Evex Analytical ( www.evex.com ) has a Jeol 840 with an Evex =3D Microanalysis system ( http://www.evex.com/prod2.htm ) and Active Digital Imaging System that we would like to replace with a new microscope.
Claudio Tarquinio Evex Analytical Microanalysis and Digital Imaging 609-252-9192 T 609-252-9091 F www.evex.com info-at-evex.com
---------- } From: Karen Zaruba {kszaruba-at-MMM.COM} } To: MSA Listserve {microscopy-at-sparc5.microscopy.com} } Subject: General: Video Printers } Date: February 29, 2000 6:45 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Any suggestions on a really good video printer to include with an SEM } purchase? We've had trouble over the years with the standard roll } printers getting paper jams, which required sending out for servicing } (i.e. lots of down time). We're willing to sacrifice some speed for } something reliable. Are there "fast" ( { 1 min. per print) video } printers that use the cut sheet paper? } } Thanks, } Karen } } P.S. vendors please respond by E-mail (not phone) or dare to face my } wrath. } -- } Karen S. Zaruba kszaruba-at-mmm.com } 3M Company, St. Paul, MN }
Karen -
We've always been happy with our little Sony UP-811 video printer. It doesn't give a very big print (about 5 inches wide), rather like a Polaroid, and not particularly high resolution or anything, but useful as a "working copy", with the actual image saved to disk. It's dead reliable, however, having been in continuous service since about 1993, with no problems that I know of. No connection with Sony, etc. (wouldn't mind owning some stock, though;-)
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada Dartmouth, Nova Scotia
I had a Mitsubishi that was reliable, but in general, my feeling is that for reproducing EM images there are NO good video printers. Between the (typical) paper cost and poor resolution, I prefer to print captured digital images. Though slow, you can get an inkjet printer these days for 1/10 the price. Faster ones for a bit more...
Colleagues.... Can anyone help this guy? It's not my area.
Nestor Your Friendly Neighborhood SysOp ------------------------------------------
Dear Dr Zaluzec
I'd be very grateful if you could help me with a problem I'm having.
I'm writing an essay in low temperature scanning electron microscopy in biology, but I'm finding it difficult to track down up-to-date source material.
If you could suggest any papers, books or book chapters related to the subject, it would be a great help. Thanks.
The following web site, http://www.emitech.demon.co.uk/, has a downloadable technical brief on low temperature EM. It talks about Emitech's products, but also has a discussion of low-temp EM in more general terms. A more dated, but very useful, reference is the book "Low Temperature Methods in Biological Electron Microscopy", which is part of the "Practical Methods in Electron Microscopy" series edited by Audrey Glauert. My copy is dated 1985. I don't know if later editions exist.
Hope this is useful.
Disclaimer: We have no financial interest in Emitech's products, other than being users of them.
Regards, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: DARRYL GRAY [mailto:ddg99-at-aber.ac.uk] Sent: Wednesday, March 01, 2000 7:53 AM To: Microscopy-at-sparc5.microscopy.com
Colleagues.... Can anyone help this guy? It's not my area.
Nestor Your Friendly Neighborhood SysOp ------------------------------------------
Dear Dr Zaluzec
I'd be very grateful if you could help me with a problem I'm having.
I'm writing an essay in low temperature scanning electron microscopy in biology, but I'm finding it difficult to track down up-to-date source material.
If you could suggest any papers, books or book chapters related to the subject, it would be a great help. Thanks.
Sorry I deleted the original question so I don't remember who posted it. Then I came across a couple of papers by John E. Scott on staining for PG's using Cupromeronic Blue. Never tried this myself, so I don't know how well it works. Here's one reference:
J.E. Scott and M. Haigh, "Identification of specific binding sites for keratan sulphate proteoglycans and chondroitin-dermatan sulphate proteoglycans on collagen fibrils in cornea by the use of Cupromeronic Blue in 'critical-electrolyte-concentration' techniques." Biochem J. (1988) 253:607-610.
-- Karen S. Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN
Does anybody know a recipe for staining the amorphous part of poly(vinylidene fluoride) and poly (e-caprolactone)? We need to view the crystalline lamellae of these polymers by TEM.
Thank you in advance for your help.
Antoine
====================== Dr Antoine GHANEM SOLVAY Research and Technology Rue de Ransbeek 310 1120 Brussels Belgium Phone 32-2-2643422 Fax 32-2-2642055 antoine.ghanem-at-solvay.com ======================
Someone in the lab needs help with hand sectioning Arabidopsis seedling stems. They are small, less than a mm in dia. and very delicate. She needs cross sections of the stem from a specific zone to do microscopy and pigment localization.
I tried it using traditional techniques, but without success. She must have sections of fresh material for her stains, she says frozen sections don't work. Any ideas?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Suspend them in some stiff agarose and try a vibratome. There is also the old elder pith trick, if you haven't already tried it. There are only a few of us ancient enough to have ever heard of that one.
At 10:57 AM 03/01/2000 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
} } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } We have made some headway in a related problem by encasing the fresh tissue in low melting point agarose, around 3%. The idea here is to make a cube of agarose with the stem oriented in side. You can then cut the whole cube with a razor. The agarose doesn't impede or interfere with most stains. This didn't work perfectly, but it was better than nothing.
Hope this helps, Tobias Baskin _ ____ ^ __ ____ Tobias I. Baskin / \ / / \ / \ \ University of Missouri / | / / \ \ \ Biological Sciences /___/ /__ /___ \ \ \__ 109 Tucker Hall / / / \ \ \ Columbia, MO 65211-7400 USA / / / \ \ \ voice: 573-882-0173 / /____ / \ \__/ \____ fax: 573-882-0123
Here are the most recent posts. I plan to use the calendar from brownbear but have not actually implemented it yet.
http://www.brownbearsw.com
Date: Thu, 26 Aug 1999 14:45:39 -0400 Reply-To: Confocal Microscopy List {CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU} Sender: Confocal Microscopy List {CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU} } From: Kristi DeCourcy {decourcy-at-MAIL.VT.EDU}
At 12:04 PM -0600 3/1/0, Karen Zaruba wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************************ You can also try any number of cationic stains (ruthenium red, alcian blue. safranin-O, etc)at concentration of about 0.1-0.5%in the primary fixation step. These dyes are electron dense and will bind to the negatively charged PG's. You can also track down references from the 1970's & early 1980's by Simeonescu & Simeonescu who did a lot of work with these types of dyes. I have used them (years ago) to look at the extracellular matrix in heart muscle.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I have a set of samples giving me fits here. This person is trying to visualize the ruthenim doped into a polymer. I have not had much luck working with this stuff and was wondering if any of you had any suggestions to pass along. A description of the sample follows.Here is what I have tried so far.
She originally brought this in on a glass slide and it is about 100µm thick. I tried to use backscattering to visualize the Ru metal in the FESEM. She thought it might be in little islands. No luck. It was too thick to shoot a beam through with the TEM so I embedded some in cold LR-White resin and sectioned. It offered enough support to get some areas thin enough to view through, but I did not see any metal. I suggested she take the grids to the Materials EM unit here on campus as the have EDS and are better equipped for this, but they saw nothing either.
I then had her try to spin coat slids,grids,and mica to float off the film, but she cannot get it to work so now I am back to the slides, scraping material off, and cold embedding. The last set was too rubbery though and even in epoxy on a good diamond I was unable to obtain a section of the material. It just moved out of the way and popped back up with nice resin sections floating away. I don't get any polymers here and we are almost exclusively a biologic unit so do any of you have any suggestions??
} Date: Wed, 1 Mar 2000 11:28:21 -0500 (EST) } From: Joanne Bedlek {bedlek-at-chem.ufl.edu} } To: Scott Whittaker {sdw-at-biotech.ufl.edu} } Subject: Re: your mail } } Scott, } } The polymer is a polydimethylsiloxane. The metal (Ru) is dissolved in } methylene chloride and added to the silicone fluid. As the polymer cures } it "hardens" to a rubbery material. } } The TEM images you took give us an idea of the polymer distribution. With } the new films I have, we are interested in the polymer distribution again. } I } suspect we will see the same droplet pattern as before. This will be a } characterization technique for a paper submission. } } As for the dispersion technique, MAIC tried EDS. They were not able to } detect the Ru. They did tell me that there is a higher content of Si on } the outer sides of the droplets as opposed to the interior areas. This is } interesting. } } I will drop off the new samples either next week or the week after. It } depends when I get back next Friday. } } I redid the one sample that was giving you so much trouble in slicing. I } hope it has cured better this time. } } } } This may be more of a dispersion thing. Materials should be able to see it } } if it is in there. You might get with them and see if they have done } } anything similar as we had no luck with that last set you brought in. I } } could also post a discussion to the microscopy listserver and see what } } comes up. I just need to know more about the composition of the samples } and } } stuff like that } }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I agree with using the agar. Try 3% and go up if you need to. If the stem is soft, you'll be able to cut it with a "Vibratome" or vibrating microtome. If it is hard, you won't.
Sara
On Wed, 1 Mar 2000, Jon Krupp wrote:
} Date: Wed, 1 Mar 2000 10:57:57 -0800 } From: Jon Krupp {jmkrupp-at-cats.ucsc.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: LM, help with hand sectioning } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings: } } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Question: I read a lot of information about Naturpaths using Dark Field Microscopy for live blood analysis and blood crystallization testing. They claim they can see nutritional deficiencies as well as disease process through this type of testing. Is this possible and how accurate are these tests? Thanks.
We have the opportunity to purchase a used Zeiss 960 SEM for use by undergraduate biology students in a number of courses and for research.
I would be interested in any comments, advice, etc. about the instrument and its appropriateness for this type of application, such as maintenance, resistance to abuse, and ease of use.
Elder pith being in short supply you might try supporting it in a fresh carrot. I have had some luck with that in a hand microtome.
I came across the method in a book from the 30's.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 } From: "Greg Erdos" {gwe-at-biotech.ufl.edu} } } } Suspend them in some stiff agarose and try a vibratome. There is also the } old elder pith trick, if you haven't already tried it. There are only a } few of us ancient enough to have ever heard of that one. } } } } Someone in the lab needs help with hand sectioning Arabidopsis seedling } } stems. They are small, less than a mm in dia. and very delicate. She needs } } cross sections of the stem from a specific zone to do microscopy and } } pigment localization. } } } } I tried it using traditional techniques, but without success. She must have } } sections of fresh material for her stains, she says frozen sections don't } } work. Any ideas? } } } } Jonathan Krupp } } Microscopy & Imaging Lab } } University of California } } Santa Cruz, CA 95064 } } (831) 459-2477 } } jmkrupp-at-cats.ucsc.edu } } } } } } Gregory W. Erdos, Ph.D. Ph. 352-392-1295 } Assistant Director, Biotechnology Program } PO Box 110580 Fax: } 352-846-0251 } University of Florida } Gainesville, FL 32611 } } }
Jon, it seems that project is beyond hand sectioning. There is a whole series of instruments available that use a oscillating or vibrating knife and the best can section down to 10 micrometers. Buy, beg, borrow don't steal one of those instruments. The mid to lower price range should do your job. Disclaimer: ProSciTech sells these instruments Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, March 02, 2000 4:58 AM, Jon Krupp [SMTP:jmkrupp-at-cats.ucsc.edu] wrote: } } Greetings: } } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } }
} Question: I read a lot of information about Naturpaths using Dark Field } Microscopy for live blood analysis and blood crystallization testing. They } claim they can see nutritional deficiencies as well as disease process } through this type of testing. Is this possible and how accurate are these } tests? Thanks. } } --------------------------------------------------------------------------- }
You CAN see red cells. But the Naturopaths are only using this as a hook to give (pseudo)scientific validity to their diagnosis. It all COMPLETE HOKUM. } } }
We are looking for a high voltage measurement cable (cable which links the high voltage transformer to the measurement resistances) in working order and dedicated to a Siemens Elmiskop 102 TEM (manufacturing model number 2810)
Here are the specifications of the cable we are looking for : 125 kV - 1.5 m length
Would it be possible to also give us a cost estimation ?
Thanks in advance
Best regards
\\ _// X-FERT ! -(-at- -at-)- -------------------oOO--(_)--Ooo------------ Philippe Drouillon Solvay Research and Technology Electron Microscopy and Image Analysis - IT Coordinator Rue de Ransbeek, 310 B-1120 Brussels (Belgium) phone : (00 32) 2 264 24 47 mailto:philippe.drouillon-at-solvay.com
A modern substitute for elder pith is expanded polystyrene foam. The dense EPS strips sold by Diatome for cleaning diamond knives are excellent for sectioning very small items like Arabidopsis stems. Chris Jeffree
} From: Gordon Couger {gcouger-at-rfdata.net} To: Jon Krupp {jmkrupp-at-cats.ucsc.edu} , Microscopy-at-sparc5.microscopy.com, Greg Erdos {gwe-at-biotech.ufl.edu}
Dear Jon, I have had great success using LR White Medium grade methacrylate for such specimens as iris ovules, mistletoe/host tissues, and a variety of young plant material. Although time consuming, these tissue were processed with osmium tetroxide, rinsed in buffer, and exposed to a 0.025% tannic acid buffer to retain lipids, starch granules, and other items frequently lost during dehyration in EtOH.
One other advantage is the variety of staining procedures one can use with material embedded in LR White. Furthermore, in addition to lipid stabilization, osmium and tannic acid provide additional contrast to the sections.
Hope this helps. Any questions? Email me at tiekotte-at-up.edu
Cheers! -Ken
---------- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97303
On Wed, 1 Mar 2000, Jon Krupp wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings: } } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
} trying to } visualize the ruthenim doped into a polymer. I have not had much luck... } Scott, Sounds like a job for cryo-ultramicrotome thin-sectioning at approximately the glass transition temperature of the polymer. This then to be followed by TEM imaging and EDS analysis of thin-sections, or FESEM-EDS examination of the "faced-off" polymer block cross-section.
Many rubbery polymers can be stained "enblock" before sectioning, for example by submersion in a dilute ruthenium tetroxide (RuO4) solution. This "staining" often aids the sample prep, by hardening the polymer (actually it initiates some cross-linking) and allowing better sectioning and sometimes actually enabling ambient microtomy. It sounds like you will not be able to make use of this technique since you are looking for the Ru distribution in the polymer.
A great polymer reference is the book, Polymer Microscopy, by Linda Sawyer and David Grubb (Chapman and Hall). Good Luck! Brad Huggins BP Amoco, Naperville, IL Microscopy and Microanalysis Lab 630 420-3668
} ---------- } } From: Scott Whittaker[SMTP:sdw-at-biotech.ufl.edu] } Sent: Wednesday, March 01, 2000 4:12 PM } To: microscopy-at-sparc5.microscopy.com } Subject: TEM polymer help } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have a set of samples giving me fits here. This person is trying to } visualize the ruthenim doped into a polymer. I have not had much luck } working with this stuff and was wondering if any of you had any } suggestions } to pass along. A description of the sample follows.Here is what I have } tried so far. } } She originally brought this in on a glass slide and it is about 100µm } thick. I tried to use backscattering to visualize the Ru metal in the } FESEM. She thought it might be in little islands. No luck. } It was too thick to shoot a beam through with the TEM so I embedded some } in } cold LR-White resin and sectioned. It offered enough support to get some } areas thin enough to view through, but I did not see any metal. I } suggested } she take the grids to the Materials EM unit here on campus as the have EDS } } and are better equipped for this, but they saw nothing either. } } I then had her try to spin coat slids,grids,and mica to float off the } film, } but she cannot get it to work so now I am back to the slides, scraping } material off, and cold embedding. The last set was too rubbery though and } even in epoxy on a good diamond I was unable to obtain a section of the } material. It just moved out of the way and popped back up with nice resin } sections floating away. I don't get any polymers here and we are almost } exclusively a biologic unit so do any of you have any suggestions?? } } } } Date: Wed, 1 Mar 2000 11:28:21 -0500 (EST) } } From: Joanne Bedlek {bedlek-at-chem.ufl.edu} } } To: Scott Whittaker {sdw-at-biotech.ufl.edu} } } Subject: Re: your mail } } } } Scott, } } } } The polymer is a polydimethylsiloxane. The metal (Ru) is dissolved in } } methylene chloride and added to the silicone fluid. As the polymer cures } } it "hardens" to a rubbery material. } } } } The TEM images you took give us an idea of the polymer distribution. } With } } the new films I have, we are interested in the polymer distribution } again. } } I } } suspect we will see the same droplet pattern as before. This will be a } } characterization technique for a paper submission. } } } } As for the dispersion technique, MAIC tried EDS. They were not able to } } detect the Ru. They did tell me that there is a higher content of Si on } } the outer sides of the droplets as opposed to the interior areas. This } is } } interesting. } } } } I will drop off the new samples either next week or the week after. It } } depends when I get back next Friday. } } } } I redid the one sample that was giving you so much trouble in slicing. I } } hope it has cured better this time. } } } } } } } This may be more of a dispersion thing. Materials should be able to } see it } } } if it is in there. You might get with them and see if they have done } } } anything similar as we had no luck with that last set you brought in. } I } } } could also post a discussion to the microscopy listserver and see what } } } comes up. I just need to know more about the composition of the } samples } } and } } } stuff like that } } } } } } } } } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { } GO GATORS } Scott D. Whittaker 218 Carr Hall } EM Technician Gainesville, FL 32610 } University Of Florida ph 352-392-1184 } ICBR EM Core Lab fax 352-846-0251 } sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ } The home of " Tips & Tricks " } } } } } }
We used High- and Low-iron diamine staining in our lab for looking at glycosaminoglycans several years ago. This is principally EM level. They can be enhanced with Thiocarbohydrazide-silver proteinate staining. I still have the protocols that we used. My reference database is at another location but the ones below should help you get an idea of what is involved and who did the work. For HID/LID check into the work by Sam Spicer, I believe he pioneered it. For TCH-SP, the work is by Thiery and speaking French would be a plus or Sannes.
Hope this helps
Chuck
REFERENCESREFERENCESREFERENCES FOR IRON DIAMINE AND TCH-SP STAINING:
IRON DIAMINE STAININGIRON DIAMINE STAININGIRON DIAMINE STAINING:
Gad, Adel, and B. Sylven. On the nature of the high iron diamine method for sulfomucins. J Histochem Cytochem. 17:156-60 (1968).
Lev and Spicer. Am. J. Pathol. 46:23 (1965).
Sorvari, Tapani E. Histochemical observations on the role of ferric chloride in the high-iron diamine technique for localizing sulphated mucosubstances. Histochem. J. 4:193-204 (1972a).
Sorvari, Tapani E. Binding of ferric ions to nuclei and other tissue sites in sections stained for sulfated mucosubstances by the high-iron diamine methods. Stain Technol. 47:245-48 (1972b).
Sorvari, T. E. and H. S. Arvilommi. Some chemical, physical, and histochemical properties of three diamine fractions obtained by gel chromatography from the high-iron diamine staining solution used for localizing sulphated mucosaccharides. Histochem. J. 5:119-30 (1973).
Spicer, S. S. The use of various cationic reagents in histochemical differentiation of mucopolysaccharides. Am. J. Clin. Pathol. 36:393-407 (1961).
Spicer, S. S. Diamine methods for differentiating mucosubstances histochemically. J Histochem Cytochem. 13:211-34 (1965).
Spicer, S. S., et al. Ultrastructural visualization of sulphated complex carbohydrates in blood and epithelial cells with the high-iron diamine procedure. Histochem. J. 1O:435-52 (1978).
Spicer, S. S. and M. H. Jarrel. Histochemical reaction of an aromatic diamine with acid groups and periodate engendered aldehydes in mucopolysaccharides. J Histochem Cytochem. 9:368-79 (1961).
Takagi, Minoru, et al. Ultrastructural localization of acidic glycoconjugates with the low iron diamine method. J Histochem Cytochem. 30:471-6 (1982).
Denys, Francis R., et al. An improved technique to enhance high-iron diamine reactive sites with thiocarhohydrazide-silver proteinate. J. Nihon Univ. Sch. Dent. 25(2):103-6 (1983).
Sannes, P.L., et al. Ultrastructural localization of sulfated complex carbohydrates with a modified iron diamine procedure. J. Histochem. Cytochem. 27:1108-11 (1979).
Takagi, M., et al. J. Histochem. Cytochem. 30:1179 (1982).
Thiery, J.P. Mise in evidence des polysaccharides sur coupes fines en microscopie electronique. J. Microscop. 6:987-1018 (1967).
------------------------------------- Name: Charles Gilbert VOC: (704) 355-5261 Carolinas Medical Center FAX: (704) 355-8424 Dept of Pediatric Research digPager: (704) 355-4088 : 2058 PO Box 32861 Charlotte, NC 28232-2861
Does anyone know of a way to label for peptidoglycan at the light or electron microscope level?
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
I am new in this discussion groud, my background is in scanning probe microscopy and mass spectroscopy. At this time I am interested in analysing plastic materials such as those used in semiconductor packaging. I would like to use FT-IR. Is this an appropriate direction to take given that some of my samples are going to be small (tens micrometers). What are the material limitations of the technique. Thank you in advance Jocho
Jochonia N. Nxumalo Material Science Engineer
Chipworks Inc. 3685 Richmond Road, Suite 500, Ottawa, Ontario, K2H 5B7 Tel:(613) 829-0414 Fax:(613) 829-0515 mailto:jnxumalo-at-chipworks.com
Dear Professor Luchtel My apologies for the confusion - I was writing without reference to Diatome's literature. EPS is just short for Expanded PolyStyrene but their polystyrol rods would be the same thing. I have found these Diatome rods useful for a number of applications, and that their density, which is a little higher than usual for commercial polystyrene foam, provides good support to the specimen when hand sectioning. However, if you want to try it out on the cheap, why not cut strips off an insulated polystyrene box or insulation tile.
By the way, the best type of blade for hand sectioning used to be the Blue Gillette double-edged carbon steel razor blades, which many of you may be too young to know about. The new, doubtless improved, stainless steel type never seems to be so sharp, and their edges dull faster. Is there still a manufacturer of carbon steel razor blades left in the world, or am I doomed to mourn forever?
Good luck Chris Jeffree
Date sent: Thu, 2 Mar 2000 07:56:53 -0400 To: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } From: Dan Luchtel {dluchtel-at-u.washington.edu}
} } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } } Jon -
After you get the stem supported (you've gotten lots of good suggestions), try taping 2 razor blades together, with a thin spacer (thick paper, filecard, etc.). Cheaper than a vibratome...
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Dear Ken, You've spurred a question that is related, yet not exactly on the same track. Does tannic acid affect the antigenicity of the tissue? I am just about to embed some tissue in LR White for immunocytological studies, and though it is established that osmium is a problem for immuno work, I have heard nothing about tannic acid. I would love to find a way to enhance the contrast of my tissue, as it often has a surreal, ghostly appearance. Thanks, Kristen
At 01:20 AM 3/2/2000 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I received a sample of DNA microarray on a glass slide and I was asked to do SEM because the investigator wants to see single and double stranded DNA. Does anyone know if this is possible. My previous exposure to EM of DNA samples was in TEM. I would appreciate any help, comments or suggestions.
Thank you,
Corazon D. Bucana ******************************************************* Corazon D. Bucana, Ph.D. Department of Cancer Biology U.T. M.D. Anderson Cancer Center 1515 Holcombe Blvd. Box 173 Houston, Texas 77030 Phone: (713) 792-8106 FAX: (713) 792-8747 Email:bucana-at-audumla.mdacc.tmc.edu FAX: (713) 792-8747
The following equipment is surplus to requirements and is available on the basis that you are responsible for all removal and transport costs. If there are competing bids they will be sold to the highest bidder. There is no guarantee that the equipment is in operational or serviceable condition although, to the best of our knowledge, it was functional when last operated.
----- Original Message ----- } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } By the way, the best type of blade for hand sectioning used to be } the Blue Gillette double-edged carbon steel razor blades, which } many of you may be too young to know about. The new, doubtless } improved, stainless steel type never seems to be so sharp, and } their edges dull faster. Is there still a manufacturer of carbon steel } razor blades left in the world, or am I doomed to mourn forever?
I miss the old blades as well. I cured the shaving problem I quit. For hand sections I have better luck with a more rigid blade. Razor blades and straight razors tend to dig in for me. I have a lot better luck with a 2.5 inch wood chisel sharpened on glass with silicone carbide grit. It is a lot of trouble to get sharp but it works better for me than razor blades.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } Yes, carbon blades are still made - one of the makers is a Japanese company - I have a box in my hand, but unfortunately I can't read Japanese - the only English words on it is "Feather". There may also be many other makers of these in Asia and Eastern Europe.
