slopes or changes)in the sample. The gradients can come from either changes in topography or changes in refractive index (ex: due to changes in concentration, gelation, etc.). In the most sensitive DIC settings, the background is usually pearly gray; one slope will be bright and the slope on the opposite side of the feature will be dark. Since you mind interprets these bright/dark cues in terms of differences in height, DIC images often appear to have increased structural or 3-dimensional information; they need to be interpreted very carefully. Sometimes adding an internal standard (ex: tiny glass beads) is helpful in understanding what is really happening. A second artifact comes from materials which respond to polarized light. Typically, they will "scramble" the DIC information. What you will see will be beautiful Pol colors, but not the true gradient information from the DIC. To test, just compare one side of the feature to the other. If the feature shows the same response on all sides (as opposed to one side being bright, the other being dark), it is probably responding to the Pol component of the system, not the DIC components.
For more on all these topics, might I suggest our book "Optimizing Light Microscopy for Biological and Clinical Laboratories"? Details are available at our website:
Thank you to those who responded to my inquiry after clean-room chairs.
We located several fitting the description in the Lab Safety Supply catalog (800-356-0783) made by a company called BioFit. The chair seats are vinyl (and look comfortable) and dissipate static by means of a brass chain fitted to the bottom of the chair, which drags on the ground. The difference between this chair and regular vinyl chairs appears to be the brass grounding chain.
No doubt other vendors carry similar products, and I have no commercial interest (of which I know) in Lab Safety Supply Co.
Thanks everyone for helping me try and build or get a microscope. My feelings are that it is better and cheaper to buy a microscope, ie - 600x from India or China for $70, (one of the plans I saw was to make a 100x for $100, but it seems outdated) but I still want to make my own eventually someday. Could someone direct me to some sources for acquiring these microscopes? Perhaps an online source or catalog. I want to use the scope to view slides with a light source from underneath, and also be able to view the surface of small objects (like within an inch in heigth or bigger).
We do not believe you would need a turbo system. Ladd sells a carbon coater (diffusion pump) that is a high vacuum system (10 to the minus 6 or 7) and we believe this would be enough for you. We could always add a turbo pump option that would add about $6,000 to the base price, which I would be happy to quote you if you are interested.
John Arnott Chairman --
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
Rick Felten 07/20/98 04:25 PM I love my Stylus Color 800 and need another printer. Has anyone compared a Epson Stylus Photo 700 to a Stylus Color 800 for printing high resolution black and white images? Ric Thanks
What do you mean by IC optics: DIC (Differential Interference Contrast or Nomarski) or ICS (Infinity Corrected System (Zeiss's infinity optics)?
Infinity correction will help if you are putting anything into the light path between the objective and the Telen (projection) lens (which focusses the light rays properly for the ocular lens.
Generally apochromatic lenses have higher numerical apertures and hence higher resolving capabilities than neofluars or achromatic lenses.
As to which microscope is better for your darkfield, I think you just have to try out the different microscopes for your particular application. This may be difficult if you're trying to buy a used one, but some of the microscope companies do sell used ones.
Matt Schibler
} ---------- } From: Stephen[SMTP:smd-at-capecod.net] } Sent: Sunday, July 19, 1998 3:37 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Zeiss vs. Leitz + } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Good day to everyone who is reading this, } } I am writing to request some help please. I am a Chiropractic } Physician } who has taken two training courses in Enderleinian Darkfield } Microscopy. I } am searching for a used microscope to use for that purpose. I would } like } your opinion comparing the quality of the Zeiss Universal to a Leitz } Orthoplan. Also, is IC optics a significant help with darkfield? Is } the } image from an Axioskop any better than that obtained from a Universal } or } ORthoplan? Finally, which type of objectives - plan Achromat. } plan-Neoflour, or Plan apochromat would suffice/be recommended? } Thank you very much. } } } } Stephen M. Driscoll }
May I suggest an HP 890C ($399). I've really been impressed with the quality when using the both the special paper and normal paper. I bought the cheaper version of this printer, the 722C ($299) for home and have been happy with it also. Both of these printers have the PhotoREt II feature and the Kodak Image Enhancements. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Rick Felten 07/20/98 04:25 PM I love my Stylus Color 800 and need another printer. Has anyone compared a Epson Stylus Photo 700 to a Stylus Color 800 for printing high resolution black and white images? Ric Thanks
I was wondering if anyone could help me analyse some results I have obtained using acridine orange. I am using rat uterus in my experiments, and attempting to detect apoptosis in the tissue. My main problem is interpreting the changes in DNA and RNA content in the tissue; so if there are any experts out there who could possibly give me some assistance, that would be great.
Thanks
Richard
Dr Richard Stump Rm 221 Anatomy and Histology Anderson Stuart Bldg. F13 University of Sydney NSW 2006
} } Thanks everyone for helping me try and build or get a microscope. My feelings } are that it is better and cheaper to buy a microscope, ie - 600x from India or } China for $70, (one of the plans I saw was to make a 100x for $100, but it } seems outdated) but I still want to make my own eventually someday. Could } someone direct me to some sources for acquiring these microscopes? Perhaps an } online source or catalog. I want to use the scope to view slides with a light } source from underneath, and also be able to view the surface of small objects } (like within an inch in heigth or bigger). } } Danny -
Here's a list of sources of compound scopes in your price range. a 3 objective monocular with condenser will cost at least $150. You'll find several books for adult hobbiests in the Project MICRO bibliography (address below).
3-OBJECTIVE MONOCULAR COMPOUND SCOPES
There is a much greater selection in this category; follow the advice in "Microscopic Explorations", and shop around. Every major school supplier offers a selection. Here are a few possibilities: Carolina Biological Supply 800-334-5551 Edmund Scientific 800-728-6999 Educational Teaching Aids 800-445-5985 Frey Scientific 800-225-FREY Insights Visual Productions 800-942-0528 Lakeshore Learning Materials 800-421-5354 Nasco 800-558-9595 Sargent-Welch 800-727-4368 Southern Precision Instruments (dealer) 800-678-7768
Caroline Schooley Educational Outreach 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/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I am posting this message for another microscopist. Your reply direct to his email or the listserver will be appreciated. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes *********************** www.proscitech.com.au *****
} I am looking for a staining method for looking at slime } deposits from } paper mills and biofilms in general using light microscopy. } I came } across some methods that stain bacterial capsules but none } that mentions } biofilms or slime layers (particularly levulose polymers). } } Bill van Eijk } email: whvanei-at-ibm.net
Dear Barbara, } } The chair at my microscopy bench has a naugahyde cover. Would that } help? } If one has the spiffy new synthetic-fabric lab coats (rather than the old-fashioned cotton ones), naugahyde is not a solution to the static- electricity problem. One could almost use that combination for an emer- gency HT source ;-). Yours, Bill Tivol
I'm sorry it's taken so long to summarize the responses I received to my query on how long to store tissues in glutaraldehyde fixative. The results are about even regarding whether or not to do it. Some people strongly believe that it is perfectly fine to store tissues for extended periods of time in glutaraldehyde--a much better way to store tissues than in buffer--even with preservatives and anti-.fungals. Several laboratories responded that they store tissues in glutaraldehyde all the time with no adverse effects. One laboratory mentioned that on occasion, they do see some membrane whorling in their tissues following long storage periods, but it is minor. One laboratory described very negative effects of extended storage in glutaraldehyde, such as loss of density and contrast. Another individual mentioned a paper written a few years ago that dealt with different formulations of fixatives and found that storage in a fixative called 4F1G (4% formaldehyde/1% glutaraldehyde in PO4 buffer) over a 5 year period showed no adverse effects, but storage in higher concentrations of glutaraldehyde may cause some artifacts. One laboratory reported that extended fixation in glutaraldehyde makes the tissue difficult to section due to hardening of the tissue. Additionally, they report occasional leaching of detail and less intense staining. Finally, an elegant paper from the South African Journal of Botany ( Coetzee J & van der Merwe C F - 1986. The influence of processing protocol on the ultrastructure of bean leaf cells. SA J Bot 52 (2) 95-99) describes several fixation schedules, vaying fixation times, buffers and periods in buffer wash. The authors suggest that maximum fixation times in Na-Cacodylate should not exceed 16 hr, phosphate buffers should not exceed 2 days, and PIPES-buffered glutaraldehyde fixation should not exceed 4 hours. The most accepatble fixations with all the bufferes were obtained with one hour fixation schedules. Those of us who have worked with plant material are well aware that plant material is much less forgiving than animal tissue, which may account for the varying accounts and opinions. The bottom line from just about everybody, however, is that it is best simply to not store tissues at all and immediately process them up to the block stage (though we all know that this is not always possible). I hope this information helps those who requested a summary. I greatly appreciate everybody's input (and it was great to hear from some old friends again!). Thanks again! Cheers, Laura
Laura M. Patrone, Ph.D. Wyeth-Ayerst Research Biomedical Imaging 641 Ridge Road Chazy, NY 12921 (518) 846-6318 e-mail: patronl-at-war.wyeth.com
Thank you very much for taking the time to put that info out there. I would be interested in what kind of weather to expect also if anyone has that kind of information.
Hi all, I am organizing the Materials Science Tutorials for the 1999 M&M Meeting and earlier I had asked for suggestions on topics from MAS and MSA members and other possible attendees of the conference. Below is a list of what I received in the way of suggestions. I am asking those of you who have the time to vote for up to three of the subjects. This will help me make the final decision on the topics for next year. If you have other suggestions or comments please let me know.
} 1. "How about GIF and perhaps PEELS?" } } 2. Defect recognition and analysis in crystalline materials. } } 3. Accessing and using on-line crystallogrphic databases } } 4. SEM Techniques } } 5. Optimizing the microscope for XEDS and WDS detection } } 6. XEDS imaging and interpretation, the right ways and the wrong ways. } } 7. Automation and Remote Control } } 8. More sample preparation (cross section of ALL methods) } } 9. Cross section samples with the FIB & FIB in general. } } 10. Spectrum imaging
I should note that FIB was a very popular choice and I have pretty much decided to do that one anyway, but I would still like to hear some yeahs or nays.
Thanks.
John M.
Note new Area Code (734)
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Does anybody out there know how to get Leica to respond? We bought a camera system for one of our microscopes and the parts are not compatable. We have been trying to talk to our Rep. and he will not return the phone calls. I also sent an email to Leica with no response. Any ideas will be appreciated.
Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
} 9. Cross section samples with the FIB & FIB in general.
} 10. Spectrum imaging
and
EBSP
Thanks for your efforts in organizing these tutorials.
Scott
F. Scott Miller Electron Microscopy Lab smiller-at-umr.edu University of Missouri-Rolla 223 McNutt Hall voice: 573 341 4727 Rolla, MO 65409 USA fax: 573 341 6934
Can anyone tell me for sure that Tannic acid is Ok to be used in Phos buffer (at about 6.9 pH)? (for fixation after glutaraldehyde). We have always used Cac bfr, but cannot in this instance. If you just answer "yes" or "no", it is good enough. Thanks, Hildy
While on assignment, I supported a lab which did a lot of work with biofilms which formed everywhere from cooling towers for power plants to beer facilities. They used DiI (UV excitation).
For the best reference for all sorts of stains, dyes, and probes:
Molecular Probes Handbook of Fluorescent Probes & Research Chemicals,
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Does anyone have experience with the nanoplast resin? I would like to know if it possess a column contamination problem when used in the TEM or SEM. Also, are the results equal to or better than conventional resins or other resins that can tolerate some water remaining in the specimen material prior to infiltration. The intended use is for isolated starch granules. These granules are often hydrated during preparation or when used in food products. When hydrated, they are much softer and somewhat swollen so resins may be able to penetrate easier. Also, it may be of interest to compare the hydrated state with the dried morphology.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
by news.mtu.edu (8.8.8/8.8.8) with ESMTP id PAA25397; Tue, 21 Jul 1998 15:42:43 -0400 (EDT) Received: from mmserver.mm.mtu.edu (mmserver.mm.mtu.edu [141.219.66.30]) by mtu.edu (8.8.8/8.8.8) with ESMTP id PAA04040; Tue, 21 Jul 1998 15:42:43 -0400 (EDT) Received: from backscatter (backscatter.my.mtu.edu [141.219.67.144]) by mmserver.mm.mtu.edu (8.8.7/8.8.7/mturelay-1.2) with SMTP id PAA25134; Tue, 21 Jul 1998 15:42:41 -0400 (EDT) X-Authentication-Warning: mmserver.mm.mtu.edu: Host backscatter.my.mtu.edu [141.219.67.144] claimed to be backscatter Message-Id: {3.0.5.32.19980721153734.0097d360-at-mmserver.mm.mtu.edu} X-Sender: opmills-at-mmserver.mm.mtu.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.5 (32)
A colleague at Tulane University asked that I submit this to the list. Please do not reply to me.
Cheers, Owen
} LAB SUPERVISOR (TEM/SEM) } } } Tulane University's Coordinated Instrumentation Facility seeks an } individual to create its EM facility consisting of 2 SEM's (Amray 1600T, } Jeol 820) and TEM (Philips 410). The successful candidate will possess } experience in operating within a multi-user environment serving biological } and physical sciences clientele. Experience with basic } biological/materials techniques is required including, histochemistry, } ultramicrotomy and thin foil preparation, electron diffraction, x-ray } microanalysis (EDS & WDS), photographic and darkroom procedures, digital } imagery practices. Experience is desired in the following: repair of } electron microscopes, high vacuum systems and accessory instrumentation and } basic computer skills (word process, spreadsheet, digital image } manipulation, HTML). B.S. physical/engineering required, M.S. preferred. } } Qualified applicants submit complete packet of cover letter and resume in } our office on or before August 31, 1998 to: } } Tulane University } Human Resources } Collins C. Diboll Complex } New Orleans, LA 70118 } } Tulane University is an AA/EOE. } }
Presuming a hurricane does not hit the week of the ICEM meeting, there are two words which accurately describe the weather in Cancun the first week of September: Steam Bath.
Dress (or undress) accordingly...
Larry
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Responding to the message of {n1311092614.19691-at-btny.purdue.edu} from "Microscopy Center" {emcenter-at-btny.purdue.edu} :
I have one grad student in my lab who is using it for gold labeling studies on algae. He is using it because he doesn't have to completely dehydrate the cells in organic solvents before infiltrating them.
I don't know if there is any problem with contamination of TEM column, at least none that I'm aware of........yet.
One important clue when using this resin, is blocks tend to come out VERY hard if you use the recommended amount of catalyst B-52, such that you can't even trim it. We use 1/4 the amount specified in the instructions that come with the resin. So advise you try a few curring runs of blocks with no sample just to work out the amount of catalyst to use such that you get blocks soft enough to trim and cut.
A thread on nanoplast ran on this listserver in March. I compiled them into a file and will send it to you off-line as an attachment.
Good luck!
Gib
} Does anyone have experience with the nanoplast resin? I would like to know } if it possess a column contamination problem when used in the TEM or SEM. } Also, are the results equal to or better than conventional resins or other } resins that can tolerate some water remaining in the specimen material prior } to infiltration. } The intended use is for isolated starch granules. These granules are } often hydrated during preparation or when used in food products. When } hydrated, they are much softer and somewhat swollen so resins may be able to } penetrate easier. Also, it may be of interest to compare the hydrated state } with the dried morphology. } } Debby Sherman, Manager Phone: 765-494-6666 } Microscopy Center in Agriculture FAX: 765-494-5896 } Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu } Purdue University or: emcenter-at-btny.purdue.edu } 1057 Whistler Building } West Lafayette, IN 47907-1057 } } } .
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
May I correct Steven Slap's comment on the Leica knifemaker - +ACI-as opposed to the Leica, which is designed for 6.4mm glass+ACI-.
The Leica EM KMR2 is designed to produce glass knives using a balanced break technique from 6.4, 8.0 and 10.0 mm glass. If you would like more details - contact the friendly folks at Leica.
Philip Hyam Product and Marketing Manager - Sample Preparation Leica Microsystems Canada
I've had similar experiences in my area. You might try to find your rep's boss's name, or try the management chain in headquarters at Deerfield IL (800-248-0123).
Leonard R. Corwin Fort Dodge Animal Health Princeton, NJ 08543-0400
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Does anybody out there know how to get Leica to respond? We bought a camera system for one of our microscopes and the parts are not compatable. We have been trying to talk to our Rep. and he will not return the phone calls. I also sent an email to Leica with no response. Any ideas will be appreciated.
Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
by imo21.mx.aol.com (IMOv14_b1.1) id RYPZa12330; Tue, 21 Jul 1998 17:27:06 -0400 (EDT) Message-ID: {34b297c0.35b507ac-at-aol.com}
In a message dated 98-07-21 12:18:37 EDT, mmr7001-at-axe.humboldt.edu writes:
{ { Does anybody out there know how to get Leica to respond? We bought a camera system for one of our microscopes and the parts are not compatable. } }
Marty,
I agree with Leonard Corwin, Leica Customer Service at (800) 248-0123 should be able to help you. I don't know which e-mail address or web page you tried, but your request may be floating in cyberspace somewhere in Germany.
Did you purchase from Leica directly or through one of their regional dealers? Territories and reps change, and it's possible your area has been reassigned. Anyway, just give Customer Service your Zip code, and they can take it from there.
Hope this helps! Bob ***************************************** Robert (Bob) Chiovetti rchiovetti-at-aol.com E. Licht Company / 1-800-865-4248 Colorado/Utah/Wyoming/Arizona/ New Mexico/West Texas Representing Leica Since 1967 *****************************************
I have used LR Gold for embedding samples destined for EM immunocytochemistry on many occassions with no trouble. In my latest embedding, the blocks are not behaving in a typical fashion. Everytime I go to section them, they pull the water out of the boat and prevent thin sectioning. I can cut 0.5 um LM sections with effort and the sections look okay. Has anybody else had this experience and, more importantly, do you know how to get around it? TIA, tom phillips
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 (573)-882-4712 (voice) (573)-882-0123 (fax)
} From all the fixations I did many years ago with Tannic acid after glutaraldehyde, my short answer is
YES.
P.S.: Try to use the lightest molecular weight mixture of tannic acids you can find. Ones from Turkish nutgalls, not Chinese nutgalls. Mallinkrodt #1763 or #1764 are what I used.
} ---------- } From: HILDEGARD CROWLEY[SMTP:hcrowley-at-du.edu] } Sent: Tuesday, July 21, 1998 9:43 AM } To: postmessage } Subject: Tannic acid+Phos bfr????? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } } Can anyone tell me for sure that Tannic acid is Ok to be used in Phos } buffer (at about 6.9 pH)? (for fixation after glutaraldehyde). We have } always used Cac bfr, but cannot in this } instance. If you just answer "yes" or "no", it is good enough. } Thanks, } Hildy }
Tom, you may have already considered these solutions. They apply to = sectioning: 1. maintain a lower than normal water level in the boat, so the level is = slightly concave but maintains contact with the edge of the knife; and = 2. use a faster cutting speed. These are effective in reducing that = notorious water "pulling" by the LR's. Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
} The sample is embedded in a Spurr's resin. I am currently using an } Iodine solution but I was wondering if there was anything better? } } Robert } } Robert.Dickson-at-kcl.fi
You should be able to stain Spurr's with PAS provided the 1% periodic acid step is long enough, e.g. 30 minutes. If you want the complete procedure get back to me.
The staining should be a lot more intense than with Iodine, but other cell components may also stain up. Presumably you are looking at plant tissue? Some cell wall polysaccharides are likely to stain, also some phenolics. Nuclei may also stain - see Litwin, JA & Kasprzyk, JM (1974) Acta Histochemica, vol 50, pp 222-227 "Some remarks on the PAS reaction in semi-thin sections of Epon-embedded tissues". If your tissue was embedded in methacrylate resin or similar you could do a control of sorts by digesting the section with amylase before staining, but this may not work on Spurr's.
O'Brien and McCully recommend aniline blue black as a counterstain: 1% aniline blue black in 7% acetic acid for 10 min at 50 deg.C, rinse in 7% acetic acid. (Yes, I've got a copy of their out-of-print book, and it's not for sale).
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
Philip Hyam wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } May I correct Steven Slap's comment on the Leica knifemaker - +ACI-as opposed to the } Leica, which is designed for 6.4mm glass+ACI-. } } The Leica EM KMR2 is designed to produce glass knives using a balanced break technique } from 6.4, 8.0 and 10.0 mm glass. If you would like more details - contact the } friendly folks at Leica. } } Philip Hyam } Product and Marketing Manager - Sample Preparation } Leica Microsystems Canada
Right Philip! But then again, if you are on a limited budget, the "old" LKB Knife Makers still do a great job for 6,4 mm glass. They can be reconditioned (by us) at a very reasonable cost and reconditioned units are available. Markus F. Meyenhofer Microscopy Labs micro-at-mail.superlink.net
I really dislike "me too" postings, but I have a similar complaint to = Marty's. I would like to suggest to any Leica personnel out there to = make your Head office aware of the need for greater accessibility of = your applications dept. This means no disregard to regional offices, but = it is often helpful to talk to people closely involved in the = development of a particular instrument.
James Wesley-Smith Electron Microscope Unit University of Natal,=20 Durban, South Africa
----------
Does anybody out there know how to get Leica to respond? We bought a camera system for one of our microscopes and the parts are not = compatable. We have been trying to talk to our Rep. and he will not return the phone calls. I also sent an email to Leica with no response. Any ideas will = be appreciated.
Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
I have just heard from Prof. Sitte that, following his retirement, this year's workshops will be the last. So, having been a speaker in them since 1986, I thought I would say a few words.
For those who do not know, the workshops have been running in Seefeld (Tirol) since 1973 and, before that, in the University of Homburg, Germany. Over 2,000 biomedical and materials scientists and technicians from around the world have participated in them.
The workshops are renowned for their technical content, social intercourse, gastronomic experience, cultural interest and scenic beauty (being on a high plateau in the Alps). Somehow, Sitte has perfected the art of being the gracious host who combines long hours of learning with enjoyment.
The courses are taught by various experts in their fields: some are the developers of ultramicrotomy and cryo-equipment, some are immuno experts and others have made fundamental contributions to the field.
The details I have are given below, although the costs may be negotiable! Students, those from former Eastern Europe and Third World countries are HALF price. Further (definitive!) details are available from Prof. Sitte: FAX Austria (0043 from UK) 5212 22 16 22 Mail address: Prof. Sitte, Reitherspitzstrasse 166, A-1600 Seefeld in Tirol, Austria. _______________________________________________________
The language of these workshops is English (Workshop 64, in the German language, runs from 14 to 29 October).
Workshop 63 E-Bio & E-Mat (Biomedical and Materials) Introductory sectioning workshop (without cryo or immuno) Begin 9.30 am 24 Sept. 98, finish 9 pm 27 Sept. 98 (course fee AS 7,000).
Workshop 63 F-Bio Cryoultramicrotomy, cryomethods, immuno-gold-cytochemistry Begin 9.00 am 4 Oct, finish 9 pm 8 Oct (course fee 13,000).
Workshop 63 A-Bio (combined E & F workshops, course fee AS 16,000). Workshop 63 A-Mat (combined E & F workshops, course fee AS 14,000).
Workshop B-Bio Cryofixation, freeze substitution, freeze drying, progressive lowering of temperature technique and low temperature embedding (without cryoultramicrotomy and immuno) Begin 9.00 am4 Oct, finish 9 pm 8 Oct (course fee AS 8,000).
Workshop 63 F-Mat Cryoultramicrotomy, diamond knife sectioning and preparation of hard/inhomogeneous samples. Begin 9.00 am 27 Sep, finish 9 pm 2 Oct (course fee AS 13,000).
Hi everyone, I am looking for a course on basic SEM theory, operation and hands on training. The course should not last longer than 5 days and should take place during this year. All suggestions are welcomed. Thanks! Gabriel Rojano rojanog-at-macom.com Reliability Engineering AMP- M/A-COM
I am curious as to the best way to determine the thickness of the car= bon evaporation technique being deposited on the substance to be analysed= . It would be beneficial to know the method by which a knowledge of thi= s informationcan be attained within a few Angstroms. Joshua McCaig. (mccaigjm-at-corning.com)=20 --=20 MZ=90
Hi listers, I have to fix 18.5 dpc mouse embryo in formalin or paraformaldehy for immunostaining. Does anyone out there knows the procedure for paraffin processing and embedding? We had tried several times unsuccessfully. I like to know specifically how long and in what temperature those huge embryo should be fixed (fixative penetration issue), and keep the antigenicity as well. Thanks.
******************************************************************* To see what is in front of one's nose requires a constant struggle.
George Orwell
Dorothy Zhang Harvard School of public Health Building 2, CVLAB 677 Huntington Ave, Boston, MA 02115 Phone# 617-432-6981 Fax# 617-432-2980
You may leave the water at slightly lower level even though you may not see the sections right after cutting.
Good luck !
Ming
On Tue, 21 Jul 1998, Tom Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have used LR Gold for embedding samples destined for EM } immunocytochemistry on many occassions with no trouble. In my latest } embedding, the blocks are not behaving in a typical fashion. Everytime I } go to section them, they pull the water out of the boat and prevent thin } sectioning. I can cut 0.5 um LM sections with effort and the sections look } okay. Has anybody else had this experience and, more importantly, do you } know how to get around it? TIA, tom phillips } } 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 } (573)-882-4712 (voice) } (573)-882-0123 (fax) }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * #1074B Dentistry Pharmacy Building * * University Of Alberta. * * Edmonton, Alberta, Canada T6G 2N8 * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
To the lung experts, What do you do with floating lung tissue (} 2mm) that is fixed by immersion. The options are: 1)leave in a refrig overnight until they sink (swirling did not work). 2)place in a vac (15 psi) while still in the fix. 3)process the floating samples hoping they eventually sink. I am currently in the frig (opting for #1) but any hints or suggesstions would be welcome. Thanks.
We have an elderly, incomplete Leitz manual microtome about the place here. Although a possible candidate for a boat anchor, weighing perhaps 50 lb, it contains wonderful German machining and has many mysterious knobs that invite twiddling. It appears to increment (click!) after doing a slice. It is under consideration for use to section plant tissues for light microscopy at medium magnification to look at fungal pathogens at the cell level. (Not by me - I do chemical microscopy so I rarely slice.) The unit is about two feet long, resembles an infertile cross between a lathe and a bologna slicer, and the carriage is connected to a wide ribbon apparently to smooth the slicing action. It is painted gray and might plausibly date from the 1960s. It appears that the sample to be cut is intended to be clamped below, while there are clamps spanning a rather large gap (about 8 inches) which could plausibly hold a blade, although the span seems rather large (flexion). There is nothing resembling a blade holder with the microtome.
To get to the point: Can anyone supply a photocopy of a manual or pertinent parts of a manual describing the function of the parts? (fax 609-275-5239) Or perhaps be prepared to discuss how this works by telephone? Do you think it likely to be worth having a blade holder fabricated, or should we post a notice to the boat owners?
Leonard R. Corwin Fort Dodge Animal Health Cyanamid Agricultural Research Center PO Box 400 Princeton, NJ 08543-0400 609-716-2278; fax 609-275-5239 corwinl-at-pt.cyanamid.com
} 2) place in a vac (15 psi) while still in the fix.
I'd reccommend this route, specifically as follows:
Take a 2-hole stopper (or make one if you can't find one), and put the wide end of a cut-off pasteur pipette, or a length of glass tubing, into one hole. Choose a size of stopper which will seal the top of the container you're using for the fix. Use tygon tubing to connect the glass tube to house vacuum, or connect it to a similar device made with a one-hole stopper which fits onto a side-arm flask--this will prevent any oil in the line from getting to the sample. Turn on the vacuum, seal the fixing container with the two-hole stopper, place your thumb over the empty hole until the residual gas emerges from the tissue, then release the pressure. Repeat until the gas has escaped from the tissue. Note that the one-hole and two-hole stoppers can be interchanged, so if you have to hold the stopper onto the fixing container, it can be better secured if you don't have to worry about rocking the device with your thumb. Good luck. Yours, Bill Tivol
In this paper Professor Lindsay of ASU characterizes biomolecular nano-wire electron transfer using scanning probe microscopy. This characterization requires the samples are held in an environment that is free of moisture and molecular oxygen.
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; SPM Technology Leaders for Environmental / In Situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
I have cross-sectioned through gold fibers in animal tissue in the past, using a diamond knife. I did not see any problem or increased "knife marks" from these very thin ( {30 um diam.) fibers. However, I have also sectioned particles in tissue such as cobalt or titatium and one can clearly see in the photographs that there is no "knife mark" prior to hitting the particle but a HUGE one continuing beyond it, of the same thickness as the particle. By the end of sectioning a few such particle samples, that area of the knife is basically useless.
In my experience the gold doesn't dull the knife any more than animal skin, but I've never done plant tissue so I don't know how that relates.
Karen -- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
I'm in dire need of a part for my Technics Hummer III sputter coater (the plastic nut at the base of the specimen pedestal has stripped threads and cannot hold a vaccum). I can't find a current address or phone number anywhere. Can someone please reply direct??
TIA and cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
MICHAEL DELANNOY wrote: } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: } 1)leave in a refrig overnight until they sink (swirling did not work). } 2)place in a vac (15 psi) while still in the fix. } 3)process the floating samples hoping they eventually sink.
{snip}
I never thought about it but I guess I am an expert (24 years)in TEM of lung. You did not specify whether embedding will be for LM or EM. If plastic embedding for EM, } 2mm is too thick and they should be sliced to not exceed 2mm. Other dimensions are not important as long as they will fit in an upside down Beem capsule with the pyramid end cut off.
First put them under vacuum in fix to pull out the air (the tissue will still be at the surface while in the vac) which will optimize contact of fix with tissue (I interpret your term immersion to indicate that the tissue was not first perfusion fixed or insufflated with fix).
Then they can go in the fridge or stay at room temp and should sink.
They will definitely sink during subsequent steps with osmium and acetone dehydration for TEM.
My experience does not extend to paraffin processing of lung.
Tom Christensen Pathology Boston University Medical Center
Try decreasing the water level in your boat (surface will be concave; make sure the edge is wet--if you have trouble with wetting, email back.)
Also, you can decrease the clearance angle of your knife.
Finally you might speed up the cutting stroke.
Be careful in aligning, that you don't wet the block face or the back of the knife. Don't allow the knife to stop directly at the block face; always go through a cutting stroke when checking distance, etc. The water has a way of "jumping" onto the hydrophillic block.
Good luck.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} I am curious as to the best way to determine the thickness of the carbon } evaporation technique being deposited on the substance to be analysed. } It would be beneficial to know the method by which a knowledge of this } informationcan be attained within a few Angstroms. Joshua McCaig. } (mccaigjm-at-corning.com) } -- } MZ
By using a Digital Thickness Monitor you should be able to measure carbon depositon within a few Angstroms. Since carbon is directional, it is important to locate the sensor close to the specimen. In our carbon substrate production we are able to replicate deposition within 8 to 10 Angstroms. For this it is vital that you duplicate the time, amperage, vacuum and location of the substrates. It is also important that you are consistent in your choice of carbon or graphite.
Hope this helps, John Arnott
Disclaimer: Ladd Research sells Vacuum Evaporators, Digital Thickness Monitors and substrates.
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
Mike, Please send me your web site address in order that I might see what you have to offer. Thanks again for the info on Leitz, Zeiss, and Olympus. Stephen
I have noticed this same problem with LR White when using a diamond knife. My conclusion was that after prolong cutting the knife edge becomes coated with plastic residue, and this causes the sections to bind up, causing a series of cutting problems (block wetting, sections not coming off the edge, compression). Try cutting the block with a glass knife, if this is the problem the block will cut fine, if not....back to the drawing board.
William R.McManus Supervisor Electron Microscope Facility Department of Biology Logan UT 84322-5305 billEMac-at-cc.usu.edu 435-797-1920 Fax 435-797-1575
Sometime ago, I had a Hummer III. Memory at this point could be failing, but I think the nut at the pedestal base (the one which could be loosened to raise/lower the specimen on mine) was a "bonnet" nut - either an electrical feed through bushing or a plastic tube fitting nut. Such a replacement would be common and cheap from a building supplies vendor.
Again, if memory serves.... Hummer is (was?) sold by Anatec(k?), in the Washington D.C. vicinity.
_Woody_
shAf wrote: } } I'm in dire need of a part for my Technics Hummer III sputter coater } (the plastic nut at the base of the specimen pedestal has stripped } threads and cannot hold a vaccum). I can't find a current address or } phone number anywhere. Can someone please reply direct?? } } TIA and cheerios, shAf } } {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} } Michael Shaffer, R.A. - ICQ 210524 } Geological Science's Electron Probe Facility - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
-- de Woody, WB4QXE
Work: Electron Microscopist/Microanalysist Balance: Ham radio "homebrewing", computers , shade tree mechanic "the Gravel Garage".
On the www page: Scanning Electron images and Ham Radio Homebrewing stuff. http://www.geocities.com/capecanaveral/3722 .
We run courses "in house", in your own laboratory on your own equipment, = on all aspects of electron microscopy. With the world the way it is this mea= ns we spend more than 85% of our time with SEM on - specimen preparation, instrument optimisation, x-ray analysis, maintenance and consultancy. =
Please take a look at our web site for more information, we are able to tune a course to any requirement.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
On Wed, 22 Jul 1998 12:31:20 -0400 (EDT) MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: 1)leave in a refrig overnight until } they sink (swirling did not work). } 2)place in a vac (15 psi) while still in } the fix. } 3)process the floating samples hoping } they eventually sink. } I am currently in the frig (opting for #1) but any hints or } suggesstions would be welcome. Thanks. } } Mike D }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
A friend had this problem with plant material. She kept the samples submerged under a piece of wire mesh.
Michael Shaffer asked: } I'm in dire need of a part for my Technics Hummer III sputter coater } (the plastic nut at the base of the specimen pedestal has stripped } threads and cannot hold a vaccum). I can't find a current address or } phone number anywhere. Can someone please reply direct??
The Hummer range of sputter coaters is manufactured by Anatech Ltd in Springfield, VA. The phone number is 800-752-7629 and the fax number is 703-941-8077. George Barr, the President, can certainly help you. His e-mail address is gbarr-at-anatechltd.com
Best regards, Steven Slap
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
6621 Electronic Drive Springfield, VA 22151 (800)-PLASMA-9 or 703-941-8860 FAX: 703-941-8077
Her ext. is #104.
She fixed my Technics Hummer V promptly and will assist in over the phone consultations.
Ginger
Ginger Baker EM Lab Manager OMS Secretary/Treasurer Research Dept., OCOM 1111 W. 17th St. Tulsa, OK 74107 Phone: (918) 561-8232 FAX: (918) 699-8629 http://osu.com.okstate.edu/dept/research/content/gbaker.htm lizard-at-osucom-fs02.ocom.okstate.edu
-----Original Message-----
I'm in dire need of a part for my Technics Hummer III sputter coater (the plastic nut at the base of the specimen pedestal has stripped threads and cannot hold a vaccum). I can't find a current address or phone number anywhere. Can someone please reply direct??
TIA and cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I have an old Vickers-Armstrong hardness tester. To jog your memory, it has a foot pedal load actuator and a microscope mounted on a hinged bracket, which is moved in place over the indentation for measurement. Anyway, the eye piece tube and the attached micrometer ocular has been damage by an unknown user; this is a multi-user tester. Can anyone tell me who is in the business of servicing such a beast, Canadian rep. if possible. This is a very good, solid hardness tester and I would like to keep it. They don't make them like this anymore.
Thanks in advance. David Chow National Research Council Canada David.Chow-at-NRC.CA
Microscopy/Microscopy Education also specializes in customize, on-site courses. We have over two dozen consultants here in the US who have experience on a wide variety of electron microscopes and related analytical techniques.
For more information, see our website: { {http://www.MME-Microscopy.com/education}
In regards to the answer concerning the Technics address and phone number, I hit "reply to all" instead of "reply" so email may bounce on the listserv. Sorry all.
Ginger
You need to contact Lisa Jackson at Anatech, Ltd.
6621 Electronic Drive Springfield, VA 22151 (800)-PLASMA-9 or 703-941-8860 FAX: 703-941-8077
Her ext. is #104.
She fixed my Hummer V promptly and will assist in over the phone consultations.
Ginger Baker EM Lab Manager OMS Secretary/Treasurer Research Dept., OCOM 1111 W. 17th St. Tulsa, OK 74107 Phone: (918) 561-8232 FAX: (918) 699-8629 http://osu.com.okstate.edu/dept/research/content/gbaker.htm lizard-at-osucom-fs02.ocom.okstate.edu
Ginger Baker EM Lab Manager OMS Secretary/Treasurer Research Dept., OCOM 1111 W. 17th St. Tulsa, OK 74107 Phone: (918) 561-8232 FAX: (918) 699-8629 http://osu.com.okstate.edu/dept/research/content/gbaker.htm lizard-at-osucom-fs02.ocom.okstate.edu
I need information about video printers for printing SEM images and need = to contact sales representatives from various companies. Please contact = me at 217-782-0898. Thanks. Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il 62794-1220
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} This prompts me to ask the obvious question. Does anyone have a foolproof } indicator to check the activity of an osmium solution, ideally without } processing tissue and viewing in the microscope? } } I 'm sure at some time we have all picked up a bottle of expensive osmium } and thought do I use it or make fresh up. This happened to me recently and I } tried soaking some osmium into a piece of cocktail stick, which appeared to } darken, but I was wrong. } } Malcolm Haswell
Malcolm, I would put a drop of corn oil (maize oil on your side of the puddle) on a slide and some of the osmium solution in a small dish (size so that the slide acts as a lid for the dish). If the osmium is good, vapors from it will blacken the oil droplet. (Any polyunsaturated oil will do.) Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
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TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Hi, All.
I am looking for a secondary detector tube for JEOL JSM 35 in a good condition. Willing to pay a fair price for it. Please reply at my email address. Many many thanks for your anticipated co-operation.
Jitu Shah
Dr.Jitu Shah H.H. Wills Physics Laboratory, University of Bristol, Royal Fort, Tyndall Avenue, Bristol BS8 1TL. UK email: jss-at-siva.bristol.ac.uk Tel: 44 117 9288719 Fax: 44 117 9255624
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Petra:
With PE/PS I have gotten good results if I first cut and then stain. I do use cryo sectioning and I cool the sample to about -100C (room temperature will not work because the Tg of PE is too low). I use folding grids to collect my sections (dry) and then I stain with RuO4 which will stain the PS. You can also stain the block first and then cut but then you are limited to sections near the surface of the specimen (the stain does not penetrate too far). Even then, you might need to re-stain your sections. Finally, I deposit carbon.
I hope this helps,
Jordi Marti ----------------------------------
Hello polymer experts,
new to the field of EM polymer blends I would like to ask you for the best way to prepare PE/PS blends for structure determination with TEM.
Of course the material has to be stained and cut, but:
- what is the best staining agent (OsO4, RuO4, or something else)? - is it better, first to stain and then to cut or the other way round? - which temperature is the best for cutting, liquid nitrogen or room temperature?
I would appreciate ever tip & trick you can give on handling this kind of material.
Tanks in advance
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Dear All, I find oil on the EDX detectors in both my SEM's, but if you think of the conditions inside the SEM, it is almost inevitable. The majority of the molecules remaining in the vacuum of an average SEM at 10 -5 torr consists of diffusion pump oil. This oil will gradually condense on all surfaces in the SEM, but on the coolest surface first. In most SEM's this is the EDX snout. One EDX manufacturer has solved the problem by warming up the snout with a small heater. The oil just goes somewhere else, but if it forms a thin film over all surfaces it is not so visible or worrying. The cleaning method recommended by the manufacturer of my EDX's is to gently run clean-grade iso-propanol over the snout, then allow to air-dry. They used to recommend Freon, but that is no longer permitted or available. Hope this helps, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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POST-DOCTORAL RESEARCH FELLOW Princeton University
A post doctoral research fellow position is available in the Princeton Materials Institute and the Princeton Center for Complex Materials at Princeton University. The appointment is for one year initially with the possibility of renewal. It involves the use of a Philips CM-200 FEG transmission electron microscope equipped with a Gatan Image Filtering System in studying carbon nanotubes, and other nanostructured materials of interest. Candidates should have a Ph.D. in the physical sciences, and with strong EM background (HREM, Electron diffraction, and EELS). Experience in image analysis and UNIX workstations, and a strong math background are highly desirable.
Applicants should send a detailed curriculum vitae, copies of 2 selected publications, and names and addresses of three referees by June 15, 1997 to:
Nan Yao Princeton Materials Institute Princeton University Bowen Hall, 70 Prospect Avenue Princeton, NJ 08540
Princeton University is an equal opportunity employer.
=============================================================== Nan Yao Princeton Materials Institute Princeton University Bowen Hall, 70 Prospect Ave. Princeton, New Jersey 08540-5211
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} To All } I have been doing TEM since 1970, and have always used NaOH pellets } (CP grade - which do contain about 0.5 - 1% Na2CO3). I make up a 10N } solution in small aliquots (~20 ml), and dump it when I see crystals } of sodium silicate (NaOH dissolves glass). I use this 10N solution } to add to the powdered Pb citrate in dH2O. I make up Pb citrate in } 100 ml quantities and dump it when it turns cloudy. Usually it keeps } for months, unless someone forgets to tighten the cap, which is not } uncommon in a central service lab. What I have learned in nearly 30 } years of TEM, and many years in other microscopy techniques is that } there are very few empirical rules. I have been in very fussy labs } that used aged Pb citrate in bottles that were so coated with } precipitate that they looked out of a sunken pirate ship, and have } been in other labs that make up Pb citrate fresh, and filter it. } Whatever works. } Bruce Cutler, Microscopy Lab, Univ. Kansas, Lawrence KS
I store our freshly made-up concentrated Reynolds solution in 3ml aliquots in Eppendorff tubes at 4C. This way there is minimal exposure to CO2 and no danger of someone leaving the container open. A tube of stain is only used once, anything left over in the tube being discarded. The stain keeps well for months in these tubes.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377 Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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If I am off-base with this response, I apologize. I have used nothing but carbon tape ever since it became readily available through regular EM suppliers but I always dreaded having to clean it off my sample holders because it was so sticky and tore so easily. Being a simple minded person, I just took a 3/8" wooden dowel, sharpened one end(like a chisel) and used it to roll the tape up into a wad that I could remove with my fingers. All the adhesive seems to comes off with the tape, but if any should be left behind it should easily come off with a mild solvent. (I always polish my holders, so I'm not sure about residual adhesive.) I also cut a piece of latex tubing to fit on the end of the rod to cushion my hand. This works for the carbon tape, I'm not sure about the other types of adhesive tapes. For what it's worth.
Bill -- ============================================================= Bill Chissoe III Electron Microscopist,University of Oklahoma E-mail: wchiss-at-ou.edu Ph. (405)325-4391 ============================================================= Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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You wrote: " I am looking for the fax number or email address of Scanning Microscopy Int. (Chicago) - can anybody help?
Thanks in advance Fiona Graham Electron Microscope Unit University of Natal, Dalbridge, 4041, South Africa tel: +27 31 260 2174 fax: +27 31 261 6550 email: GRAHAM-at-ph.und.ac.za"
I have used 73211.647-at-compuserve.com within the last couple of months and it was OK.