Grazyna Tokarczyk gmtokarc-at-is.dal.ca Dalhousie University Halifax, Canada } } ----- Original Message ----- } } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } } By the way, the best type of blade for hand sectioning used to be } } the Blue Gillette double-edged carbon steel razor blades, which } } many of you may be too young to know about. The new, doubtless } } improved, stainless steel type never seems to be so sharp, and } } their edges dull faster. Is there still a manufacturer of carbon steel } } razor blades left in the world, or am I doomed to mourn forever? } } I miss the old blades as well. I cured the shaving problem I quit. } For hand sections I have better luck with a more rigid blade. } Razor blades and straight razors tend to dig in for me. I have } a lot better luck with a 2.5 inch wood chisel sharpened on glass } with silicone carbide grit. It is a lot of trouble to get sharp but } it works better for me than razor blades. } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00 } } } }
Dear all, We have a complete Balzers 301 freeze fracture unit with thickness monitor, many ancillary accessories and spare parts that is in need of a new home. No guarantees that the equipment is in operational or serviceable condition but, to the best of our knowledge, it was functional when last used. Please contact me directly for additional information. Cheers, Mark **************************************************** Mark A. Sanders University of Minnesota Program Director Twin Cities Campus Imaging Center St. Paul, MN 55108 23-35 Snyder Hall ph: 612-624-3454 1475 Gortner Ave. fax: 612-624-1799 http://biosci.umn.edu/imagingcenter President: Minnesota Microscopy Society http://resolution.umn.edu/MMS/
Dear colleagues, We are in the process to get a confocal microscope for a multi-user facility. We are not decided yet between different apparatus such as Wallac ultraview, zeiss LSM 510 or Biorad. We would very much appreciate any advice helping us for this choice.
Many thanks.
Pierre-Yves Sizaret Electron Microscopy Facility University of Tours France
Twice now, I have had customers in Europe refer to a technique as REM. The sample prep sounds the same as SEM, and the first time I passed it off as a typo. Now, with this second reference, I'm not so certain. Is it possible that these customers are referring to SEM, possibly with their native language? Or is it a whole different technique/instrument? Could someone please help me clear up this confusion?
Thanks in advance,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Raster Elektronen Mikroskopie = Scanning Electron Microscopy
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Robert J. Palmer Jr., Ph.D. Oral Infection and Immunity Branch National Institute of Dental and Craniofacial Research Bldg. 30, Room 308 National Institutes of Health Bethesda MD 20892 Ph 301-594-0025 FAX 301-402-0396
Has anyone heard of a fluorescent agar or a way to make agar fluorescent? We want to make sure the agar can be fluorescently imaged, but the dye cannot diffuse out into the surrounding media. Thanks- Dave
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Dear Colleagues In a microstructurally uniform alloy, can one expect to get the chemical composition of the alloy through microprobe (assuming all the corrections are applied correctly)? Would the composition tally with the one determined by chemical analysis? TIA Anita
Confocal issues are discussed on another listserver--see below
The best solution is to get one of each manufacturer's confocal systems! Each one has problems and each one has solutions. In my experience the Wallac system is not sensitive enough for some of our samples but is great for live imaging. The BioRad 1024 is difficult because of their interface to the Nikon TE300 and their use of an old OS2 operating system (the migration to Windows NT is still in the future) but they have a good service organization in our area and there are many other users that can help with advice. I do not have hands on experience with the Leica, Zeiss, Noran and other systems. They each have attractive features. A recent discussion of Zeiss and Leica will be sent directly to you. Make a list of features/advantages/disadvantages and weight them--the results may surprise you and guide your decision.
to subscribe to the confocal newslist send an email to:
{LISTSERV-at-LISTSERV.ACSU.BUFFALO.EDU}
in the body of the message write:
SUBSCRIBE CONFOCAL your name
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
Scanning Electron Microscopy (English) RasterElektronenMikroskopie (German)
(It's 3 long words in English, one monumental word in German. I once gave a talk titled: "Rasterelektronenmikroskopische Untersuchungsmethoden fuer prozessinduzierte Halbleiterdefekte".... German is not for the timid ;-)
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: "David_Bell-at-Millipore.com"-at-sparc5.microscopy.com[SMTP:"DAVID_BELL-at-MILLIP ORE.COM"-at-SPARC5.MICROSCOPY.COM] } Sent: Friday, March 03, 2000 9:02:50 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Question Re: REM } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Colleagues,
Twice now, I have had customers in Europe refer to a technique as REM. The sample prep sounds the same as SEM, and the first time I passed it off as a typo. Now, with this second reference, I'm not so certain. Is it possible that these customers are referring to SEM, possibly with their native language? Or is it a whole different technique/instrument? Could someone please help me clear up this confusion?
Thanks in advance,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
For those who are 'hand sectioning', try the teflon coated single edged blades. I believe Ted Pella and EMS carry them. They are alittle more money, but they slide through tissue with great ease.
Cheers, Ken
On Thu, 2 Mar 2000, Gordon Couger wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } ----- Original Message ----- } } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } } By the way, the best type of blade for hand sectioning used to be } } the Blue Gillette double-edged carbon steel razor blades, which } } many of you may be too young to know about. The new, doubtless } } improved, stainless steel type never seems to be so sharp, and } } their edges dull faster. Is there still a manufacturer of carbon steel } } razor blades left in the world, or am I doomed to mourn forever? } } I miss the old blades as well. I cured the shaving problem I quit. } For hand sections I have better luck with a more rigid blade. } Razor blades and straight razors tend to dig in for me. I have } a lot better luck with a 2.5 inch wood chisel sharpened on glass } with silicone carbide grit. It is a lot of trouble to get sharp but } it works better for me than razor blades. } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00 } } } }
The issue might only be are all the elements "visible" to the x-ray analyzer. I don't trust the C or N numbers that my system spits out. Since I am using EDS, I will usually trust my careful analyses to a few tenths of a percent. I can see down to about a tenth of a percent, but the relative errors are large. But it is usually well sufficient for determining the alloy.
Now just because the alloy is microstructurally uniform, do not count on the composition to be uniform. I have seen zoning in grains and changes in composition with length within single crystal ingots. And if you have multiphase microstructure, it can be challenging to determine to overall composition.
Warren S.
At 01:55 PM 3/3/2000 -0500, you wrote:
} Dear Colleagues } In a microstructurally uniform alloy, can one expect to get the chemical } composition of the alloy through microprobe (assuming all the corrections } are applied correctly)? Would the composition tally with the one } determined by chemical analysis? } TIA } Anita
In reply to David Bell's inquiry, I know some Eujropeans refer to an SEM as a Raster Electron Microscope.
Sam Purdy National Steel Technical Center Trenton MI
} ---------- } From: "David_Bell-at-Millipore.com"-at-sparc5.microscopy.com } Sent: March 2000 11:02 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Question Re: REM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Colleagues, } } Twice now, I have had customers in Europe refer to a technique as REM. } The } sample prep sounds the same as SEM, and the first time I passed it off as } a } typo. Now, with this second reference, I'm not so certain. Is it } possible } that these customers are referring to SEM, possibly with their native } language? Or is it a whole different technique/instrument? Could someone } please help me clear up this confusion? } } Thanks in advance, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation } 80 Ashby Road } Bedford, MA 01730 } (781) 533-2108 } }
I have in my hot little hand a box of carbon steel single edge razor blades from Ted Pella. For the few times that I needed to to make a transparent section (being a LM metallographer), I found these razor blades to be effective. Usual disclaimers
Sam Purdy } ---------- } From: Grazyna M Tokarczyk } Sent: March 2000 926 } To: Gordon Couger } Cc: c.jeffree-at-ed.ac.uk; Dan Luchtel; microscopy-at-sparc5.microscopy.com } Subject: Re: hand sectioning - carbon blades } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Thu, 2 Mar 2000, Gordon Couger wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } Yes, carbon blades are still made - one of the makers is a Japanese } company - I have a box in my hand, but unfortunately I can't read Japanese } - the only English words on it is "Feather". There may also be many other } makers of these in Asia and Eastern Europe. } } Grazyna Tokarczyk } gmtokarc-at-is.dal.ca } Dalhousie University } Halifax, Canada } } } } ----- Original Message ----- } } } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } } } By the way, the best type of blade for hand sectioning used to be } } } the Blue Gillette double-edged carbon steel razor blades, which } } } many of you may be too young to know about. The new, doubtless } } } improved, stainless steel type never seems to be so sharp, and } } } their edges dull faster. Is there still a manufacturer of carbon steel } } } razor blades left in the world, or am I doomed to mourn forever? } } } } I miss the old blades as well. I cured the shaving problem I quit. } } For hand sections I have better luck with a more rigid blade. } } Razor blades and straight razors tend to dig in for me. I have } } a lot better luck with a 2.5 inch wood chisel sharpened on glass } } with silicone carbide grit. It is a lot of trouble to get sharp but } } it works better for me than razor blades. } } } } Gordon } } } } Gordon Couger gcouger-at-couger.com } } } } Stillwater, OK www.couger.com/gcouger } } 405 624-2855 GMT -6:00 } } } } } } } } } }
I am looking for information on an instrument that I recently acquired. The item is a McArthur Blood Cell Counter, which is a McArthur portable microscope with a number of mechanical additions and a special stage, apparently for doing blood count. It is in a neatly bundled case, very portable, but lacking instructions. If anyone one is familiar with the specific application of the set I would appreciate comments, particularly if you have ever used such an instrument.
There's also the technique of Reflection Electron Microscopy, which is the imaging analogue of RHEED (reflection high energy electron diffraction). I am sure your reference is to the SEM though, as the other respondents indicate.
Just for interest, REM (like RHEED) is a surface sensitive technique and can give beautiful images of monolayer surface steps, eg on the sapphire surface, and this can all be done in the TEM. The sample is simply rotated 90 degrees from the usual TEM imaging position and so electrons are 'reflected' from the surface at glancing angle as they pass through the sample chamber.
My best wishes to all,
Mark
%%%%%%%%%%%%%%%%%%%%%%%%%%% Mark Yeadon Senior Research Fellow Institute of Materials Research and Engineering 3 Research Link Singapore 117602
Assistant Professor Department of Materials Science National University of Singapore Singapore 119602
The original question has been answered well. However, I would like to add the historical footnote that von Ardenne's seminal 1938 paper was entitled "Das Elektronen-Rastermicroskop". A raster is a pattern of horizontal lines. The German word is derived form the Latin "rake". I guess a rake's pattern is a raster, a bit coarse for an SEM though. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Saturday, March 04, 2000 2:03 AM, "David_Bell-at-Millipore.com"-at-sparc5.microscopy.com [SMTP:"David_Bell-at-Millipore.com"-at-sparc5.microscopy.com] wrote: } } } Hello Colleagues, } } Twice now, I have had customers in Europe refer to a technique as REM. The } sample prep sounds the same as SEM, and the first time I passed it off as a } typo. Now, with this second reference, I'm not so certain. Is it possible } that these customers are referring to SEM, possibly with their native } language? Or is it a whole different technique/instrument? Could someone } please help me clear up this confusion? } } Thanks in advance, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation } 80 Ashby Road } Bedford, MA 01730 } (781) 533-2108 }
I have recently encountered several mentions of this type of thing recently, in the oddest places. One was a presentation by the motivational speaker, Tony Robbins, who, in front of 13,000 people at the Hartford Civic Center, described how his recent feelings of fatigue and lack of energy had been diagnosed by darkfield microscopy as due to chlamydia in his blood. He cited work done by Dr. Robert Young at InnerLight.
As for crystal testing, I am still a strong supporter of Polarized light work for that. (Interestingly, it looks a lot like darkfield and, perhaps, is mis-cited by Naturpaths who might not know the difference).
Hope this is helpful.
Caveat: MME has no connection with either Mr. Robbins, Dr. Young, or InnerLight.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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} ******************************************************
At 06:33 PM 3/1/00 -0600, pfieber-at-elite.net"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We did a bit of market and applications research into this area for the 3 years before it emerged (as well as launching a company). These microarrays are typically analyzed on larger scale using instruments which are derivatives of confocal. If you are looking for individual DNA molecules first make sure that they are really likely to be there (i. e, get a full history of how this microarray was prepared and later used). I would then suggest looking at it with AFM. ThermoMicroscopes, Digital, and Molecular Imaging have all had experience in this area. Let me know if you need specific contact info.
Best regards Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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}
At 01:19 PM 3/2/00 -0600, Corazon Bucana wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Not only have I had my hands on several McArthurs, I had the privilege of visiting the inventor at home. Your comment about the "neat bundle" is very accurate. Dr. McArthur showed me a number of accessories (including fluorescence!) for this system. Although I did not have a chance for an extended use, my recollection was that all of them worked passably well.
There were several generations of McArthur microscopes. The early ones were made of metal and were very durable and usable. A somewhat later generation was made of white plastic, I think for the BBC Open University. The general opinion of some of my colleagues who knew this system indicated that the earlier generation was a better bet.
I was not aware that anyone was commercially making this microscope today. Dr. McArthur was advanced in years when I visited him over a decade ago and, at that time, their construction was very much a cottage industry. Is this a new system or are you buying it second hand? (I suspect the latter, since you mentioned that the instructions are missing). It is hard to comment on the bits and pieces without further information. I am planning to attend a number of meetings (Experimental Bio, M&M, Cell Bio, Neuro as well as Semicon West, Materials Solutions, and ISTFA) over the next 6 months. If you are going to any of them or will be in the area, I would be delighted to take a look at what you have and see if I can dredge up any old memories of how it works.
Finally, if you can send pictures, I can forward them to colleagues in the UK who own McArthurs.
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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}
At 07:39 PM 3/3/00 EST, MICROFAB-at-aol.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have recently encountered several mentions of this type of thing recently, in the oddest places. One was a presentation by the motivational speaker, Tony Robbins, who, in front of 13,000 people at the Hartford Civic Center, described how his recent feelings of fatigue and lack of energy had been diagnosed by darkfield microscopy as due to chlamydia in his blood. He cited work done by Dr. Robert Young at InnerLight.
As for crystal testing, I am still a strong supporter of Polarized light work for that. (Interestingly, it looks a lot like darkfield and, perhaps, is mis-cited by Naturpaths who might not know the difference).
Hope this is helpful.
Caveat: MME has no connection with either Mr. Robbins, Dr. Young, or InnerLight.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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} ******************************************************
At 06:33 PM 3/1/00 -0600, pfieber-at-elite.net"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Amazing. Definitely a major feat of dark field light microscopy. To be able to see and distinguish an organism that is between 0.1u and 0.5u is truly amazing.
Did he say what magnification was realized in this DF system? The only DF condensers I know of for compound LMs are not aplanatic. So is begs the question of how one could resolve, much less see, such a tiny bacteria using LM.
Perhaps the sample was specially treated?
gary g.
At 01:09 PM 3/5/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
REM = 'Raster Elektronen Mikroskopie' in German (SEM in english) but also Reflection Electron Microscopy, which also needs not a lot of specimen preparation. It looks very much like preparation for SEM in deed. Regards Gerhard Frank
"David_Bell-at-Millipore.com"-at-sparc5.microscopy.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Colleagues, } } Twice now, I have had customers in Europe refer to a technique as REM. The } sample prep sounds the same as SEM, and the first time I passed it off as a } typo. Now, with this second reference, I'm not so certain. Is it possible } that these customers are referring to SEM, possibly with their native } language? Or is it a whole different technique/instrument? Could someone } please help me clear up this confusion? } } Thanks in advance, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation } 80 Ashby Road } Bedford, MA 01730 } (781) 533-2108
-- Gerhard Frank UNIVERSITAET ERLANGEN-NUERNBERG Phone: +49 9131 85 28606 Institut fuer Werkstoffwissenschaften VII Fax: +49 9131 85 28602 Cauerstr. 6, D-91058 Erlangen, Germany frank-at-ww.uni.erlangen.de
Just a reminder that darkfield is detection limited, not resolution limited. It only takes 2-3 photons, scattered from a feature, to indicate that it is there.
Chlamydia is must be much bigger than the range you specified. We are in the process of sending out a special mailer for a new spectral imaging system, with chlamydia as the "cover shot". I think that this image was acquired with a 40x objective, but will check. Data is intentionally not provided on the card because we are encouraging recipients to go to the web page.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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}
At 04:03 PM 3/5/00 -0600, Dr. Gary Gaugler wrote: } Amazing. Definitely a major feat of dark field light microscopy. } To be able to see and distinguish an organism that is between } 0.1u and 0.5u is truly amazing. } } Did he say what magnification was realized in this DF system? } The only DF condensers I know of for compound LMs are } not aplanatic. So is begs the question of how one could } resolve, much less see, such a tiny bacteria using LM. } } Perhaps the sample was specially treated? } } gary g. } } } } At 01:09 PM 3/5/00 , you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings, To respond to a comment made by Gary Gaugler, and without in ANY WAY seeking to validate or support the claims of naturopaths, darkfield microscopy has imaged individual microtubules, diam 25 nm. Same with Nomarski optics, and others. How is that possible? It is important to keep in mind what the famous "resolution limit" means, it means telling the difference between two and one object. It has nothing to say about the smallest absolute size of an object that can be detected.
If you imagine a perfectly black slide with a pinhole in it made by a perfect iris that can shrink right down to infinitely small (I did say a perfect iris), there is no limit in theory to how small that pinhole can be and still be imaged. Once the diameter of the pinhole becomes less than a certain length, set by diffraction, then the size of the image of the pinhole will no longer get any smaller. Instead, the contrast in the image will go down. But if you can put enough light through and have a good enough detector, you can see that image.
Going back to our microtubules, take a photo of the darkfield image and measure the diameter of the microtuble in the image: it will be around 250 nm or thereabouts.
For the record, the person who I think has published the most dark field images of microtubules is called Hotani, he works in Japan, I am not sure where.
Of course, doing this in practice is not easy, but it is certainly possible.
As ever, Tobias
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An earlier thread on this listserver (May-June 1999, and also summarized in the January/February 2000 issue of Microscopy and Microanalysis) asked about the availability of software for the processing and analysis of 16 bit per channel images, which are obtained from flat bed scanners, cooled digital cameras, and some microscopes (especially AFMs). Following the encouragement of members of this list, we've written a set of image processing and measurement routines for these images. Those interested can get information at http://members.aol.com/FoveaPro/
Disclaimer: Please note that Fovea Pro is a commercial product sold by Reindeer Games. My connection with the company is through my son, Chris, who wrote the programs, and my own role in writing the accompanying tutorial.
We have a Tracor Northern 5500 EDX unit that we would like to give away. It comes with a manual, mainframe, and monitor. If anyone is interested please contact:
Dr Brian Andrews NIH, NINDS Bethesda MD 301-435-2796
sba-at-helix.nih.gov
Thanks Chris :):):):):):):):):):):):):):):):):):):):):):):):):):):):):):)
Christine A. Brantner, Ph.D.
Biologist/Lab Manager
National Institutes of Health National Institute of Neurological Diseases and Stroke Lab of Neurobiology Section of Analytical Cell Biology Bldg 36, room 2A21 phone 301-435-2803 36 Convent Dr fax 301-480-1485 Bethesda, MD 20892-4062
Hello: I will add just a few comments to those of Tobias Baskin. When we first developed Video Microscopy in the early eighties, we were visulizing microtubules and other structures smaller than the ca. 200 nm resolution limit using DIC and computer image enhancement. This is discussed in the early papers from 1981 and 1983 (R.D. Allen and N.S. Allen 1983, J. of Microscopy),and more easily available in the Inoue and Spring second edition of Video Microscopy, Plenum Press. The important thing to remember is that you can visualize items smaller than 100 nm, but you are not resolving them. With the new method Polarization Modulation DIC we get very good images of microtubules and other fine structures (Holzwarth, G., Webb, S.J., Kubinski, D.J., N.S. Allen. 1997. Improving DIC Microscopy with Polarization Modulation. Jour. of Microscopy 188:249-254). Contrast is doubled with this method and it is worth a try. It is commercially available from Hamamatsu or you can build your own ,Nina Allen } Greetings, } To respond to a comment made by Gary Gaugler, and without in } ANY WAY seeking to validate or support the claims of naturopaths, } darkfield microscopy has imaged individual microtubules, diam 25 nm. } Same with Nomarski optics, and others. How is that possible? It is } important to keep in mind what the famous "resolution limit" means, } it means telling the difference between two and one object. It has } nothing to say about the smallest absolute size of an object that can } be detected. } } If you imagine a perfectly black slide with a pinhole in it } made by a perfect iris that can shrink right down to infinitely small } (I did say a perfect iris), there is no limit in theory to how small } that pinhole can be and still be imaged. Once the diameter of the } pinhole becomes less than a certain length, set by diffraction, then } the size of the image of the pinhole will no longer get any smaller. } Instead, the contrast in the image will go down. But if you can put } enough light through and have a good enough detector, you can see } that image. } } Going back to our microtubules, take a photo of the darkfield } image and measure the diameter of the microtuble in the image: it } will be around 250 nm or thereabouts. } } For the record, the person who I think has published the most } dark field images of microtubules is called Hotani, he works in } Japan, I am not sure where. } } Of course, doing this in practice is not easy, but it is } certainly possible. } } As ever, } Tobias } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Nina Stromgren Allen Professor, Department of Botany; Director, Cellular and Molecular Imaging Facility Box 7612, Department of Botany North Carolina State University Raleigh, NC 27695-7612 Phone: 919-515-8382 (Office), 515-3525 (Lab), 515-2727 (Department Secretary) Fax: 919-515-3436
Is anyone familiar with a printer(s) that can provide equivalent detail to a polaroid print or to the images that can be collected from the scanning electron microscopes? I have looked at QMS Color Magic II printers that have 2400 dpi x 600 dpi, 257 shades of gray, with a cell size of 150 cells per inch (lines per inch). This is comparable to a Codonics 300 line dye-sub printer that I have seen. However, the dye-sub printers tend to smear fine details to the point of not being visible. I saw an add for an Alps printer with a dot size of 10 microns at 2400 dpi. The add did not indicate the shades of gray so I have no way of knowing what the numer of cells per inch are. I suspect that it would have 257 shades of gray which would give a cell size of 16x16 and a cells or lines per inch of 150. Does anyone have user information about these or any other printers that might work? No one talks about the cells per inch (lines per inch) in any of the documentation. For me, cells per inch has been the best measure of resolution. For the printers that we have, the extra dots (300 dpi matrix 4x4, 600 dpi matrix 8x8, 1200 dpi matrix 12x12, 2400 dpi matrix 16x16) serve to provide more dynamic shades of gray. A 300 dpi with 17 shades of gray and a 600 dpi with 65 shades of gray both have 75 cells per inch. The only difference is more contrast in the 600 dpi with 65 shades of gray. A simple formula for this is dpi/cell matrix size= cells (lines) per inch, 2400 dpi/16=150 cells per inch and a maximum # of shades of gray = 1+(dpi/lpi)squared. This tid bit of information came from The Image Processing Handbook, 2nd ed. by John C. Russ. I would appreciated any help that I can get in finding a printer that will do the job. Thanks.
In a message dated 3/6/00 5:55:31 PM, Vicki.L.Baliga-at-pmusa.com writes:
.."A 300 dpi with 17 shades of gray and a 600 dpi with 65 shades of gray both have 75 cells per inch. The only difference is more contrast in the 600 dpi with 65 shades of gray. A simple formula for this is dpi/cell matrix size= cells (lines) per inch, 2400 dpi/16=150 cells per inch and a maximum # of shades of gray = 1+(dpi/lpi)squared. This tid bit of information came from The Image Processing Handbook, 2nd ed. by John C. Russ...."
I guess it is worth pointing out that the calculation Vicki has quoted here is a bit optimistic. Most printers have dots that overlap somewhat, and the dots may spread a bit on the paper. Plus the dots are large enough to be resolved by the human eye, which creates interesting effects when they start to touch or overlap. And the shades of grey that perfect dots might produce would be linear rather than log, as human vision perceives brightness. It would be conservative to estimate that a printer that seems technically capable of producing (e.g.) 65 shades of grey in a given printing cell size will actually produce images with about half that number of really useful shades of grey. But that is still OK since that is about all the distinct shades that a person can detect on a print (and more than most Polaroid prints have, although the negatives are much better).