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 617-275-4695 FAX: 617-275-4695 Alternate FAX: 617-271-0252
email dmrelion-at-world.std.com
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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JOINT SPRING MEETING
MISSOURI-ILLINOIS-KANSAS MICROBEAM ANALYSIS SOCIETY & CENTRAL STATES MICROSCOPY SOCIETY
Presentations will be in the Arthur Mag Conference Center (Mag Center) of Midwest Research Institute, Kansas City, MO
APRIL 25, 1997 ====================================================== PROGRAM
8:30-9:00 Registration, Vendor Display Setup in the Mag Center.
9:00-9:10 Welcome by Dr. William P. Duncan, Director, Midwest Research Institute.
9:10-9:25 Dr. Peter Moroz, Jr., G.S. Technologies, Kansas City, MO "Microstructure of Metal"
9:30-9:45 Harold McCormick, C-K Engineering Inc., St. Louis, MO "Wearever of Metals"
9:50-10:05 Garry Crabtree, TechSpec Inc., Raytown, MO "Material Response to Deep Cryogenic Tempering"
10:10-10:30 Morning Break-Refreshments from BAR ROMA (cash cart).
10:30-10:45 Dr. Ody Maningat, Midwest Grain Products, Inc., Atchison, KS "Wheat Starch and Wheat Gluten Research"
10:50-11:05 Dr. Diane Durham, KU Medical Center, Kansas City, KS "Regeneration of Sensory Hair Cells in the Avian Cochlea-SEM Analysis"
11:10-11:25 Dr. Peter Smith, KU Medical Center, Kansas City, KS "Light and Electron Microscopic Investigation of Nervous System Plasticity"
11:30-11:45 Dr. Amit Mukherjee, KU Medical Center, Kansas City, KS "Electron Microscopic Analysis of the Polymerization of FtsZ, an Essential Cell Division Protein of E-Coli"
12:00-1:15 BUFFET LUNCH Generously provided by HITACHI, catered by Nance's Deli and Catering. Buffet includes vegetable manicotti, garden salad, garlic bread, soft drinks, etc.
1:15-1:35 Business Meetings
1:35-1:55 Paul Benson, The Nelson-Atkins Museum of Art, Kansas City, MO "Scientific Technique as Applied to Art Objects"
2:00-2:15 Marv Hart, Century Lubricants, Kansas City, KS "Lubrication and Grease Technology"
2:20-2:35 Garth Kristoff, Allied Signal, Kansas City, MO "Overview of the Technical Transfer and Solvent Substitute for Electronic Cleaning"
2:40-3:00 Afternoon Break-Refreshments from BAR ROMA (cash cart)
3:00-3:30 Michael Saba, Digital Instruments, Eden Prairie, Minnesota. "Applications in Scanning Probe Microscopy"
3:35-3:50 John Wilson, Mo. Regional Criminalistic Lab, Kansas City, MO "Capabilities of Crime Scene Investigation"
****************************************************** Complimentary lunch will be provided for all attendees by HITACHI. However, we need to know how many will be attending the luncheon, prior to the meeting. Please phone or email one of the following:
Larry Irwin 913-268-9009 Dan Kremser 314-935-5605 dkremser-at-levee.wustl.edu
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Peter Jordan wrote: } } Hi: } I have received two E-mail letters from Philip Koeck over the past week } or so through this forum. Both letters I can not read or delete. } Whenever I try I get an error message saying that the program has } performed an illigal action and turns netscape off. Anybody having the } same problem or any solutions? Thank you, Peter Jordan .......................... I also use Netscape 3.0 and have the same problem. I don't have a solution, but I do have a way to get rid of the "poison message":
(1) read, and delete or transfer all the rest of the mail in the Inbox, but don't try to read or delete the "problem" message. (This is tricky, and you may have to "error-out" and reload Netscape a couple times.) (2) When the Inbox is empty except for this message, exit from Netscape. (3) Go to the NETSCAPE\NAVIGATOR\MAIL subdirectory, and locate the file "Inbox". Delete it (or rename it if you want to be cautious). (4) The next time you load Netscape, it will recreate an empty Inbox file.
This is ugly, but it's the only way I've found to get rid of the darn thing!
I hope somebody who doesn't have this problem will help us out by sending a message to the originator to find out what he is doing! I can't open his messages to get a return address. -- Fred Schamber ....................... mailto:fhscham-at-SGI.NET Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Hi, Everyone,
My background is in EM of biological, medical and food samples. I now have the opportunity to expand my skills and work on a set of materials samples.
My question is:
What are the standard methods for assessing surface degradation of materials samples (especially plastic polymers)? Are there macroscopic as well as microscopic methods which can give statistically "good" descriptions of the extend and type of degradation of such surfaces?
Thanks for any help (methods, references, review papers, etc.) you can provide. Please contact me offline.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist 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 e-mail: allanwojtasp-at-em.agr.ca Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Keith: I can't help you with sonicating water, but there is only one thing to do with flat beer, bin it. But then you people in the West Country have no idea what constitutes good beer. Come to Cambridge for a pint of Greene King Abbot or, better still Adnams. Both will knock your socks off
PatrickOn Wed, 16 Apr 1997, Keith Ryan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Ian } } I really appreciate the tip about sonicating water to degas it, I would not } have believed it. Now, do you have any suggestions for flat beer? } } Regards - Keith Ryan } Plymouth Marine Lab., UK } } }
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Would freshly distilled water be equivalent to degassing via sonication ?
Leo
On Wed, 16 Apr 1997, Ian Montgomery wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } Good morning Reynolds users } } } } In nearly 30 years of preparing lead citrate according to } } Reynolds I have experienced no problems in using a stock } } sodium hydroxide solution (1M) which was made up from pellets. } } However, this solution must be freshly made up. Is it because I } } always use Reynolds in its concentrated form that I do not } } experience any problems? } } } } Rob } } } Like Rob I've been making Reynold's lead citrate since I was a boy. } I use NaOH pellets but degas the distilled water by sonicating it for a few } minutes before making the stain. } } Ian. } } } Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
Peter Jordan wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi: } I have received two E-mail letters from Philip Koeck over the past week } or so through this forum. Both letters I can not read or delete. } Whenever I try I get an error message saying that the program has } performed an illigal action and turns netscape off. Anybody having the } same problem or any solutions? Thank you, Peter Jordan
Yes I am. It's very frustrating. I'd appreciate any help.
Thanks.
Eric Eric H. Metzler 1241 Kildale Sq. N. Columbus Ohio 43229-1306 USA
Phone: 614 888 3642 E-mail: spruance-at-infinet.com Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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} For Sale: JEOL JSM T-200 scanning electron microscope. Purchased in } 1984; used about 50 hours, total time. Includes chiller and a small } sputter coater. $1500 OBO. Contact Dr. Jon Martin at 912/752-4060. } Located at Mercer University School of Medicine, Macon Georgia.
E-mail contact: HORST_MN-at-Mercer.EDU ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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If you have "copies of copies" which work ok, and you have suitable blank media, there is no reason you cannot copy these again to generate archive disks. If you have a "verify" command/switch - use it. ...Or am I missing something??
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We have a NORAN (Tracor Northern) 5400 Series II EDS system (Model Number TN-5402/BBAA) acquired in 1987. It does not have a hard drive and operates off of two floppy drives. It requires 5.25" double sided, high density floppy disks, the originals of which are no longer readable. They have not been in use other than to be the source of copies made periodically but apparently they have simply deteriorated with time. We have copies of copies that are working pretty well but I'm becoming concerned that they may crash/die and we'll have no good source from which to make new copies. NORAN does not have replacement masters of this vintage so I'm hoping to find someone who has original master disks in good working order who would be willing to loan them to be copied.
We need two disks of the set from the November, 1987 Release 1C. What will work with our system is very specific and the original label on the master disks reads:
SERIES II SOFTWARE RELEASE 1C NOV., 1987, TRACOR NORTHERN, INC.
Of the disks in this release, we need:
SQ/SSQ/PRZ/ZAF Master NOT BOOTABLE
and
MSCAN2 Master
Thanks in advance for any help or suggestions you may have.
___________________________________________________________________________ Barbara Reine, Botany Dept. Box 351330 Univ. of Washington, Seattle, WA 98195-1330 e-mail: reine-at-u.washington.edu; ph: (206) 543-1955 ____________________________________________________________________________ Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
id NAA42426 (8.8.6/50); Thu, 23 Jul 1998 13:02:14 -0500 Message-Id: {199807231802.NAA42426-at-mail1.doit.wisc.edu} X-Sender: steve-at-facstaff.wisc.edu (Unverified) X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
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To all who responded to my plea for help: Thank you very much. Sandwiching the "bad" files between two "good" files allows you to move them into the trash bin and then to delete them. For all computer greenhorns like me this is done by clicking on the good file, then holding down the shift key clicking on the other good file. All files between the clicked files will be marked and then can be moved. It was realy nice to get all these respones. Thanx again, Peter Jordan Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Hello all,
We work as consultants for a computer imaging hardware and software manufacturer's rep, and have had a rather unusual request. Perhaps someone, either end users or vendors can assist.
We are looking for the equivalent of a MacBeth Color Chart, typically used in video, but this needs to be very small, translucent, and mounted on a standard histo glass slide. Ideally, 1/32" square (1 mm x 1 mm OK) and it would contain at least the primary colors, incl. black and white. I suspect there may be a problem with ?translucent black and white. I think episcopic illumination would also suffice for the chart, however so it could be opaque. At this time, I do not have more detail on their exact application.
We have checked with Munsell and MacBeth to no avail, so we would appreciate any assistance, and even a referral as it's likely this is a custom application. You can e-mail me directly if you like.
Thanks in advance!!
Daryl Martin dmartin-at-ic.net
(313) 213-8444
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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} What are the standard methods for assessing surface degradation of } materials samples (especially plastic polymers)? Are there macroscopic } as well as microscopic methods which can give statistically "good" } descriptions of the extend and type of degradation of such surfaces? } Dear Paula, I recently ran across a good review "Electron Microscopy in Polymer Science" by G.H. Michler, Applied Spectroscopy Reviews, vol 28(4), pp 327-384 (1993). This paper discusses surface characteriza- tion and many other topics. Additionally, I suggest STM or AFM as possibilities, but I don't have any referrences for them. Perhaps comparing the specular vs diffuse reflectivities of polymer surfaces before and after damage would be informative for degradation features ~ 1 micrometer in size. Good luck. Yours, Bill Tivol Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Meeting Announcement -
MAMAS (Mid-Atlantic Microbeam Analysis Society) and the Surface and Microanalysis Science Division, NIST Meetin at the National Institute of Standards and Technology, Gaithersburg, MD on Thursday, May 15, 1997, 10:30 am- 3:00 PM Lecture Room D, Administration Bldg.
10:30am Coffee and Doughnuts
10:45am Prof. David R. Veblen, Dept. of Earth and Planetary Science, Johns Hopkins University "Transmission Electron Microscopy of Minerals"
12 noon Lunch
1:15pm Dr. Michael Kersker, TEM/STM Project Manager, JEOL USA "200 KV FEG: Refried Beans with a Fiery New Salsa"
For more information, contact Ryna Marinenko (301)975-3901, FAX(301)417-1321, email:ryna.marinenko-at-nist.gov
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There is another option for scanning film negatives. A Leaf MicroLumina camera can be coupled with a high frequency light box and a copy stand to provide a simple and highly flexible solution to the problem of scanning film and transparencies.
The camera can be racked up and down on the stand to provide a field of view from 1" X 1.5" up to 9" X 12". The camera has a resolution of 2700 X 3380 pixels and can capture 12 bit grayscale and 36 bit color.
This configuration is currently offered by our company to the electron microscopy community under the name TEMSCAN.
If anyone is interested in this product, please contact us by phone: 516-773-4305 or e-mail: sales-at-electroimage.com. Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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PROPOSED IMMUNOCYTOCHEMISTRY NEWSGROUP
This is THE FINAL URGENT REQUEST for your comments, suggestions, etc about the 3RD RFD for my PROPOSED NEW NEWSGROUP SPECIALIZING IN IMMUNOCYTOCHEMISTRY/IMMUNOHISTOCHEMISTRY/OTHER RELATED AFFININITY METHODS called "sci.bio.immunocytochem" now posted in "news.groups"
The latter contains material of a rather varied nature (!) but it is necessary to use this unmoderated group to hold the discussions about all the different proposed new groups.
So, if you are connected to Usenet, and you are keen to see A NEW IMMUNOCYTOCHEMISTRY NEWSGROUP, then please go to "news.groups", ignore all the rubbish, look for articles posted on 20.4.97 or 24.4.97 (or later) and you should find one of my postings "3RD RFD: sci.bio.immunocytochem". Select "follow-up article" (or equivalent) from your newsreader menu, and post your message so that it appears under the 3RD RFD.
If you don't have access to Usenet, you can read the proposal at the Royal Microscope Society web site {http://www.rms.org.uk} , or at the Introduction to Immunocytochemistry (Center for Cell Imaging Department of Cell Biology Yale University School of Medicine) web site {http://info.med.yale.edu/cellimg/CCIimmuno.html}
MANY THANKS to all of you who have already posted your responses to my RFDs! BUT WE STILL NEED MORE DISCUSSION in news.groups please! SOON I shall be posting my "CALL FOR VOTES" to the people in charge of Usenet newsgroups. When you see "CFV: sci.bio.immunocytochem", you will be able to e-mail your vote to the vote-taker.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Message on behalf of Richard Easingwood; } } We are looking at the various options for large-format (4 x 5 inch) } negative scanners and wonder whether we could get some feedback from actual } users. } I have heard that the Leaf 45 scanner (which I understand was the Rolls } Royce/Cadillac of scanners) is discontinued and no longer available - can } anyone confirm this? } } Does anyone have experiences (good or bad) with the Polaroid SprintScan 45? } } We want to use the scanner for getting high quality resolution scans from } our sheet film TEM micrographs. } } Thanks in advance for any feedback. } } Regards, } } Richard } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences } University of Otago } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } e-mail: richard.easingwood-at-stonebow.otago.ac.nz } } }
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Good morning Reynolds users
In nearly 30 years of preparing lead citrate according to Reynolds I have experienced no problems in using a stock sodium hydroxide solution (1M) which was made up from pellets. However, this solution must be freshly made up. Is it because I always use Reynolds in its concentrated form that I do not experience any problems?
Rob
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377 Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Thanks to the several people who responded to my question.
The following was received directly from Bart Cannon and has some useful detail. I thought others might be interested.
} Arthur Wehnelt (1871-1944) was the German physicist who invented the } "Hot Cathode" electron tube which employed what was called a "grid" to } direct a stream of electrons. It corresponds fundamentally to the } electron gun used in oscilloscopes, TVs, and electron microscopes. Grid } and wehnelt have been interchangable terms to some degree.
-- Fred Schamber ....................... mailto:fhscham-at-SGI.NET Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Dear Mark,
We buy our plastic storage pages from the University bookstore and also from local photography stores. They are archival quality and come in a variety of sizes. They are made by Print File Archival Preservers, PO Box 607638, Orlando Fl, Ph:407-886-3100; Fax 407-886-0008. The type I use for 4x5 polaroid prints is product #45-8P (which are a heavier weight than the negative preservers).
Hope this helps.
Rosemarie Rosell Assistant Research Scientist Dept. of Plant Sciences University of Arizona Tucson,AZ 85721 Ph: 520-621-1230 Fax: 520-621-8839
On Wed, 30 Apr 1997, Mark Blackford wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } } I am trying to find a source of plastic pages for three-ringed binders } which have pockets large enough to hold polaroid prints (these are 131mm x } 106mm). Ideally each page should have 4 pocket and be able to fit two } prints back-to-back so images are visible from both sides of the page. } } I would really appreciate it if someone could give me an } address/phone/fax/email of a supplier of these items (if, indeed, these } things are made). Cheers, } } Mark Blackford } TEM Group } Materials Division, ANSTO } PMB 1, } Menai, N.S.W. } Australia } 2234 } Phone 61 2 9717 3027 } Fax 61 2 9543 7179 } } Disclaimer: } The views expressed in this E-mail message do not necessarily represent the } official views of ANSTO from which this message was conveyed. } } } Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Hi there,
I'm looking for carbon EM calibration grid from SPI model 411CG-AB but couldn't find any information about SPI.Does anyone knows they phone # or www address? Thanks.
**************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * mxq-at-u.washington.edu * * (206)543-1514(phone) * * (206)543-3100(fax) * ****************************
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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At 04:46 PM 4/29/97 +1200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, this is true. However, you may be able to locate a refurbished Leaf 45 Scanner by contacting a local digital photography distributor.
} } } } Does anyone have experiences (good or bad) with the Polaroid SprintScan 45? } } } } We want to use the scanner for getting high quality resolution scans from } } our sheet film TEM micrographs. } } } } Thanks in advance f snip, snip
A new option which I have not seen discussed on this forum is a drum scanner. The company ScanView which has made high-quality drum scanners for the pre-press market has now begun to market cheaper drum scanners at a relatively low price. We recently purchased the Scanview 3000 (3000 dpi, ~$15,000) which has an optical density range of 3.6, color resolution of 3 x 12 bit, 4096 levels, and a scanning area of 8.5" x 11.5". Some advantages of this scanner over the Leaf 45 are that the 3000 dpi resolution is available over the entire drum and the scan speed is much faster at the same resolution since it is a single pass scan. So, enlargements of an area of the negative which is off center are easy to do. You can also mount up to 6 negatives at once and set up batch jobs which will free up some of your time. We have been very pleased with the results so far. I believe it will deliver all possible information from the negative since the resolution of negatives are not much better than 3000 dpi in most circumstances. ScanView also makes two cheaper drum scanners the Scanmate Plus II (2600 dpi, o.d.=3.6) and the Scanmate Magic (2000 dpi, o.d.=3.0). I suggest that you contact your local digital photography dealer for a demonstration.
I have no financial interest in ScanView. I am just a satisfied user.
******************************************************* David F. Teter Los Alamos National Laboratory Materials Science and Technology: Metallurgy (MST-6) Mail Stop: G755 Los Alamos, NM 87545 ph: (505)665-0160 fax: (505) 665-0657 e-mail: teter-at-lanl.gov *******************************************************
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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NYMA Inc., an aerospace engineering company serving NASA at Lewis Research Center, is seeking to fill the following two challenging positions;
SCANNING ELECTRON MICROSCOPIST
Required to operate and maintain three scanning electron microscopes, and provide materials characterization support. The individual must also be able to assist and instruct research staff in the principles and operation of scanning electron microscopy.
Requirements: B.S. degree with 3 years experience or 7+ years of extensive hands on experience in electron microscopy. Must have a thorough working knowledge of x-ray energy/wavelength dispersive spectroscopy, preparation equipment, and vacuum systems. A working knowledge of x-ray diffraction and electronics is a plus. Excellent communication and interpersonal skills are required.
TRANSMISSION ELECTRON MICROSCOPIST
Extensive experience in operating, and maintaining a 200 KeV transmission electron microscope in support of material characterization of advanced high temperature materials. The successful candidate will work with materials researchers to understand materials' properties.
Requirements: Ph.D. in material science with 3+ years extensive experience operating a transmission electron microscope using SAED, CBED, XEDS, EELS, and PEELS to analyze a wide range of materials. The individual we seek must have extensive experience in sample preparation using electro-polishing, ion-milling, PIPS, and PIMS. A working knowledge of a dedicated STEM and x-ray diffraction is a plus. Excellent communication and interpersonal skills are required.
Qualified candidates should submit a resume to : Todd Leonhardt, NYMA, Inc., Mail Stop 105-1, 2001 Aerospace Parkway, Brook Park, OH 44142 or by E-mail to todd.a.leonhardt-at-lerc.nasa.gov
Posted Date: April 28,1997 Closing Date: Open until filled
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Fellow microscopists I am doing some TEM of cultured lymphocytes. All membranes seem to be absent. There are halos where membranes should be but no membranes. So far I have tried 2.5 glut in 0.1M cacodylate 2.5 glut in 0.2M cacodylate (caused shrinkage) 4 paraformaldehyde 1 glut in 0.1M cacodylate.
Post fixing in Osmium dehydrating in ethanol and embedding in Spurr
I am going to try making up the fix in the culture medium next.
Has anyone any thoughts of anything else to try. I would be particularly interested in the use of Ca+ Mg+ and sucrose.
Many thanks
Chris Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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G'day Colleagues...
Just a quick note to let you all know that that Microscopy Listserver Archives are now on-line. I have made accessible all Email postings covering the period Oct.1993 through Mar.1997 and will update the archive monthly.
The index is not directly searchable, however, it is chronologically sorted by Month and Year.
If you download a given month's postings then you can search the downloaded WWW page for any keyword/phrase that you wish by using the native search/find option of your WWW Browser. (In NetScape this is located in the Edit Pull Down Menu and is called FIND).
You may access the archive at the MSA WWW site.
http://www.msa.microscopy.com
just follow the links to the Reference/Educational Activities Page.
I'll get around to putting together a completely searchable index sometime in the forseeable future, but for now this will go a reasonable way to letting everyone find "old messages and postings" and all the other miscellaneous requests I receive for information.
Cheers...
Nestor Your Friendly Neighborhood SysOp.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Aloha, y'all (I've just been in Texas)
Below is a request for instructions and/or parts for a cryostat from some colleagues.
Thanks in advance for your help!
Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* **************************************************************************** Message:
We have recently obtained a surplus cryostat, a Milles Scientific Microtome Model 4553. The unit chills well and the microtome is mechanically in good shape. Unfortunately, it does not have instructions, an antirolling plate or specimen stubs. Does anyone know a source for these items? We have been unable to locate a phone number or address for Milles Scientific. Perhaps they have gone out of business or merged with another company. Perhaps someone has surplus parts for this model we could acquire. Your assistance would be appreciated.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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I am working with distarch phosphates (carbohydrate chains covalently linked together by Phosphorous), created with POCl3. I was hoping that someone can tell me whether it would be possible to use some type of probe (maybe with fluorescence?) to be able to detect the phosphorous based crosslinks. I have used SEM with EDS and this technique is not sensitive enough to detect the low level of phosphorous in the starch.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Negafile has polaroid 4x5 pages 4 per sheet. I'm not sure if they have gone to one size now or if they still have the two 4x5 sizes. You used to have to specify an "L" in the part number when ordering, but the number on the sheet itself, 4504, was the same for both. Check with them. I do not have their phone number, but their address is P.O. Box 78, Furlong, PA 18925, USA
- -Scott Walck
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UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Dear all
I may be dragging the thread away from contamination, but I thought that most electron microscope users avoided silicon based oils in diff pumps etc
because they are extremely difficult to remove from the interior of an electron microscope and any contamination would normally have electrical insulating properties (catastrophic in an e.m.). Perhaps I am wrong but I would welcome any comments. After all this is one of the reasons why Santovac oils and their relatives became so popular (despite their costs).
If I am labouring under a mis-apprehension then I apologise.
Malcolm Haswell University of Sunderland UK
All of the silicon based DP fluids I am aware of (which may not be all that are on the market) will break down under an electron beam and deposit a layer similar to glass on the nearest cool surface in the column. I don't know of any EM manufacturers that would recommend their use because they are almost impossible to remove if they do get in the column. We don't even use silicon based fluids in our vacuum evaporator.
K. A. Brackett, Ph.D. TN & Assoc./USEPA
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
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Petra;
I have a good reference that you might want to check out:
"Ruthenium Tetraoxide Staining of Polymers for Electron Microscopy", by J.S. Trent, et al, Macromolecules Vol. 16, #4, pp. 589- 1983.
This article is VERY informative.
Regards,
Bob *********************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com ***********************************
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Hello polymer experts,
new to the field of EM polymer blends I would like to ask you for the best way to prepare PE/PS blends for structure determination with TEM.
Of course the material has to be stained and cut, but:
- what is the best staining agent (OsO4, RuO4, or something else)? - is it better, first to stain and then to cut or the other way round? - which temperature is the best for cutting, liquid nitrogen or room temperature?
I would appreciate ever tip & trick you can give on handling this kind of material.
Tanks in advance
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
id NAA41274 (8.8.6/50); Thu, 23 Jul 1998 13:02:43 -0500 Message-Id: {199807231802.NAA41274-at-mail1.doit.wisc.edu} X-Sender: steve-at-facstaff.wisc.edu (Unverified) X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
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Ultrasonication has been a standard technique for degassing fluids in our labs for many years. (Before that we boiled where possible or pulled a gentle vacuum on a closed container. Main purpose was to degas solutions to be passed through automatic light blockage partcle counters. (Air bubbles count very nicely as particles.) Nice thing about sonication is that you can safely sonicate oils and other fluids that you wouldn't want to boil or pull into a vacuum system.
Bob Holthausen Pall Corporation
} } On Wed, 16 Apr 1997, Ian Montgomery wrote: } } } Like Rob I've been making Reynold's lead citrate since I was a boy. } } I use NaOH pellets but degas the distilled water by sonicating it for a few } } minutes before making the stain. } } } } Ian. } } Leo, If my memory serves I got sonication from the Technical Hints and Tips in the Proceedings of the RMS. I cover my options by sonicating the water whether fresh or hours, days old. Ian.
Steve Limbach Associate Researcher Bock Research Lab. 1525 Linden Dr.
UW-Madison Madison, Wisc. 53706
TEL 608 263-2582 FAX 608 262-4570 EMAIL slimbach-at- facstaff.wisc.edu
I would like to thank everyone who responded to my request of information about digital cameras. I am sorry that I did not reply to you individually. No more information, please!
Thank you very much for your kind help.
Hong You Dept. of Cell Biology University of Cincinnati
You may want to check with Bruce D. Newell, Scientific Imaging Systems, Eastman Kodak Co., 343 State Street, Rochester, NY, 14652. In his talk at Microscopy and Microanalysis '98, in Atlanta (9 AM, Tuesday, 14 July 1998), he showed a slide of a tiny Macbeth chart meant to be viewed under a light microscope. I don't think it is being produced commercial yet (I asked at the Kodak Exhibitor's booth).
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
On Thu, 23 Jul 1998 steve-at-facstaff.wisc.edu wrote:
} Hello all, } } We work as consultants for a computer imaging hardware and software } manufacturer's rep, and have had a rather unusual request. Perhaps someone, } either end users or vendors can assist. } } We are looking for the equivalent of a MacBeth Color Chart, typically used } in video, but this needs to be very small, translucent, and mounted on a } standard histo glass slide. Ideally, 1/32" square (1 mm x 1 mm OK) and it } would contain at least the primary colors, incl. black and white. I suspect } there may be a problem with ?translucent black and white. I think episcopic } illumination would also suffice for the chart, however so it could be } opaque. At this time, I do not have more detail on their exact application. } } We have checked with Munsell and MacBeth to no avail, so we would appreciate } any assistance, and even a referral as it's likely this is a custom } application. You can e-mail me directly if you like. } } Thanks in advance!! } } Daryl Martin } dmartin-at-ic.net } } (313) 213-8444 } } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu }
You may want to check with Bruce D. Newell, Scientific Imaging Systems, Eastman Kodak Co., 343 State Street, Rochester, NY, 14652. In his talk at Microscopy and Microanalysis '98, in Atlanta (9 AM, Tuesday, 14 July 1998), he showed a slide of a tiny Macbeth chart meant to be viewed under a light microscope. I don't think it is being produced commercial yet (I asked at the Kodak Exhibitor's booth).
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
} Hello all, } } We work as consultants for a computer imaging hardware and software } manufacturer's rep, and have had a rather unusual request. Perhaps someone, } either end users or vendors can assist. } } We are looking for the equivalent of a MacBeth Color Chart, typically used } in video, but this needs to be very small, translucent, and mounted on a } standard histo glass slide. Ideally, 1/32" square (1 mm x 1 mm OK) and it } would contain at least the primary colors, incl. black and white. I suspect } there may be a problem with ?translucent black and white. I think episcopic } illumination would also suffice for the chart, however so it could be } opaque. At this time, I do not have more detail on their exact application. } } We have checked with Munsell and MacBeth to no avail, so we would appreciate } any assistance, and even a referral as it's likely this is a custom } application. You can e-mail me directly if you like. } } Thanks in advance!! } } Daryl Martin } dmartin-at-ic.net } } (313) 213-8444
We are working with Bruce to provide the color chart commercially and look to have a product this fall. We will put a posting at our website when it is available:
{ {http://www.MME-Microscopy.com/education}
Please check in September. ... Thanks for the interest.
We are working with Bruce to provide the color chart commercially and look to have a product this fall. We will put a posting at our website when it is available:
{ {http://www.MME-Microscopy.com/education}
Please check in September. ... Thanks for the interest.
-----Original Message----- From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] Sent: Thursday, July 23, 1998 7:52 PM To: MICHAEL DELANNOY Cc: microscopy-at-Sparc5.Microscopy.Com Subject: Re: Floatin lung tissue
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On Wed, 22 Jul 1998 12:31:20 -0400 (EDT) MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: 1)leave in a refrig overnight until } they sink (swirling did not work). } 2)place in a vac (15 psi) while still in } the fix. } 3)process the floating samples hoping } they eventually sink. } I am currently in the frig (opting for #1) but any hints or } suggesstions would be welcome. Thanks. } } Mike D }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
A friend had this problem with plant material. She kept
the samples submerged under a piece of wire mesh.
Dave
Mike,
Place gauze over the top of the fixative and tissue, this prevents the tissue from rising to the surface and drying out.
-----Original Message----- From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] {mailto:[mailto:David.Patton-at-uwe.ac.uk]} Sent: Thursday, July 23, 1998 7:52 PM To: MICHAEL DELANNOY Cc: microscopy-at-Sparc5.Microscopy.Com {mailto:microscopy-at-Sparc5.Microscopy.Com} Subject: Re: Floatin lung tissue
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On Wed, 22 Jul 1998 12:31:20 -0400 (EDT) MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu {mailto:delannoy-at-welchlink.welch.jhu.edu} } wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } -----------------------------------------------------------------------. } } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: 1)leave in a refrig overnight until } they sink (swirling did not work). } 2)place in a vac (15 psi) while still in } the fix. } 3)process the floating samples hoping } they eventually sink. } I am currently in the frig (opting for #1) but any hints or } suggesstions would be welcome. Thanks. } } Mike D }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk {mailto:David.Patton-at-uwe.ac.uk} "University of the West of England"
A friend had this problem with plant material. She kept
the samples submerged under a piece of wire mesh.
Dave
Mike,
Place gauze over the top of the fixative and tissue, this prevents the tissue from rising to the surface and drying out.
-----Original Message----- From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] {mailto:[mailto:David.Patton-at-uwe.ac.uk]} Sent: Thursday, July 23, 1998 7:52 PM To: MICHAEL DELANNOY Cc: microscopy-at-Sparc5.Microscopy.Com {mailto:microscopy-at-Sparc5.Microscopy.Com} Subject: Re: Floatin lung tissue
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On Wed, 22 Jul 1998 12:31:20 -0400 (EDT) MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu {mailto:delannoy-at-welchlink.welch.jhu.edu} } wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } -----------------------------------------------------------------------. } } } To the lung experts, } What do you do with floating lung tissue (} 2mm) that is fixed } by immersion. The options are: 1)leave in a refrig overnight until } they sink (swirling did not work). } 2)place in a vac (15 psi) while still in } the fix. } 3)process the floating samples hoping } they eventually sink. } I am currently in the frig (opting for #1) but any hints or } suggesstions would be welcome. Thanks. } } Mike D }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk {mailto:David.Patton-at-uwe.ac.uk} "University of the West of England"
A friend had this problem with plant material. She kept
the samples submerged under a piece of wire mesh.
Dave
Mike,
Place gauze over the top of the fixative and tissue, this prevents the tissue from rising to the surface and drying out.
I have got some requests on a summary of the answers concerning my recently posted question. I wish to express my thanks for the three answers I received: --- } Try the very recent reference in } Journal of Histochemistry and Cytochemistry vol.46, No.3 (March 1998) } } "TUNEL Apoptotic Cell Detection in Tissue Sections: Critical Evaluation and } Improvement" by Labat-Moleur,Francoise et. al. in Grenoble, France. } } Raymond Koelling } Immunex Corp. } Dept. of Molecular Immunology } 51 University Street } Seattle, WA 98101 --- } Check out Boehringer Mannheim's website http://biochem.beohringer.com } It has a complete Guide to Cell Proliferation and Apoptosis Methods. } } Ronnie Houston } Cytochemistry & Molecular Pathology } Texas Scottish Rite Hospital for Children } 2222 Welborn Street } Dallas, TX 75219 --- } This is a protocol that I used a few years ago to check for } apoptosis. My samples were embedded in paraplast plus. } } TUNEL assay for paraffin embedded samples } } 1) Deparaffinze & rehydrate to buffer } 2) Proteinase K digestion 15 min } 3) Distilled water rinses 3X 5 min } 4) Air dry } 5) Reaction Mixture (8-10 ul under coverslip), humid, 37C 60 min } 6) Distilled water to remover the coverslip } 7) Distilled water wash 20 min } 8) 0.1M Sodium pyrophophate (make fresh) 10 min } 9) 0.1M Periodic acid (make fresh) 10 min } 10) Distilled water 5 min } 11) 2% BSA 20 min } 12) Horse radish peroxidase-avidin 30-45 min } 13) Tris buffered saline 3X 5 min } 14) DAB or AEC } } Counterstain with Methyl Green } } The reaction mix came from a Boehringer Mannheim Terminal transferase kit. } The Proteinase K was 10ug Pro.K/ml 50mM TRIS, pH 8.0, use 100 ul per section. } I used either the HRPO-avidin at 1:1000 with 2% BSA + 0.5M NaCl or a ready } to use solution of streptavidin-HRP from BioGenex. } The AEC came from a BioGenex substrate kit. } Reaction Mixture (use 10ul per section): } use a glass coverslip to cover the slide & incubate at a humid 37 degrees C } 100ul 5x TdT buffer (labelling buffer) } 100ul 1:4 diluted 25 mM CoCl2 } 10ul 1mM biotin-16-dUTP } 7.5ul TdT } 282.5ul distilled water } } This used to work very well for me, if you have any questions please feel } free to contact me. } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207
Thank you very much again.
-- Mit freundlichen Gruessen Yours sincerely ************************************************************** * PD Dr. T. J. Filler | specialist in anatomy * * Westfalian Wilhelms-Univ.| phone: *49 (0) 251 83 552 26 * * Institute of Anatomy | fax: *49 (0) 251 83 552 41 * * Vesaliusweg 2-4 | e-Mail: filler-at-uni-muenster.de * * D-48149 Muenster Germany | filler-at-medsnt01.uni-muenster.de * ****** http://medweb.uni-muenster.de/institute/anat **********
Hello out there all you overworked & underpaid microscopy people. Hey, if you think you have it bad what about us poor students? Rice and lentils aren't exactly a good diet you know! Anyway - can anyone tell me if it is possible to carry out a decent X-ray microanalysis of an SEM sample of tissue? I am trying to see if 'calcified' tissue really does contain calcium, and can't find out whether the electron penetration into the tissue will be deep enough to let me know if there is any calcium in there or not. I mean, will I simply get info regarding the surface cells and not the actual interstitium (which is where the good stuff is hiding). I can't strip the cells off either as this is valuable PM material and I have to do routine SEM on it as well.... Thanks for the help in advance!
Alex ---------- Alex Black Department of Anatomy National University of Ireland, Galway Ireland
I have a Mitsubishi P78U, large format black & white video printer connected to my Kevex 8005. It works well and has been trouble free. The catch is, when
most video printers are working well, the quality (resolution/tonal range) is sub-standard for demanding SEM imaging. If you are capturing NTSC video, the
vertical resolution will never be greater than about 500 lines for the entire image. Most digital systems capture 1024 lines (pixels) or more. My polaroids are typically exposed at 2000 lines. Horizontal video resolution is related to system bandwidth, but is seldom greater than the vertical res.
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I need information about video printers for printing SEM images and need to contact sales representatives from various companies. Please contact me at 217-782-0898. Thanks. Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il 62794-1220
by generic.axs2000.net (8.9.1/8.8.8) with SMTP id IAA15279 for {Microscopy-at-MSA.Microscopy.com} ; Fri, 24 Jul 1998 08:56:38 -0400 Message-Id: {199807241256.IAA15279-at-generic.axs2000.net} To: MICROSCOPY BB {Microscopy-at-Sparc5.Microscopy.Com}
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Maoxu Qian wrote: =================================================== I'm looking for carbon EM calibration grid from SPI model 411CG-AB but couldn't find any information about SPI.Does anyone knows they phone # or www address? ==================================================== I think I can reply with some element of authority to this question!
The above mentioned part number was for our now discontinued product, the carbon composite grid. Now I am sure I will be corrected quickly should I be wrong, but the one person in the world who could make this product stopped making it some several years ago. So these grids are just not available any more. Note: If anyone has any that are not going to be used, we would love to have them back since we do still get requests for them. PS : It was never intended to have any applications as a "calibration grid".
We did acquire a pretty good idea of why one would want to use this grid and have offered on our website a list of substitute grid products. So far as we know, one or more of the substitute products seems to have been quite acceptable (or better or cheaper) in just about any application to our knowledge that has been presented.
You can find us from the information given below. Hope this helps!
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 ############################ ==================================================
While some folk out there are talking knifemakers it might be useful for the LKB 7800 owners to have a look at our 1992 publication in J. Microscopy. In this short technical note we describe a series of simple modifications which transformed the performance of our two resident LKB's.
To quote from the conclusion;
'Analysis of the performance of an LKB 7800 series KnifeMaker revealed that its random failure was largely due to an accumulation of manufacturing tolerances, and of wear in the mechanism. After the playy was eliminated it was still necessary to adjust the cutter wheel positio accurately in order to obtain acceptable knives. With the modifications and adjustments outlined above, the number of acceptable knives that could be obtained from one strip of glass was improved byy at leasdt 100%.'
Don't throw it away - fix it !
Ref; Journal of Microscopy, Vol.168, Pt 1, October 1992, pp 111-114.
We had a, later, short paper on the advantages of applying tungsten coating to glass knives but I am sure that this is well documented.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Private Bag X01, Scottsville 3209, KwaZulu-Natal, South Africa. Tel +27 (0)331 260 5155, Home +27 (0)331 962676 Fax +27 (0)331 260 5776 email: bruton-at-emu.unp.ac.za
This bouncing mail is originating at the University of Wisconsin, Madison, they were also the culprits the last time a loop got started about 2 years ago.
I am attempting to contact the individual, however, looking through the mail it appears that some of these messages are at least a year old and are being resent from slimbach-at- facstaff.wisc.edu for unknown reasons. It is NOT something that I have direct control over as it originates the University of Wisconsin. Neither of the two addresses I have identified as the source are current subscribers to the Listserver.
In any event please DONOT send any mail or reply to to the addresses below as it may be reflected to the server
My grad. school diet was PBR and pizza... all of the basic food groups...:-)
-- Begin original message --
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Hello out there all you overworked & underpaid microscopy people. Hey, if you think you have it bad what about us poor students? Rice and lentils aren't exactly a good diet you know! Anyway - can anyone tell me if it is possible to carry out a decent X-ray microanalysis of an SEM sample of tissue? I am trying to see if 'calcified' tissue really does contain calcium, and can't find out whether the electron penetration into the tissue will be deep enough to let me know if there is any calcium in there or not. I mean, will I simply get info regarding the surface cells and not the actual interstitium (which is where the good stuff is hiding). I can't strip the cells off either as this is valuable PM material and I have to do routine SEM on it as well.... Thanks for the help in advance!
Alex ---------- Alex Black Department of Anatomy National University of Ireland, Galway Ireland
-- End original message -- regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
A Philips EM 301 is available as a gift to anyone who is willing to take responsibility for removing it (including packing and shipping costs). The instrument has been in use until recently and is basically in good shape. It does have a couple of minor problems which would require a routine service visit. I do not have the exact age of the instrument but Philips stopped making this model about twenty years ago.
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 7 - October 15, 1998
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2050 (Includes room and board)
Application Deadline: August 4, 1998
Admission application and information: Carol Harnel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratio-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual research problems with the faculty.
Software for Logon and Booking of Electron Microscopes.
Lehigh University Microscopy Center wishes to replace the software and hardware it currently uses for logging on and off the microscopes, and for booking sessions, keeping records and so on.
Please let me know if you are aware of any good system for this purpose. Windows preferred but a Mac system would be quite acceptable.
(People with a long memory may recall that I made a similar posting a few years ago while still in my previous job at Illinois. The result of that posting was a lot of interesting information but we did not locate a system which really seemed to do everything one would wish. We hope there has been a lot of progress in the last few years.)
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
At 12:35 PM 7/24/98 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alex,
Rice and lentils will keep you alive forever, especially if washed down with good Irish stout.
I assume you are doing your x-ray analysis in an SEM? If so, you will get information both from the surface and from some distance down into the tissue, depending on the accelerating voltage (kV) you use. The higher the voltage, the deeper you penetrate the tissue.
Unless you are using a spot analysis mode, your x-ray spectrum will give you all the elements in the visual field of view. (This is apparently what you plan, since if you can't see the precise areas you want to measure, you can't put a beam precisely on them.) The quantity of calcium in the tissue will be a major factor in whether or not you can see it. If it is there in tiny amounts (say less than 1%), coupled with the fact that it may be under an unknown thickness of non-calcium bearing cells, you may not be able to see it at all.
Definitely don't coat your tissue with Au or Au/Pd. If coating is necessary, use carbon. Or, if you have variable pressure capability, try using the lowest pressure (1-5 Pa, for example) that allows you to stop the charging effects. You lose capability for high-resolution imaging, but for low mag work this may not matter.
Finally, if your system can do element mapping, it might be fun to run a map for calcium. It just might see it and give you a visual reference to its location. Can't hurt to try.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Corneliu Sarbu National Institute for Materials Physics POBox MG-7 R-76900 Bucharest-Magurele Romania
are being returned as undeliverable. Does anyone have another email address for him? Or, if Dr. Sarbu is reading this, can you confirm your email address please: csarbu-at-alpha1.infim.ro ?
Thanks, Phil
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
Thanks for the generous offers of information, not only by e-mail but by fax and telephone. I have passed your replies on to the prospective user of the instrument (Mark Schmitt).
Leonard R. Corwin Fort Dodge Animal Health Cyanamid Agricultural Research Center PO Box 400 Princeton, NJ 08543-0400 609-716-2278; fax 609-275-5239 corwinl-at-pt.cyanamid.com
I have a student in the lab who needs to double label some TEM sections. I've never done this before so I'm turning to y'all for expert advice. He needs to do two different kinds of double labelling. 1) Label with two antibodies both raised in mice. 2) Label with one mouse & one rabbit anitibody.
We're not clear as to how to proceed so if anyone out there has a protocol that they could let him use or at least tell us what to do, that would be great! At least we have the different size gold secondaries.
Seeing double in Berkeley,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207
As I told you in a message to this group a few weeks ago, we are looking = at new SEMs and would like to look at fixed but wet tissue i.e. Fallopian = tubes to see if their microvilli and cilia are in good enough condition to = continue with more preparation or in the case of hair cells in the rat = cochlea, to see if they are sufficiently exposed or if part of the lateral = wall still needs to be dissected away. Surely some people out there use variable pressure or environmental = microscopes to look at wet, fixed tissues without introducing artefacts = from drying or freezing. I have been in contact with several microscope = representatives and am getting conflicting answers about what will and = will not work and how long a wet sample can be looked at in the scope = without drying out. I need to hear from people who have these microscopes = and work with wet tissues and cold stages. This is a big event for us. Our current SEM scope is 23 years old, so we = don't get the chance to make a purchase like this often. We want to be as = well informed as possible and at this point, I am very confused. Please = help me answer these questions: How long can the wet tissue be looked at in either type scope? Is a cold stage necessary and how long can cold, wet samples be viewed? What adverse affects on the tissue result from the freezing temperature of = the cold stage? Can tissue be looked at in either of these scopes and then be processed = for high vacuum SEM and observed at high vacuum and high resolution? Thank you very much for any advice you can give.