Halftone printing is never going to be as good as something that really covers the resolution cell with a uniform shade of grey (or color). Dye sub and a few ink jet printers do a much better job than laser printers in that regard.
I sent an email to LISTSERV-at-LISTSERV.ACSU.BUFFALO.EDU as you suggested with SUBSCRIBE CONFOCAL in the body of the message.
I got this message back: No LISTSERV list by the name of "CONFOCAL," is known to exist. Note that lists can be marked "confidential" and that the existence of such lists is usually known only to the server that is actually hosting it.
I'm very interested in finding a venue that is more oriented toward confocal microscopy. Any further suggestions?
Catherine Ripley ***************************************** Albert Einstein College of Medicine 1300 Morris Park Avenue Bronx, NY 10461 ***************************************** *****************************************
If you take a drop of blood from a pinprick and put it on a slide, you will see (at rather lower magnification) what the "live cell" analysts purport to see. You will see red cells starting to agglutinate. Thats all. It is nearly impossible to find a white cell. If you find a bacterium you are probably near death. Using dark field adds mystery but no more useful information.
Check out these websites for three different perspectives on live cell microscopy
1. http://www.hcrc.org/faqs/livecell.html
Critical evaluation pointing out the supposed features naturopaths claim as showing pathological change are natural variation.
Gushing plug for the technique including claiming ability to detect poor nutrition, vitamin deficiency etc. See the extract below.
None of these claims that illness can be diagnosed by observing coverslipped blood have any scientific validity at all.
"Live cell analysis can determine cellular nutritional status. Nutritional status is essential to aid in curbing and reversing the free radical cascade of destructive, deteriorating cell structures.
This unique analysis of peripheral blood can reveal a good prospectus of the immune system. By observing the morphology of the white corpuscles and their activity in contrast to the extent of active foreign antigens and microbes within the serum and red blood cells, the strength of the immune system can be judged. Weakness and dysfunction in any one of these three major components and influence the strength and function of the other two. This can be indicative of a progressive disease process."
A photographic negative and/or print is continuous tone. Digital printers (ink jet, laser, etc.) attempt to reproduce continuous tone via modulation of the size of the dots or the number of dots in an area. Continuous tone digital printers typically use dye sublimation to produce photographic quality results. In this regard, there really isn't a true correspondence to dpi. Alternatively, some ink jets use CMYK or even 5 color jets rather than RGB or RGB+B. Some of the new generation do a rather good job of color printing.
But since you are talking about SEM images, these are gray scale images rather than color. So, what are the options for b/w?
600dpi and 1200dpi laser printers do a rather good job of printing SEM images. One must adjust the toner density to achieve maximum quality results. But regardless, they are not continuous tone prints. The closest thing to continuous tone b/w is the Orion (I think that is the name) printer. It is an effective 300dpi and prints on expensive paper--which is par for the course, considering the cost of the printer (about $25K). And the printer is sloooooow.
I use a HP Laserjet 4M Plus and am very pleased with the results. But the printed images are not archival and are not continuous tone. But they are very inexpensive.
If you can describe what your actual end purpose is, perhaps others could offer some good options for you to consider. Are you talking about proofing, archiving, presentations, etc?
gary g.
At 11:20 AM 3/6/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sir Could you please supply me details on "transmission electron microsocopy" for my 16yr old daughter at high school, or alternatively where to look on the Internet. Thanking you Peter Jupe
Splitting hairs on detecting microtubules vs. detecting pathogenic organisms...
I guess I will have a go at this and muddy the waters...
As a background...The inability of Chlamydiae to produce metabolic energy restricts these obligate 'parasites' to an intracellular existence.
The infectious component (elementary body) is about 3um in diameter, well within the resolvable range of the OLM. Once acquired in the host cell through phagocytosis, a vaculated 'initial body' (0.5 -1.0um) is formed: huge by OLM standards!
'Elementary bodies' and 'initial bodies' possess distinctive staining properties using Giemsa staining and Macchiavello's staining techniques, not be mention detection by immunocytochemistry.
One can perhaps differentiate Chlamydia induced inclusion bodies juxtaposed to the nucleus of the host cell using Darkfield illumination. However, differential diagnosis of Chylamydia by Darkfield alone is questionable.
If one considers Darkfield and the ability of a 'substance' of a given refractive index to scatter light with respect to the R.I. of the adjacent material, we must accept the notion of "detection vs. resolution' (B. Foster ' Optimizing Light Microscopy for Biological and Clinical Laboratories', pg. 57): therefore, we may detect an inclusion body, but can we identify the pathogen of the inclusion body? Perhaps Phase Contrast would be more sensitive to refractive differences?
As the devil's advocate, I see the issue of using Darkfield as a definite diagnostic tool being dilute to 'mine is smaller than yours'. Not being up on the naturopathic literature, I question how only using Darkfield Illumination can definitely dx Chlamydia?
However, comparing various organisms and the use of Darkfield Illumination, 'Treponema pallium' can easily be detected by Darkfield; an organism of 0.2 um wide, but 5-15 um in length! Furthermore, the inclusion body of Rabies can easily be detected by Darkfield and Brightfield; however, we i.d. the total sum of virons: an example of 'detection vs. resolution'.
Yes, we as microscopists scoff at unconventional claims. Perhaps, Mr. Tim Robbins should present a blood sample for us to examine under the light of mutual cooperation and enlightenment'!!
I present my premise for your delicate evisceration...
I also tried to subscribe to the confocal list when Larry suggested it and it worked like a charm. I cut and pasted the email address right from his posting.
LISTSERV-at-LISTSERV.ACSU.BUFFALO.EDU
Try again. Cheers, John
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-- C. John Runions, Ph.D. Department of Plant Sciences University of Cambridge Cambridge CB4 1YE UK
email at work cjr41-at-cam.ac.uk email at work runions-at-ntlworld.com Phone (01223) 766 545
In the earlier days {25 years ago} of medical diagnosis, before the advent of the rapid, highly specific methods of identifying pathogenic organisms, darkfield microscopy was routinely applied to rule out specific organisms. In particular, the diagnosis of syphilis was aided with this mode of imaging. When fluids from a suspect lesion were applied to a slide and imaged by darkfield, the characteristic shape of the spirochaete organisms was clearly seen. More recently, darkfield examination of synovial fluids from arthritic joints was used to rule out or diagnose Lyme disease, before specific testing became available.
Is there a short, general reference, like a review, that explains immuno EM, it's uses, major techniques, and shortfalls? I would like to use it to send to investigators that are thinking about using the technique but don't know much about labeling at the EM level. I have a lot of specific references and it would take a lot of time to reference or copy them. If the investigator wants to know more I can direct them to those, otherwise a general one would get them and me on the same page when we start talking specifics on their project. Thanks
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Rick Vaughn writes: Is there a short, general reference, like a review, that explains immuno EM, it's uses, major techniques, and shortfalls? I would like to use it to send to investigators that are thinking about using the technique but don't know much about labeling at the EM level. I have a lot of specific references and it would take a lot of time to reference or copy them. If the investigator wants to know more I can direct them to those, otherwise a general one would get them and me on the same page when we start talking specifics on their project. Thanks Rick Vaughn rlvaughn-at-unmc.edu
The best reference you can ask for is the 1993 book by Gareth Griffith: "Fine Structure Immunocytochemistry." Springer Verlag. In it you will find all the major techniques, pitfalls, antibody use, fixatives and even quantitative methods for estimating antigen number. My copy has been replaced many times, I think because someone else needed it more than I did!
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I know this has been covered periodically, but as I've not needed the technique, I've not paid attention. I need some methods of embedding cells cultured on glass coverslips. Obviously, the most important issue is removing the coverslip from the polymerized block.I've looked through my own reference materials and the only method I found involved using Thermanox coverslips (upon which these particular cells will not grow, I'm informed).Please reply dircectly to my email address as well as to the Listserver (I haven't received postings for several days). Thank you,Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.eduFay and Carl Simons Center for Biology of Hearing and DeafnessCentral Institute for the Deaf4560 Clayton AvenueSt. Louis, MO 63110voice: 314-977-0257 fax: 314-977-0030
I need to fix cells for viewing with SEM. However, the fixation method needs to be consistent with my experimental methods:
1) I need to fix the cells in less than 1 minute and due to this time limitation, I cannot spin the cells into a pellet and must add the cell suspension to the fixative; and
2)I am looking for "effects" to the cell membrane caused by my experiment, so I need to keep any possible fixative damage/effects at a minimum.
I have fixed cells using plunge freezing and viewed them with TEM, this is time consuming and I ran into a lot of freeze damage. I would like to try chemical fixation and SEM. It has been suggested that I try glutaraldehyde + buffers, then postfix with osmium tetroxide and dehydrate in an ethanol series.
Is this the best method? I am open to suggestions. I am using suspensions of prostate cancer cells in RPMI (+serum) growth media; I can perform the experiment in buffered saline and use a high concentration of cells.
Thanks, Robyn
Robyn K. Schlicher, M.S. Laboratory for Drug Delivery Institute for Bioengineering and Biosciences Georgia Institute of Technology 315 Ferst Drive Atlanta, GA 30332 rkslick-at-yahoo.com
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hi } i am doing research at university of massachusetts lowell.i am working on } charaterization of Polytetrafluoroethylene(PTFE).While doing TEM i have } stained the PTFE with osmium tetroxide.Can any one enlighten me on the } mechanism oF OsO4 with PTFE? } thanks } ________________________________________________ } Amit A.Kaulgud } Research Assistant. } University of Massachusetts Lowell } Department of Chemical/Materials Engineering. } 170, Riverside Street,#3 } Lowell MA -01854. } Phone/Voice:978-459-3248. } email: amit_kaulgud-at-hotmail.com
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Below is a question from one of my former students who is now working in a hopsital histotech lab. It concern the selflife of glutaraldehyde fixative. I always mix my fix fresh, so some other perspectives might be useful!! Here's the Question:
} My boss wants to know how long a working solution of glutaraldehyde } remains stable (2.5% in cacodyalate buffer). How long can we keep it in } the refrigerator? } Bob Robert J. Schmitz Electron Microscope Lab Department of Biology, CNR Building University of Wisconsin, Stevens Point Stevens Point, WI 54481 phone (715) 346-2420 FAX (715)346-3624 email rschmitz-at-uwsp.edu http://biology.uwsp.edu/faculty/RSchmitz/home.html
In addition, I would point that Paul's web site at http://www.hei.org/htm/cryo.htm is not bad source, too.
} Date: Wed, 08 Mar 2000 00:40:11 -0800 } From: Paul Webster {pwebster-at-hei.org} } Subject: TEM - ImmunoEM reference } To: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } Reply-to: Paul Webster {pwebster-at-hei.org} } X-Mailer: QuickMail Pro 1.5.4 (Mac) } X-MIME-Autoconverted: from quoted-printable to 8bit by ultra5.microscopy.com id } CAA01345 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
At the Fall MRS this year there will be a symposium dedicated to environmental SEM and low vacuum SEM, invited speakers include Eric Doehne from Getty Conservation Institute, David Joy from the University of Tennessee and Matthew Phillips from The University of Technology in Sydney. Contributed papers from all environmental SEM and low vacuum SEM users are encouraged, the Preliminary Call for Papers follows:
Preliminary Call for Papers , Session for MRS 2000 (Boston) http://www.mrs.org/meetings/fall2000/cfp/symposia.html
Low Vacuum SEM/ESEM in Materials Science "Wet SEM: the Liquid Frontier of Microscopy"
Abstract Deadlines: June 5th for abstracts sent by mail and June 19th for abstracts submitted via the MRS Web Site (http://www.mrs.org).
Scanning electron microscopy and x-ray microanalysis can now be performed at pressures as high as 1 - 5000 Pa (~0.01 to 40 torr) in the class of instruments known variously as "low vacuum", "elevated pressure", and "environmental" SEMs. This represents an increase of 3 to 6 orders of magnitude in pressure compared to the operating conditions of ordinary high vacuum SEMs. At the upper end of this pressure range, water can be maintained in the liquid state at temperatures from 3 - 10 deg. C. This remarkable situation has opened up broad new areas of opportunity in many fields, especially materials science. Dynamic processes can be observed with many of the familiar strengths of conventional SEM: high resolution, large depth of focus, and a variety of imaging signals that reveal compositional, topographic, electrical and other specimen properties. X-ray microanalysis for elemental characterization is also possible, although somewhat compromised. Chemical, physical, and mechanical experiments can readily be performed in user-specified environments. The microscope can thus be thought of as an appendage to an experimental chamber. Papers are solicited in all aspects of materials science in which this new microscopy is used.
Co-organizers:
Dr. Brad Thiel Polymers & Colloids Group Cavendish Laboratory University of Cambridge Madingley Road Cambridge, CB3OHE, UK 44 1223 337 272 44 1223 337 000 (fax) bt202-at-cam.ac.uk
Dr. John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 (734) 936-3352 (734) 763-2282 FAX jfmjfm-at-engin.umich.edu
Dr. Dale Newbury Surface and Microanalysis Science Division National Institute of Standards and Technology Gaithersburg, MD 20899-8371 USA 301-975-3921 301-417-1321 (fax) dale.newbury-at-nist.gov
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42#161# 16' 48" Long. 83#161# 43' 48"
I have used fluoronanogold for a couple of times and it did not work on the EM level. Then I realized that this batch has been already expired. Does anyone know if upon aging fluoronanogold could lose gold leaving only FITC molecule attached to Ig? I did see a fluorescence signal before running silver enhancement though. Currently I will try regular ultrasmall gold probe to clear up this problem but I wonder what was the trick.
thanks for your help
Kyrill
********************** Dr. Kyrill Ukhanov Institute for Zoophysiology, University of Potsdam, Lennestrasse 7a, D-14471 Potsdam, Germany phone +49-0331-9774859 fax +49-0331-9774861
Dear Jaci, The problem of cell embedding after their cutivation on plastic or galss is rather simple. You do not need to use Thermanox. You can use glass. The trick is that immediately after polymerization you have place glasses at -20 degree C and your samples will be detached. If thsi scheme does not work you can use liquid nitrogen. However, do not insert smplaes into it (only glass). If this scheme does not work you can use commercial solution of HF (acid). It dissolve glass efficiently. For instance, coverslips are dissolved in 30 min. Then you have to wash samples in buffer (pH7.4) for a long time and water (several hours) otherwise you will have problems with contrasters. We usually glue samples after detachment from glass and work with this sandwich. For this you have to use incomplete polymerization of Epon on chamber slides (12 hours). Then samples can be detached at -20 and then glued with the fresh resin for 24 hours. If you use full polymerization time on glass or any polymerization on plastic you will not be able to glue samples.
Sincerely yours, Alexander Mironov Italy
On Wed, 8 Mar 2000, Jaci Lett 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 know this has been covered periodically, but as I've not needed the } technique, I've not paid attention. I need some methods of embedding } cells cultured on glass coverslips. Obviously, the most important } issue is removing the coverslip from the polymerized block.I've looked } through my own reference materials and the only method I found involved } using Thermanox coverslips (upon which these particular cells will not } grow, I'm informed).Please reply dircectly to my email address as well as } to the Listserver (I haven't received postings for several days). Thank } you,Jaclynn M. Lett, Research Assistant } jmlett-at-cid.wustl.eduFay and Carl Simons Center for Biology of Hearing and } DeafnessCentral Institute for the Deaf4560 Clayton AvenueSt. Louis, } MO 63110voice: 314-977-0257 fax: } 314-977-0030 } } } }
A few moments in liquid nitrogen and the coverslip will come off. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, March 09, 2000 10:10 AM, Jaci Lett [SMTP:jmlett-at-cid.wustl.edu] wrote: } } } I know this has been covered periodically, but as I've not needed the } technique, I've not paid attention. I need some methods of embedding } cells cultured on glass coverslips. Obviously, the most important } issue is removing the coverslip from the polymerized block.I've looked } through my own reference materials and the only method I found involved } using Thermanox coverslips (upon which these particular cells will not } grow, I'm informed).Please reply dircectly to my email address as well as } to the Listserver (I haven't received postings for several days). Thank } you,Jaclynn M. Lett, Research Assistant } jmlett-at-cid.wustl.eduFay and Carl Simons Center for Biology of Hearing and } DeafnessCentral Institute for the Deaf4560 Clayton AvenueSt. Louis, } MO 63110voice: 314-977-0257 fax: } 314-977-0030 } }
I presume all microscopists use somewhat arbitrary shelf-life assumptions, because we know from experience that within those constraints there normally is no trouble. UA in water } 1 month, made up Spurr's in freezer } 3 months, Lead Citrate } 1 year. For working strength GA I used quite conservatively 1 week.
GA forms an undesirable polymer with time and temperature. So freezing the bulk stock (ampoules can stand a fridge-freezer) results in a very long life, certainly a couple of years and just refrigerated its several months. This is arbitrary too since there is no definite cut-off when the stuff suddenly becomes useless.
I saw a publication many years ago that claimed the "undesirable polymer" protected cells against osmotic shock and in structural studies some polymer may be desirable (the amount can be determined simply in a spectrometer, please don't ask for the peak locations. I don't carry that info!) Apparently its different for immunological studies, for that the freshest GA should be used.
Soon after Sabatini advocated GA for TEM use, I guess now over 30 years ago, as noted, some studies related polymer to storage time/temperature. I don't think that these looked at working strength solutions, but I expect that there would be no difference in relative polymer concentrations.
The reason for shorter shelf-life for working solutions was assumed and probably originated with the common use of additives like sucrose or phosphate buffers. Sucrose in an Os fixatives leads to obvious oxidation within hours. I doubt that cacodylate in GA would be a problem and expect that it would remain useable for several months when refrigerated. I have never bothered to test this, however, and stuck with an assumed short shelf-life for working strengths GA. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, March 09, 2000 10:09 AM, Schmitz, Robert [SMTP:rschmitz-at-uwsp.edu] wrote: } } } Below is a question from one of my former students who is now working in a } hopsital histotech lab. It concern the selflife of glutaraldehyde fixative. } I always mix my fix fresh, so some other perspectives might be useful!! } Here's the Question: } } } My boss wants to know how long a working solution of glutaraldehyde } } remains stable (2.5% in cacodyalate buffer). How long can we keep it in } } the refrigerator? } } } Bob } Robert J. Schmitz } Electron Microscope Lab } Department of Biology, CNR Building } University of Wisconsin, Stevens Point } Stevens Point, WI 54481 } phone (715) 346-2420 } FAX (715)346-3624 } email rschmitz-at-uwsp.edu } http://biology.uwsp.edu/faculty/RSchmitz/home.html } }
Email: puusimak-at-paju.oulu.fi Name: Päivi Maria Susanna Uusimäki
School: the university of Oulu
Question: What is a suitable fixation-method for my specimens? I am trying to find antigen-antibody interactions on the bacterial membrane. My bacteria are different strains of LABs, my special interests are lipoteichoic acids and I am using confocal and fluorescence microscopes. The big problem has been : specimens tend to "float away" under was- hings !
I have recently been asked to participate in an effort at Los Alamos concerned with Electron Tomography of polymers. I haven't done the required image reconstruction before. Does anyone know of any commercially available software that will reconstruct several 2D images taken at various tilt angles into a single 3D reconstruction of an object? I suppose it would be analogous to the software used for CAT scans? Also, if you've done this before, what kind of hardware requirements are there? I've got a JEOL 2010 configured at the moment to allow only ±30 degrees of tilt. Is that enough? I assume I also need some way of automating the specimen tilting process and will need beam blanking to avoid specimen damage. Can anyone confirm or deny all this?
Thanks all. It was just some anomaly. It went through the second time.
Catherine Ripley ***************************************** Albert Einstein College of Medicine 1300 Morris Park Avenue Bronx, NY 10461 ***************************************** *****************************************
EuroFE 2000 is a forum to bring together interested parties in the field of Field Emission Technologies. This includes microscopists working with materials used for field emission sources (DLC, Nanotubes etc.) and those developing field emission based sources for SEM & TEM.
There is also a strong emphasis on links with other display technologies such as LCD's, Light Emitting Polymers and Phosphors.
The conference and workshop sessions will build on the success of EuroFE '99 & will be held in the UNESCO World Heritage city of Segovia.
The aim of this conference will be to focus on the European dimension of Field Emission, on Industrial Problems and to bring together in a Forum the various groups (from Industry and Public Institutions) working in this area. In addition to the normal program of keynote speakers, oral presentations and poster sessions, it is also planned to hold workshops on topics such as "overcoming obstacles to industrial production".
In common with EUROFE'99 conference, there will be an emphasis on the applications of field emission technologies, as well as basic scientific research. A crucial issue during a conference is time allowed to discussions between participants in order to exchange ideas and therefore possibly define strategies in order to solve real problems. During EUROFE'2000, following each session, large breaks will be set-up in order to allow these discussions.
One of the major sessions will be related to Field Emission applications such as future big FE flat panel displays (1m2). Companies such as Motorola, Samsung, PixTech, Candescent and Saint-Gobain will participate actively in this session. The "BigFED" project coordinated by EUROFE will be presented at the conference in order to define strategies to be able to present projects to the EU within this objective.
The key topics of interest will be:
Industrial applications (Flat panel displays, etc.) and related problems (reliability, lifetime, etc.) Field Emission from diamond, DLC and nanotubes FE simulation and modelling Understanding Field Emission Space applications Device characterisation (surface analysis, etc.) Novel cathodes - technology and fundamentals Other related areas: (Phosphors, LCDs, OLEDs, CRTs, New materials, Novel devices, etc.).
An important issue during conferences is to make possible student participation in order to form them and allow initiation of contacts with either industry or public institutions. For this reason, this year, the EUROFE2000 organisation set-up a special reduced registration fee for students including also the accommodation.
Regards
Tim
****************************************************************** Tim E. Harper EuroFE Network Co-Chairman EuroFE is a network of the European Science Foundation Phone +34 91 640 71 85 Fax +34 91 640 71 86
Responding to the message of {v03007805b4ec9d6d6eae-at-[206.69.208.21]} from R schlicher {rkslick-at-yahoo.com} :
} I need to fix cells for viewing with SEM. However, } the fixation method needs to be consistent with my } experimental methods: } } 1) I need to fix the cells in less than 1 minute and } due to this time limitation, I cannot spin the cells } into a pellet and must add the cell suspension to the } fixative; and
snip!
I would like to try chemical } fixation and SEM. It has been suggested that I try } glutaraldehyde + buffers, then postfix with osmium } tetroxide and dehydrate in an ethanol series.
Robyn,
You could try simultaneous glutaraldehyde + osmium fixation, which should work well for cells in suspension in particular. Mix up seperate solutions of each, such that when mixed in equal amounts, you get the concentrations of fixes and buffers you want. Its usually recommended that you do this at lower temperatures, like about 4C, so pre-cool the solutions and vials first, to prevent the glut and osmium from fighting with each other, but even for "less than a minute", as you put it, maybe you could still work at room temperature. Try mixing the two fixatives together without cells first, to determine the maximum time until discoloration occurs, which means the osmium is precipitating out or whatever goes on there.
Your first rinse could just be a large dilution, to seperate the warring fixatives, then spin down and rinse a few more times before going into the dehydration series.
Give me a few hours to dig out references for this technique, will send them out later.
Gib
Gib Ahlstrand 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.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
Here are two references for the simultaneous glut/osmium technique I mentioned earlier.
1. Franke, W.W., Krien, S. & Brown, J.R.M. (1969) Simultaneous glutaraldehyde osmium tetroxide fixation with post osmication. Histochemie, 19, 162-164.
2. Hirsch, J.G. & Fedorko, M.E. (1968) Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and post fixation in uranyl acetate. J. Cell Biol. 38, 615-627.
Good luck!