Donna Wagahoff SIU School of Medicine PO Box 19230 Springfield, Il. 62794-1220 217-782-0898 fax 217-524-3227
Would the person who posted the notice about spare Philips 300 parts please resend it with a valid email address. I'm interested, but replys get returned undeliverable.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Alex Black wrote: ================================================== ........ can anyone tell me if it is possible to carry out a decent X-ray microanalysis of an SEM sample of tissue? I am trying to see if 'calcified' tissue really does contain calcium, and can't find out whether the electron penetration into the tissue will be deep enough to let me know if there is any calcium in there or not. I mean, will I simply get info regarding the surface cells and not the actual interstitium (which is where the good stuff is hiding). I can't strip the cells off either as this is valuable PM material and I have to do routine SEM on it as well.... ==================================================== Assuming you have a large enough sample that you are not going to get good data by SEM/EDS from the interior of the sample, there is a technique that can be used that involves embedding, sectioning and then plasma etching the resin away in a plasma etcher. The analysis is done by analytical TEM.
You can see the end result of that kind of a sample prep on our website URL http://www.2spi.com/catalog/instruments/etchers4.html
The technique is fairly straight-forward: You will have to embed your sample, thin section it and pick it up on a silicon dioxide filmed grid. You then expose the (unstained) section to an oxygen plasma in a small low power (not more than 100 watts) plasma etcher, to remove all organics, leaving the non-organics scattered around, showing the structure almost as a "ghost" image. However, the beauty of the technique is that you have removed the carbon that would contribute to high Bremmstrahlung background radiation, increasing greatly then the sensitivity for what it is that you do want to detect.
But this will enable you to get closer to if not actually on the target of your objectives.
After removal of the organics, and this is all supported on a SiO2 filmed grid, the inorganics essentially outline a "ghost" structure of the original cell structure. So to whatever degree you need maximum sensitivity, your sample is now in that form where you could have the best shot at detracting what you want to detect.
There have been publications using the technique, I just don't have any of them at my fingertips at the moment.
Disclaimer: SPI Supplies manufactures the plasma etchers and produces the silicon dioxide filmed grids on a custom basis all of which can be seen on our website below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We are in the process of developing just such a chart in conjuction with Kodak. Prototypes have already been constructed (including a gray scale, which should satisfy your black and white requirements).
I will forward your request; perhaps we can arrange for you to test one.
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I suggest using some tool software to do such thing. Assuming that you have proper hardware, you can:
1. Dupe the original disk by Disk-duping software(such as HD-Copy) as a backup; 2. Use a disk diagnostic software(such as Norton's Norton Disk Doctor) to repair the original disk; 3. Then try to dupe the repaired original disk again to get a usable one.
But from my point of view, it would be no problem if you have "copies of copies" that can work well. However, disk-duping software is recommended because some commercial disk may be encrypted between disk tracks, which can not be replicated by COPY command.
Regards.
Dr Yanping Zhou(Celia)
Electronic Microscopy Group Shanghai Institute of Ceramics Chinese Academy of Sciences 1295 Dingxi Road, Shanghai 200050 P. R. China } } } We have a NORAN (Tracor Northern) 5400 Series II EDS system (Model } Number TN-5402/BBAA) acquired in 1987. It does not have a hard drive and } operates off of two floppy drives. It requires 5.25" double sided, high } density floppy disks, the originals of which are no longer readable. They } have not been in use other than to be the source of copies made } periodically but apparently they have simply deteriorated with time. } We have copies of copies that are working pretty well but I'm becoming } concerned that they may crash/die and we'll have no good source from } which to make new copies. NORAN does not have replacement masters of this } vintage so I'm hoping to find someone who has original master disks in } good working order who would be willing to loan them to be copied. } } We need two disks of the set from the November, 1987 Release 1C. What will } work with our system is very specific and the original label on the master } disks reads: } } SERIES II SOFTWARE RELEASE 1C } NOV., 1987, TRACOR NORTHERN, INC. } } Of the disks in this release, we need: } } SQ/SSQ/PRZ/ZAF Master } NOT BOOTABLE } } and } } MSCAN2 Master } } } Thanks in advance for any help or suggestions you may have. } } ___________________________________________________________________________ } Barbara Reine, Botany Dept. Box 351330 } Univ. of Washington, Seattle, WA 98195-1330 } e-mail: reine-at-u.washington.edu; ph: (206) 543-1955 } ____________________________________________________________________________ } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu } }
While investigating a problem last week with the aluminum foil backing on our manufacturing plant's ceiling tiles, I thought it would be expedient to put a relevant question to an "Ask the Experts" group I found through SPI's website. To the metallurgists of that group I asked why fine abrasive action on most if not all metals creates a black substance. I asked if that would be the reduced form of the metal. The problem our plant is having is that in handling these tiles, fingers become black and then smudge the white surface. I received a short reply from the webmaster that my question was outside the scope of their interest and would normally be deleted without comment.
So, microscopists, does anyone have an idea about the cause of the blackening? Reflected light at 100x or so isn't very illuminating.
While investigating a problem last week with the aluminum foil backing on our manufacturing plant's ceiling tiles, I thought it would be expedient to put a relevant question to an "Ask the Experts" group I found through SPI's website. To the metallurgists of that group I asked why fine abrasive action on most if not all metals creates a black substance. I asked if that would be the reduced form of the metal. The problem our plant is having is that in handling these tiles, fingers become black and then smudge the white surface. I received a short reply from the webmaster that my question was outside the scope of their interest and would normally be deleted without comment.
So, microscopists, does anyone have an idea about the cause of the blackening? Reflected light at 100x or so isn't very illuminating.
Dear Dave, } } To the metallurgists of that group I asked why fine abrasive action on most } if not all metals creates a black substance. I asked if that would be the } reduced form of the metal...
No, it is not reduced metal--at least in the case of aluminum, which oxidises readily on exposure to air or water. } } So, microscopists, does anyone have an idea about the cause of the blackening? My guess is that the size and shape of the particles does not re- flect light. This guess could be tested by microscopic examination to characterize the particle size and shape. This problem may have already been solved by the particle experts (among whom McCrone Associates are frequent contributers to the list).
} Reflected light at 100x or so isn't very illuminating. } Due to the low albido. :-) Yours, Bill Tivol
On Thu, 23 Jul 1998 steve-at-facstaff.wisc.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Good morning Reynolds users } } In nearly 30 years of preparing lead citrate according to } Reynolds I have experienced no problems in using a stock } sodium hydroxide solution (1M) which was made up from pellets. } However, this solution must be freshly made up. Is it because I } always use Reynolds in its concentrated form that I do not } experience any problems? } } Rob } } } Robin H Cross } Director : EM Unit, Rhodes University, Grahamstown, South Africa } eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377 } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu } } There are probably a number of things you are doing RIGHT. However, if the NaOH solution is not of the correct normality (1.0), you will have a shift in staining. If it is far off to the alkaline side, you will get essentially no staining. Too "acid" and you get at first intense stain and then dumping. I have 4 years worth of data on this. If you get the correct normality with pellets, fine. Just remember the fact (listed in the original Reynold's paper that variations in pH cause variations in stain intensity.) All you need to know is this concept. How you get there is really immaterial. Bye, Hildy
P.S. I personally take the liberty of "diddling" with the pH of the Reynolds lead citrate. I know exactly how "acid" I can go to increase my intensity, if I want. I do this easily with a commercial source of NaOH which is titrated to the exact known normality.
On Thu, 23 Jul 1998 steve-at-facstaff.wisc.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Would freshly distilled water be equivalent to degassing via sonication ? } } Leo } } On Wed, 16 Apr 1997, Ian Montgomery wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } -----------------------------------------------------------------------. } } } } } } Good morning Reynolds users } } } } } } In nearly 30 years of preparing lead citrate according to } } } Reynolds I have experienced no problems in using a stock } } } sodium hydroxide solution (1M) which was made up from pellets. } } } However, this solution must be freshly made up. Is it because I } } } always use Reynolds in its concentrated form that I do not } } } experience any problems? } } } } } } Rob } } } } } Like Rob I've been making Reynold's lead citrate since I was a boy. } } I use NaOH pellets but degas the distilled water by sonicating it for a few } } minutes before making the stain. } } } } Ian. } } } } } } } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu } } Hi,
Guess what folks? I have been making Reynolds Pb citrate for 7 years without boiling, degassing water, etc. If you take care to chealate the Pb and citrate correctly, carbon dioxide just does not turn up as a problem. Now, end of Pb discussion! So long, Hildy Crowley
P.S. I never let students make Pb which I plan to use. I always make my own, because too many "inventions" without having studied the Reynold's paper lead to morasses.
I would need a 2.5 x microscope objective to fit a Diavert (a discontinued Leitz model; mine is about 12 years old). The tube length is 170 mm, with a DIN (45 mm) distance between slide and turret. If any of you on the list can supply this accessory, I would be interested in purchasing.
When a metal is finely divided the particles form a black powder. This is solely an optical problem, and not a materials problem. You might want to coat the aluminum with a thin coat of polymer to eliminate the abrasion.
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} While investigating a problem last week with the aluminum foil backing on our } manufacturing plant's ceiling tiles, I thought it would be expedient to put a } relevant question to an "Ask the Experts" group I found through SPI's website. } To the metallurgists of that group I asked why fine abrasive action on most if } not all metals creates a black substance. I asked if that would be the reduced } form of the metal. The problem our plant is having is that in handling these } tiles, fingers become black and then smudge the white surface. I received a } short reply from the webmaster that my question was outside the scope of their } interest and would normally be deleted without comment.
My understanding of this phenomenon (whereby any bright, shiny metal- including platinum-when ground finely enough gives a black rather than shiny powder) is not due so much by oxidation but to the fact that the metals have been ground in the micrometer range. Such particles no longer reflect the light in an ordered manner (like a mirror) back to the eye but in fact bend it in so many directions that the light is reflected mostly away from the eyes and gives the impression of absorbing rather than reflecting light. A good example of this is the photographic negative. The negatives have metallic (reduced) silver but it is of such a fine order (micrometers) that the negatives look very dark rather than shiny. When one rubs their hands over soft metals (like aluminum) you abrade off very fine flecks of the metal (micrometer range) that give your hands the blakened appearance. Harder metals (stainless steel for example) would do the same except that they do not abrade as easily.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 ####################################################################
Steve, Use either the Mg or Ca (*2-3-mM) in the fix and buffers (for maintaining membrane tonicity and keeping those myelin figures to a minimum). Also try some reduced osmium, it may make the membranes stand out, as will a good Ua/Pb (preferable citrate and nitrate) section stain.
I apparently accidently forwarded some junk email to the microscopy server. If you received any junk email from this address, MY SINCEREST APOLOGIES.
I realized I had accidentally hit the "forward" button on my email program, and hit the reset button ASAP. I guess I didn't catch it in time. I am not affiliated with this group that originally sent the mail, and I hate spam as much as the next guy. SORRY.
} I suggest that it may be necessary for } Nestor to configure the software that is used for this list to prevent } posting from non list members. } } What do other people think. Ian:
Well put, Ian. I agree. The sex post was the pits.
Blystone in Texas
Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu} Professor of Biology Trinity University San Antonio, Texas 78212 210.736-7243 210.736-7229 FAX
I am looking for a person with extensive FIB and TEM sample prep experience
to fill a position at the IBM Almaden Research Center. The work would involve extensive collaboration with the groups at Almaden and in the IBM Storage Systems Division that are developing next-generation heads and disks, photoresists, low dielectric constant materials, and MRAM.
Candidates should have experience in as many of the following areas as possible: FIB, TEM sample prep - especially tripod polishing and micromanipulation, TEM, and SEM.
Please contact me directly. Submit your resume to: Robby Beyers K19/D1 IBM Almaden Research Center 650 Harry Road San Jose, CA 95120-6099
I am finishing my master (a pre-embedding study on serotonin) and before=20 i am trying to localise serotonin in a cnidarian tissu by post-embedding.= =20 My tissues are embedded in lowycril and are well preserved. I have no=20 problem in cutting them and I recolt them on nickel grids with formvar.=20 THE PROBLEM is the immuno in itself where I have way to much back ground=20 noise everywhere even on the formvar. My antibody is diluted 1 in 500 or=20 in 1000 or in 2000 for 4 hours and the secondary 1 on 75 for 1 hour,=20 centrifuged before use.=20 Those solution are rinsed and diluted in PBS .1M, BSA .5%, and .05%tween=20 20 (I have tried 1% cold water gelatin), and for the pre-incubation I=20 add10% normal goat serum. I simply transfer my grids from one droplet to=20 the other. As well, before preincubating, I rinse my tissues in water and= =20 sodium metaperiodate saturated for 30 min.
However whatever I try I don't get rid of this background noise. =20 HELP!!!!!!!!!
THANKS
NATACHA BENRIMOH UNIVERSIT=C9 DE MONTREAL DEPARTEMENT DE BIOLOGIE (514) 343-6111 #1052 benrimon-at-ere.Umontreal.ca
Dave, the black material is aluminum oxide that forms on all untreated aluminum, which is being wiped off by handling and transferred to the cosmetic part of the tile. Possible solutions would be another handling method or overcoating the aluminum somehow. We use disposable cotton glove liners to keep our hands clean(er), but that probably won't keep the tiles clean. Best of luck,
Vern
Vern Rieck Quality Engineer Aavid Thermal Products, Inc. rieck-at-aavid.com
-----Original Message----- From: David_R_Stadden-at-Armstrong.com [SMTP:David_R_Stadden-at-Armstrong.com] Sent: Monday, July 27, 1998 9:05 AM To: Microscopy-at-sparc5.microscopy.com Subject: Black metal particles
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
While investigating a problem last week with the aluminum foil backing on our manufacturing plant's ceiling tiles, I thought it would be expedient to put a relevant question to an "Ask the Experts" group I found through SPI's website. To the metallurgists of that group I asked why fine abrasive action on most if not all metals creates a black substance. I asked if that would be the reduced form of the metal. The problem our plant is having is that in handling these tiles, fingers become black and then smudge the white surface. I received a short reply from the webmaster that my question was outside the scope of their interest and would normally be deleted without comment.
So, microscopists, does anyone have an idea about the cause of the blackening? Reflected light at 100x or so isn't very illuminating.
steve-at-facstaff.wisc.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear all } } I may be dragging the thread away from contamination, but I thought that } most electron microscope users avoided silicon based oils in diff pumps etc } } because they are extremely difficult to remove from the interior of an } electron microscope and any contamination would normally have electrical } insulating properties (catastrophic in an e.m.). Perhaps I am wrong but I } would welcome any comments. After all this is one of the reasons why } Santovac oils and their relatives became so popular (despite their costs). } } If I am labouring under a mis-apprehension then I apologise. } } Malcolm Haswell } University of Sunderland } UK } } All of the silicon based DP fluids I am aware of (which may not be all that } are on the market) will break down under an electron beam and deposit a } layer similar to glass on the nearest cool surface in the column. I don't } know of any EM manufacturers that would recommend their use because they } are almost impossible to remove if they do get in the column. We don't } even use silicon based fluids in our vacuum evaporator. } } K. A. Brackett, Ph.D. } TN & Assoc./USEPA } } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu
Malcom, I've been watching this thread with considerable interest, also. ETEC sold almost every one of its microscopes with Transene Vacoil (later Vacoil-S) which I believe is redistilled Dow-Corning 705. I have never had any problems with it in 21 years of servicing these instruments. Even a burped DP (and I've dealt with my share) has never been a problem beyond the fact that you have to clean everything. Many ETECs were sent out with the water-cooled baffle plumbed in just before the DP rather than before the power supplies. Those instruments will condense MP oil on their EDS detectors until they get replumbed. but silicone doesn't seem to present any problems.
Ken Converse owner Quality Images third party SEM service Delta, PA
steve-at-facstaff.wisc.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } I find oil on the EDX detectors in both my SEM's, but if you think of the } conditions inside the SEM, it is almost inevitable. The majority of the } molecules remaining in the vacuum of an average SEM at 10 -5 torr consists } of diffusion pump oil. This oil will gradually condense on all surfaces in } the SEM, but on the coolest surface first. In most SEM's this is the EDX } snout. One EDX manufacturer has solved the problem by warming up the snout } with a small heater. The oil just goes somewhere else, but if it forms a } thin film over all surfaces it is not so visible or worrying. } The cleaning method recommended by the manufacturer of my EDX's is to gently } run clean-grade iso-propanol over the snout, then allow to air-dry. They } used to recommend Freon, but that is no longer permitted or available. } Hope this helps, } Mary } } Mary Mager } Electron Microscopist } Metals and Materials Eng., UBC } 6350 Stores Rd. } Vancouver, B.C. V6T 1Z4 } CANADA } tel:604-822-5648, fax:604-822-3619 } e-mail: mager-at-unixg.ubc.ca } } Steve Limbach } Associate Researcher } Bock Research Lab. } 1525 Linden Dr. } } UW-Madison } Madison, Wisc. 53706 } } TEL 608 263-2582 } FAX 608 262-4570 } EMAIL slimbach-at- facstaff.wisc.edu
Mary, If you system is properly trapped, you shouldn't get any condensation, or at least very little. My experience has been that the oil is often mechanical pump oil, not DP oil, and may arise from valving over at too low a pressure, allowing considerable backstreaming of MP oil.
Ken Converse owner Quality Images third party SEM service Delta, PA
Our Hitachi 2460N variable pressure SEM has always been condensing oil on our detector snout. Typically, we have a large drop form every month. It has bothered me (a lot of things do these days) but everyone says that it is to be expected. I don't think so.
} If you system is properly trapped, you shouldn't get any condensation, } or at least very little. My experience has been that the oil is often } mechanical pump oil, not DP oil, and may arise from valving over at too } low a pressure, allowing considerable backstreaming of MP oil.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I suspect that much of your problem is due to the Formvar. Try doing the immunolabeling on naked gold grids. You can get more support by using a fairly high hexagonal mesh (300 mesh). Formvar is *very* sticky for antibodies and tracers!
If background is still high, try using a hypertonic saline rinse, about 5X normal (750 mM NaCl) every time you need to rinse. Just be sure to use a normal saline rinse or two before you continue with the labeling protocol.
Good luck, let us know how things work out!
Best regards,
Bob ***************************************** Robert (Bob) Chiovetti rchiovetti-at-aol.com E. Licht Company / 1-800-865-4248 Colorado/Utah/Wyoming/Arizona/ New Mexico/West Texas U.S.A. Representing Leica Since 1967 *****************************************
In my service days pretty well all the data that we gained by carrying ou= t an analysis on the detector oil droplet was that it is rotary pump fluid.= I should say even now I hardly see a microscope without this droplet on the=
EDX snout.
It was no good just changing to another fluid as the complete vacuum syst= em would be "contaminated" by the fluid. To start again required every part=
of the system to be cleaned, even rotary pump lines.
A system that has not been used could be cleaned up from day one using activated charcoal in a fore line trap. Try to help a used system withou= t a 100% clean up and you gain nothing.
Steve Chapman Senior Consultant =
Protrain for electron microscopy training and consultancy world wide see http://ourworld.compuserve.com/homepages/protrain
A student in my lab will be travelling to the UK from Australia to attend a conference next year in march. He would like to visit other labs who are working on citrus. He is making a study of oleocellosis a disorder related to damage to oil glands in the rind of the fruit during postharvest handling. He is currently undertaking a developmental study of oil gland development. Anyone able to help with interpretation of sectioned material would be ideal. He is preparing material for LM and TEM, hopes to use confocal for 3D reconstructions too.
replies to tknight-at-waite.adelaide.edu.au (or to me)
thanks
Meredith Wallwork
Dept Horticulture Viticulture and Oenology Waite Campus University of Adelaide South Australia AUSTRALIA
22 years ago I arrived at a new lab that had just bought a new Siemens Elmiskop102 TEM (their last model produced) and an ETEC Autoscan. Siemens, to the end recommended Silicon fluid for the pump system. By then it was "common" knowledge that silicone fluids caused column contamination problems. I changed both instruments fairly soon to Santavac 5. It is news to me that the Etec also contained silicone, it was just simpler to use one fluid in both instruments.
Point is some manufacturers are slow changing to new realities, and Old EMists spent horrendous time just keeping instruments in reasonable conditions. A lot more time was spent cleaning apertures and columns and adjusting the scope to be astigmatic twenty and more years ago. Instrument maintenance now is (comparatively) a snap. The ETEC is now quite geriatric, but if you are using a silicone based fluid it's time to change and lower maintenance time and increase average performance. It's easy to test if your vacuum fluid contains silicone; just put a drop onto a SEM stub, centre the stage and give it a few seconds analysis under an EDS. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************* www.proscitech.com.au *****
At 06:31 PM 7/27/98 -0600, John J. Bozzola wrote: } Our Hitachi 2460N variable pressure SEM has always been condensing oil on } our detector snout. Typically, we have a large drop form every month. It } has bothered me (a lot of things do these days) but everyone says that it } is to be expected. I don't think so. } } } If you system is properly trapped, you shouldn't get any condensation, } } or at least very little. My experience has been that the oil is often } } mechanical pump oil, not DP oil, and may arise from valving over at too } } low a pressure, allowing considerable backstreaming of MP oil.
We have a 2460N in our lab. Our standard procedure is to leave the system sucking air at 40 Pa during its idle times. This serves to maintain viscous flow conditions (proper term?) under which the oil is swept from the system. We have not yet had to clean our EDS detector during the several years we have been running it, and we still get good low energy peaks.
Meanwhile, we have a JEOL 840A with light element EDS at the other end of the hall and we see droplets form on it quite quickly. We lose more than half of our O intensity over a 6 month period and need to reclean.
BTW, I understand the viscous flow principle is the idea behind Ronald Vane's system for keeping columns clean. We haven't bought his system yet, but would definitely recomend considering it if the cleaning gets to be too bothersome.
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
A few days ago I posted a metallurgical question to the group in which I referred to an "Ask the Experts" group I'd found "through SPI's website". My message indicated a lack of success with that forum, but I want to make it clear that this was simply a link that I'd arrived at through SPI's pages and was not the SPI folks themselves. I've always been impressed with SPI (and certainly their web presence) and in no way wanted to reflect adversely on them.
Thanks to all who responded to my question; your feedback was a big help.
Dave Stadden Armstrong World Industries, Inc. Testing & Analysis Lab
Since the subject of Reynold's stain has come up recently I have a question. My stain performs well (I think) but with time more and more crystals precipitate out of solution onto the botton of the flask. Is this bad? Am I doing something wrong? The stain is made in the traditional way and is stored in a volumetric flask sealed with Parafilm in the 'fridge.
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 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Has anyone tried using foreline traps to reduce oil buildup on EDS detectors? I have been thinking about trying one of the units made by Taylor Engineering that use copper maze absorbent. Stanley L. Flegler Center for Electron Optics Michigan State University
It seems that there is some confusion over my previous post. Let me try to clarify here.
At 12:59 PM 7/28/98 -0500, someone wrote: } I have a question in respect of your comment: } } Meanwhile, we have a JEOL 840A with light element EDS at the other end of } } the hall and we see droplets form on it quite quickly. We lose more than } } half of our O intensity over a 6 month period and need to reclean. } } Do you use the 40Pa flow of "air" (not N2?) on the 840A as well? If not, is } there any particular reason not to do it? If yes, why do you think it is } worse than your 2460N?
Our Hitachi 2460N is a "leaky" or environmental SEM (hence the N designation), while our JEOL 840A is conventional "high vacuum-only" SEM. Both are working just as designed in their day. So while we have a choice of pressure and gas for the sample chamber of the Hitachi, we have no such choice of vacuum level for the 840A. I rather wish we did, but at this time that would require a third party solution, hence my prior reference to XEI.
As far as why air and not N2, we normally run our 2460N chamber with helium for the reduced scattering that it affords at a given vacuum level. We simply pull the He hose from the SEM inlet filter barb when we are ready to leave the scope for the night. I suppose were we purists or wealthier we might just leave the He hooked up all the time.
I hope this helps clarify. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
After you make up your Reynolds, aliquot it into 1.5 ml eppy/microfuge = tubes and store in the frig. When you need it centrifuge a tube at = medium to high speed for 5 or so minutes and use. The solution when = stored in microfuge tubes will last several months. The lead citrate = solution from what I understand reacts with the glass somehow and ppt = outs.=20
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
} Has anyone tried using foreline traps to reduce oil buildup on EDS } detectors? I have been thinking about trying one of the units made by } Taylor Engineering that use copper maze absorbent. } Stanley L. Flegler } Center for Electron Optics } Michigan State University
FWIW, we have a trap on the foreline of the rotary pump on our JEOL 840A for a couple of years. It may have helped some, but it has in no way stopped the oil condensation on our detector.
Is it possible to set up darkfield illumination without the expense of a whole different condenser? I have tried knife edge illumination (lying a sharp-edged on the (in my case) light source condenser on the base of the microscope, so that the knife (or card stock?) edge runs exactly through the axis of the light path). I thought of painting a black disk on the center of a transparent filter.
I'd be grateful for any useful suggestions. (I am using a Zeiss standard 16 with an na 1.35 condenser).
Alan -- Alan E. Davis Marianas High School (Science Department) AAA196, Box 10001 adavis-at-netpci.com http://www.saipan.netpci.com/~adavis Saipan, MP 96950 15.16oN 145.7oE GMT+10 Northern Mariana Islands
Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
Dear Sirs: I would like to apply for a post-doctor position in your laboratory. Now I have graduated as a Ph.D in Institute of Physics, Chinese Academy of Sciences. My study has focused on:
(1) The High Resolution Electron Microscopy investigation on the structure of NiFe/Mo and Fe/Mo multilayers (relating to GMR effect).
(2) The microstructure inverstigation on C ions-implanted silicon by HREM.
I would appreciate it very much if you would supply such a position for me in your laboratory.
} Since the subject of Reynold's stain has come up recently I have a } question. My stain performs well (I think) but with time more and more } crystals precipitate out of solution onto the botton of the flask. Is } this bad? Am I doing something wrong? The stain is made in the } traditional way and is stored in a volumetric flask sealed with Parafilm } in the 'fridge. } } Geoff McAuliffe, Ph.D.
---------------------------------------------------------------------------- -------- The problem is with atmospheric carbon dioxide. Every time you open the flask to remove some stain you introduce CO2. This reacts with the Lead Citrate in solution to give Lead Carbonate which is insoluble.
When you make up Lead Citrate it is important to use CO2 free water, either freshly produced from a high quality deioniser or boil distilled/deionised water and allow it to cool. Sonication will not remove dissolved CO2 as the gas has formed a low concentration of Carbonic acid in the water :- CO2 + H2O = H2CO3 ( H+ & HCO3 - )
To keep Lead Citrate, aliquot it into smaller containers like small vials or tubes and store in the fridge. Then use a new one each time you stain.
We have found that boil off from liquid nitrogen is by far the best venting material. It is dry, it is clean, it is cheal.We hane the whole lab' plumbed with a line going back to a 100litre Dewer which if I remember lasts for 3-4 weeks. You obviosly have to be careful with pressure. There is nothing like getting a 100lb SEM stage whislting out of the microscope and landing on your lap. Really brtings tears to your eyes.
Patrick Echlin Multi-Imaging Centre University of Cambridge.
On Wed, 29 Jul 1998, Trevor Sewell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } } }
Either proposition will work. Both means require a bit of maintenance. I would be inclined to use nitrogen; it's drier and cleaner. The cost is mostly in the rental of the cylinder and not the gas. I used a full N2 cylinder to run some pneumatic EM valves and instrument backfilling. When the high pressure cylinder ran low, it was switched to serve for gas agitation during film development. The low pressure cylinder was always available to exchange for a full cylinder when required without interrupting EM operations. Over-pressurising is avoided by not relying on a timer to backfill the column but the use of a demand valve (from a diver shop) this type of valve delivers gas only while the diver, or the microscope "sucks". A "T" junction with a needle outlet near the cylinder could also be useful to dust-off particles from microscope parts etc. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
Hi Trevor, This is a great topic for discussion as there are quite a few factors = that come into play. Basically the main criteria for any vacuum system is to keep the = atmosphere in it constant and clean. By venting and opening the door of = the chamber in a SEM to change samples obviously throws that criteria = out the window.=20 As is always the case with microscopy we have to make compromises. The = best solution would be to fit an airlock to the chamber. In this way you = would maintain a better vacuum, however then you are limited to sample = size and it's expensive to buy an airlock. The second option is to back fill with dry nitrogen gas. Nitrogen being = the gas that is most prevalent in the air that we breath and that vacuum = manufactures have therefore designed their vacuum pumps and gauges = around.=20 By back filling with dry nitrogen you are maintaining a fairly constant = atmosphere to pump after venting and changing the sample. Maintaining = the positive pressure in the chamber, as you open the door, keeps the = surrounding atmosphere out. Yes, the negative side to this is the danger = of putting to much pressure on the window of the ED detector and = therefore blowing it. This can be prevented by simply removing the latch = that hold the door closed.=20 The surface area of the door is much larger than the area of the window, = so the door will be forced open far before the pressure on the window of = the ED detector suffers. In fact on normal W filament systems where you = vent the entire column, I have seen the gun popping up and down due to = dry nitrogen backfilling, when the door latch is still on. No damage to = the ED detector. As mentioned the vacuum components are also suited to Nitrogen. Pirrani = and penning gauges are calibrated on Nitrogen pressures, so by using = nitrogen you have a better chance of attaining the same readings each = time you pump again. The Turbo molecular pump, in particular, prefers to = pump nitrogen and is thus more efficient. Water vapour is particularly = difficult for a turbo pump to pump. So if your EM is situated near the = coast or you have a lot of heavy breathers using the EM, the humidity in = the lab will change on an hourly, daily and seasonal basis. On a bad = day, with high humidity, the pumping speed of the chamber to reach VAC = OK status, could decrease in terms of 5 to 10 minutes from a good dry = day.=20
Thus in conclusion ( yes I eventually got there), to backfill with dry = nitrogen improves pump down speed, cleanliness of the system, stability = and repeatability of the required vacuum and prolonged life of the = vacuum pumps. This will be very helpful if you require high resolution = work or very accurate ED analysis from your system on a regular basis. = There is also no need to purchase an air lock for your system. On the down side, as there always is in microscopy, there is the cost of = the nitrogen, ensuring the door latch is not left on to prevent ED = window damage ( however I am not aware of anyone where this has = happened. ), changing samples in a hurry so as not to waste too much = Nitrogen and the hassle of making sure you have a supply of Nitrogen = handy all the time.=20 In my opinion, a SEM that is totally vented each time you need to change = samples, i.e. vent the column , gun and chamber, and is situated near = the coast or any other high humidity area, should definitely use dry = nitrogen backfilling. If you are simply venting the chamber but wish to do high resolution = work or stable ED analysis, I would still look at dry nitrogen = backfilling. If you are analysing or looking at really foul samples at low mag any = way, I would not bother.
That's it from me. I hope it helps and good luck.
Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
-----Original Message-----
Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
I've had some problems sending the microscope stage to positions already visited and stored using the ISIS "autorun" program. Indeed, I almost observe a "shift" of about 20 microns which means that the stage never comes back to its original position.
Suppose the stage is located by X1 Y1 and I then move to another location X2 Y2. If I then come back to X1 Y1, the image has shifted by about 20 microns to X3Y3. However if I change location and want to come back to X1Y1, the stage accurately comes back to X3Y3 each time...
Though I have never quantified it, using dry nitrogen seems to speed pumpdown somewhat, likely by minimizing water entry into the vacuum system. It is of no help, of course, when I get the ocassional specimen which outgasses significantly.
I have bottled nitrogen in the lab, but instead use the gas take-off provided on the LN2 cryo which is also in the lab.
Over pressure is not a problem so long as I don't hold the chamber door shut when venting....
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
(SMTP Gateway for microscopy-at-sparc5.microscopy.com); Wed, 29 Jul 1998 09:03:29 -0400 Message-Id: {199807291303.AA11170-at-gateway.ppg.com} Received: by gateway.ppg.com (Protected-side Proxy Mail Agent-1); Wed, 29 Jul 1998 09:03:29 -0400
----------
Position Open -- Immediately
Microscopy and Microanalysis Research Assistant
An immediate position is open for an SEM and TEM microscopist at the North Carolina State University Analytical Instrumentation Facility (AIF) as a retirement replacement.
Duties and responsibilities include: Teaching of laboratories; user training and assistance; and instrumentation development, modification and maintenance; and analysis of a wide variety of samples. An MS or equivalent experience in a materials related discipline along with three
or more years hands on experience with SEM and/or TEM is required. Required skills include: extensive hands on experience with SEM, TEM and
related techniques and accessories (e.g. specimen preparation and associated tools such as evaporators, etc.); teaching/user training; and
familiarity with modern electronics, computer systems 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 118A EGRC; 1020 Main Campus Drive; Raleigh, NC 27695-7531 or email PRUSSELL-at-NCSU.EDU.
North Carolina State University is an Equal Opportunity, Affirmative Action Educator and Employer. Minority and Female Applicants are especially encouraged.
Inrormation about our lab can be found at http://spm.aif.ncsu.edu/aif/
-- Phillip E. Russell Professor, Materials Science and Engineering Director, Analytical Instrumentation Facility Box 7531, Room 318 EGRC 1010 Main Campus Drive North Carolina State University Raleigh, NC 27695-7531
Hi Alan, I have made perfectly acceptable darkfield-like micrographs by simply introducing a central stop in the light pathway at the condenser level exactly as you suggest. One time for a course I was taking, I used a dime to block the central part of the collumated light beam. This was a long time back so I don't recall exactly where I placed it but what-the-heck. Experiment a little. John
________________________ C. John Runions, Ph.D. Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
I agree that boil off from LN2 is the best backfill. I used to use it extensively when I was working with UHV systems. There is absolutely no H2O in it. It is the driest nitrogen that you can get and there is also very little O2. LN2 is usually very available in an electron microscopy lab that has EDS detectors on the instruments and it is cheap. However, you don't have to have the boil off from a large tank and the plumbing that goes with it or the overpressure danger. An open container of LN2 will suffice. It is at atmospheric pressure and therefore will not overpressure your vacuum system. Make sure that the tube that you stick in is long enough so that the liquid that is drawn out is converted to gas before entering the vacuum system. Also, make sure that the tube doesn't go to the bottom of your dewar, otherwise it will suck up debris.
Incidentally, for safety reasons, you should not use a heater in the bottom of a dewar that can be pressurized. You should also not pressurize a LN2 dewar with air. Liquid oxygen can form at the bottom of the dewar over time. But following this thread, avoid overpressuring the vacuum system. The open LN2 approach does that.
Nitrogen is pumped fairly well by all vacuum pumps, especially ion pumps. The rated pump speeds are all referenced to N2. There are other gases in filtered air that become significant partial pressures in the ultimate vacuum because of their lower pump speeds.
Use LN2, your vacuum system will love you for it.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
We have found that boil off from liquid nitrogen is by far the best venting material. It is dry, it is clean, it is cheal.We hane the whole lab' plumbed with a line going back to a 100litre Dewer which if I remember lasts for 3-4 weeks. You obviosly have to be careful with pressure. There is nothing like getting a 100lb SEM stage whislting out of the microscope and landing on your lap. Really brtings tears to your eyes.
Patrick Echlin Multi-Imaging Centre University of Cambridge.
On Wed, 29 Jul 1998, Trevor Sewell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } } }
I've always recommended backfilling with N2 at about 1-2 lbs. over atmospheric pressure. I'm from the better-safe-than-sorry school and prefer to keep my chamber as clean as possible. As far as overpressurization, it hasn't been a problem with our Amray scopes because the stage/door is not locked to the chamber. At atmospheric pressure, there is sufficient leakage past the O-ring seal to prevent damage to our thin window EDS. I can only recall one incident in the past 11 years (more than 100 microscope-years) when N2 broke a window. That SEM was used by many people, was not well maintained, had undiagnosed problems with the N2 purge (more flow! decrease cycle time!) and a door that latched to the chamber.
My 2 cents
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
How often are you venting the chamber ? We have found on our LEO 440 that venting with nitrogen takes a long time (up to 30 minutes !!) (Maybe we're doing something wrong, Luc?). Although the pump down time decreases, you're probably going to lose more time venting than you'll gain with the decreased pump down time if you're changing samples often. Venting with nitrogen will keep the SEM cleaner, though, if time is not a problem.
Charles Bushell Mintek
} -----Original Message----- } From: Luc Harmsen [SMTP:anaspec-at-icon.co.za] } Sent: 29 July 1998 02:35 } To: 'Trevor Sewell'; 'MSA listserver'; 'MSSA listserver South Africa' } Subject: RE: Venting SEM chambers with bottled nitrogen or air } } Hi Trevor, } This is a great topic for discussion as there are quite a few factors that } come into play. } Basically the main criteria for any vacuum system is to keep the } atmosphere in it constant and clean. By venting and opening the door of } the chamber in a SEM to change samples obviously throws that criteria out } the window. } As is always the case with microscopy we have to make compromises. The } best solution would be to fit an airlock to the chamber. In this way you } would maintain a better vacuum, however then you are limited to sample } size and it's expensive to buy an airlock. } The second option is to back fill with dry nitrogen gas. Nitrogen being } the gas that is most prevalent in the air that we breath and that vacuum } manufactures have therefore designed their vacuum pumps and gauges around. } } By back filling with dry nitrogen you are maintaining a fairly constant } atmosphere to pump after venting and changing the sample. Maintaining the } positive pressure in the chamber, as you open the door, keeps the } surrounding atmosphere out. Yes, the negative side to this is the danger } of putting to much pressure on the window of the ED detector and therefore } blowing it. This can be prevented by simply removing the latch that hold } the door closed. } The surface area of the door is much larger than the area of the window, } so the door will be forced open far before the pressure on the window of } the ED detector suffers. In fact on normal W filament systems where you } vent the entire column, I have seen the gun popping up and down due to dry } nitrogen backfilling, when the door latch is still on. No damage to the ED } detector. } As mentioned the vacuum components are also suited to Nitrogen. Pirrani } and penning gauges are calibrated on Nitrogen pressures, so by using } nitrogen you have a better chance of attaining the same readings each time } you pump again. The Turbo molecular pump, in particular, prefers to pump } nitrogen and is thus more efficient. Water vapour is particularly } difficult for a turbo pump to pump. So if your EM is situated near the } coast or you have a lot of heavy breathers using the EM, the humidity in } the lab will change on an hourly, daily and seasonal basis. On a bad day, } with high humidity, the pumping speed of the chamber to reach VAC OK } status, could decrease in terms of 5 to 10 minutes from a good dry day. } } Thus in conclusion ( yes I eventually got there), to backfill with dry } nitrogen improves pump down speed, cleanliness of the system, stability } and repeatability of the required vacuum and prolonged life of the vacuum } pumps. This will be very helpful if you require high resolution work or } very accurate ED analysis from your system on a regular basis. There is } also no need to purchase an air lock for your system. } On the down side, as there always is in microscopy, there is the cost of } the nitrogen, ensuring the door latch is not left on to prevent ED window } damage ( however I am not aware of anyone where this has happened. ), } changing samples in a hurry so as not to waste too much Nitrogen and the } hassle of making sure you have a supply of Nitrogen handy all the time. } In my opinion, a SEM that is totally vented each time you need to change } samples, i.e. vent the column , gun and chamber, and is situated near the } coast or any other high humidity area, should definitely use dry nitrogen } backfilling. } If you are simply venting the chamber but wish to do high resolution work } or stable ED analysis, I would still look at dry nitrogen backfilling. } If you are analysing or looking at really foul samples at low mag any way, } I would not bother. } } That's it from me. I hope it helps and good luck. } } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za } } } -----Original Message----- } From: Trevor Sewell [SMTP:sewell-at-uctvms.uct.ac.za] } Sent: 29 July 1998 09:18 } To: Microscopy-at-sparc5.microscopy.com } Subject: Venting SEM chambers with bottled nitrogen or air } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } }
Another risk to the EDS thin window from venting which I haven't seen mentioned here is ballistic damage -- particulates getting blasted off the sample and punching a pinhole in the window. We see this problem probably more often than implosion failures. Be aware of the geometry of the venting flow pattern relative to the detector if you're considering a customized venting system and analyze powders or other particulates.
The whole concept behind Darkfield is that you eliminate the background light by putting some sort of device in the front focal plane of the condenser (where the iris is located) so that the light approaching the objective comes in at such a high angle, it misses the objective entirely. You can certainly make your own darfield patch stop ... and your message indicates that your thinking is headed in the right direction. Here are the instructions:
1. Set the microscope up for regular brightfield, Koehler illumination
2. Remove the eyepiece and look down into the "back focal plane" of the objective (all the way down the tube). You should be able to see the image of the condenser iris. (You can test by watching as you adjust the iris setting)
3. Open the condenser iris until it just disappears from the objective back focal plane.
4. Very carefully (so as not to disturb the iris setting), remove the condenser from its mount. Measure the diameter of the opening. That is the diameter of the circle you will need to pain on the center of your transparent "filter"
Note that the condenser iris setting is very dependent on the numerical aperture of the objective. This home-made approach will work best with low mag objectives (4x, 10x, 16x).
The alternative to going to all this trouble is to buy a simple "patch stop" from Zeiss. (Other suppliers, like Edmund Scientific at www.edsci.com, or Carolina Biological may also carry them). These patchstops are made from shim metal and work well just by opening the condenser iris fully then carefully inserting the stop just shy of the iris. They may need a bit of tape (smooth black electrical tape works well) to hold them in place but be careful not to get the adhesive on the blades of the iris.
Another variation on this theme is Rheinberg illumination. Instead of blocking the center with black (and getting darkfield), you can block it with a colored filter. The center of the filter will color the background and the ring around the edge will color the features in your specimen. EX: blue center stop with yellow ring produces yellow objects against a blue background. This technique works beautifully 10x objectives and with moderately scattering objects like filamentous mold and fairly flat diatoms. We offer sets of Rheinberg filters (I think there are over a dozen rings per sheet)for modest price. See our web site: { {http://www.MME-Microscopy.com/education} . While you are there, check out my book, "Optimizing Light Microscopy for Biological and Clinical Laboratories". It has lots of little experiments which might be of value to both you and your students.
adavis-at-netpci.com wrote: } } Is it possible to set up darkfield illumination without the expense of a whole different condenser? I have tried knife edge illumination (lying a sharp-edged on the (in my case) light source condenser on the base of the microscope, so that the knife (or card stock?) edge runs exactly through the axis of the light path). I thought of painting a black disk on the center of a transparent filter. } } I'd be grateful for any useful suggestions. (I am using a Zeiss standard 16 with an na 1.35 condenser). }
Alan:
I have used a cork-borer to cut dark field stops of various sizes out of heavy black paper. A machinist could punch out metal stops of various sizes from a sheet of aluminum.
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 e-mail: mcauliff-at-umdnj.edu ***************************************************************
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One possibility is the use of a few drops of mineral oil (like PCR oil or similar) on the top of each aliquot - this protects the solution from carbon dioxide.
Michal
Michal Jarnik Lab of Structural Biology Research, NIAMS, National Institutes of Health, Bethesda, Maryland 20892. Ph: (301) 435-2587
Hi, I am trying to determine the theoretical minimum to the spot size of a laser (gaussian) beam at the focus of a 100X/0.85 (air) microscope objective for uniform illumination of the objective. Does the diffraction limit criteria for incoherent illumination also apply in the case of coherent gaussian beams?