Gib Ahlstrand
Responding to the message of {v03007805b4ec9d6d6eae-at-[206.69.208.21]} from R schlicher {rkslick-at-yahoo.com} :
} Hello, } } I need to fix cells for viewing with SEM. However, } the fixation method needs to be consistent with my } experimental methods: } } 1) I need to fix the cells in less than 1 minute and } due to this time limitation, I cannot spin the cells } into a pellet and must add the cell suspension to the } fixative; and } } 2)I am looking for "effects" to the cell membrane } caused by my experiment, so I need to keep any } possible fixative damage/effects at a minimum. } } I have fixed cells using plunge freezing and viewed } them with TEM, this is time consuming and I ran into } a } lot of freeze damage. I would like to try chemical } fixation and SEM. It has been suggested that I try } glutaraldehyde + buffers, then postfix with osmium } tetroxide and dehydrate in an ethanol series. } } Is this the best method? I am open to suggestions. } I am using suspensions of prostate cancer cells in } RPMI (+serum) growth media; I can perform the } experiment in buffered saline and use a high } concentration of cells. } } Thanks, } Robyn } } } Robyn K. Schlicher, M.S. } Laboratory for Drug Delivery } Institute for Bioengineering and Biosciences } Georgia Institute of Technology } 315 Ferst Drive } Atlanta, GA 30332 } rkslick-at-yahoo.com
Gib Ahlstrand 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.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
At 6:09 PM -0600 3/8/0, Jaci Lett wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Your are left with a mirror-finish block face that is perfectly intact. Even with all the safety precautions, I find this method preferable and more reliable in my hands than the "heat and snap-off" method that many people use.
Just don't let your students do it!
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Dear microscopists, Can anyone offer advice on doing immunofluorescence at the light level? I have cryopreserved plant tissue that I am contemplating embedding and using for immunolocalization. I have a protocol that uses butyl, methyl methacrylate. I am wondering several things. Why butyl, methyl methacrylate? Are there other resins/media that are sufficient? I'd appreciate any advice anyone has. Kristen Lennon Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
I have a question that is not a typical microscopy question but I thought I would call upon the extensive knowledge of the readers of this list server.
Does anyone know who manufacturers elemental boron?
We are using boron as a substrate for particle microanalysis (especially carbonaceous particles). Elemental boron can be purchased in the form of “lumps” or “nuggets” several centimeters in dimension from chemical supply houses such as Alfa Aesar and Aldrich. These companies however will not reveal their suppliers and so any questions that I want to ask the manufacturer have to go through the third party supply house. In addition to being silly, this procedure is extremely slow and inefficient. I would prefer to bypass the third party altogether.
So if anyone has any idea who manufacturers elemental boron I would greatly appreciate hearing from you.
Disclaimer: Any opinions expressed are my own and not those of my employer, The Federal Government.
Eric Windsor
Eric S. Windsor Physical Scientist NIST 100 Bureau Dr. Stop 8371 Gaithersburg, MD 20899-8371 (301) 975-3930 Fax: (301) 417-1321 Eric.Windsor-at-NIST.Gov
One of the researchers here is trying to do silver and/or gold enhancement of an immunolabel in brain slices, but is getting amazing background. We switched to the gold enhancement because everyone was saying how much cleaner it is....didn't help. I've seen one report in which the researchers pre-washed their samples in EDTA, because they felt that they were getting background from Mg in the tissue that was nucleating silver deposition - anyone else have any thoughts/experience on this? Anyone else doing enhancement in tissue slices?
He is using kits to do the enhancement - I'm not sure which companies are the suppliers, but they aren't supposed to be light-sensitive (my first culprit). The samples are washed extensively in water before enhancement, and even the no-antibody controls are "lighting up".
I do not have the information you need but I am old enough to remember seeing one of the first papers on tomography. It chose a cute title, something like: "Algorithm for Reconstructive Tomography or ART" Which was fine until the next issue of the journal had a paper (giving an improvement on the method) called: "Fast Algorithm for Reconstructive Tomography."
a transmission electron microscope is in principle the same as a transmission light microscope, except that it uses high energy electrons instead of light. This allows a much higher resolution (it is possible to see atoms or cyrstalline structures), but necessitates much bigger instruments, as there is vacuum technology, high voltages and X-ray generation involved. Many different techniques are available on these machines.
Rather than explaining all to you, you could do a search on the web looking for "transmission electron microscopy" or "TEM". Another starting point would be to go to
http://ncmi.bcm.tmc.edu/
and select one of the sites under "Links". Or go to the MSA site
http://www.msa.microscopy.com/
and find your way from there. I am sure people will help you here if you have specific questions.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: JUPE Peter[SMTP:JUPE.PETER-at-HDH.COM.AU] } Sent: Monday, March 06, 2000 8:35:46 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Transmission electron microsocopy } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sir Could you please supply me details on "transmission electron microsocopy" for my 16yr old daughter at high school, or alternatively where to look on the Internet. Thanking you Peter Jupe
Dear Jaci. If you wish to have access to all the cells on the coverslip, the best way is to flat embed the whole coverslip in a Chang monolayer mold (20mm or 10mm) available from EMS. Be sure to drain the coverslips well and wipe excess resin from the bottom of the coverslip. Invert over filled mold and polymerize overnight. Remove from oven and use metal file to file down edges of embedded coverslip, then immerse in concentrated hydrofluoric acid (use plastic forceps, work in fume hood, wear nitrile glove sand rinse everything well with water). It will dissolve the glass in 15-30 mins. It works every time and you have a complete embedded coverslip. You can then stain it with toludine blue, find the identified cell, mark it with a Ladd marking objective. I then punch out the cell(s) of interest using a hole punch and attach the 5mm circles to blank stubs using quick setting epoxi-patch bond (hardens in 5-10 minutes). This is available from Dexter Corporation 1-603-474-5545 and works extremely well. I sometimes have low contrast so Alexander Mironov's washing method may solve this problem. Otherwise, stain for 30 mins in 5% aqueous UA and 3 mins in Lead citrate for good results. Good luck, JoAnn Buchanan
When I stepped outside of the science building tonight, I ran into, literally, our university president who said he was just walking over to try to find me. He cautioned me about overreacting and then proceeded to tell me to go ahead and get my best quote on a new SEM and service contract!!!!!!
Now that I've got both feet back on the ground, I need any help that any of you can provide, based on your experience and knowledge. We're looking for a new SEM that will support undergraduate courses, be a good training machine for students, support student and faculty research and have a good track record for reliability and being reasonably user friendly. It should be state of the art, but not be so expensive to operate and keep up that it would limit how much students and faculty could use it.
I was trained on an Amray in the 80's and ran our TEM until it became cost ineffective a few years ago. But I know things have changed considerably since my pre-computer controlled everything SEM's and would appreciated the benefit of any experience any of you might have had with new SEMS.
Anything to recommend? Models? Features to insist on or avoid? Reliability, service costs and frequency? Companies to go with or stay away from? What new models do you have and would you recommend them to others? Why or why not?
I'd really appreciate any and all insight you can provide. When your president walks to YOUR office and says the equipment grant proposal HE made to a major foundation was just approved and that he needs me to get the best quote on an SEM to him ASAP, I want to have something to go on, and reasons to go along with it based on user input, without delay.
Thanks in advance for any assistance you can provide.
Lee Hadden Professor and Chair, Department of Biology Wingate University Wingate, NC 28174
I am only using distilled Glutaraldehyde - and only up to 7 days after making up the fixative (Karnovsky's) and storing it in the fridge.
UV Spectra of pure Glutaraldehyde show a single absorbance peak at 280 nm (monomeric ) - whereas the commercial material shows an additional stronger peak at 235nm. This is assumed to be due to the formation of oligomer and polymer. The ratio of absorbance at 235 and 280 nm can be used as a measure of the quality of the GA.
Monomeric-polymeric mixtures yield good ultrastructural preservation when the ratio of monomeric to polymeric forms is 1:1 or 1:2. Ratios outside this limits may prove unsatisfactory. (Weakley 1974) (unfortunately I do not have the exact source/ Paper handy where I got that from.)
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)181 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
I have 95% success detaching glass coverslips embedded with epon inverted on beem capsules described by Leona by simply placing the polymerized coverslip on a hotplate ( } 85.C) with the beem capsule up and gently prying them apart with a straigtedge razor. Cells stay where you want them. I found by that instead of using the beem capsules I just use the lid of one or the lid of a microfuge tube ( makes a better platform). Once separated put the top on a slide and the cells are easily visualized with brightfield (osmicated) or phase (non osmicated). Hank Adams Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, TX
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Reply to: Re: methods of embedding cells cultured on glass coverslips Growing cells on coverslips, fixing them in situ and processing them into epoxy resin is fairly straight forward. Remember that the cells are a monolayer and are only about 30 microns thick so fixation, rinsing, dehydration and infiltration times can be cut down considerably.
The final step of removing the coverslip from the polymerized block is not, however, as trivial as everyone makes it out to be. Yes, dipping the warm block in liquid nitrogen and then re-warming it, will eventually take the glass off the plastic (leaving the cells in the block), but only if the thin layer of plastic that always seems to appear on the back of the glass is removed. The block and glass are also more easily removed if the resin is polymerized with the glass resting cell-side down on resin. The alternative way, of immersing the coverslip cell-side up, and letting the resin cover the cells, is safer, but leads to problems in removing the glass.
If there is a problem with removing the glass, making scratches with a diamond scribe can help and sometimes bending the whole block and glass coverslip can be useful ( if this is possible).
Don't worry about immersing the block in the nitrogen either, there really doesn't seem to be just one way that this happens. If the process is able to remove the glass, the glass may shoot off so violently you may wish you had put on your safety glasses. The next block may not work.
One other method I heard of was to put the block, glass-side down, on a hot plate. Wait until the glass gets hot and I'm told it will easily slide off. I never tried it. Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Alexander Mironov wrote: } } Dear Jaci, } The problem of cell embedding after their cutivation on plastic or galss } is rather simple. You do not need to use Thermanox. You can use glass. The } trick is that immediately after polymerization you have place glasses at } -20 degree C and your samples will be detached. If thsi scheme does not } work you can use liquid nitrogen. However, do not insert smplaes into it } (only glass). If this scheme does not work you can use commercial solution } of HF (acid). It dissolve glass efficiently. For instance, coverslips are } dissolved in 30 min. Then you have to wash samples in buffer (pH7.4) for a } long time and water (several hours) otherwise you will have problems with } contrasters. } We usually glue samples after detachment from glass and work with } this sandwich. For this you have to use incomplete polymerization of Epon } on chamber slides (12 hours). Then samples can be detached at -20 and then } glued with the fresh resin for 24 hours. If you use full polymerization } time on glass or any polymerization on plastic you will not be able to } glue samples. } } Sincerely yours, Alexander Mironov } Italy } } } On Wed, 8 Mar 2000, Jaci Lett wrote: } } } I know this has been covered periodically, but as I've not needed the } } technique, I've not paid attention. I need some methods of embedding } } cells cultured on glass coverslips. Obviously, the most important } } issue is removing the coverslip from the polymerized block.I've looked } } through my own reference materials and the only method I found involved } } using Thermanox coverslips (upon which these particular cells will not } } grow, I'm informed).Please reply dircectly to my email address as well as } } to the Listserver (I haven't received postings for several days). Thank } } you,Jaclynn M. Lett, Research Assistant } } jmlett-at-cid.wustl.eduFay and Carl Simons Center for Biology of Hearing and } } DeafnessCentral Institute for the Deaf4560 Clayton AvenueSt. Louis, } } MO 63110voice: 314-977-0257 fax: } } 314-977-0030 } }
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id PAA04949 for dist-Microscopy; Thu, 9 Mar 2000 15:14:56 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id PAA04946 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 9 Mar 2000 15:14:26 -0600 (CST) Received: from newton.wadsworth.org (newton.wadsworth.org [199.184.18.6]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id PAA04939 for {microscopy-at-sparc5.microscopy.com} ; Thu, 9 Mar 2000 15:14:15 -0600 (CST) Received: from wadsworth.org (ithaca [172.16.1.89]) by newton.wadsworth.org (8.8.8/8.8.8) with ESMTP id QAA20303 for {microscopy-at-msa.microscopy.com} ; Thu, 9 Mar 2000 16:09:09 -0500 (EST) Sender: tivol-at-wadsworth.org Message-ID: {38C8115C.5905598E-at-wadsworth.org}
Chris Adams wrote:
Dear Chris,
} I have recently been asked to participate in an effort at Los Alamos } concerned with Electron Tomography of polymers. I haven't done the } required image reconstruction before. Does anyone know of any } commercially available software that will reconstruct several 2D } images taken at various tilt angles into a single 3D reconstruction } of an object?
Yes. SPIDER is commercially available, and will do 3D reconstructions by any of several methods. Check it out at www.wadsworth.org; click on Resources (pretty far down on the left-hand side); then click on SPIDER.
} I suppose it would be analogous to the software used } for CAT scans?
Some of the reconstruction methods, at least, are used in CAT scans, but others (e.g., projection onto convex sets) may not be--I don't know the latest software for CAT scans.
} Also, if you've done this before, what kind of } hardware requirements are there?
We run on an Onyx using unix; I don't know all the available versions of SPIDER.
} I've got a JEOL 2010 configured at } the moment to allow only ±30 degrees of tilt. Is that enough?
No. +/- 50 deg is adequate, +/- 60 deg is better.
} I assume I also need some way of automating the specimen tilting } process and will need beam blanking to avoid specimen damage. Can } anyone confirm or deny all this? }
Automation is optional, but beam blanking is mandatory-- especially for cryo-specimens. If your goniometer is sufficiently accurate, you can change the tilt setting with the beam blanked, then adjust the position and focus quickly after the beam is back on. If you don't have a sufficiently good goniometer, you need to have either a low-dose kit (assuming you can recognize both your area of interest and a nearby area for focusing) or a very sensitive video-rate camera, such as the intensified CCD we have on the HVEM, so you can find and focus the area of interest with minimal exposure of the specimen.
Hi Kristen, I have published several papers on using butylmethylmethacrylate for immuno at the light level, including a paper where cryofixation and freeze substituion were used, on plant material. Perhaps the protocol you have is by me?
To answer your questions, a mixture of butyl and methyl methacrylate gives nice blocks and preserves the tissue well. I have tried using pure butyl or pure methyl and the sectioning or preservation was worse. These resins, alone or in combination, have the crucial advtange of being mostly removable post embedment with a brief incubation in actetone. This improves the access of your antibody to your antigen greatly. It is important when doing immuno work at the light level on sections to remember that if an antibody can penetrate say 15 nm into the plastic (I am guessing at this number), this is most of the thickness of an ultra thin section but only a teeeny bit of a semithin section. So, for light work especially, being able to remove the embedment is crucial. The usual way to do this is to embed in paraffin and this is fine if you just want tissue level distinctions. But if you want subcellular localization or for other reasons you want your stuff to be better preserved, then plastic gives much nicer results. DOes this help? I will be quite happy to help you further with butylmethyl methacrylate, as I have a professional fondness for the stuff. Good luck, Tobias
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America _ ____ ^ __ ____ Tobias I. Baskin / \ / / \ / \ \ University of Missouri / | / / \ \ \ Biological Sciences /___/ /__ /___ \ \ \__ 109 Tucker Hall / / / \ \ \ Columbia, MO 65211-7400 USA / / / \ \ \ voice: 573-882-0173 / /____ / \ \__/ \____ fax: 573-882-0123
Dear Tamara Is he washing in de-ionized water? Brain is always absorbent, so timing and reagent volumes may need adjustment. Gold and silver enhancement are tricky so I would protect from heat and light as much as possible. Good Luck. Dana
We are looking for someone to provide analytical services on analyzing surface topography on biomedical implants specifically using a TOPSCAN confocal microscope. It needs to be performed on this instrument for comparison with data previously taken on such an instrument. Does anyone know of anyone who has this instrument and who might entertain performing such an analysis? Your help on this will be greatly appreciated.
Charles R. Anderson, Ph.D. President Anderson Materials Evaluation, Inc. 1450 South Rolling Road Halethorpe, MD 21227 (410) 455-5698 Fax (410) 455-5679
An independent materials analysis laboratory for failure analysis, quality control, product and process development. We offer small-spot XPS, Auger microprobe, AFM, SEM, optical metallographic microscopy, white light interference microscopy, mass spectroscopy, and thermal analysis (TGA, DSC, TMA, DMA) services.
To All: We have a Jeol 100CX II and experiencing a constant moving of the Objective Aperture moving when scanning grids, especially in Posistion # 2 Grid. Any solution or ideas? We changed apertures,cleaned holder etc. Thank you, Peter Stolzenberg, PESTO INC. pestoem-at-aol.com
We just bought a new top-end type microscope (Field emission semi-inlens SEM). I evaluated four companies. I would have been happy with any of the microscopes that I looked at. In my opinion, all of the microscopes available today are damn good throughout the product range. You will probably be happy with any microscope that you buy. If your budget is limited and defined as ours was, that is going to limit you as to what machine that you will buy. You can use that to see what the manufacturers will put into your package. There are other intangibles that go into the decision. The most important is service, in particular, the service in your area. Ask around! I did and got some interesting comments. Then there are some of the others that you have to judge for yourself. Things like the user interface -you will be training students to use it, you want a versatile machine, but one that is easy to learn to use; output capabilities; digital/film or both; networkability; etc.
If performance is the issue, then the best thing that you can do is to take a number of your samples and take them to the application labs and run them while you are there. You should take samples that are representative of the types of samples that you will be working with. Make sure that you compare the same conditions on all your samples and that the output that you get back can be compared at the same magnifications. Standardize the output that you want the images to be in, for example 8 bit grayscale TIFF images or whatever. You should also do the samples in the same order from lab to lab. Print the results from digital images on the same printer. Then get a group of experienced users to help you judge the results.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
--
} -----Original Message----- } From: hadden-at-wingate.edu [ mailto:hadden-at-wingate.edu {mailto:hadden-at-wingate.edu} ] } Sent: Thursday, March 09, 2000 2:51 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: suggestions for new SEM?? } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } -------------------------------------------------------------- } ---------. } } } When I stepped outside of the science building tonight, I ran into, } literally, our university president who said he was just } walking over to } try to find me. He cautioned me about overreacting and then } proceeded to } tell me to go ahead and get my best quote on a new SEM and service } contract!!!!!! } } Now that I've got both feet back on the ground, I need any } help that any } of you can provide, based on your experience and knowledge. We're } looking for a new SEM that will support undergraduate } courses, be a good } training machine for students, support student and faculty } research and } have a good track record for reliability and being reasonably user } friendly. It should be state of the art, but not be so expensive to } operate and keep up that it would limit how much students and faculty } could use it. } } I was trained on an Amray in the 80's and ran our TEM until it became } cost ineffective a few years ago. But I know things have changed } considerably since my pre-computer controlled everything } SEM's and would } appreciated the benefit of any experience any of you might } have had with } new SEMS. } } Anything to recommend? Models? Features to insist on or avoid? } Reliability, service costs and frequency? Companies to go } with or stay } away from? What new models do you have and would you } recommend them to } others? Why or why not? } } I'd really appreciate any and all insight you can provide. When your } president walks to YOUR office and says the equipment grant } proposal HE } made to a major foundation was just approved and that he } needs me to get } the best quote on an SEM to him ASAP, I want to have } something to go on, } and reasons to go along with it based on user input, without delay. } } Thanks in advance for any assistance you can provide. } } Lee Hadden } Professor and Chair, } Department of Biology } Wingate University } Wingate, NC 28174 } } hadden-at-wingate.edu } http://www.wingate.edu {http://www.wingate.edu} } 704-233-8238 } } }
Dear Tamara: To figure out what went wrong with your investigator's experiment, we need more detail about how his experiment was conducted. Would you ask him to contact me and email his protocol to me? I believe that I can help him.
Hong ============== Hong Yi Emory Neurology hyi-at-emory.edu
On Thu, 9 Mar 2000, Tamara Howard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings! } } One of the researchers here is trying to do silver and/or gold enhancement } of an immunolabel in brain slices, but is getting amazing background. We } switched to the gold enhancement because everyone was saying how much } cleaner it is....didn't help. I've seen one report in which the } researchers pre-washed their samples in EDTA, because they felt that they } were getting background from Mg in the tissue that was nucleating silver } deposition - anyone else have any thoughts/experience on this? Anyone else } doing enhancement in tissue slices? } } He is using kits to do the enhancement - I'm not sure which companies are } the suppliers, but they aren't supposed to be light-sensitive (my first } culprit). The samples are washed extensively in water before enhancement, } and even the no-antibody controls are "lighting up". } } Any suggestions will be greatly appreciated! } } Tamara Howard } CSHL } }
The topic of reducing background has been raised a number of times by our customers, and we have collected together several approaches and their references, and I hope some of the following are helpful:
One possible mechanism for "background" signal is hydrophobic interactions between gold particles and tissue components, and if your samples can tolerate them, additives which break these interactions down and solubilize hydrophobic species might help in reducing background. Washes which effectively solubilize gold compounds and other hydrophobic species include:
* 0.6 M triethylammonium bicarbonate buffer (prepared by bubbling CO2 into an aqueous suspension of triethylamine with stirring; for a reference, see Safer, D.; Bolinger, L., and Leigh, J. S.; J. Inorg. Biochem., 26, 77 (1986)).
* A small amount of detergent, such as Tween-20 or Triton X-100, or an amphiphile such as benzamidine or 1,2,3-trihydroxyheptane.
* If you are using the gold enhancer, raising the ionic strength of the solution is sometimes helpful - we have found that raising the sodium chloride concentration in the actual gold enhancement mixture to 0.5 M can actually reduce the background. Alternatively, lowering the pH of the gold enhancement mixture by about 0.5 pH units may help.
Acetonitrile coordinates quite well to silver ions, so if your background problem is due to the interaction of silver from the enhancement reagents with a component of your tissues, treatment with acetonitrile or a wash solution containing a proportion of acetonitrile may help (provided the sample can withstand it).
To reduce the background for silver enhancement, one procedure which has been found to be effective is to wash with 0.02 M sodium citrate buffer, pH 7.0, several times immediately before silver enhancement - this has been used to reduce background in experiments with the combined fluorescein and gold probe FluoroNanogold when used with HQ Silver (Nanoprobes). When the Danscher formulation of silver enhancer was used instead, 0.02 M sodium citrate buffer at pH 3.5 was found to be most effective. In immunoblots, we have also observed that washing with 0.05 M disodium EDTA, pH 4.6, before silver enhancement also results in low background. (Reference: Powell, R. D.; Halsey, Carol M. R.; Spector, D. L.; Kaurin, S. L.; McCann, J.;, and Hainfeld, J. F. A covalent fluorescent-gold immunoprobe: "simultaneous" detection of a pre-mRNA splicing factor by light and electron microscopy. J. Histochem. Cytochem., 45, 947-956 (1997)).
A number of methods have been described for stopping the silver enhancement reaction or for "back-developing" to remove extraneous deposited silver. These prevent the continuation of the reaction in the specimens after development is complete (for example, if the silver is only slowly removed from the tissue), and may help reduce your background signal.
Sodium thiosulfate (1 % aqueous solution, freshly made) is a good "stop" reagent for both silver and gold deposition (Van Driel, D. 1997. Gold toning for silver enhanced immunogold reacted tissue. Micros. Today, 97-7, 28), so it is reasonably safe to assume that there will be no further deposition after it is applied. However, one caveat: you should use caution with gold enhancement because there have been reports that sodium thiosulfate can remove the enlarged gold particles. This might be a good choice to begin with (1-2 minute incubation after enhancement and washing with water; after the sodium thiosulfate, rinse thoroughly again with deionized water).
Other reaction stop and back development methods include:
(i) 1% acetic acid (Scopsi, L. 1989. Silver-enhanced colloidal gold method., p. 260. In M. A. Hayat (ed.), In Colloidal Gold: Principles, Methods, and Applications, vol. 1. Academic Press, San Diego, CA).
(ii) 1% acetic acid followed by photographic fixer (Agefix, Agfa-Gevaert, or Ilfospeed 200, Ilford) (Scopsi, L. 1989., same reference as (i)).
(iii) direct photo fix, using those just mentioned (Burry, R.W. 1995. Pre-embedding immunocytochemistry with silver-enhanced small gold particles, p. 217-230. In MA Hayat (Ed.). Immunogold silver staining: Principles, methods and applications. CRC Press, Boca Raton).
(iv) brief rinse in 2.5% sodium chloride (Scopsi, L. 1989., same reference as (i)).
(v) 15-25% aqueous sodium thiosulfate plus 15% sodium sulfite (Danscher, G. 1981. Histochemical demonstration of heavy metals. A revised version of the silver sulphide method suitable for both light and electron microscopy. Histochemistry 71, 1-16).