It has been asked if crystals at the bottome of Reynold's Pb citrate is undesirable.
Answer: Yes. Something is wrong. Probably the chealation is not complete, allowing the crystals to settle out. The shaking (by hand and not by stirrer, rotator or shaker) and inversion for 25 minutes is critical. It is boring, but necessary. Pb is best not kept in glass. Transfer it to a disposable plastic syringe. It is worth looking into, since there is a problem lurking somewhere which might result in sudden, unexpected loss of grids. Bye, Hildy Crowley {hcrowley-at-du.edu}
Trevor, We vent all columns with nitrogen that is syphoned off a liquid nitrogen storage tank. This nitrogen gas is also run through a tube with a drying agent to make sure it doesn't pick up moisture from the lines.
We have scuba valves on the lines that will close as soon as you reach atmospheric pressure. This eliminates the problem of accidentally pressurizing the microscope column.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-aux.btny.purdue.edu Purdue University or: emcenter-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Do folk have strong opinions about the desirability of venting SEM chambers with either air (passed through a filter and desiccant column) or bottled nitrogen?
In the case of LEO SEMs with LaB6 filaments it strikes me as being an unnecessary expense to vent with bottled nitrogen as the column region is seldom vented. Also venting with bottled nitrogen adds complexity as precautions must be taken not to over pressurize the chamber and damage the EDS window.
I would appreciate comments for my guidance.
Best regards,
Trevor Sewell
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for the following problem always indicate your position as to direction and when you record where you were and wish to return to there always return in the same direction. If you head to it in a differnt direction you will also add an amout of space that includes the backlash in the stage.... Does any of this make sense? Let me know if I need to phrase it in a differnt manner. Thanks Ed Sharpe, Archivist SMECC
P.S. It is an old trick I learned when I was learning about tool and die making as an expansion of my mind. Dear all,
I've had some problems sending the microscope stage to positions already visited and stored using the ISIS "autorun" program. Indeed, I almost observe a "shift" of about 20 microns which means that the stage never comes back to its original position.
Suppose the stage is located by X1 Y1 and I then move to another location X2 Y2. If I then come back to X1 Y1, the image has shifted by about 20 microns to X3Y3. However if I change location and want to come back to X1Y1, the stage accurately comes back to X3Y3 each time...
Something is wrong with your system. For some reason, your flow is down. It shouldn't take any longer to vent with N2 than with air. Check the you still have gas, that you don't have any kinks in your supply hose, and that your valve is open. As a check, take the hose off at the inlet to your vacuum system. Moisten your lips and have the gas flow over your lips. You should just feel a slight cooling sensation. That is a good flow rate for venting a system.
How often are you venting the chamber ? We have found on our LEO 440 that venting with nitrogen takes a long time (up to 30 minutes !!) (Maybe we're doing something wrong, Luc?). Although the pump down time decreases, you're probably going to lose more time venting than you'll gain with the decreased pump down time if you're changing samples often. Venting with nitrogen will keep the SEM cleaner, though, if time is not a problem.
Charles Bushell Mintek
} -----Original Message----- } From: Luc Harmsen [SMTP:anaspec-at-icon.co.za] } Sent: 29 July 1998 02:35 } To: 'Trevor Sewell'; 'MSA listserver'; 'MSSA listserver South Africa' } Subject: RE: Venting SEM chambers with bottled nitrogen or air } } Hi Trevor, } This is a great topic for discussion as there are quite a few factors that } come into play. } Basically the main criteria for any vacuum system is to keep the } atmosphere in it constant and clean. By venting and opening the door of } the chamber in a SEM to change samples obviously throws that criteria out } the window. } As is always the case with microscopy we have to make compromises. The } best solution would be to fit an airlock to the chamber. In this way you } would maintain a better vacuum, however then you are limited to sample } size and it's expensive to buy an airlock. } The second option is to back fill with dry nitrogen gas. Nitrogen being } the gas that is most prevalent in the air that we breath and that vacuum } manufactures have therefore designed their vacuum pumps and gauges around. } } By back filling with dry nitrogen you are maintaining a fairly constant } atmosphere to pump after venting and changing the sample. Maintaining the } positive pressure in the chamber, as you open the door, keeps the } surrounding atmosphere out. Yes, the negative side to this is the danger } of putting to much pressure on the window of the ED detector and therefore } blowing it. This can be prevented by simply removing the latch that hold } the door closed. } The surface area of the door is much larger than the area of the window, } so the door will be forced open far before the pressure on the window of } the ED detector suffers. In fact on normal W filament systems where you } vent the entire column, I have seen the gun popping up and down due to dry } nitrogen backfilling, when the door latch is still on. No damage to the ED } detector. } As mentioned the vacuum components are also suited to Nitrogen. Pirrani } and penning gauges are calibrated on Nitrogen pressures, so by using } nitrogen you have a better chance of attaining the same readings each time } you pump again. The Turbo molecular pump, in particular, prefers to pump } nitrogen and is thus more efficient. Water vapour is particularly } difficult for a turbo pump to pump. So if your EM is situated near the } coast or you have a lot of heavy breathers using the EM, the humidity in } the lab will change on an hourly, daily and seasonal basis. On a bad day, } with high humidity, the pumping speed of the chamber to reach VAC OK } status, could decrease in terms of 5 to 10 minutes from a good dry day. } } Thus in conclusion ( yes I eventually got there), to backfill with dry } nitrogen improves pump down speed, cleanliness of the system, stability } and repeatability of the required vacuum and prolonged life of the vacuum } pumps. This will be very helpful if you require high resolution work or } very accurate ED analysis from your system on a regular basis. There is } also no need to purchase an air lock for your system. } On the down side, as there always is in microscopy, there is the cost of } the nitrogen, ensuring the door latch is not left on to prevent ED window } damage ( however I am not aware of anyone where this has happened. ), } changing samples in a hurry so as not to waste too much Nitrogen and the } hassle of making sure you have a supply of Nitrogen handy all the time. } In my opinion, a SEM that is totally vented each time you need to change } samples, i.e. vent the column , gun and chamber, and is situated near the } coast or any other high humidity area, should definitely use dry nitrogen } backfilling. } If you are simply venting the chamber but wish to do high resolution work } or stable ED analysis, I would still look at dry nitrogen backfilling. } If you are analysing or looking at really foul samples at low mag any way, } I would not bother. } } That's it from me. I hope it helps and good luck. } } Luc Harmsen } Anaspec, South Africa } International technical support on microscopy. } Tel: +27 (0) 11 476 3455 } Fax:+27 (0) 11 476 7290 } anaspec-at-icon.co.za } } } -----Original Message----- } From: Trevor Sewell [SMTP:sewell-at-uctvms.uct.ac.za] } Sent: 29 July 1998 09:18 } To: Microscopy-at-sparc5.microscopy.com } Subject: Venting SEM chambers with bottled nitrogen or air } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } }
I too have been planning to go to the ICEM in Can Cun, Mexico, and have been concerned about matters of safety there. However, I have talked with several people who have been there recently, and a couple who go there often on business, and have been informed that the area near the Conference Center, where the meetings will be held, is populated primarily by hotels and other commercial institutions, and is highly 'touristized', and so is probably no more 'dangerous' than any other major tourist center. However, I have received several warnings about being careful with luggage, wallets, briefcases, etc. in airports in Mexico, and on tours outside the Can Cum tourist area.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Barbra Foster's advice on dark field is exactly right. However, there is another way. Dark field is one of the few contrast methods in light mocroscopy that does not use Koehler illumination. You can place an opaque circular stop in the middle of the top lens of your condenser and adjust the height of the condenser while observing a focused specimen until a dark field image is formed. (Note, I do not mean on top of a flip-in lens if there is one but on top of the main condenser lens with any flip-in lens out.) Start with a stop about 4 to 5 mm in diameter. You may have to experiment with the size. This will work for 10 X and 20 X objectives. The thinner and more opaque the stop the better.
Bob
_____________________________________________________________ C. Robert Bagnell Jr., Ph.D., Res.Assoc.Prof. Microscopy Services Laboratory Department of Pathology & Laboratory Medicine CB #7525 UNC-CH, Chapel Hill, N.C. 27599 ph 919-966-2413 fx 919-966-6718
Dear Frank, } } I've had some problems sending the microscope stage to positions } already visited and stored using the ISIS "autorun" program. Indeed, I } almost observe a "shift" of about 20 microns which means that the } stage never comes back to its original position. } Our positioning program on the IVEM takes backlash into account and always drives to a set distance at a given orientation with respect to the end position. The last adjustment is then made slowly and always in the same direction. One chooses an initial position and goes away then returns several times until the stage returns reliably to the chosen position. Thereafter, moving to new positions and returning to old ones is automatic. I don't know who wrote our program (or yours), so I can't say how you might incorporate this procedure. Good luck. Yours, Bill Tivol
Regarding the discussion on darkfield imaging, I sometimes image specimens in pseudo darkfield using a 10X objective combined with the phase-3 (100x) phase ring of our condenser assembly. The darkfield image is by no means perfect but is often useful nonetheless. The ph3 condenser ring directs a cone of light through the specimen, which passes to the outside of the objective (darkfield). Put a small beaker of coffee on top of a slide to see this for yourself. Specimen visualization is dependent on light refracted into the objective by the specimen.
I can feel some of you nodding your heads...try it, you might be surprised!
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
Hi Jayesh, The eq for diffraction limited spot size is D(lim)=2.44 x Lambda x focal length/D(a) where D(a) is the diameter of the beam incident the lens. Remember this is the "theoretical diffraction limited spotsize" (assumes paraxial approximation) For input beams that are not collimated, but have a full divergence angle of A, the focal spot diameter will be approximately D(lim)=fA Most optical equations assume a paraxial approximation for the incident rays & a TEM 00 (Gaussian) E-field. There is also the issue of chromatic aberrations which would limit your (broad band) spot size. Diffraction limited spot size is wavelength dependent. Even if you were provided with a diffraction limited spot size or spot resolution by your objective manufacture, somewhere in fine print it is probably at a specified wavelength or 2. There is also an inverse dependence on the diameter of the beam incident on the lens. If the manufacture specified a Dlim.spot size , they are probably assuming the "clear aperture " of the objective is illuminated. The focal length is also wavelength dependent although a 100x objective hopefully has some chromatic correction. Your laser beam may not fill the clear aperture or it may, this brings in a another problem. Theoretically a Gaussian beam (E-field) extends forever. In practice to minimize Fresnel fringing (rings) your clear aperture should be 5x the beam radius at the lens. Fresnel fringes are an optical interference effect. Coherent light is required for optical interference to occur. Coherence is generally ignored in broad band applications. FYI, In reality a incandescent source has a coherence length of ~10um. Another point worth making. Unless you have a many 100K or few M$ lens, you theoretical diffraction limited spot size is no better than 1/2um. A microscope with a ~$100, 40x objective can resolve a 1um period. Using a lower mag. objective will give you a greater depth of field. With a 40X obj. depth of field is still only a few um. Where do you measure the beam waist? The focal plane is not the focal length from the end of the objective & it is not the working distance (although it will be close). It is the focal length measured from a plane known a A2H2 (a principle plane). Your objective supplier may be able to supply the approximate distance from some location on the objective. Since I have no idea what laser you are using, I'll point out that optical coatings can be damaged by lasers. If you have enough power, a focused beam will ionize the air (usually limited to pulsed laser applications). Laser beams can burn dust on the lens & in doing so damage the AR coatings. Also since you are interested in small spot sizes. UV(} 300nm) is heavily attenuated by most non reflective objectives.
hope this helps,
Bruce Brinson Rice U.
Jayesh C. Jasapara wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } I am trying to determine the theoretical minimum to the spot size of a } laser (gaussian) beam at the focus of a 100X/0.85 (air) microscope } objective for uniform illumination of the objective. Does the diffraction } limit criteria for incoherent illumination also apply in the case of } coherent gaussian beams? } } Thanks, } Jayesh
The big problem here is water vapor. Nitrogen backfill is primarily used to prevent the incursion of water vapor into the chamber since most vacuum systems have a hard time dealing with it.
The use of a desiccant column is well advised for any instrument. In the long run however, there are two problems. The first is the regeneration of the desiccant. You have to pay attention and bake the water out of it at regular intervals. The second is a long term accumulation of desiccant dust. Over time, the desiccants appear to produce a very fine dust that will eventually clog the chamber vent valve, resulting in a service call.
Nitrogen systems can produce their own problems. The need to have a constant source of nitrogen can be eased by using the vapor from a large storage dewar if liquid nitrogen is always available in the lab due to EDS or other cryogenic systems. Generally, the nitrogen is cheap but the demmurage charges for the dewar are not. If you use a little more nitrogen per month, the cost is minimal if you already have the need for the large dewar.
The savings is basically in time. The primary volume in an SEM vacuum system is in the sample chamber. It is not system cleanliness that is in question here, it is basically the time involved in pumping the chamber to required levels. If one can exclude water vapor from entering the sample chamber during sample changes, there will be an appreciable improvement in pump down times. A well airconditioned room is a start, desiccants next and a nitrogen backfill is best. Airconditioning is a necessary first step, since a warm and humid environment can also cause other problems such as the condensation of water on the exterior of water cooling lines used for diffusion pump and electronics cooling.
Most system contamination comes not from gases not pumped out, but from air leaks and pump oils. By the time a system reaches operational vacuum, the overwhelming majority of environmental gases, including water vapor, will be gone.
} Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Plenty. You should first determine if the mechanical assembly is operating properly. When dealing with the magnifications involved in microscopy, backlash is a primary concern. This refers to the difference in position caused by approaching the desired position from different directions. In many optical microscope stages, there are no anti-backlash measures taken to reduce the effect. The gear teeth, or leadscrew threads, are allowed to rest against the opposing, driving teeth or threads. If the teeth or threads are not tightly meshed, then the position attained can vary by the amount that the teeth or threads don't mesh.
In your case, you are probably coming from a low position to the first, X1 Y1, position. Then going upward to the X2 Y2 position. Don't be surprised when you move downward back to the X1 Y1 position to find the ultimate position offset by the amount of backlash. As a simple expedient, move back to X1 Y1 by first dropping well below that position, and moving back up to it. If you always come into a given position in the same direction, you should see a great improvement.
In the case of a simple stage movement, without anti-backlash provisions, the best you can do is clean the gears and leadscrews carefully, lubricate them properly and ensure that they are adjusted to provide a good tooth/thread mesh without binding.
Anti-backlash gears and leadscrew followers are essentially devices that bias the gear/leadscrew to always favor one side of the tooth/thread through spring action. Regardless of the direction of movement, they should always 'push' up against the same side of the tooth or thread. Basically they are comprised of two gears or leadscrew followers (nuts) that are spring loaded in opposing directions. One gear or follower is attached to the mechanism being moved, the other simply 'floats' to provide the bias.
This problem is referred to as 'repeatability' and can often vary a great deal from the accuracy spec that most people pay attention to. In the case of anti-backlash movements, proper cleaning and lubrication should be all that is needed to maintain a good repeatability. If not, then the separate parts should be inspected for wear.
} Dear all, } } } I've had some problems sending the microscope stage to } positions already visited and stored using the ISIS "autorun" } program. Indeed, I almost observe a "shift" of about 20 microns } which means that the stage never comes back to its original } position. } } Suppose the stage is located by X1 Y1 and I then move to } another location X2 Y2. If I then come back to X1 Y1, the image } has shifted by about 20 microns to X3Y3. However if I change } location and want to come back to X1Y1, the stage accurately } comes back to X3Y3 each time... } } } Any suggestions? } } F Allen R. Sampson Advced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I have been responsible for a number of demonstration laboratories in the=
past and I have always insisted upon using a dry gas rather than dirty ol= d (English) wet air.
Why?
1. Pump down time is improved considerably to ~50% in some cases, but = it takes months for this to happen.
2. Column cleanlyness improves so if you work at low kV (and you all should try it) you will get a longer life from your column and its apertures between cleaning.
3. Contamination is the biggest killer of high resolution microscopy and=
this is also dramatically reduced.
Regards
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the studies of the three-dimensional structures in biology, medicine and material sciences. The three-dimensional analysis and representation are crucial steps to assess the information. These conferences are the most efficient meeting points between developers and users in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation and in particular to the role of the green fluorescent protein.
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
For about five years we have vented our Zeiss DSM 962 and TEM 900 using a nitrogen generator. It remains permanently switched on and takes nitrogen from the atmosphere; purifies it to remove moisture and other gases then transfers it to a tank to provide gas on tap. The main advantage of this system is that it is almost maintenance free and avoids the problem of changing cylinders. The venting time on the DSM is around 1minute 30 and the vacuum levels on both instuments are very good.
Frieda Christie, Electron Microscopist, Scientific & Technical Services, Royal Botanic Garden, 20A Inverleith Row, Edinburgh, EH3 5LR
I have been very surprised by the tone of several postings on the subject of safety in Cancun. Let us be quite clear about this. A visit to Cancun will be a lot safer than a visit to New York or any other major US city. If your bags are mishandled or stolen, it is much more likely to occur in Miami than in Cancun.
Cancun is a fine place. It is a great resort, suitable for an excellent vacation. The Congress promises to be most successful.
No one should be deterred from attending by these implied slights.
Alwyn Eades . Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I could not agree more! I have not visited Cancun, but I have been to Cozumel a couple of times and it is no different to any other Carribean vacation destination. In fact it is a custom built vacation city, built with the foreign visitor in mind. I am not going to the meeting for financial reasons, but no one should be deterred from visitng for safety reasons!!!
At 7:44 AM -0500 07/30/1998, Alwyn Eades wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
________________________________ Please note the new Area Code ________________________________
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
I forgot to add a neat part of the experiment. If you rotate the objectives out of position (you may have to remove one to get enough room) then place a piece of white paper (like a business card)on edge on the stage, over the light as it emerges from the condenser, you can see how you have changed it by any of the adjustments we discussed. Try it first with plain Koehler. Open and close the condenser iris and you can see how that affects the angle of light approaching the sample. Then close the condenser iris and open and close the field iris. You will see that it does, indeed, control the size of the illuminated field. Then try the experiment with (a) your darkfield patch stop and (b) Rheinberg filters. It's a neat demonstration.
If you have access to the High School shop and if they make screwdrivers as part of their curriculum, have them cut some of the handle stock (usually slighly turbid yellow plastic) into about 1" lengths. Oil one of these little "columns" to a microscope slide with a drop of immersion oil (Nujol or mineral oil from the pharmacy works well, too). Try the same experiments.
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At 10:14 AM 7/29/98 -0400, Barbara Foster wrote:
} } } }
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Dear Alan,
The whole concept behind Darkfield is that you eliminate the background light by putting some sort of device in the front focal plane of the condenser (where the iris is located) so that the light approaching the objective comes in at such a high angle, it misses the objective entirely. You can certainly make your own darfield patch stop ... and your message indicates that your thinking is headed in the right direction. Here are the instructions:
1. Set the microscope up for regular brightfield, Koehler illumination
2. Remove the eyepiece and look down into the "back focal plane" of the objective (all the way down the tube). You should be able to see the image of the condenser iris. (You can test by watching as you adjust the iris setting)
3. Open the condenser iris until it just disappears from the objective back focal plane.
4. Very carefully (so as not to disturb the iris setting), remove the condenser from its mount. Measure the diameter of the opening. That is the diameter of the circle you will need to pain on the center of your transparent "filter"
Note that the condenser iris setting is very dependent on the numerical aperture of the objective. This home-made approach will work best with low mag objectives (4x, 10x, 16x).
The alternative to going to all this trouble is to buy a simple "patch stop" from Zeiss. (Other suppliers, like Edmund Scientific at www.edsci.com, or Carolina Biological may also carry them). These patchstops are made from shim metal and work well just by opening the condenser iris fully then carefully inserting the stop just shy of the iris. They may need a bit of tape (smooth black electrical tape works well) to hold them in place but be careful not to get the adhesive on the blades of the iris.
Another variation on this theme is Rheinberg illumination. Instead of blocking the center with black (and getting darkfield), you can block it with a colored filter. The center of the filter will color the background and the ring around the edge will color the features in your specimen. EX: blue center stop with yellow ring produces yellow objects against a blue background. This technique works beautifully 10x objectives and with moderately scattering objects like filamentous mold and fairly flat diatoms. We offer sets of Rheinberg filters (I think there are over a dozen rings per sheet)for modest price. See our web site: { {http://www.MME-Microscopy.com/education} . While you are there, check out my book, "Optimizing Light Microscopy for Biological and Clinical Laboratories". It has lots of little experiments which might be of value to both you and your students.
Well, I hope you enjoy Cancun, but...... Several years (approx 5? ?) the American Chemical Society held a meetin= g in Mexico city. An ACS member was killed in his hotel room when he surpri= sed a thief when he returned unexpectedly to his room. I just read about a M= exican police officer who kidnapped three teenagers, took them to a police sta= ble, raped the two younger girls (while the old one locked herself in a room= to prevent the assault). The three escaped later. All the travel protect= ion web sights have lists an arm long discussing what not to do in Mexico (like= don't take any taxi that comes along). Let me see, I don't speak the language (my fault) I don't have family connections to their society I can't carry a effective means of self defense I don't understand the society while I am far from wealthy, to them I am extremely wealthy. Take some advice from my friend Mas Ayoob: "Don't go were you are not w= anted."
jae5-at-lehigh.edu on 07/30/98 08:59:43 AM To: microscopy-at-Sparc5.Microscopy.Com -at- INTERNET cc:
I have been very surprised by the tone of several postings on the subje= ct of safety in Cancun. Let us be quite clear about this. A visit to Ca= ncun will be a lot safer than a visit to New York or any other major US city= . If your bags are mishandled or stolen, it is much more likely to occur = in Miami than in Cancun.