(vi) 1% acetic acid, washes in acetate buffer, toning in 0.05% HAuCl4 3-10 min, with excess silver removed with 3% sodium thiosulfate. We found that Nanogold-labeled proteins run on a polyacrylamide gel kept low backgrounds when stopped with 10% acetic acid with 10% glucose in water, as opposed to just a water stop (Takizawa, T. and J.M. Robinson. 1994. Use of 1.4-nm immunogold particles for immunocytochemistry on ultra-thin cryosections. J. Histochem. Cytochem. 42, 1615-1623).
(vii) A modified Farmer's solution was used for the reversal (0.3 ml 7.5% potassium ferricyanide, 1.2 ml of 20% sodium thiosulfate, 60 ml water) (Danscher, G.: Histochemistry, 71, 1-16 (1981)). Application of this solution briefly to your sample before gold toning may help to remove background silver deposition.
Hope this helps,
Rick Powell
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Please help this gentleman. He needs inexpensive EDS system suitable for Philips TEM (horizontal entrance detector) complete, and old EDAX-9800 or similar, computer unit only. Reply to Bill Lawry at eht-at-stlnet.com or (314)531-9868. Thank you.
Vitaly Feingold Scientific Instruments and Applications (770)232-7785 ph. (770)232-1791 fax.
Dear Peter, I have a 7 holed, self-cleaning, gold foil aperture in my TEM. The hole diameters are 30um and 60um. At 60kV, the aperture does tend to wander abit until it gets warmed up. After an hour or so of operation, I generally have to tweak the xy alignment and then I am ready to continue.
I am always amazed when the instrument is used by other investigators. Frequently the aperture is halfway into the the beam, i.e., the crescent of the aperture is covering 1/3 of the field of view. As I stated, I am aware the gold foil aperture tends to drift slightly over time exposed to the beam, and consequently adjust accordingly.
I too would be keen to hear any additional comments on this 'trait' of gold foil apertures.
-Ken
On Thu, 9 Mar 2000 PESTOEM-at-aol.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To All: } We have a Jeol 100CX II and experiencing a constant moving of the Objective } Aperture moving when scanning grids, especially in Posistion # 2 Grid. } Any solution or ideas? We changed apertures,cleaned holder etc. } Thank you, } Peter Stolzenberg, PESTO INC. } pestoem-at-aol.com } }
I have just started on a TEM-project investigating the microstructures of different tool steels. The steels contain various types of carbides with different compositions, shapes and sizes. I have started out by making thin foils using electrolytical jet polishing. I get nice thin foils where I can study and analyse the martensitic matrix, but the carbides are off course too thick to be analysed. To overcome this problem I have considered a combination of electropolishing and ion milling, but I have also heard about an extraction replication technique. However, I haven't been able to find any literature on this technique. Have any of you tried this technique and/or could you point out some literature describing the technique? Also, if you have other suggestions to deal with the described problem I would be pleased to hear about them.
Regards, Jesper
---------------------------------------------------- Jesper Vejloe Carstensen Research Scientist, M.Sc., Ph.D. Materials Research Department Risoe National Laboratory P.O. Box 49 DK-4000 Roskilde, Denmark Phone: +45 4677 5776 Fax: +45 4677 5758 E-mail: jesper.v.carstensen-at-risoe.dk Web: http://www.risoe.dk/AFM/Personal/Jeca/jeca.htm ----------------------------------------------------
The url of the conference site seems to have been stripped out by some mail servers. Just in case it's www.cmp-cientifica.com/eurofe
Regards
Tim
****************************************************************** Tim E. Harper EuroFE Network Co-Chairman EuroFE is a network of the European Science Foundation Phone +34 91 640 71 85 Fax +34 91 640 71 86
One accessory worth considering, especially for a SEM used by students would be a ChamberScope. This is a small IR video camera mounted to the chamber which will allow students to see the position of samples in the chamber in real time. Could avoid damaging crashes. Take a look at GW Electronics for one model....
We have Carl Zeiss LSM 510 2-photon microscope armed with a T:sapphire laser. We are successful with one color imaging, but we wpould like to try two-color two-photon imaging, and of course we have problems with finding a proper set of fluorophoes. My understanding is that they should have similar excitation and different emissions. Has anyone tried using two or more fluorophores ? Which combinations are the best? What wavelengths are recommended?
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Is there a short, general reference, like a review, that explains immuno EM, it's uses, major techniques, and shortfalls? I would like to use it to send to investigators that are thinking about using the technique but don't know much about labeling at the EM level. I have a lot of specific references and it would take a lot of time to reference or copy them. If the investigator wants to know more I can direct them to those, otherwise a general one would get them and me on the same page when we start talking specifics on their project. Thanks
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In the earlier days {25 years ago} of medical diagnosis, before the advent of the rapid, highly specific methods of identifying pathogenic organisms, darkfield microscopy was routinely applied to rule out specific organisms. In particular, the diagnosis of syphilis was aided with this mode of imaging. When fluids from a suspect lesion were applied to a slide and imaged by darkfield, the characteristic shape of the spirochaete organisms was clearly seen. More recently, darkfield examination of synovial fluids from arthritic joints was used to rule out or diagnose Lyme disease, before specific testing became available.
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Catherine,
I also tried to subscribe to the confocal list when Larry suggested it and it worked like a charm. I cut and pasted the email address right from his posting.
LISTSERV-at-LISTSERV.ACSU.BUFFALO.EDU
Try again. Cheers, John
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-- C. John Runions, Ph.D. Department of Plant Sciences University of Cambridge Cambridge CB4 1YE UK
email at work cjr41-at-cam.ac.uk email at work runions-at-ntlworld.com Phone (01223) 766 545
We are in the market for two relatively inexpensive flatbed scanners. One needs to have a USB attachment and drivers for a Mac. This one is pretty straightforward. Up to about $200 (US) should suit our needs.
For the second one I am considering one with an adapter to allow scanning of negatives and 35 mm slides. I have heard a bad report of a scanner not focussing correctly on the negative or transparency. Does anyone have any comments to support or refute the focusing problem? We are able to pay up to $400 if I can find one that performs well. Obviously, we are not considering high end scanners. I would prefer USB, but could cope with another type of connection. Thank you for any help you can provide.
Sincerely, Maureen Petersen Dept Plant Pathology University of FL Gainesville, FL 32611
It has been a number of years since I've been directly involved in operating and maintaining a JEOL 100CX, but as I recall both the specimen holder and the aperture are inserted into the rather limited space between the objective pole pieces, with the specimen rod fitting in above the aperture. If the aperture consistently moves when you move the specimen holder, then I'd strongly suspect that either the end of your specimen rod has been bent downward just a bit, or the end of the aperture holder has been bent upward slightly so that the specimen rod rubs the aperture holder when it is moved thus causing the aperture to move. The clearance between these two devices is only a fraction of a millimeter, and so it wouldn't take much bending to cause this problem.
Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
} } Anita, } } I have been using the MultiPrep for TEM of semiconductors since December. It } } has made my life much much easier. I am responsible for the TEM } } preparation/imaging/data at ON Semi. Since I have started using the MultiPrep I } } have been able to consistently turn a job around in 1 day and less. I have only } } failed on 1 sample since December and that was due to my bad judgement. Once you } } get a routine for making the samples I have found the results to be very } } consistent. Do you have an Ion Mill? I use the PIPS and it contributes to } } the success of the samples I make. Although if you did not have a PIPS I think } } that you would still get very good results using the MultiPrep. } } For me the major differences between using the MultiPrep and the Tripod Polisher } } are: } } 1. Consistent amount of force placed on the samples using the MultiPrep. } } 2. Having the digital micrometer on the MultiPrep takes some of the guess work } } out of the sample prep. I am able to tell how much material I am removing } } 3. Piece of mind. Since I have developed a set routine I am able to make 2 } } samples a day and feel confident that they will not fail. Hand polishing has } } always been a crap shoot for me. } } From your signature line I am assuming you work on various materials. The work } } I have done is primarily Silicon, but I think that the results would be good for } } other materials also. I have submitted an abstract for the MSA conference this } } year. I would be happy to speak with you about the system if you will be } } attending. } } } } Leah L Dobbs } } ON Semiconductor } } CSAL TEM } } 602-244-4820 } } 1-888-637-9415 (6379415-at-skytel.com) } } s20260-at-onsemi.com } } } } My disclaimer " I do not work for Allied High Tech, and do not have vested } } interest in the companies financial success. I am merely a satisfied customer". } } } } } } ----- Original Message ----- } } From: "Anita Garg" {Anita.Garg-at-lerc.nasa.gov} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Thursday, February 24, 2000 5:55 AM } } Subject: Re: Allied Multiprep Polisher for TEM samples } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear Colleagues } } } Does anybody have experience on the Allied MultiPrep Polishing } } } System, especially in terms of getting a good TEM sample? How well do } } } the Allied's Precision Cross-Section Tool and TEM Wedge Polisher work } } } on this semi automatic multiprep polisher? } } } Also, how does their TEM Wedge Polisher compare with the South Bay } } } Tripod Polisher? } } } TIA } } } Anita } } } ******************************************* } } } Dr. Anita Garg } } } NASA Glenn Research Center at Lewis Field } } } Advanced Metallics Branch } } } Mail Stop 49-3 } } } 21000 Brookpark Road } } } Cleveland, OH 44135 } } } Phone : (216) 433-8908 } } } Fax : (216) 977-7132 } } } E-mail : Anita.Garg-at-grc.nasa.gov } } } ******************************************* } } } } } } } }
The extraction replica technique was developed by Bob Fisher at the U. S. Steel Laboratory back in the early 1950's when we were all trying to identify the precipitates in various alloy systems for the first time. I don't seem to be able to lay my hands on the original reference; however, the technique is described in reasonable detail on pages 115-117 of the book 'Techniques for Electron Microscopy', Desmond Kay, Ed., Blackwell Scientific, 1961.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Hi, we have some software freely downloadable for processing ED of protein crystals. It's at http://ncmi.bcm.tmc.edu/software.html. Look for packages auto and edp. The former is a front-end to a bunch of fortran programs for automated processing, whereas the latter is a GUI-based program.
Jaap
-- Jaap Brink, Ph.D., Biochemistry, One Baylor Plaza, Baylor College of Medicine, Rm. N420 Alkek Building, 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 Fri, 10 Mar 2000, Chengge Jiao wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } Does anybody know where I can get free software for electron doffraction and } more... } } Can we download them free ? } } Chengge JIAO } } H.H.Wills Physics Lab } University of Bristol } c.g.jiao-at-bristol.ac.uk } }
Firstly, thanks to everyone who helped out with the earlier question on image analysis. We are using NIH Image 1.62 (which has a watershed filter to separate touching particles) to measure their diameters. (Worked like a charm -- thanks John Russ).
We need a NIST particle, sized 5 µm or so, to use as a standard. Unfortunately, the standard we recently purchased was sized using light scattering (which apparently gives a higher reading than electron microscopy) and our sizing does not agree with that figure.
Does anyone know were we can find 5.0 µm polystyrene particles that have been sized by TEM and are NIST certified or traceable?
Thanks,
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Every year when I teach the liver lectures for my histology course, I explain to the students that up to 25% of the hepatocytes are binucleate and another 50% are polyploid. These textbook facts match up with what I see in the scope. But I don't understand why hepatocytes (or transitional epithelial cells) are frequently binucleate (or polyploid). Any one able to answer this structure - function question?
My second histology question concerns the gallbladder epithelium. I have a great prepared H&E 1.5 um paraffin slide from Carolina Biological. Most of the epithelial cells have a few fine red-stained granules in them. I assume these are glycoproteins and/or mucin-type glycoproteins. A few scattered cells in the epithelium have larger, intensely eosinophilic granules in them. I can't figure out what these cells are. None of the standard textbooks or atlases discuss them. Before anyone suggests they are mucin-filled goblet cells, I should point out that intestinal goblet cells are my main research focus and these are unlike any mucin secreting cell I have seen in the intestine, respiratory tract, stomach, conjunctiva, etc. They look, in fact, more like Paneth cells in the small intestine. Any ideas?
Thanks, Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
It is my understanding that many instances of so-called aperture thermal drift are actually the result of hysteresis in either the condenser or objective lenses. This seems to be more of a problem in JEOL instruments than in others that I'm aware of. It is particularly pronounced when you spend time scanning large areas with little or no changes in magnification or brightness. When the current is changed to either lens, the periphery of an aperture may enter the field of view. By realigning the aperture, you actually misalign it along the optical axis. This is why a TEM is often in such abysmal alignment after use by students and less experienced users. Our JEM 1210 has 9 lenses, with each one potentially affected by hysteresis. Most of our users know when it is time to switch off the lens power temporarily and reload the alignment values. In the case of a less automated instrument such as the 100-C series, it will often suffice to run C2, Obj, and Int lenses through their entire ranges before aligning apertures.
The problem with extraction replicas for your carbides is that they will still be too thick to analyse! Extraction replicas work better the smaller the carbides (in my humble opinion!). I would recommend mechanical thinning followed by ion-milling (we typically don't bother with the intermediate jet-polishing step). Of course, if you also have fine carbides, you will need the extraction replicas in order to be able to examine them without interference from the matrix!
Good luck,
Tony Garratt-Reed.
At 10:31 AM 03/10/2000 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6479 **
First of all you never indicated from which species this gall bladder epithelium came from. Goblet cells are characteristic in the bovine species. The cat epithelium may contain globule leukocytes. Also, endocrine cells have been reported in bovine.
Nancy A. Monteiro-Riviere, Ph.D., DABFE, DABFM Professor of Investigative Dermatology and Toxicology North Carolina State University College of Veterinary Medicine Department of Clinical Sciences Center for Cutaneous Toxicology and Residue Pharmacology 4700 Hillsborough St. Raleigh, NC 27606 Tel: 919-513-6426 FAX: 919-513-6358 email: Nancy_Monteiro-at-ncsu.edu CCTRP Homepage: http://cctrp.ncsu.edu
} ---------- } From: Tom Phillips[SMTP:PhillipsT-at-missouri.edu] } Sent: Friday, March 10, 2000 1:47 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: binucleate hepatocytes & granules in gallbladder: basic } histoquestions } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Fellow microscopists: } } Every year when I teach the liver lectures for my histology course, I } explain to the students that up to 25% of the hepatocytes are } binucleate and another 50% are polyploid. These textbook facts match } up with what I see in the scope. But I don't understand why } hepatocytes (or transitional epithelial cells) are frequently } binucleate (or polyploid). Any one able to answer this structure - } function question? } } My second histology question concerns the gallbladder epithelium. I } have a great prepared H&E 1.5 um paraffin slide from Carolina } Biological. Most of the epithelial cells have a few fine red-stained } granules in them. I assume these are glycoproteins and/or mucin-type } glycoproteins. A few scattered cells in the epithelium have larger, } intensely eosinophilic granules in them. I can't figure out what } these cells are. None of the standard textbooks or atlases discuss } them. Before anyone suggests they are mucin-filled goblet cells, I } should point out that intestinal goblet cells are my main research } focus and these are unlike any mucin secreting cell I have seen in } the intestine, respiratory tract, stomach, conjunctiva, etc. They } look, in fact, more like Paneth cells in the small intestine. Any } ideas? } } Thanks, Tom } } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax) }
I am searching for a Leitz Periplan photomicro eyepiece, 10X/18, 23mm diameter. The old Leitz part number was 519749. The current Leica part number is 11519749, although it is a discontinued item. This particular eyepiece has a threaded mount on the viewing end. The thread was used to mount eyecups on the eyepiece, but it was also used as a photoeyepiece.
If anyone has one of these photo eyepieces and would be willing to part with it, please contact me off-list.
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Your best bet is to embed in paraffin. However, if you still want to use resin for immunolocalization, you may try Immuno-Bed Embedding resin, available from Electron Microscopy Sciences. It is a fairly low viscosity media.
Soumitra
} Dear microscopists, } Can anyone offer advice on doing immunofluorescence at the light } level? I } have cryopreserved plant tissue that I am contemplating embedding and using } for immunolocalization. I have a protocol that uses butyl, methyl } methacrylate. I am wondering several things. Why butyl, methyl } methacrylate? Are there other resins/media that are sufficient? I'd } appreciate any advice anyone has. } Kristen Lennon } Kristen A. Lennon } Cell, Molecular & Developmental Biology Group } Department of Botany & Plant Sciences } University of California } Riverside, CA 92521 } kalen-at-citrus.ucr.edu
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
Here are two references more recent than the two I sent earlier regarding simultaneous glut/osmium fixation methods for single cells in suspension:
Dentler, WL (1999) Fixation of Tetrahymena cells for electron microscopy. Methods in Cell Biology 62:323-331
but is modified from
Omoto, C.K, and Kung, C. (1980). Rotation and twist of the central pair microtubules in the cilia of Paramecium . J. Cell Biol 87:33-46
Responding to the message of {v03007805b4ec9d6d6eae-at-[206.69.208.21]} from R schlicher {rkslick-at-yahoo.com} :
} Hello, } } I need to fix cells for viewing with SEM. However, } the fixation method needs to be consistent with my } experimental methods: } } 1) I need to fix the cells in less than 1 minute and } due to this time limitation, I cannot spin the cells } into a pellet and must add the cell suspension to the } fixative; and } } 2)I am looking for "effects" to the cell membrane } caused by my experiment, so I need to keep any } possible fixative damage/effects at a minimum. } } I have fixed cells using plunge freezing and viewed } them with TEM, this is time consuming and I ran into } a } lot of freeze damage. I would like to try chemical } fixation and SEM. It has been suggested that I try } glutaraldehyde + buffers, then postfix with osmium } tetroxide and dehydrate in an ethanol series. } } Is this the best method? I am open to suggestions. } I am using suspensions of prostate cancer cells in } RPMI (+serum) growth media; I can perform the } experiment in buffered saline and use a high } concentration of cells. } } Thanks, } Robyn } } } Robyn K. Schlicher, M.S. } Laboratory for Drug Delivery } Institute for Bioengineering and Biosciences } Georgia Institute of Technology } 315 Ferst Drive } Atlanta, GA 30332 } rkslick-at-yahoo.com
Gib Ahlstrand 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.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
I am going to give a course on electron microscopy including TEM, SEM, STM, AFM, etc. to graduate students. Could you give me information on textbooks about these microscopy.
Thanks
**************************************************** Prof. Jianguo Wen
Department of Physics Boston College 140 Commonwealth Avenue Chestnut Hill, MA 02467 Tel: (617) 552-3586 Fax: (617) 552-8478 Email: wenji-at-bc.edu ****************************************************
Are there any Amray-trained service people out there that are independently servicing the 1910FE system on the West Coast? I'm in Sacramento CA. I am not looking for someone who knows how to spell Amray. I am looking for someone who knows what the 1910FE is all about. Big difference. Introductory questions:
1. Can you condition the emitter/gun? How and with what? 2. How do you handle the differential vacuum aperture? 3. what is your experience in working with the Windows control s/w? 4. What do you know about the IP8? 5. Do you have any knowledge about moving from 486 ISA to P-II or P-III systems for NibbleNet and frame capture?
Since the gobbling of Amray by KLA-Tencor, Amray SEM service is dismal--more so day by day. The service people are excellent--but they are being pulled in all directions, and mostly towards the KLA big semi systems.
The Amray SEM systems were/are really good--and that is why KLA bought Amray. But KLA is not a research SEM outfit like Amray was.
If I had to get something other than this 1910FE, I don't know what it would be. The Hitachi and Philips SEMs are great/nice but ridiculously expensive to buy and maintain. And I do not care for Jeol.
Me thinks that the research SEM options are shrinking daily.
Duke Scientific should produce something like this. I know that they used a TEM to perform their sizing at one time. I'm not sure if this is the still the case. If not, maybe they can point you in the right direction. Here's Duke's website as well as a few other particulate manufacturers:
================================================ Marc W. Helvey Accounts Manager VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 phone:(408) 428-1800, ext. 108 FAX: (408) 428-9555 Mobile: (408) 307-3833 e-mail : marc.helvey-at-vlsistd.com {mailto:marc.helvey-at-vlsistd.com} internet: http://www.vlsistd.com {http://www.vlsistd.com/} ================================================
On Fri, 10 Mar 2000 19:11:40 -0500, Jianguo Wen wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am going to give a course on electron microscopy including TEM, SEM, STM, } AFM, etc. to graduate students. Could you give me information on textbooks } about these microscopy. } } Thanks } } **************************************************** } Prof. Jianguo Wen } } Department of Physics } Boston College } 140 Commonwealth Avenue } Chestnut Hill, MA 02467 } Tel: (617) 552-3586 Fax: (617) 552-8478 } Email: wenji-at-bc.edu } **************************************************** } } Prof. Wen - In 1994, I took a course on electron microscopy (for biologists) at San Francisco State University given by Gregory Antipa (my former M.A. advisor and principal investigator). The textbook he used was titled "Electron Microscopy (:) Principles and Techniques for Biologists" by John J. Bozzola and Lonnie D. Russell (eds. Jones and Bartlett Publishers, Boston .. London). Although this textbook is intended for biology students, it was used in a class that any graduate student could sign up for given his/her interests. I found the textbook to be illuminating and interesting to read. It has a historical chapter on electron microscopy, and then discusses, among many topics, fixation uses / types of / .. embedding procedures and types - TEM; similar topics - SEM; microtomy; staining procedures, types, etc. ; TEM itself; SEM itself; immunocytochemistry and EM ; autoradiography; freeze-fracture ; analytical electron microscopy (e..g., x-ray microanalysis subchapter; EELS subchapter; SAD diffraction mode sub-subchapter), and many other topics relevant to electron microscopy. Unfortunately .. it does not cover STM (Scanning-Tunneling Microscopy) nor AFM (Atomic Force Microscopy). I personally do not know of any textbooks that cover those types of electron microscopy, but I would guess that someone in this list would have suggestions. For that matter, I am not even aware if this textbook is still being published, but perhaps Gregory Antipa (e-mail: antipa-at-sfsu.edu ) can tell you, or someone else. I hope that this information may serve as a useful starting point in your search for a textbook on electron microscopy for graduate students. Nelson Conti (former graduate student) San Francisco State University 1600 Holloway Avenue San Francisco, California (CA) 94132 URL: www.sfsu.edu
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Feather makes the blades we LM histotechs use most often in our everyday sectioning. They are distributed by Allegiance (used to Baxter, used to be SP....). Just FYI. Wanda Shotsberger (HT ASCP) -----Original Message----- } From: Grazyna M Tokarczyk [mailto:gmtokarc-at-is.dal.ca] Sent: Friday, March 03, 2000 8:26 AM To: Gordon Couger Cc: c.jeffree-at-ed.ac.uk; Dan Luchtel; microscopy-at-sparc5.microscopy.com
Dear John, I worked with characterisation of latex sphere size for many years and traced them to NIST standard. You may wish to contact Interfacial Dynamics Corp. as they manufacture spheres and no doubt have the capacity to trace to NIST. I do not have their address information at hand, but they are located in Portland, OR.
If you have other questions, please contact me at me University of Portland email address.
Dear Tamara, Without more information about the procedure that was followed it is hard to suggest a solution. But since the issue of background with immunogold and enhancement is becoming a regular topic in this List, it may be worthwhile to address it more in-depth. In troubleshooting I always find it a good idea to trace down the origin of the problem logically, rather than shooting in the dark. In all the years we have been involved in immunogold detection and silver enhancement, we have always benefited from simple approaches. These are documented in troubleshooting brochures we use during our workshops and which describe step-by-step how to proceed to have the best chance to pinpoint the problem. Anyone who is interested, please send me an e-mail.
Now just some comments on some of the things you mentioned:
- working on brain slices, you are probably doing pre-embedding. One of the issues in pre-embedding is penetration. Most of the time gold conjugates need a longer time to get inside thicker specimens, but under the proper conditions they do get there. But what often is easily forgotten is that when it takes a long time for reagents to penetrate into the slice, it also takes a long time for unreacted reagents to get washed out again. So washing should be appropriate.
- you mentioned that there is no difference between silver enhancement and gold enhancement. Have these specimens been incubated with gold conjugate? What if there is background even when the gold conjugate incubation step was left out? With any enhancement system there is a risk of getting background through (i) autonucleation or (ii) by the formation of metal particles in the specimen, formed as a result of the interaction of components in the specimen (e.g.aldehydes, borohydrid) with the noble metal salts in the enhancement mix. This may also be related to the type of metal salt in the enhancement solution: noble metal salts need only a little push to become reduced to the metal again and the more noble the metal the smaller the push it needs. As for judging if autonucleation has occurred, as long as the enhancement solution is as clear as it was when the enhancement started, this should not be a real problem.
- background from magnesium. Gallyas and Danscher (and many others) have looked into many aspects of (auto)metallography, I can give references if you are interested.
- light sensitivity of enhancement reagents: light sensitivity can never be reduced to zero. There are tricks that reduce light sensitivity to a degree where it no longer seriously interferes with enhancement on particles, but again, these are noble metal salts, and they need just so much.... In any case, if light would be causing a problem, which I don't think is very likely whatever brand you used, you should have the same phenomena as with autonucleation: the mixture should become turbid or colored after a while. If this is not the case, there is no reason to be worried about a possible light sensitivity.
Jan =========================== Jan Leunissen AURION http://www.aurion.nl Costerweg 5 6702 AA Wageningen phone: (31)-317-497676 fax: (31)-317-415955 You will find more tech info on our website.
Need help on PEELS } } To all, } } I am working on grain boundary chemistry measurement } with Digipeels. Try to find out the segregation } behavior of boron, which is about 0.5at% averagely. } However, I cannot find any edge for boron in the } spectrum. As I don't know what's the optimum setting } for PEELS to check the grain boundary segregation, if } you have any idea on this, please let me know. } } Thanks, } Lung __________________________________________________ Do You Yahoo!? Talk to your friends online with Yahoo! Messenger. http://im.yahoo.com
I'd like to point out that by extraction replica you will NOT overcome the problem of the carbides' thickness: you will get only a complete separation of them from matrix. So, according to my experience, the ion-milling would be a solution in that it could lead to a THINNING of your carbides.
In case there is a possibility to partially dissolve the carbides, by chemical means, after their extraction, then the extraction replica technique will work; but I don't know about such a chemical technique for dissolvind (partially!) the carbides.
Corneliu Sarbu, Ph.D. Dept.of Metallurgy and Materials Science Catholic University of Leuven Belgium
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Dear all,
I have just started on a TEM-project investigating the microstructures of different tool steels. The steels contain various types of carbides with different compositions, shapes and sizes. I have started out by making thin foils using electrolytical jet polishing. I get nice thin foils where I can study and analyse the martensitic matrix, but the carbides are off course too thick to be analysed. To overcome this problem I have considered a combination of electropolishing and ion milling, but I have also heard about an extraction replication technique. However, I haven't been able to find any literature on this technique. Have any of you tried this technique and/or could you point out some literature describing the technique? Also, if you have other suggestions to deal with the described problem I would be pleased to hear about them.
Regards, Jesper
---------------------------------------------------- Jesper Vejloe Carstensen Research Scientist, M.Sc., Ph.D. Materials Research Department Risoe National Laboratory P.O. Box 49 DK-4000 Roskilde, Denmark Phone: +45 4677 5776 Fax: +45 4677 5758 E-mail: jesper.v.carstensen-at-risoe.dk Web: http://www.risoe.dk/AFM/Personal/Jeca/jeca.htm ----------------------------------------------------
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Dear Jesper: Extraction replicas are an old technique. I used it 30 years ago.
1. Prepare a lightly etched metallographic specimen. 2. Evaporate with carbon to form a thin film. 3. If you want increased contrast, shadow with Cr at an angle. 4. Score the surface with a razor to form 1 mm rectangles. 5. Etch again until the carbon squares float off. 6. Scoop up the rectangles and immerse in water.The square may flatten out. If so, it can be mounted on TEM grid and dried by touching the side of the grid to a piece of filter paper. 7. If the released square curls up when immersed in the rinse water. scoop it up with a grid and gently lower it into a bath of acetone. It should snap out into a flat sheet floating on top of the acetone. Again mount on a grid and dry. If this doesn't work, remove the replica from the acetone bath and lower it into the water bath so that surface tension flattens out the replica.
I'm surprised thnat you could not find a description of thsi technique in the literature. Try some of the older texts on TEM techniques.
Good luck, Sam Purdy National Steel Corp. Technical Center Trenton, MI, USA } ---------- } From: "Carstensen, Jesper Vejl¿" } Sent: March 2000 4:31 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: TEM: Carbides in tool steels } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } I have just started on a TEM-project investigating the microstructures of } different tool steels. The steels contain various types of carbides with } different compositions, shapes and sizes. I have started out by making } thin } foils using electrolytical jet polishing. I get nice thin foils where I } can } study and analyse the martensitic matrix, but the carbides are off course } too thick to be analysed. To overcome this problem I have considered a } combination of electropolishing and ion milling, but I have also heard } about } an extraction replication technique. However, I haven't been able to find } any literature on this technique. Have any of you tried this technique } and/or could you point out some literature describing the technique? Also, } if you have other suggestions to deal with the described problem I would } be } pleased to hear about them. } } Regards, Jesper } } ---------------------------------------------------- } Jesper Vejloe Carstensen } Research Scientist, M.Sc., Ph.D. } Materials Research Department } Risoe National Laboratory } P.O. Box 49 } DK-4000 Roskilde, Denmark } Phone: +45 4677 5776 } Fax: +45 4677 5758 } E-mail: jesper.v.carstensen-at-risoe.dk } Web: http://www.risoe.dk/AFM/Personal/Jeca/jeca.htm } ---------------------------------------------------- } } } }
At 06:05 PM 3/10/00 , I wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
[snip]
Ability to spell "Amray" is/was not meant to be negative. I have had bad experience with service people who say they can work on systems but in reality cannot. I have replaced my Amray 1830 LaB6 system with the Amray 1910FE. The field emission system is much more complex and horribly more expensive to replace if not properly handled/serviced. There are special procedures especially for the FEI 305FE Zr/W gun assembly that must be followed or the emitter is zapped.
Bad experience was unfortunate with the LaB6 system. I fear it would be fatal on a field emission system. That is what I was clumsy in saying.
First you are going to have to do a bit of reading. I know of one person who studied the detection limits both experimentally and theoretically in PEELS recently ( {2 years) and that is Michael Natusch (formerly Cambridge). He found current detection limits of C in steel was only about two or three atomic percent (four to six times higher than your boron average). Check out Micron volume 30 (1999) pp173-183 (Natusch, Humphreys, Menon & Krivanek) as a starting point and for references.
Even if you do see a boron peak in the spectrum you will have to work out the signal to noise ratio (SNR) of the edge. The real problem is that you may see something that isn't there i.e. noise, or conversely you don't see something that you should see i.e. poor experimental execution.
To optimise the SNR you are going to need a lot of counts from the boundary itself. One way to see if your probe is on the boundary is to look for a possible chemical shift in the core loss edges of the matrix material since the bonding character of the material is different at the grain boundary.
Good luck. ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
Dear all on the Microscopy List, I am looking for good advice concerning surface analysis of metals using a microscope. The objects are prehistoric tools, jewellery, etc. made out of bronze, silver or gold (or combinations). I am interested in traces of the tools/techniques that was used on the object. The research technique should be non-destructive. The results will be used in archaeological research. I am looking for suggestions about what materials to use, what equipment is suitable/normally available and not too expensive. Is there anybody out there who deals with these subjects in a modern, industrial context - and who could tell me some more about, how they do it ? References for litterature, companies, etc.
Off hand, I would think that BSE imagine with a SEM might be useful. The difference between SE and BSE could be revealing. Without imaging the actual specimens, I can only speculate. But based on what you are analyzing, SE, BSE and perhaps EDX analysis can be insightful.
gary g.
At 12:29 PM 12/12/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Please notice that extracting carbides from the matrix doesn't make them thinner. So, it seems that your idea about ion milling is right.
Best regards and good luck,
Witold
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
We offer training in EM maintanance either through one of our "Monitoring and Maintaining the EM" short courses or on site as required by the client.
Steve Chapman Senior Consultant Protrain - for EM Consultancy and Training World Wide Tel 44+ 1280 814774 Fax 44+ 1280 814007 www.emcourses.com
----- Original Message ----- } From: electron microscope laboratory {emlab-at-udsm.ac.tz} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, March 10, 2000 5:24 AM
a 45 degree laser line across the sample will tell a lot about the surface. Getting a fine enough beam would be fun.
If some one wants to try it I have a start on the code in C.
Gordon W5RED
G. C. Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger "You miss 100 percent of the shots you never take." - Wayne Gretzky
----- Original Message ----- } From: "Dr. Gary Gaugler" {gary-at-gaugler.com} To: "Sren Albek" {albek-at-dorit.ihi.ku.dk} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, March 12, 2000 7:53 PM
Dear Dr. Gauler,
The E.FJELD Co. has factory trained service engineers to handle conventional AMRAY SEM's (LaB6, tungsten). We offer routine service, spare parts and service agreements on AMRAY SEM's throughout the country.
Your requested is tough... It is extremely difficult, regardless of manufacturer (Hitachi, Philips, JEOL or KLA), to find experienced service people for FE technology. We do not supply such service.
If I find a source, I will keep you posted.
Regards,
Mark Reynolds
E.FJELD Co., Inc. 3 Executive Park Drive No. Billerica, MA 01862
TEL: (978)667-1416 x10 FAX: (978)667-9059
On Friday, March 10, 2000 7:06 PM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Are there any Amray-trained service people out there that are } independently servicing the 1910FE system on the West Coast? } I'm in Sacramento CA. I am not looking for someone who knows } how to spell Amray. I am looking for someone who knows what } the 1910FE is all about. Big difference. Introductory questions: } } 1. Can you condition the emitter/gun? How and with what? } 2. How do you handle the differential vacuum aperture? } 3. what is your experience in working with the Windows control s/w? } 4. What do you know about the IP8? } 5. Do you have any knowledge about moving from 486 ISA to P-II } or P-III systems for NibbleNet and frame capture? } } Since the gobbling of Amray by KLA-Tencor, Amray SEM service } is dismal--more so day by day. The service people are excellent--but they are being } pulled in all directions, and mostly towards the KLA big semi systems. } } The Amray SEM systems were/are really good--and that is why KLA bought } Amray. But KLA is not a research SEM outfit like Amray was. } } If I had to get something other than this 1910FE, I don't know what it } would be. The Hitachi and Philips SEMs are great/nice but ridiculously } expensive to buy and maintain. And I do not care for Jeol. } } Me thinks that the research SEM options are shrinking daily. } } gary g. } }
I know little about liver, but if these are fairly large cells under high demands to produce [mRNAs], the polyploidy may be a response to a biological need. In a far simpler system, the nematode, there is good evidence that ploidy gets regulated in direct correspondance to the absolute size of the cell, across many different tissues, such that the ploidy perhaps matches the needs of the cell.
see Hedgecock and White (1985) Dev Biol 107: 128-133.
Though vertebrate cells may or may not be so closely regulated, the tendency might lie along a similar continuum?
Just a thought. Best regards, David H. Hall Center for C. elegans Anatomy Department of Neuroscience 1410 Pelham Parkway Albert Einstein College of Medicine Bronx, NY 10461
Check out my web page for examples of the different modes of scanning electron microscopy. It is a simple web page and has a section showing examples of the different modes of SEM. I recently quit my job and built a SEM lab in my basement and also have x-ray microanalysis for generating elemental spectra on micro-volumes. I am an expert in the different SEM contrast mechanisms, but not a metallurgist w/ any experience in the field that you mention. SEM is non-destructive provided you can get the material into the chamber. Some SEMs have large chambers, mine is on the smaller side (perhaps samples {4" and somewhat flat sample would be best). I would be glad to run 2-3 samples for free to show you what SEM can do as a feasibility study for SEM analysis of your samples. All my output can be in the form of jpg images so that you can send the data to experts in archaeology and they can comment on the significance.
Good Luck
Ric Felten www.semguy.com
-----Original Message----- } From: S¿ren Albek [mailto:albek-at-dorit.ihi.ku.dk] Sent: Sunday, December 12, 1999 1:29 PM To: Microscopy-at-sparc5.microscopy.com
Dear all on the Microscopy List, I am looking for good advice concerning surface analysis of metals using a microscope. The objects are prehistoric tools, jewellery, etc. made out of bronze, silver or gold (or combinations). I am interested in traces of the tools/techniques that was used on the object. The research technique should be non-destructive. The results will be used in archaeological research. I am looking for suggestions about what materials to use, what equipment is suitable/normally available and not too expensive. Is there anybody out there who deals with these subjects in a modern, industrial context - and who could tell me some more about, how they do it ? References for litterature, companies, etc.
In addition to the advice you have already received you might wish to consider contacting your local Forensic Science Laboratory and talk to the Firearm and Toolmark Section. What you wish to do is to determine the Class characteristics of the tools used, as you do not have the actual tool for comparison. There is also some Toolmark analysis done at certain types of machine shops but a really experienced Toolmark examiner at your local Forensic Science Lab would be much easier to locate and probably more interested in your project (I'm currently doing some work for a project on bullets from a sight, in my free evenings and I know others have in the past). Low power stereo microscopy can tell you a lot. The examiners can point you in the right direction and provide some literature sights etc., or may be interested in doing the work for you.
Jim
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } "S¿ren Albek" {albek-at-dorit.ihi.ku.dk} 12/12/99 10:29AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all on the Microscopy List, I am looking for good advice concerning surface analysis of metals using a microscope. The objects are prehistoric tools, jewellery, etc. made out of bronze, silver or gold (or combinations). I am interested in traces of the tools/techniques that was used on the object. The research technique should be non-destructive. The results will be used in archaeological research. I am looking for suggestions about what materials to use, what equipment is suitable/normally available and not too expensive. Is there anybody out there who deals with these subjects in a modern, industrial context - and who could tell me some more about, how they do it ? References for litterature, companies, etc.
Hi, I need advice on double labeling (on-grid). I've had success doing each of the labeling experiments independently but now the researcher wants to see double labeling. I'm doing indirect labeling with a monoclonal antibody to fungal cell wall antigens followed by a gold labeled secondary. In addition, I want to label the sections with gold conjugated wheat germ agglutinin. Should I do the antibody labeling first and then the lectin? I've found one reference that I haven't had a chance to follow up on (Cailliez et al, 1988) in Immunogold Silver Staining Principles, Methods, and Applications by Hayat. Any advice would be greatly appreciated. Beth
we routinely use Lowicryl K4M. We normally buy it as a kit from EM science. They also offer a it as a MonoStep Single Component. We would like to try it. Has anybody done the comparison between the kit and the monostep stuff (quality, shelflive, handling, polymerisation)? Thanks for your time,
Dear all, thank you very much for all the kind answers, good ideas, and good suggestions. It has provided me with a good range of possibilities (I am a little overwhelmed by now :-). I must now evaluate the answers and I will most likely return with some more questions - some of these off-list, when appropriate.
Once again, thank you for the interest, and with many regards,
Dear Listers, I have a question about the cleaved glass blades used to cut cross sections of copolymers. Is there any way to quantify the sharpness of the blades? Do they have a radius of curvature, or anything like that? A client of ours has asked if the blade would be sharp enough to slice through the hard domain (polystyrene hardness) at -20 C or -80 C, or if it would just push it out of the way. This also applies to the disposable blades on a cryostat used to section the same sample at -20C prior to the cryo-ultramicrotoming. The sample is very brittle at -120C. Rosemary
Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Hello, We have an old semicaps 4000 for pc windows 3.1 system. I've been trying to use it to get scale bars burned into images, but I must say that it is very buggy. The images never burn scale bars correctly, calibration procedures are never fully written in the manual, and often minimizing and maximizing windows causes images to have very 'weird' color schemes.
After contacting semicaps I was told that there were no bug fixes to the software as it was bug free. Anyone out there have some experience with this system? Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
My post has generated several responses. All of which are very helpful. Thanks to all who responded.
From following up on what I have discovered from the response from Alex Greene, I'm rather stuck. It turns out that Amray has a lock on the FE gun/emitter that they developed and which incorporate the FEI W/Zr emitter (305FE). As such, no other party will support the Amray FE systems.
There are several folks who will and do support the Amray LaB6 systems. But none can or will support the FE systems. So for the time being, I am stuck with Amray. Be that as it may. As I write this, Amray (KLA-Tencor) is preparing to work on my FESEM under my existing maintenance contract. This is good. I guess that so long as this keeps up, I am OK. But I do expect that KLA will drop all Amray field support in the near future. Consequently, perhaps that will be a better time to seek alternative maintenance sources. I do expect at this future juncture, KLA/Amray will supply parts but will not supply warm bodies. If this is an opportunity for independents, who knows? If you are experiencing similar troubles, I would for one appreciate hearing about your situation. Otherwise...
Unless somebody has done that very same thing, one cannot know. However, I am optimistic that you could section that material with a glass knife. A new knife should run virtually to a molecular edge, although that finest edge would not survive the first section. The glass knife is sharp, the question concerns more its hardness and toughness. An LKB trainer years ago told me that a glass knife at liquid nitrogen temperature is almost as hard as is diamond at room temperature. So there is a good chance that glass could do. Try it. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, March 14, 2000 9:05 AM, Rosemary Walsh [SMTP:rw9-at-psu.edu] wrote: } } } Dear Listers, } I have a question about the cleaved glass blades used to cut cross } sections of copolymers. Is there any way to quantify the sharpness of } the blades? Do they have a radius of curvature, or anything like that? } A client of ours has asked if the blade would be sharp enough to slice } through the hard domain (polystyrene hardness) at -20 C or -80 C, } or if it would just push it out of the way. This also applies to the } disposable blades on a cryostat used to section the same sample at } -20C prior to the cryo-ultramicrotoming. The sample is very brittle } at -120C. } Rosemary } } Rosemary Walsh, Manager } The Electron Microscope Facility for the Life Sciences, } A Shared Technology Facility, The Life Sciences Consortium } 1 South FrearLab } Penn State University } University Park, PA 16802 } (814) 865-0212 } rw9-at-psu.edu } http://www.lsc.psu.edu/stf/em/home.html } }
We would like to do immunohistochemistry on rat lung tissue and we are especially interrested in CD4, CD8 and some macrophage-markers such as ED1 and ED2. But we would like to have good morphology as well. I have found some articles about IHC (rat and mice) on tissue fixed in paraformaldehyde-lysine-peroidate and emdedded in low temp paraffin. But most of these articles are rather old (1985-1990). Is anyone familliar with these techniques? Is it difficult to repeat and to get similar results?
Joost Bruijntjes TNO Nutrition and Food Research Institute Zeist Holland E-mail: J.Bruyntjes-at-voeding.tno.nl
Dear Andrew In principle, sections can be orientated using any two features in the block that run parallel to each other and normal to the knife edge. It would be nice to have reference rods included in the block, and there are various possible approaches to this, but I cannot see a) a way of meeting both your criterion of providing a dimensional reference for the assessment of shrinkage, and of orientating the section b) a method which is generally applicable to all microtomy techniques.
To measure shrinkage, I think I would be looking at making the fresh specimen a standard size, following its dimensions through processing. Two ways of making specimens a standard size occur to me: 1) Cut slices of the tissue with razor blades mounted side- by side separated by a spacer. Re-cut the tissue slice to make strips with square cross section, or dice it into cubes 2) take a tissue core with a carefully sharpened stainless steel tube (cannula, large hypodermic needle) and follow the core diameter through the process.
For section orientation, reference cores or rods can be melted with a needle (wax, gelatine) or drilled (plastic) normal to the block face. The holes may be left as voids or filled with a contrasting embedding material (e.g. coloured wax, resin, gelatine). However this is all a bit of a fiddle, and becomes either very difficult to achieve or impossible in ultramicrotomy of small block faces, ultracryotomy or cryostat sectioning with Tissue-tek or other liquid embedment.
A general principle for section orientation and alignment that is applicable to almost all microtomy techniques is to cut a mesa on the block face of rhomboidal or trapezoidal shape. The mesa is cut by removing waste with the corner of the knife (which must be square) to a little more than the thickness of the serial sequence you wish to cut. Since the edge-facets of the mesa will be perfectly aligned normal to the block face the sections can be aligned using any two corners, or if they should be lost/obscured, by means of the surviving edges.
This may fail with whole-mounts. In this situation, usually there are features on the periphery of the specimen that can act as reference marks, and you still have the option to cut the block face into a trapezoid or make a mesa, or melt holes in the embedding material with a hot needle, etc.
Finally, a number of digital image analysis packages now offer fourier-based image correlation for registering images when making stereo-pairs, for example, and montages. AnalySIS is one of these:
www.soft-imaging.de
Chris
Date sent: Fri, 3 Mar 2000 09:01:49 +1300 To: c.jeffree-at-ed.ac.uk } From: Andrew McNaughton {andrew.mcnaughton-at-stonebow.otago.ac.nz}
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Please notice that extracting carbides from the matrix doesn't make them thinner. So, it seems that your idea about ion milling is right.
Best regards and good luck,
Witold
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
To all, thank you for the many suggestions regarding the moving aperture, It is definitly not the distance between aperture & specimen holder. It was checked several times and the clearance is there. Also, the aperture was stable before and is now moving the most in the # 2 position. I would appreciate any further comments. Thank you . Peter A. Stolzenberg, Pesto Inc. " pestoem-at-aol.com"
hello, I have to prepare cross sectionning samples, and I do not know where to buy the small vise that I need to clamp the specimen when I have glued them (I have already the Mbond 610 adhesive). I would greetly appreciate any information about this thing (I am not even sure of the name of it) thank you in advance christine
} } Dear all } } We would like to do immunohistochemistry on rat lung tissue and we are } especially interrested in CD4, CD8 and some macrophage-markers such as ED1 } and ED2. But we would like to have good morphology as well. I have found } some articles about IHC (rat and mice) on tissue fixed in } paraformaldehyde-lysine-peroidate and emdedded in low temp paraffin. } But most of these articles are rather old (1985-1990). Is anyone familliar } with these techniques? Is it difficult to repeat and to get similar results? } } } Joost Bruijntjes } TNO Nutrition and Food Research Institute } Zeist } Holland } E-mail: J.Bruyntjes-at-voeding.tno.nl
********** I have had some luck with the PLP fixation you mention (McLean & Nakane, J Histochem Cytocem 22, 1974). I gives good structural preservation, and like most immuno techniques, you won't know if it works with your antigen until you try it. Are you sure you mean low temp paraffin and not resin? Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I'm looking for a source for vibration isolation tables for ultramicrotomy. The marble tables that we are using now are not adequate (except during Spring Break when no one is in the building...) I'm thinking of a system that uses compressed gas (N2?) to float the tables.
Please respond offline (vendors welcome).
thanks, all
Ann Hein Lehman EM Facility Mgr Trinity College Hartford CT 860-297-4289 860-297-2538 ann.lehman-at-trincoll.edu
I have recently performed an immunoEM labeling procedure with myelinated nerve embedded in Lowicryl, K4M resin. Unfortunately the myelin is severely altered (it appears as large clealr blebs) because I am not able to osmicate this tissue due to the UV polymerization of this resin. Does anyone have any suggestions as to preservation of lipid laden myelin without the use of osmium? I used methanol in the dehydration procedure which has probably caused this artefact....
Thanks! Karen Jensen, M.S. Associate Scientist Electron Microscope Research Imaging Core University of Rochester Medical Center Rochester, NY
I lost contact w/ a certain person. All I can remember is that they were experienced in confocal microscopy and lived in North West Ct, I think Cornwall.
Dr. Gary Gaugler wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } My post has generated several responses. All of which are very } helpful. Thanks to all who responded. } } From following up on what I have discovered from the response from } Alex Greene, I'm rather stuck. It turns out that Amray has a lock on } the FE gun/emitter that they developed and which incorporate the } FEI W/Zr emitter (305FE). As such, no other party will support the } Amray FE systems. } } There are several folks who will and do support the Amray LaB6 systems. } But none can or will support the FE systems. So for the time being, } I am stuck with Amray. Be that as it may. As I write this, Amray } (KLA-Tencor) is preparing to work on my FESEM under my existing } maintenance contract. This is good. I guess that so long as this } keeps up, I am OK. But I do expect that KLA will drop all Amray } field support in the near future. Consequently, perhaps that will } be a better time to seek alternative maintenance sources. I do } expect at this future juncture, KLA/Amray will supply parts } but will not supply warm bodies. If this is an opportunity for } independents, who knows? If you are experiencing similar } troubles, I would for one appreciate hearing about your situation. } Otherwise... } } Stay tuned. } } gary g. Gary, Your earlier comments on how good AMRAY is really puzzle me, but be that as it may, if any of their Federal Government customers are on the ball, Amray will be told, as was Perkin Elmer ETEC, that they must support the equipment for about 7 years or face lawsuits. There is an implied responsibility when one sells expensive equipment.