Cancun is a fine place. It is a great resort, suitable for an excellen= t vacation. The Congress promises to be most successful.
No one should be deterred from attending by these implied slights.
Alwyn Eades . Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Dear all, =20 =20 Thank you for the many replies I have received regarding the backlash=20 problems I recently communicated to the list. From the replies I=20 received, it appears that some of you associated the problem with the=20 ISIS software mentioned in the E-mail and more particularly the=20 AUTORUN application. However, the above software has been performing=20 very well, i.e showing excellent repeatability (double clicking on=20 stored points does recall the correct coordinates). The problem as=20 some of you correctly pointed out, is associated with the lack of=20 backlash when carrying out stagee rotations. When using translations=20 only to go from 1 point to the other (and later come back to those=20 positions) everything is fine, but as soon as a rotation is introduced= =20 the repeatability is affected.Unfortunately there is nothing I can do=20 about it other than using translations to move from one point to the=20 other or always approaching a position from the same direction =20 F
I've found this works fine with a 10X objective and the 40X phase ring, or a 20X objective and the 100X phase ring. The image-especially the 10X-40X one was as good as a true dark-field image.
Phil
} } Regarding the discussion on darkfield imaging, I sometimes } image specimens in pseudo darkfield using a 10X objective } combined with the phase-3 (100x) phase ring of our } condenser assembly. The darkfield image is by no means } perfect but is often useful nonetheless. The ph3 condenser } ring directs a cone of light through the specimen, which } passes to the outside of the objective (darkfield). Put a } small beaker of coffee on top of a slide to see this for } yourself. Specimen visualization is dependent on light } refracted into the objective by the specimen. } } I can feel some of you nodding your heads...try it, you } might be surprised! } } Doug } ---------------------- } Douglas R. Keene } Associate Investigator } Shriners Hospital Microscopy Unit } 3101 S.W. Sam Jackson Park Road } Portland, Oregon 97201 } 503-221-3434 } DRK-at-shcc.org
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
We are talking Cancun here! It is a tourist destination, harming tourists is extremely bad for business. I cannot believe you are comparing Mexico City with Cancun. Mexico City is just like New York! Or come to think of it any other huge urban sprawl!
Please stop this rampant racism and just go and enjoy the conference!
At 9:43 AM -0400 7/30/98, FRANK KARL wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Note new Area Code (734)
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
I quite agree with Alwyn. I have been to Cancun and Cozumel numerous times and have never had any problems regarding safety or theft. I have walked the streets well into the night (something I would never do in most american cities) and not had any problem.The only inconvience I have ever incurred were people trying to sell me timeshares, even then, a couple of polite but firm "No Gracias" usually sends them on their way. While there I highly recommend visiting the Mayan ruins of Chichen Izta and Cubo. Take a bus tour though, driving can be dangerous as the roads are narrow and not in the best of shape. I once had a very large bull charge our rented jeep while doing 80 kph (the jeep not the bull). If you SCUBA dive, Cozumel has some of the best sites in the western hemisphere. On a side note I always drink bottled water while there, no matter where I am staying.
(sigh) It's a shame my company won't send me to meetings outside of the USA.
Chuck Mass Bio-Medical Imaging Wyeth-Ayerst Research 641 Ridge Road Chazy NY 12921 massc-at-war.wyeth.com
Dear Frank, } } [skip] When using translations } only to go from 1 point to the other (and later come back to those } positions) everything is fine, but as soon as a rotation is introduced } the repeatability is affected.Unfortunately there is nothing I can do } about it other than using translations to move from one point to the } other or always approaching a position from the same direction } The same technique which eliminates backlash from translations will also work for rotations (i.e., always approach the new orientation from the same direction), and the additional complications due to cross- talk between the translational and rotational backlashes can also be corrected by always doing one first, then doing the other, and approach- ing from the same directions. A possible protocol would be to rotate until one is a few degrees cw of the desired orientation, next translate to a point (delta-x, delta-y) from the desired position, then progress ccw to the desired orientation, and finally translate to the desired posi- tion. The important thing here is to choose the signs of delta-x and delta-y so that the movement in the (-delta-x, -delta-y) direction will not reintroduce rotational backlash. One may have to repeat the final adjustments in certain cases to be sure that the backlashes have, indeed, been properly taken up. Yours, Bill Tivol
We are looking to expand our capabilities here at The Jackson Laboratory so that we can image either entire mice or living, attached parts (legs, ears, etc.) One of our options may be MRI. So we're looking to find out who makes MRI machines, what they cost, if they do whole, small animals, what level of resolution can be achieved (i.e. enough to detect auto-immune induced degeneration), what qualifications are needed to operate this, etc. For people who may already be doing this, we'd be interested in your experience with these techniques.
We also think that ultrasound may be another way to do imaging. Or there may be methods out there that we don't know of yet. If anyone has any experience or ideas or knows of someone (investigator or manufacturer) who is doing this, I'd really like to hear from them.
Thank you!
Lesley Bechtold Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor ME 04605
{fixed} I see we have mixed feelings about safety in Can Cun. I have travelled to many places in Mexico, both on business and pleasure. In particular I have visited the hotel region of Can Cun where ICEM is being held and I would agree with Alwyn Eades and John Mansfield that this is is a no more dangerous place that any tourist area in the U.S. However, if you propose to visit some of the marvellous tourist attactions on the Yucatan peninsula such as Tolumn, Merida, Chichen Itza etc etc and you are not going on an organized trip {bold} {underline} beware {/underline} {/bold} , especially if you do not speak Spanish and are not in tune with the Mexican mentality. Police bribery for alleged traffic offences is not uncommon (if you have rented a car), petty overcharging for gas and other items is common and crime can be problem. My advice if you are not travelling in a large organized group is:- don't carry lots of cash, just a credit card or two and carry them in a safety holster or belt and brush up on your Spanish. Can Cun and its surroundings are magnificent (if a bit hot in summer). Take appropriate precautions and enjoy yourself.
Alan Fox
Director, Center for Materials Science and Engineering
} While there I highly recommend visiting the Mayan ruins of Chichen Izta } and Cubo. Take a bus tour though,
I have been to Chichen Itza and it was well worth the visit, we flew there from Cozumel. The tour of the ruins and the round trip airfare was about $130 each five years ago. Highly recommended. Clmb the pyramid if it is still open, they were going to close it was the tourists were wearing it away!
} On a side note I always drink bottled water while there, no matter where I am staying. Oh, yes this is a MUST! Even with the most careful precautions in Acapulco we all got Montezuma's Revenge from something we ate or drank. Things to note:
1. Ice, dont use it unless you are sure it was made with purified water. You can usually tell by the shape of the cubes, if it looks like the machine packed ice you get in the US then it is usually OK, but I would always ask to make sure. If the ice is home made it may have been made with tap water and then you will be in trouble, your system is not used to the flora and fauna of that part of the world!
2. Dont eat unpeeled fruit like apples, grapes, pears, etc. they may have been washed in tap water.
3. Dont eat salad vegetables (lettuce, cucumber, tomatoes, etc.) as they are likely to be washed in tap water too. This is difficult, as many foods come with a salad garnish and simply removing it is not always enough. We had shark quesadillas in Acapulco that I think were our downfall as they were accompanied by shredd lettuce as a garnish.
4. Brush your teeth with bottled water too, the little you may swallow while doing this may make a difference.
I know this all sounds like overkill, but it can save you many hours in the little room! Drink bere or pop it is usually cheap and plentiful in Mexico and the beer is good too!
John M.
Note new Area Code (734)
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Most of the people have been describing the boil off from large tanks outside (where the plumbing is required) or the large upright (5 feet high - 2.5 feet diameter). Both of these have valves to take off the vapor above the liquid. This pressure can be quite high and they also have pressure relief valves and safety blowout plugs.
You are correct about what I was saying. I used a small dewar that is used for absorption pumps which are basically styrofoam. You can use the styrofoam that is used to protect acid bottles when they are shipped or your dewar that you use to fill the EDS detectors. (If your are in a bind, put a tube into the EDS detector dewars.) If you have an open container, just use Tygon. If you have a narrow neck, use a copper or stainless steel tube to go into the tank so that it won't go hard if it is bent and attach the Tygon tube onto the warm end of the tube. Now you have to clear the line of the air in it just like any other line that you attach to a vacuum system. Just after you put the tube into the LN2, there will be blow out at the other end of the tube until the temperature equilibrates. Position that end over the inlet to the vacuum system to flush out the inlet also. While the gas is flowing, put the tube on the inlet and that will give you a fairly clean line.
It doesn't matter what diameter(s) that you use, just length. About 8-10 feet is good enough. If you use a stainless steel tube to insert in the LN2, it will blow gas a little longer than if you use copper because of its lower thermal conductivity.
I agree that boil off from LN2 is the best backfill. I used to use it extensively when I was working with UHV systems. There is absolutely no H2O in it. It is the driest nitrogen that you can get and there is also very little O2. LN2 is usually very available in an electron microscopy lab that has EDS detectors on the instruments and it is cheap. However, you don't have to have the boil off from a large tank and the plumbing that goes with it or the overpressure danger. An open container of LN2 will suffice. It is at atmospheric pressure and therefore will not overpressure your vacuum system. Make sure that the tube that you stick in is long enough so that the liquid that is drawn out is converted to gas before entering the vacuum system. Also, make sure that the tube doesn't go to the bottom of your dewar, otherwise it will suck up debris.
Incidentally, for safety reasons, you should not use a heater in the bottom of a dewar that can be pressurized. You should also not pressurize a LN2 dewar with air. Liquid oxygen can form at the bottom of the dewar over time. But following this thread, avoid overpressuring the vacuum system. The open LN2 approach does that.
Nitrogen is pumped fairly well by all vacuum pumps, especially ion pumps. The rated pump speeds are all referenced to N2. There are other gases in filtered air that become significant partial pressures in the ultimate vacuum because of their lower pump speeds.
Use LN2, your vacuum system will love you for it.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
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(412) 820-8651 (office) (412) 820-8161 (fax)
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We have found that boil off from liquid nitrogen is by far the best venting material. It is dry, it is clean, it is cheal.We hane the whole lab' plumbed with a line going back to a 100litre Dewer which if I remember lasts for 3-4 weeks. You obviosly have to be careful with pressure. There is nothing like getting a 100lb SEM stage whislting out of the microscope and landing on your lap. Really brtings tears to your eyes.
Patrick Echlin Multi-Imaging Centre University of Cambridge.
On Wed, 29 Jul 1998, Trevor Sewell wrote:
}
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-----------------------------------------------------------------------. } } Do folk have strong opinions about the } desirability of venting SEM chambers } with either air (passed through a filter and } desiccant column) or bottled nitrogen? } } In the case of LEO SEMs with LaB6 filaments } it strikes me as being an unnecessary expense to } vent with bottled nitrogen as the column region is } seldom vented. Also venting with bottled nitrogen } adds complexity as precautions must be taken not } to over pressurize the chamber and damage the } EDS window. } } I would appreciate comments for my guidance. } } Best regards, } } Trevor Sewell } } } }
At 10:20 AM 7/30/98 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hmmmm..... People are discussing safety at a destination they are unfamiliar with and are labelled racists.
Who is overreacting?
Just a thought.
Randy Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I've followed with interest the debate on safety in Cancun. I've spent many years doing fieldwork in remote parts of Latin America and spent some time in such cities as Caracas, Montevideo, Buenos Aires, Santiago, and Lima. Without exception, I feel safer in those localities than most US cities. While in Peru this summer, I heard about:
A mass execution of two gang members and three innocent bystanders in Tacoma, WA, the nearest city to where I live.
A brutal beating on a rural highway in the small twon I live in.
Murder and mayhem in the US Capitol building.
During other trips to that region, I've read in their papers about the senseless murder of European tourists in Miami - on several occasions.
Most Latinos I've spoken with in South America express deep concern about visiting the gun-toting, violence prone society north of the Rio Grande.
I'd say that your chances of encountering crime and personal harm probably diminish when you leave US borders. So - go to Cancun and feel safer than at home!
A friend of mine needs to part with his FF instrument. Please direct your inquiries directly to Dr. Ip. (S. Samuelsson)
CRESSINGTON CFE-50 FREEZE ETCH UNIT This is an oil diffusion pumped unit with full freeze etch capability. It has a rotary stage. Deposition of Pt/C is carried out using EH5 electron beam guns and is controlled by an EB1202PC/C electron beam controller and an MTM22 quartz crystal monitor. This CFE-50 was installed in 1992 but has made less than 30 runs during its life because our laboratory changed research projects shortly after the unit was purchased. We will consider any reasonable offer.
Contact: Wallace Ip, Department of Cell Biology, University of Cincinnati College of Medicine, PO Box 670521, Cincinnati, OH 454267-0521. Tel. (513) 558-3614 Fax (513) 558-4454 Email: wallace.ip-at-uc.edu
} } I have been very surprised by the tone of several postings on the subject } of safety in Cancun. Let us be quite clear about this. A visit to Cancun } will be a lot safer than a visit to New York or any other major US city. } If your bags are mishandled or stolen, it is much more likely to occur in } Miami than in Cancun. } } Cancun is a fine place. It is a great resort, suitable for an excellent } vacation. The Congress promises to be most successful. } } No one should be deterred from attending by these implied slights. } } Alwyn Eades
I'd like to add that I've been all over the Yucatan (Chichen Itza, Uxmal, Merida, Coba, Tulum, etc, and have never encountered anything but exciting sights, fun, friendliness, good food, etc., etc. We WERE attacked by a swarm of beautiful butterflies on the road to Coba, tho...
Caroline Schooley Educational Outreach 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/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
We are using a Leica CPC unit for freezing bare grid technique. I was wondering if anyone else has one of these units yet and can give us some advice on it.
We have several problems:
The release mechanism for the plunger is threaded for what looks like a place to attach a photographic cable release. This would be great as otherwise one needs three hands to operate the unit and normally these type of units have a footswitch to control the drop of the plunger into cryogen. So, we purchased an air release (the kind with a hollow rubber ball at the end which you squeeze to release the shutter), and this turns out to be the wrong thread. I was wondering if Leica have made a part for this, and whether it is included in the price of the unit, but the local office in Canada did not know.
I think its a pretty crappy release mechanism for a 16,000 dollar unit! (it does not even include a binocular). All of the homemade ones I have seen (including the one my workshop made for me in Oxford) had an electronic release (they even made a 12 volt solenoid for me, so as to avoid sparks and ignition hazard when using ethane as a cryogen). You can buy an electronic footswitch in Tandy (UK) or Radio shack (N. America) for a few pennies.
By the way, the metal mirror attachment comes as an accessory which will cost you an extra 4000 dollars- it isnt included with the standard unit, so be warned! The brochure gives you the impression that this thing will do every type of freezing possible. Make sure they include the cost of the pump to keep the thing filled from a Dewar too, it won't work without this as its not intended to be able to fill it manually!
A big problem we have is ice contamination. The unit relies on a nitrogen "inert gas" atmosphere" from boil off. Problem with this is that you get "snow" falling into the unit from the turbulent air on top. You cant fill it with enough liquid to get your grids quickly under liquid where they are safe.
When using cryogens such as liquid ethane, the thing has to use a heater to keep the ethane from going solid. This is all very well, and it works, but it causes the ethane to evaporate very quickly, so instead of having to thaw the stuff with a metal rod, you continually have to top it up. Also you can't adjust the distance of the grid forceps from the surface of the ethane, the movement of the grid from start to stop is fixed, you cant change the length of movement or the distance it plunges, as it is the grid is dangerously close to the cold area and can pre-freeze if you are not careful. We will probably drill a few more holes so the thing can at least be mounted at different heights, but this wont give continuous adjustment. Also the forceps clamp does not hold very well and you are limited to forceps which have straight sides and are the same length as the paur supplied with the unit. A lot of the specimen holder appears to be mad out of heavy gauge hypodermic needles.!?!
So, as long as you don't need the automatic fill capability, you can make something yourself out of a styrofoam box and bottle cap and a plunger arm can be made in any workshop very cheaply- all it has to be is a cylindical metal rod running on roller bearings. A couple of clamps and stops control the height and distance of throw. Use a metal rod to thaw the cryogen. after freezing a specimen, top up so the ethane cup is under liquid nitrogen, and it will remain ice free for some minutes until you need to use it again, then just pour iff the excess nitrogen and warm the ethane with the metal rod (I mounted mine on a rubber cork, for insulation). You can use the same container for metal mirror, all you need is blocks of metal and if gravity is not fast enough for you, attach some heavy duty rubber bands.
Dr Timothy F. Booth Canadian Food Inspection Agency National Centre For Foreign Animal Disease Suite T2300 1015 Arlington St. Winnipeg Manitoba R3E 3M4 CANADA http://www.hc-sc.gc.ca/main/lcdc/web/bmb/fedlab_e.html#toc email tbooth-at-em.agr.ca Tel 204 789 2022 Fax 204 789 2038
There is nothing wrong with NYC either. It's a nice city; however, you just must be aware of where you are and remember not to act carelessly with yourself or your belongings. I reccommend being aware of your surroundings and of scammers that you often find in tourist traps and airports etc., but *please*, if you live your whole life in *fear* you'll have no fun at all.
Andrea
On Thu, 30 Jul 1998 10:20:30 -0400 jfmjfm-at-engin.umich.edu (John F. Mansfield) wrote:
} } } } I have been very surprised by the tone of several postings on the subject } } of safety in Cancun. Let us be quite clear about this. A visit to Cancun } } will be a lot safer than a visit to New York or any other major US city. } } If your bags are mishandled or stolen, it is much more likely to occur in } } Miami than in Cancun.
I'd like to add that I've been all over the Yucatan (Chichen Itza, Uxmal, Merida, Coba, Tulum, etc), and have never encountered anything but exciting sights, fun, friendliness, good food, etc., etc. We WERE attacked by a swarm of beautiful butterflies on the road to Coba, tho...
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Hi Bruce, Thanks for your help. I happen to be a novice in the field of microscopy - my specialization being femtosecond laser systems. I have a couple more questions for you. The only information I have on the objective is that it is a 100X and the N.A. is 0.85 . I am using this objective to focus down femtosecond pulses centered at 800nm wavelength and when I measure the spot size in the focal plane using the edge scan method, it measures to be about 350nm for full aperture illumination. Is this small a spot size possible for a wavelength of 800nm?
The location of the focal plane has been located accurately by looking for the steepest rise for the edge scan. The beam hitting the objective is not collimated by a telescope - it is a diverging beam.
The way I calculate the focal length of the objective from the given N.A. is by the following formula : f = D(a)/(2*tan(arcsin(N.A.))) = 1.55mm, where D(a) is the diameter of the incident beam on the lense. I am not sure about the validity/accuracy of this expression. Could you tell me how good or bad this expression works out to be?
If I use the formula assuming the paraxial case approx. holds, then the theoretical limit to the spot size is 500nm. But from the diverging beam formula Dlim = fA, I get a smaller theoretical spot size of about 395nm. This is closer to what I measure experimentally. The objective I use is of good quality (from Zeiss ) - so it should be corrected for chromatic aberration. Also the mean power being focussed is low - and so there is no danger of damaging coatings.
Thanks, Jayesh
} Hi Jayesh, } The eq for diffraction limited spot size is D(lim)=2.44 x Lambda x focal } length/D(a) } where D(a) is the diameter of the beam incident the lens. Remember this is } the "theoretical diffraction limited spotsize" (assumes paraxial } approximation) } For input beams that are not collimated, but have a full divergence angle } of A, the focal spot diameter will be approximately D(lim)=fA } Most optical equations assume a paraxial approximation for the incident } rays & a TEM 00 (Gaussian) E-field. There is also the issue of chromatic } aberrations which would limit your (broad band) spot size. Diffraction } limited spot size is wavelength dependent. Even if you were provided with a } diffraction limited spot size or spot resolution by your objective } manufacture, somewhere in fine print it is probably at a specified wavelength } or 2. There is also an inverse dependence on the diameter of the beam } incident on the lens. If the manufacture specified a Dlim.spot size , they } are probably assuming the "clear aperture " of the objective is illuminated. } The focal length is also wavelength dependent although a 100x objective } hopefully has some chromatic correction. } Your laser beam may not fill the clear aperture or it may, this brings in } a another problem. Theoretically a Gaussian beam (E-field) extends forever. } In practice to minimize Fresnel fringing (rings) your clear aperture should } be 5x the beam radius at the lens. Fresnel fringes are an optical } interference effect. Coherent light is required for optical interference to } occur. Coherence is generally ignored in broad band applications. FYI, In } reality a incandescent source has a coherence length of ~10um. } Another point worth making. Unless you have a many 100K or few M$ lens, } you theoretical diffraction limited spot size is no better than 1/2um. A } microscope with a ~$100, 40x objective can resolve a 1um period. Using a } lower mag. objective will give you a greater depth of field. With a 40X obj. } depth of field is still only a few um. } Where do you measure the beam waist? The focal plane is not the focal } length from the end of the objective & it is not the working distance } (although it will be close). It is the focal length measured from a plane } known a A2H2 (a principle plane). Your objective supplier may be able to } supply the approximate distance from some location on the objective. } Since I have no idea what laser you are using, I'll point out that } optical coatings can be damaged by lasers. If you have enough power, a } focused beam will ionize the air (usually limited to pulsed laser } applications). Laser beams can burn dust on the lens & in doing so damage the } AR coatings. Also since you are interested in small spot sizes. UV(} 300nm) } is heavily attenuated by most non reflective objectives. } } hope this helps, } } Bruce Brinson } Rice U. } } Jayesh C. Jasapara wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hi, } } I am trying to determine the theoretical minimum to the spot size of a } } laser (gaussian) beam at the focus of a 100X/0.85 (air) microscope } } objective for uniform illumination of the objective. Does the diffraction } } limit criteria for incoherent illumination also apply in the case of } } coherent gaussian beams? } } } } Thanks, } } Jayesh } } } }
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