Perhaps supplying parts and detailed instructions will cover them legally, in which case we third party folks may be able to help out. If anyone has any more detailed info on Amray service, I would love to know because I have serviced them off and on over the years and would be willing to help out here in the Northeast/Mid-Atlantic area.
Ken Converse owner Quality Images third party SEM service 16 Creek Rd. Delta, PA 17314
Have a look at: Toda, Y et al: Application of tyramide signal amplification system to immunohistochemistry: A potent method to localize antigens that are not detectable by ordinary method. Pathology International 1999; 49: 479-483. They mention CD4 specifically in addition to other antigens and outline several antigen retrieval methods.
South Bay Technology, Inc. manufactures both a cross sectioning vise (formerly produced by VCR Group) and also an XTEM clamp. Both tools are used for the same purpose you describe although they do it a little bit differently. I am sending you a data sheet covering each one as an attachment to a separate e-mail. We also offer a complete cross section sample preparation kit which provides all of the bits and pieces that you would need to make the cross section.
David Henriks Vice President TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Leroux christine } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
hello, I have to prepare cross sectionning samples, and I do not know where to buy the small vise that I need to clamp the specimen when I have glued them (I have already the Mbond 610 adhesive). I would greetly appreciate any information about this thing (I am not even sure of the name of it) thank you in advance christine
I am looking for a backscattered electron detector (for elemental contrast) to attach to a JEOL JSM-5800 SEM. This is for a university, so low-cost or donation would be most desirable. Thanks for your help!
Hi, Leroux: I think a negative tweezle will do, provided that your sample is small. Just make sure that you do not heat your sample too fast. Actually I designed a spring vice especially for cross section samples, it is very good for both big and small samples, the glue line can go thinner than 100nm easily, but normally I do not use it since almost all of my samples are small---those film growers are really stinge:) Anyway, if you need, I can send a schematic to you.
Good Luck.
Hao Li
On Tue, 14 Mar 2000, Leroux christine wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } hello, } I have to prepare cross sectionning samples, and I do not know where to } buy the small } vise that I need to clamp the specimen when I have glued them (I have } already the } Mbond 610 adhesive). } I would greetly appreciate any information about this thing (I am not even } sure of the } name of it) } thank you in advance } christine } } ************************************** } Christine leroux } Lab. M.M.I } Universite de Toulon-Var, Bat.R } B.P.132 } F-83957 La Garde Cedex } tel: 00 33 (0) 4 94 14 25 07 } fax: 00 33 (0) 4 94 14 21 68 } } ************************************** } }
Does anyone have a protocol for making Toluidine blue stain to be used for staining thick sections of resin embedded biological samples ?
Thanks a lot,
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
Can anyone suggest name of an individual or independent company in New Mexico, West Texas or Arizona who can fix/service research level compound microscopes ?
Thanks in advance,
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
We have an old PGT System 4plus (no detector) sitting here that won't boot. Apart from that it was working fine. If anyone would be interested in it for parts (or whatever else you want), they are welcome to it. Not interested in cash, just pay for your shipping and its yours. We are located near Ottawa, Canada.
Please contact Mark Chambers at 599-6500 ext. 4269 for more info, or if you are interested.
You can buy a nice little vise from South Bay Technology. This vise was previously made by VCR. It can be put on a hot plate and has Teflon jaws and a Teflon slide base so that the epoxy doesn't stick. I have two of these. They work great. I put a small dab of epoxy on t he Teflon jaw to determine when the epoxy is cured and scraped it off with a glass microscope slide. SBT also has two spring loaded clamping devices that can be used, but I recommend the small vise. You can also buy a spring loaded clamping device from Fischione and from Gatan. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Leroux christine [mailto:leroux-at-univ-tln.fr] } Sent: Tuesday, March 14, 2000 9:45 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM cross section preparation } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.} html } } } -------------------------------------------------------------- } ---------. } } } hello, } I have to prepare cross sectionning samples, and I do not } know where to } buy the small } vise that I need to clamp the specimen when I have glued } them (I have } already the } Mbond 610 adhesive). } I would greetly appreciate any information about this thing } (I am not even } sure of the } name of it) } thank you in advance } christine } } ************************************** } Christine leroux } Lab. M.M.I } Universite de Toulon-Var, Bat.R } B.P.132 } F-83957 La Garde Cedex } tel: 00 33 (0) 4 94 14 25 07 } fax: 00 33 (0) 4 94 14 21 68 } } ************************************** } }
Here is the web site for TMC: http://www.techmfg.com/vibrasolutions.html and http://www.techmfg.com/65floorplatfrm.html -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Lehman, Ann [mailto:Ann.Lehman-at-trincoll.edu] } Sent: Tuesday, March 14, 2000 9:37 AM } To: 'MSA Listserver' } Subject: Vibration isolation tables } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Dear Listers, } } I'm looking for a source for vibration isolation tables for } ultramicrotomy. } The marble tables that we are using now are not adequate } (except during } Spring Break when no one is in the building...) I'm thinking } of a system } that uses compressed gas (N2?) to float the tables. } } Please respond offline (vendors welcome). } } thanks, all } } Ann Hein Lehman } EM Facility Mgr } Trinity College } Hartford CT } 860-297-4289 } 860-297-2538 } ann.lehman-at-trincoll.edu }
} Dear Microscopists, } } Does anyone have a protocol for making Toluidine blue stain to be used for } staining thick sections of resin embedded biological samples ? } } Thanks a lot, } } Soumitra } }
Toludine Blue Stain
100g Toludine Blue 100g Borax (sodium borate) 1000 mls ddw.
Older stain works better than newer. Filter before use.
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
Hello all, We are trying to get a project started that involves imaging the microstructure of mineral oils and polyalphaolefins as they solidify. The solid temperature range for materials like these is approximately -20 to -70 C. Does anyone have experience with these or similar materials. We are hoping to image the particle structure (crystals) by SEM. Any feedback from those with similar projects/experiences would be very welcome (on or off the list). Even better would be references to any published reports of this kind of material exam.
Thanks, Brad Huggins BPAmoco, Electron Microscopy Lab hugginbj-at-bp.com
At 01:13 PM 3/14/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
[snip]
} Gary, } Your earlier comments on how good AMRAY is really puzzle me, but be that } as it may, if any of their Federal Government customers are on the ball, } Amray will be told, as was Perkin Elmer ETEC, that they must support the } equipment for about 7 years or face lawsuits. There is an implied } responsibility when one sells expensive equipment. } } Perhaps supplying parts and detailed instructions will cover them } legally, in which case we third party folks may be able to help out. If } anyone has any more detailed info on Amray service, I would love to know } because I have serviced them off and on over the years and would be } willing to help out here in the Northeast/Mid-Atlantic area. } } Ken Converse } owner } Quality Images } third party SEM service } 16 Creek Rd. } Delta, PA 17314 } } 717-456-5491
I take it that you don't think that Amrays are very good. What aspect of them do you find lacking? I would tend to agree that the early models are no comparison to their generations of the last 6-10 years. The FE systems are particularly good. My 1910FE does a great job being an FE and having the flat lens. The 1800 series with conical lenses are not as good, but are required when doing IC wafers.
You have a great point about US Gov users. I know several of them. When does the 7 years start? From what event or timepoint? Amray has not stopped providing service. But the word is that they most likely will in about 2 years....based on their takeover by KLA-Tencor.
One fellow pointed out the proprietary nature of the FE gun. Without access to this, third party providers cannot work on the systems....at least not on the gun assembly.
My second topic for the day is FESEM or TEM imaging of the crystalline/amorphous microstructure of poly(ethylene terephthalate), PET. I have on many occasions had success achieving minimal contrast of such domains in certain unstained PET polymers in the SEM and FESEM. Recently we have "almost" been able to observe the crystalline domains with enough clarity to size them, this by direct examination of polished (microtomed) surfaces. I would like to pursue the microstructural characterization a step further by using staining or etching techniques (or some other trick that some one may have up their sleeve!). In Polymer Microscopy (L Sawyer, D Grubb) aminolysis appears to be (cautiously) recommended. Staining with PTA is also mentioned. I am looking to image crystalline domains in the range of approximately 50 to 150 nm.
Does anybody on the List have first hand experience with these or other techniques to enhance the direct (or indirect) observation of these domains in PET. I'm not looking to re-invent the wheel here, so any help will be greatly appreciated. Get back to me direct or on the List.
Again, Thanks in advance, Brad Huggins BPAmoco, Electron Microscopy Lab hugginbj-at-bp.com
} hello, } I have to prepare cross sectionning samples, and I do not know where to } buy the small } vise that I need to clamp the specimen when I have glued them (I have } already the } Mbond 610 adhesive). } christine
You haven't told us what equipment you will use to prepare your "cross sectionning samples", so it's hard to provide advice!
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I would suspect Elaine is giving a 10X stock solution?
We use 1% Toluidine Blue dissolved in 1% sodium borate. Filter before use. Diluting 10-fold with 1% sodium borate, to 0.1% toluidine blue, allows lightly counterstaining sections developed for autoradiography.
Refs. Mercer, E.H., J Royal Microscopy Society, 81:179 (1963) Burns, W.A., "Thick Sections: Technique and Applications", Diagnostic Electron Microscopy, Chapter 4, B.F. Trump and R.J. Jones, eds., John Wiley and Sons, New Yourk , 1978.
Regards, Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
} } Dear Microscopists, } } } } Does anyone have a protocol for making Toluidine blue stain to be used for } } staining thick sections of resin embedded biological samples ? } } } } Thanks a lot, } } } } Soumitra } } } } } } } Toludine Blue Stain } } 100g Toludine Blue } 100g Borax (sodium borate) } 1000 mls ddw. } } Older stain works better than newer. Filter before use. } } } Dr. Elaine Humphrey } Biosciences Electron Microscopy Facility } University of British Columbia } 6270 University Blvd, mail-stop Botany } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-unixg.ubc.ca } } } }
} We have an old semicaps 4000 for pc windows 3.1 system. } I've been trying to use it to get scale bars burned } into images, but I must say that it is very buggy. ...
First, I need to admit no experience with this system ... but even with my software which does produce accurate scale bars, I still prefer to annotate the images with an image editor like Photoshop.
If you have a magnification standard, then you should be able to determine an instrumental constant which can be assumed doesn't change with magnification, but may change with the SEM's working distance. You should be able to end up with this equation:
microns per pixel = constant/magnification
This may need to be determined for each fixed working distance, or you may be able to determine how to incorporate WD into the equation (my own software compensates with the objective lens setting so the mag is relatively accurate). If your image acquisition options include pixel dimentions (e.g., 256x256, 512x512, 1024x1024), then this needs to be a factor as well (in the denominator).
I then created a spreadsheet for the pixel dimentions of common scale bars for different magnifications, printed it, and then made it available at the Photoshop workstation.
I can provide an example of the spreadsheet to anyone interested.
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/
Thanks Glen, you are quite right. It is easier to make up 10g Toluidine Blue, 10g sodium borate and 1000mls water. Elaine
} I would suspect Elaine is giving a 10X stock solution? } } We use 1% Toluidine Blue dissolved in 1% sodium borate. Filter before use. } Diluting 10-fold } with 1% sodium borate, to 0.1% toluidine blue, allows lightly } counterstaining sections developed for autoradiography. } } Refs. } Mercer, E.H., J Royal Microscopy Society, 81:179 (1963) } Burns, W.A., "Thick Sections: Technique and Applications", Diagnostic } Electron Microscopy, Chapter 4, B.F. Trump and R.J. Jones, eds., John } Wiley and Sons, New Yourk , 1978. } } Regards, } Glen } } } Glen MacDonald } Research Scientist } Hearing Research Laboratories of the } Virginia Merrill Bloedel Hearing Research Center } Box 35-7923 } University of Washington } Seattle, WA 98195-7923 } (206) 616-4156 } glenmac-at-u.washington.edu } } } } Dear Microscopists, } } } } } } Does anyone have a protocol for making Toluidine blue stain to be used for } } } staining thick sections of resin embedded biological samples ? } } } } } } Thanks a lot, } } } } } } Soumitra } } } } } } } } } } } } Toludine Blue Stain } } } } 100g Toludine Blue } } 100g Borax (sodium borate) } } 1000 mls ddw. } } } } Older stain works better than newer. Filter before use. } }
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
We have a FEI CM20 (TEM) and XL30 (FEGSEM) and are in the process of reviewing service contracts vs insurance. The service contract runs a min of 40K for both instruments. We currently have a tech that can do minor service, yearly maintenance, and alignments.
I'd appreciate any input as to how other organizations have dealt with this.
} } Dear Collegues } } I'm having a problem with LR-White polimerization. } } I use to polimerise the LR-white resin at 60¼C without any aditives and it } usually works without problems. A new batch of the resin failed to } polimerise, and the manufacturer claims that it is necessary to use the } catalyst. } } I understand that the "accelerator" should not be used except for cold cure, } and even then may be harmfull since the specimen may be subject to too much } heat even under those conditions. } } I would like your opinion on: } 1. is it necessary to use the catalyst for heat cure? } } 2. If it is, is there any difference among the resins that may justify the } apparent success of the old protocol (good cure without catalyst?) } } Thanks in advance } } Dr. A.P. Alves de Matos } } }
Full-time The Dow Chemical Company, Midland, Michigan with possible relocation to Dow's Freeport, Texas facility Equal opportunity employer offering a competitive compensation and benefits package including 401k, stock purchase, performance incentives, and educational assistance
Responsibilities: Support existing businesses using SPM and develop new customers for SPM as appropriate. Calibrate and maintain SPM instrumentation base consisting of Digital Instruments' Multimode SPM, D3000 SPM, Hysitron Nanoindentation and TA Instruments Micro-TA SThM. Participate in business aligned, original research programs through internal development funding. Bring in new technology through previous experience or external contacts as appropriate.
Qualifications: MS degree in Polymer Science, Materials Science or Chemistry. Ph.D. degree is preferred. Experience in scanning probe microscopy characterization of polymeric materials. Experience with Digital Instruments Nanoscope is desired. Routine experience with a variety of imaging modes (e.g. TappingMode, phase, friction) as well as unscanned modes (force curve, nanoindentation) is also desired. Leading research in the polymer SPM area.
Send Resumes by April 10, 2000 to: The Dow Chemical Company, P. O Box 1655, Workforce Planning Department Job 00-94/MLK, Midland, Michigan, United States, 48641-1655 or e-mail: R&D-at-dow.com. Email respondents must list Job 00-94/MLK and their last name as the first and second items on the Subject line. Only those selected for an interview will be contacted.
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Weight 0.5 g Tolyidine Blue in some bottle (I am using 50 ml polypropylene tissue culture tubes), add 50 ml 1x PBS and heat it in boiled water for 20-30 min. When hot, filter it using Whatman filter paper (Cat No 1001 090).
Staining: Plastic sections up to 0.5 microns thick on the microscope slide cover with several drops of the stain and heat on heat-plate, 60oC for 5-10 min. Wash away stain with tap water, wash in distilled water, dry and enjoy the view.
Godd luck, Sergey.
} Date: Tue, 14 Mar 2000 13:18:28 -0700 } From: Soumitra Ghoshroy {ghoshroy-at-nmsu.edu} } Subject: Toluidine blue stain } X-Sender: ghoshroy-at-cnmailsvr.nmsu.edu } To: Microscopy-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
We have a FEI CM20 (TEM) and XL30 (FEGSEM) and are in the process of reviewing service contracts vs insurance. The service contract runs a min of 40K for both instruments. We currently have a tech that can do minor service, yearly maintenance, and alignments. We have not had a service contract for the last 2 years and have had no problem getting FEI to come out ASAP when needed.
I'd appreciate any input as to how other organizations have dealt with this.
I have a (possibly) similar problem with JEOL 100SX microscope. Very often the beam keeps tilting in a motion that displaces it relative to the objective aperture. The effect is the same as moving the aperture. I suspect of circuit instability but the assistance was unable to find any trouble in that area. Another possibility is charge accumulation inducing electric fields and displacing the beam. Since you are lucki to have the problem mostly in one position, I would suspect of some kind of electric isolation of the aperture holder either in that hole or produced when the entire piece goes into that position. The charge effects could be also related to the specimen itself. Jeol seems to be particularly sensitive to this. If you are looking at epon sections try to coat them with a thin layer of carbon (after staining, evaporate directly into the sections) - it gives much improvement for me.
Dr. A.P. Alves de Matos
To all, thank you for the many suggestions regarding the moving aperture, It is definitly not the distance between aperture & specimen holder. It was checked several times and the clearance is there. Also, the aperture was stable before and is now moving the most in the # 2 position. I would appreciate any further comments. Thank you . Peter A. Stolzenberg, Pesto Inc. " pestoem-at-aol.com"
Most LR White is sold with catalyst already added. This will cure at 60 degrees and must be stored refrigerated. If you purchased uncatalysed (the only way we supply LR White because we are a long way from the UK) then prior to first use the catalyst must be added to the whole bottle and well mixed to dissolve. Thereafter store the catalysed LR White refrigerated.
If you want to cold cure you need to additionally add accelerator. Accelerator is not supplied with a bottle of LR White, but when you purchase a "kit". Instruction notes are linked online from the LR White. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, March 15, 2000 11:27 AM, A.P. Alves de Matos [SMTP:apmatos-at-ip.pt] 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 Collegues } } } } I'm having a problem with LR-White polimerization. } } } } I use to polimerise the LR-white resin at 60oC without any aditives and it } } usually works without problems. A new batch of the resin failed to } } polimerise, and the manufacturer claims that it is necessary to use the } } catalyst. } } } } I understand that the "accelerator" should not be used except for cold } cure, } } and even then may be harmfull since the specimen may be subject to too } much } } heat even under those conditions. } } } } I would like your opinion on: } } 1. is it necessary to use the catalyst for heat cure? } } } } 2. If it is, is there any difference among the resins that may justify the } } apparent success of the old protocol (good cure without catalyst?) } } } } Thanks in advance } } } } Dr. A.P. Alves de Matos } } } } } } } }
I do not know what the objective aperture strip looks like on a 100 CX so I am sort of guessing.
I have seen some aperture rods where the end consists of say three or four holes of identical diameter and small Pt/Mo discs with different sized holes etched in. If the No2 aperture is moving the most then this could possibly explain it. This disc maybe more loosely held or has a worse connection to ground than the others. Can you check these with a multimeter or something?
What does the movement look like:- continuous slow/ slow with fast jumps/ rapid shaking/random direction/ one direction: along the rod or side to side? And does the total beam current affect the motion greatly?
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
I need to embed some complex glass mouldings which also contain components made of plastics (HDPE, PP, PC) in (for preference) clear resin for sectioning with a diamond saw. This has turned out to be much more problematical than I had expected, for two reasons:
1) Very considerable shrinkage of the resins occurs during curing (applies to acrylic, epoxy or polyester types) . This causes separation of the resin from the glass and plastics components inside partially-enclosed voids, so that the original spatial relationships are lost.
2) bonding of the resins to the glass is quite variable, but typically poor. Bonding to plastics is poor. When the bond-strength to glass is high, particularly with epoxies, the stresses induced by the shrinking resin during curing are large enough to fracture the glass.
Is there such a thing as a non-shrinking resin? How can I improve bond-strength between glass and resin? How can I improve bond-strength between resin and HDPE or PP?
Any tips gratefully received Yours Chris ===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
To stain resin, e.g. Spurr's, toluidine blue must be prepared at high pH.
0.5% toluidine blue in 0.1%sodium carbonate (Na2CO3) (pH 11.1) works well. Stain sections on slides on a hot plate at 60C for 30sec. to 2 min. and rinse with water. Some resins and plastics are much easier to stain than Spurr's, e.g. JB-4, GMA etc. so I also use 0.1% tol blue in 0.01% sodium carbonate (pH 6.6). The basic protocol stays the same, you vary concentrations to adjust pH and therefore staining intensity. Hope this helps, John.
Soumitra Ghoshroy wrote:
} Dear Microscopists, } } Does anyone have a protocol for making Toluidine blue stain to be used for } staining thick sections of resin embedded biological samples ? } } Thanks a lot, } } Soumitra } } ***************************************************************** } Soumitra Ghoshroy Ph.D. } Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3600 } Fax: 505-646-5665 } e-mail: ghoshroy-at-nmsu.edu } http://confocal.nmsu.edu/eml
-- C. John Runions, Ph.D. Department of Plant Sciences University of Cambridge Downing St. Cambridge CB2 3EA UK
From your comments I would suspect that you have a charging problem. As it seems to be dependent on the aperture chosen it may be on the aperture mechanism. Try cleaning it, check that there is electrical continuity along the rod and that the mechanism is properly grounded. Do you have a touch switch? If so then ensure that it is grounded through the resistor properly.
Good luck, Ron
On Tue, 14 Mar 2000 PESTOEM-at-aol.com-at-sparc5.microscopy.com wrote:}
} thank you for the many suggestions regarding the moving aperture, It is } definitly not the distance between aperture & specimen holder. It was checked } several times and the clearance is there. Also, the aperture was stable } before and is now moving the most in the # 2 position. } I would appreciate any further comments. } Thank you . Peter A. Stolzenberg, Pesto Inc. " pestoem-at-aol.com" } }
=========================================================================== 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 know oil based fuels have been imaged at low temperature. Suggest you read (better still, Buy) my book "Low Temperature Microscopy and Analysis" Plenum Press New York 1992. I would modestly suggest it has a lot of information about how to prepare, observe and analyse specimens at low temperature. My own current interests revolve around low volyage, low temperature x-ray microabalysis of bio-organic materials. Let me know if I can be of any further help
Patrick Echlin University of Cambridge UK
pe13-at-cus.cam.ac.uk
On Tue, 14 Mar 2000, Huggins, Bradley J wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } We are trying to get a project started that involves imaging the } microstructure of mineral oils and polyalphaolefins as they solidify. The } solid temperature range for materials like these is approximately -20 to -70 } C. Does anyone have experience with these or similar materials. We are } hoping to image the particle structure (crystals) by SEM. Any feedback from } those with similar projects/experiences would be very welcome (on or off the } list). Even better would be references to any published reports of this } kind of material exam. } } Thanks, } Brad Huggins } BPAmoco, } Electron Microscopy Lab } hugginbj-at-bp.com } } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You have fallen into the same trap I did some time back. Some suppliers ship the LR White resin with the catalyst already included. Others do not, sending the catalyst along with instructions to add it prior to using the resin. The reason to not mix the ingredients prior to shipping is that if the resin is subject to hot temperatures or prolonged shipping time, the resin mix could be compromised. This is a good point which is certanly valid when shipping resin in the summer or to remote locations.
However, most of us do not run into these problems and are used to receiving the resin in the complete mixed form. In my case,I ordered from a different supplier than normal. The labeling of the bottles was not clear and I did not realize that the enclosed "extra" ingredient was the catalyst, not the accelerator used for cold curing polymerization, until I also ran into the problem of the resin not polymerizing as expected.
Moral of the story....always read the fine print...both in the catalog and after receiving the product, especially when ordering from a different supplier than in the past.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
On Tuesday, March 14, 2000, A.P. Alves de Matos {apmatos-at-ip.pt} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id IAA06960 for dist-Microscopy; Wed, 15 Mar 2000 08:16:34 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id IAA06955 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 15 Mar 2000 08:16:04 -0600 (CST) Received: from mailgw.cc.uga.edu (mailgw.cc.uga.edu [128.192.1.101]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id IAA06947 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 15 Mar 2000 08:15:53 -0600 (CST) Message-Id: {200003151415.IAA06947-at-ultra5.microscopy.com} Received: from calc.vet.uga.edu by mailgw.cc.uga.edu (LSMTP for Windows NT v1.1b) with SMTP id {0.01A98475-at-mailgw.cc.uga.edu} ; Wed, 15 Mar 2000 9:07:39 -0500 Received: from CALC_SERVER/SpoolDir by calc.vet.uga.edu (Mercury 1.40); 15 Mar 100 09:10:35 EST Received: from SpoolDir by CALC_SERVER (Mercury 1.40); 15 Mar 100 09:10:33 EST
I have recently performed an immunoEM labeling procedure with } myelinated nerve embedded in Lowicryl, K4M resin. Unfortunately the } myelin is severely altered (it appears as large clealr blebs) because I } am not able to osmicate this tissue due to the UV polymerization of } this resin. Does anyone have any suggestions as to preservation of } lipid laden myelin without the use of osmium? I used methanol in the } dehydration procedure which has probably caused this artefact....
Karen,
It sounds like the myelin is being partially extracted during dehydration because of lack of OsO4 stabilization. You might try acrolein...it is the only fixative other than OsO4 that will preserve and retain lipids. Make sure that you read all the precautions about it...its pretty nasty stuff.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 USA http://www.vet.uga.edu/vpp/wls/steffens.html
At 1:18 PM -0700 3/14/0, Soumitra Ghoshroy wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
After your sections have been dried onto the slide, cover them with a drop of stain for 30 sec - 1 min. and wash off with water. Dry.
If its too light, repeat.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
} We are trying to get a project started that involves imaging the } microstructure of mineral oils and polyalphaolefins as they solidify. The } solid temperature range for materials like these is approximately -20 to -70 } C. Does anyone have experience with these or similar materials. We are } hoping to image the particle structure (crystals) by SEM. Any feedback from } those with similar projects/experiences would be very welcome (on or off the } list). Even better would be references to any published reports of this } kind of material exam. } }
Dear Bradley, Doug Dorset has done extensive work with electron crystallography of waxes and lipids. He does electron diffraction with a TEM, but perhaps his results will be useful to you. Yours, Bill Tivol
for methacrylate sections we use 0.5 % Toluidinblue in 0.1 mol phosphate buffer (it should also work just with water) and for epoxide sections 0.1% Toluidinblue in 2% Na-bicarbonate. staining time depends on temperature, thickness, and material. After staining wash with water and dry the sections.
regards, Anne
Soumitra Ghoshroy schrieb:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Microscopists, } } Does anyone have a protocol for making Toluidine blue stain to be used for } staining thick sections of resin embedded biological samples ? } } Thanks a lot, } } Soumitra } } ***************************************************************** } Soumitra Ghoshroy Ph.D. } Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3600 } Fax: 505-646-5665 } e-mail: ghoshroy-at-nmsu.edu } http://confocal.nmsu.edu/eml
-- Dr. Anne Heller, AG Elektronenmikroskopie, Institut fŸr Botanik (210), UniversitŠt Hohenheim, Garbenstra§e 30 D-70593 Stuttgart
I forgot another cheaper alternative that I have used in the past. You can use a small, medium, or large binder clip to clamp samples together depending on their size. You can use Teflon tape on the surface to prevent sticking to the clamp. If you make a little Teflon jig to hold the sample it works quite well. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Leroux christine [mailto:leroux-at-univ-tln.fr] } Sent: Tuesday, March 14, 2000 9:45 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM cross section preparation } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.} html } } } -------------------------------------------------------------- } ---------. } } } hello, } I have to prepare cross sectionning samples, and I do not } know where to } buy the small } vise that I need to clamp the specimen when I have glued } them (I have } already the } Mbond 610 adhesive). } I would greetly appreciate any information about this thing } (I am not even } sure of the } name of it) } thank you in advance } christine } } ************************************** } Christine leroux } Lab. M.M.I } Universite de Toulon-Var, Bat.R } B.P.132 } F-83957 La Garde Cedex } tel: 00 33 (0) 4 94 14 25 07 } fax: 00 33 (0) 4 94 14 21 68 } } ************************************** } }
Ann, I'm also looking for vibration isolation tables for ultramicrotomy. I've been looking at Vibraplane (http://www.kineticsystems.com) for isolation tables and bench-top isolation platforms, which are also sold through stereo equipment companies (i.e. http://www.soundsofsilence.com http://www.audioodyssey.com). I need a height-adjustable isolation table - other vibration sensitive equipment in our lab is successfully being used on the isolation platforms on top of separate height-adjustable tables. I would appreciate hearing of any information you receive offline about the isolation tables.
Teresa Boes Analytical Services Lab Hewlett-Packard Corvallis, OR 541-715-7055 teresa_boes-at-hp.com
-----Original Message----- } From: Lehman, Ann [mailto:Ann.Lehman-at-trincoll.edu] Sent: Tuesday, March 14, 2000 6:37 AM To: 'MSA Listserver'
Dear Listers,
I'm looking for a source for vibration isolation tables for ultramicrotomy. The marble tables that we are using now are not adequate (except during Spring Break when no one is in the building...) I'm thinking of a system that uses compressed gas (N2?) to float the tables.
Please respond offline (vendors welcome).
thanks, all
Ann Hein Lehman EM Facility Mgr Trinity College Hartford CT 860-297-4289 860-297-2538 ann.lehman-at-trincoll.edu
} Date: Wed, 15 Mar 2000 11:15:59 -0500 } To:Soumitra Ghoshroy {ghoshroy-at-nmsu.edu} } From:sherwood-at-HELIX.MGH.HARVARD.EDU (Peggy Sherwood) } Subject:Re: Toluidine blue stain } } } Soumitra, } } We use a 0.5% Toluidine Blue in Borate Buffer (1 gram Toluidine Blue, 1 } gram sodium borate and 200ml water). Tousimis (and maybe other suppliers) } sells a Methylene Blue- Toludine Blue stain for epoxy stain (Cat. # 4166B) } which we have used: (consists of 0.2% methylene blue & 1.0% toluidine } blue certified stains in Na-Borate buffer). Either way--we always filter } our stains (using a syringe with filter unit attached) right onto } sections. Then we put slide ( super frost plus, so sections will adhere) } on hot plate (temp. -at- 60 decrees C). We rinse sections with water and } then check stain--time is arbitrary: depending upon tissue type & section } thickness as to how dark or light your stain result. } } When I worked in Ophthalmology research, I used a wonderful dichromatic } stain: methylene blue-azure II-basic fuchsin stain. It was similar to H } & E but for epoxy sections. (made great 2X2 slides). The reference is: } Humphrey, C.D. and Pittman, F.E. (1974) Stain Technology 42:9-14. I } actually have a copy that came as an LKB application note 303 which I } could fax to you. Feel free to contact me off line. Good luck! } } Peggy Sherwood } Wellman Labs of Photomedicine } Lab Associate-Photopathology Lab } Massachusetts General Hospital } 50 Blossom Street } Boston, MA 02114 } 617-726-6983 } 617-724-4839 (voice mail) } 617-726-3192 (fax) } sherwood-at-helix.mgh.harvard.edu } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all of you who responded to my question about the stain.
This is a great microscopy resource.
Best wishes.
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
Soumitra, We use the following recipe for toluidine blue:
These stock solutions can be made in large volumes and stored. .5 % toluidine blue in distilled water .25 % sodium borate in distilled water
Mix equal parts of the above. Stain dried resin sections on hot plate (low setting) until edge of stain begins to evaporate (5-10 secs) Rinse, blot excess water from bottom of slide and dry on hot plate.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
Patrick, Are you telling me that in your book, you have a reference to some work where some oil based fuels were cooled to a solid, and them imaged in that frozen state? If so, this is precisely the kind of information that I would like to see. If not, do you have a reference? I do not have a copy of your book, but I have seen it, some years ago. In any case, thanks for the reminder on that resource.
And even more specifically, if anyone has knowledge of any similar work done on mineral oil and like materials... it sure would save me some work on the development side...
Thanks for the feedback, Brad
} ---------- } From: Dr P. Echlin[SMTP:pe13-at-cus.cam.ac.uk] } Sent: Wednesday, March 15, 2000 6:43 AM } To: Huggins, Bradley J } Cc: Microscopy listserver } Subject: Re: Cryo-microscopy of solid mineral oils } } Dear Bradley: } } I know oil based fuels have been imaged at low temperature. Suggest you } read (better still, Buy) my book "Low Temperature Microscopy and } Analysis" Plenum Press New York 1992. I would modestly suggest it has a } lot of information about how to prepare, observe and analyse specimens } at low temperature. My own current interests revolve around low volyage, } low temperature x-ray microabalysis of bio-organic materials. Let me } know if I can be of any further help } } Patrick Echlin } University of Cambridge UK } } pe13-at-cus.cam.ac.uk } } } On Tue, 14 Mar 2000, Huggins, Bradley } J wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hello all, } } We are trying to get a project started that involves imaging the } } microstructure of mineral oils and polyalphaolefins as they solidify. } The } } solid temperature range for materials like these is approximately -20 to } -70 } } C. Does anyone have experience with these or similar materials. We } are } } hoping to image the particle structure (crystals) by SEM. Any feedback } from } } those with similar projects/experiences would be very welcome (on or off } the } } list). Even better would be references to any published reports of this } } kind of material exam. } } } } Thanks, } } Brad Huggins } } BPAmoco, } } Electron Microscopy Lab } } hugginbj-at-bp.com } } } } } } }
I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order to study the dislocations substructure. But I have some problems, for example, I can not focus the image due to the magnetism of the ferrite.
I am looking for suggestions to avoid this problem. What could I do?
Thank you in advance.
Dr. J. M. Manero Universidad PolitŽcnica de Catalunya. E.T.S.I. Industriales de Barcelona. Spain. e-mail: manero-at-cmem.upc.es
Ben, If you have other microscopes on campus you might consider forming a consortium of all the users and hiring your own on-site service engineer to take care of all the instruments. We've been doing this since 1981 and it's worked very well. One man's salary is considerably less than service contracts on many instruments. You also have the luxury of instant service. Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
1) Start (before final thinning) with the thinnest sample you can support ( {50 microns). Pure iron will be quite soft so you'll have to be very careful not to bend it - especially since you're looking at dislocation structure. You could support it on a copper key-grid to minimize damage. If you have to electropolish while it is on the copper key-grid, you'll need to lacquer the copper prior to electropolishing and then clean it off before putting it into the microscope. 2) If you still can't focus, try taking it out of the microscope and rotating it in the holder. Sometimes a different orientation really helps. 3 Plan on doing alignment every time you move or tilt. It is a given with magnetic samples. You'll need to adjust the beam tilt and the astigmatism continually. I always used the "dark-field" beam tilts as they are stronger than the bright field set. You'll be amazed at how good you get at aligning after a few months of magnetic work. Later work with non-magnetic samples will be a breeze. 4) One final warning. The objective lens can actually suck your sample out of the holder during insertion of the sample. If you have a really thin sample, it can break off chunks. These chunks/sample will ALWAYS stick to the lens and make every other user miserable. Your service guy will be forced at some point to clean off the lens of your sample (or sample bits) and you will be reviled. A simple solution is to always turn off the objective lens when you are inserting and removing your sample.
Good luck - magnetic work is possible and fun once you get the hang of it.
Robin Griffin
-----Original Message----- } From: Jose Maria Manero [mailto:manero-at-cmem.upc.es] Sent: Wednesday, March 15, 2000 11:59 AM To: Microscopy-at-sparc5.microscopy.com
Dear all,
I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order to study the dislocations substructure. But I have some problems, for example, I can not focus the image due to the magnetism of the ferrite.
I am looking for suggestions to avoid this problem. What could I do?
Thank you in advance.
Dr. J. M. Manero Universidad PolitŽcnica de Catalunya. E.T.S.I. Industriales de Barcelona. Spain. e-mail: manero-at-cmem.upc.es
We've been having stability problems with our mercury vapor lamps on our Reichert Polyvar microscope. We have been told by one source that the power supply is likely the problem and another source tells us that the bulbs are the problem. The source illumination, when imaged, can be seen to flicker at random intervals and causes the illumination to vary. Does anyone have experience with this microscope, and if so which brand of lamps do you use? This microscope uses a 220W/4 L1 lamp. Thanks.
David O'Neil National Research Council of Canada Institute for Marine Biosciences 1411 Oxford Street Halifax, NS B3H 3Z1 ph: (902)426-8258 fax: (902)426-9413 e-mail: david.o'neil-at-nrc.ca
I certainly agree that EM of magnetic samples is quite doable, but I'm not sure I'd agree that it is "fun".
Perhaps the best tip I can give is to reduce the amount of magnetic material within the polepiece. I have cut my magnetic samples down to less than the standard 3mm size and glued them securely onto a Cu slot grid. The magnetic effects are much less of a problem.
Cheers, Henk
At 12:13 PM 3/15/00 -0600, rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com wrote: } {snip}
} Good luck - magnetic work is possible and fun once you get the hang of it. } } Robin Griffin
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Soumitra, We use a Toluidine Blue and Methylene blue staining solution:
Dissolve 0.5gm Borax in 95 ml DH2O, add 0.5gm of methylene blue, mix then add 5 ml of a 2% Toluidine blue solution in H2O.
Filter the stain just before staining. Stain for 1 minute on a hot plate set slightly lower than where the stain starts to bubble. Rinse with DH2O and dry.
We like this stain because it is fast, very clean, and very pretty. We prefer this stain to other Toluidine Blue solutions.
George
George Lawton Chief Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75390-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
A lab on our floor is having trouble with their B & L spectrophotometer. does anyone know who, hopefully in New England, might be able to service the unit?
1. Reduce the sample size as much as possible, glue a tiny Fe piece on a Cu or other nonmagnetic grids, for example. 2. Turn off the objective lens as you load your sample. 3. Do beam alignment as many times as necessary. -cy Rodel Inc. 451 Bellevue Road Newark, Delaware 19713 USA
-----Original Message----- } From: Jose Maria Manero [mailto:manero-at-cmem.upc.es] Sent: Wednesday, March 15, 2000 12:59 PM To: Microscopy-at-sparc5.microscopy.com
Dear all,
I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order to study the dislocations substructure. But I have some problems, for example, I can not focus the image due to the magnetism of the ferrite.
I am looking for suggestions to avoid this problem. What could I do?
Thank you in advance.
Dr. J. M. Manero Universidad PolitŽcnica de Catalunya. E.T.S.I. Industriales de Barcelona. Spain. e-mail: manero-at-cmem.upc.es
A simple construction we used for years with a heavy microscope was a wooden frame standing on a tennis balls. The microscope was on the 2nd floor of a building by a busy road. Never a shake was recorded on film!
Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
As an independent service provider of several SEM manufacturers for over 20 years, I think I can also comment & agree with Ken on several fronts concerning Amray. However, I don't want to get into a "bashing" of any given manufacturer, at least online. I will say that every Company (SEM or otherwise) has their own way of doing things. Each Company is different, depending upon the people running the operation.
Many manufacturer's think they have a "lock" on the customer and on the service because of their FE guns or perhaps on another part. This reminds me of ETEC just before they got out of the SEM service business. ETEC had an epoxy encapsulated high voltage power supply that only ETEC manufactured. I inquired with ETEC about the cost of one of these units. ETEC responded with a price tag of $50,000.00. Yes $50,000.00 in 1982! ETEC service also let it be known that they were the only source for this part. Furthermore, customers considering a third party for service were informed (and threatened) by ETEC about this situation. Customers were stuck paying ETEC's exorbitant maintenance costs.
The ETEC customers were hesitant to go to a third party service until the high voltage issue had been resolved. I spent many days (and nights) re-engineering another high voltage power supply until I had it perfectly duplicated. When customers would ask about the high voltage issue, I would show them my version & invite them to use it in their system. Within two years I had over 50% of ETEC's service business at 60% of ETEC contract costs.
Today several manufacturers still think they have a "lock" on service. I have been informed by one manufacturer that this is their way of "putting you (third party maintenance) out of business". If I had the time and the inclination, I would spend it duplicating the FE gun some of these manufacturer's think they have a "lock" on. I have considered such a move because no one has a "lock" on technology.
I have had several requests but have not responded because the time & effort necessary is not viable for only three customer requests.. Fortunately for the manufacturer, I am busy and, quite frankly, not as "hungry" as I used to be. Maybe it comes with age. If I ever get "unhungry" or not as busy, I would take these manufacturer's up on their smugness and beat them at their own game by duplicating their FE guns. The ultimate winner is the customer.
The only manufacturer that has a great gun that can be serviced by the customer is Hitachi. Hitachi has the patent on an internal heater for their gun which makes exchanging FE guns obsolete. Hitachi FE guns can be re-conditioned by the customer in-house at a cost of $750.00 instead of $7,000.00 for Amray or $17,000.00 for JEOL. Hitachi is doing well and doesn't need to worry about "locking" third party maintenance or anyone else out. They have too many sales to worry about & don't have the time to be petty.
Great thing about the free enterprise system is that real talent always has customers "beating a path" to their door. Marginal talent needs to cajole customers to their way of thinking. Great talent has customers calling on them.
Regards,
Earl Weltmer Scanservice Corporation Third Party Maintenance
} Gary, } Your earlier comments on how good AMRAY is really puzzle me, but be that } as it may, if any of their Federal Government customers are on the ball, } Amray will be told, as was Perkin Elmer ETEC, that they must support the } equipment for about 7 years or face lawsuits. There is an implied } responsibility when one sells expensive equipment. } } Perhaps supplying parts and detailed instructions will cover them } legally, in which case we third party folks may be able to help out. If } anyone has any more detailed info on Amray service, I would love to know } because I have serviced them off and on over the years and would be } willing to help out here in the Northeast/Mid-Atlantic area. } } Ken Converse } owner } Quality Images } third party SEM service } 16 Creek Rd. } Delta, PA 17314 } } 717-456-5491
I take it that you don't think that Amrays are very good. What aspect
of them do you find lacking? I would tend to agree that the early models are no comparison to their generations of the last 6-10 years. The FE
systems are particularly good. My 1910FE does a great job being an FE and having the flat lens. The 1800 series with conical lenses are not as good, but are required when doing IC wafers.
You have a great point about US Gov users. I know several of them. When does the 7 years start? From what event or timepoint? Amray has not stopped providing service. But the word is that they most likely will in about 2 years....based on their takeover by KLA-Tencor.
One fellow pointed out the proprietary nature of the FE gun. Without access to this, third party providers cannot work on the systems....at
We are trying to examine oil inclusions trapped in quartz under UV light. The oil fluoresces and the quartz does not. Sounds easy. Unfortunately the epoxy that the sample is embedded in also fluoresces quite strongly making identification difficult. So I was wondering if there are any non-fluorescent epoxies available on the market, or alternatively if there are any pigments/dyes that can be added to the epoxy to reduce its fluorescence. If not then perhaps an alternative to embedding in epoxy might be the best option....
Any suggestions warmly welcomed. :-)
Michael Joss Quantitative Microscopist CSIRO Division of Petroleum Resources Fluid History Analysis Group Phone: 9490 8148 Fax: 9490 8467
} We are trying to examine oil inclusions trapped in quartz under UV light. } The oil fluoresces and the quartz does not. Sounds easy. Unfortunately the } epoxy that the sample is embedded in also fluoresces quite strongly making } identification difficult. So I was wondering if there are any } non-fluorescent epoxies available on the market, or alternatively if there } are any pigments/dyes that can be added to the epoxy to reduce its } fluorescence. If not then perhaps an alternative to embedding in epoxy } might be the best option.... } } Any suggestions warmly welcomed. } :-) } } Michael Joss } Quantitative Microscopist } CSIRO Division of Petroleum Resources } Fluid History Analysis Group } Phone: 9490 8148 } Fax: 9490 8467 }
} We are trying to examine oil inclusions trapped in quartz under UV light. } The oil fluoresces and the quartz does not. Sounds easy. Unfortunately the } epoxy that the sample is embedded in also fluoresces quite strongly making } identification difficult. So I was wondering if there are any } non-fluorescent epoxies available on the market, or alternatively if there } are any pigments/dyes that can be added to the epoxy to reduce its } fluorescence. If not then perhaps an alternative to embedding in epoxy } might be the best option.... } } Any suggestions warmly welcomed. } :-) } } Michael Joss } Quantitative Microscopist } CSIRO Division of Petroleum Resources } Fluid History Analysis Group } Phone: 9490 8148 } Fax: 9490 8467 }
The tennis balls will isolate vibration nearly perfect. We used this approach a lot in Russia simply because we had no other choice. But tennis balls have tendency to deflate under pressure of the weight of the machine. The deflation will become noticeable in a few moths. Then the entire frame has to be lifted in order to replace tennis balls. Much better approach is to use pneumatic antivibration mounts by Barry Controls. They come in different sizes for loads from 100 lb. to several tons, and available from a number of suppliers including McMaster-Carr. Unless, of course, Australian tennis balls are superior to Russian ones... Cheers.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax -----Original Message----- } From: Diana van Driel {dianavd-at-eye.usyd.edu.au} To: MicroscopyList {microscopy-at-sparc5.microscopy.com}
I have read the previous suggestions and would agree with them - the message is get as much of that magnet out of your microscope as possible. However other tips that you may find useful:
If you have a side entry stage you will find it very difficult to set the eucentric height with a magnetic specimen. Either note the objective lens current at eucentric focus and set that or focus a non magnetic specimen at eucentric and then insert your secimen. Having done that focus the specimen using the eucentric height control.
Try to have as small a hole as possible in your specimen. This will reduce the effect of having a magnet one side of the beam and not the other. Of course the best specimen would not have a hole but just uniform thin area.
You will have to realign the beam every time you move the specimen.
When out of focus you may see the Fresnel image of Domain walls, when they change contrast from black to white (or vice versa) then you are in focus.
Good luck, Ron
On Wed, 15 Mar 2000 18:59:22 +0100 Jose Maria Manero {manero-at-cmem.upc.es} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order } to study the dislocations substructure. But I have some problems, for } example, I can not focus the image due to the magnetism of the ferrite. } } I am looking for suggestions to avoid this problem. What could I do? } } Thank you in advance. } } Dr. J. M. Manero } Universidad Politécnica de Catalunya. } E.T.S.I. Industriales de Barcelona. Spain. } e-mail: manero-at-cmem.upc.es } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I've read most of the replies but found no mention of differentiation. This is one of the most attractive features of the toluidine stain as it can give a two colour, pink and blue rendition. This shows cell features similar to a double staining with Haematoxylin & Eosin.
After staining on a hotplate add a drop of acid alcohol and then rinse the slide with distilled water. The acid alcohol can destain partially or even completely, but a brief application brings forth the two colours.
Make acid alcohol by adding to 25ml of 50% Ethanol a couple of drops of 1N HCl. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, March 15, 2000 6:18 AM, Soumitra Ghoshroy [SMTP:ghoshroy-at-nmsu.edu] wrote: } } Dear Microscopists, } } Does anyone have a protocol for making Toluidine blue stain to be used for } staining thick sections of resin embedded biological samples ? } } Thanks a lot, } } Soumitra } } ***************************************************************** } Soumitra Ghoshroy Ph.D. } Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3600 } Fax: 505-646-5665 } e-mail: ghoshroy-at-nmsu.edu } http://confocal.nmsu.edu/eml
We are examining Staphylococcus epidermidis on the surface of acrylic bone cement. We have problems with the marking of bacteria. Artefacts of particles from the Polymethylmethacrylate (bone cement) look nearly the same than the bacteria. We fix with glut, dehydrate, do critical point drying and coat with gold afterwards. Does anyone have experience with marking bacteria in SEM to surely identify them? Thank you in advance, Christian Heisel, M.D. University of Heidelberg Dep.of Orthopedics Schlierbacher Landstra§e 200 A 69118 Heidelberg Germany Phone:0049-6221-965- Fax:0049-6221-966121 e-mail:christian.heisel-at-ok.uni-heidelberg.de
} I have recently performed an immunoEM labeling procedure with } } myelinated nerve embedded in Lowicryl, K4M resin. Unfortunately the } } myelin is severely altered (it appears as large clealr blebs) because I } } am not able to osmicate this tissue due to the UV polymerization of } } this resin. Does anyone have any suggestions as to preservation of } } lipid laden myelin without the use of osmium? I used methanol in the } } dehydration procedure which has probably caused this artefact....
Karen,
Try to switch to cryosections. This reference could help in solving your problem. Liou W, Geuze HJ, Slot JW Histochem Cell Biol 1996 Jul;106(1):41-58 -- Alexander Mironov Jr. Division of Tumor Biology, H4 The Netherlands Cancer Institute Plesmanlaan 121 1066 CX Amsterdam
One concern raised by a granting agency about our proposal for new equipment for a multi-user SEM lab was our lack of a management plan (primarily for time allocation among users and for conflict resolution). We have been operating under a first come-first served policy, and conflicts have not been a problem. I would greatly appreciate insights from managers of multi-user facilities of any sort as to your approaches to allocation of access to instrumentation, and methods of resolving conflicts among researchers.
fernando =================================================== Fernando D. Balducci Laboratorio de Microscopia Electr—nica Facultad de Ingenier’a - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
The technique we now use on Fe and ferritic steels allows getting an excellent behaviour in the TEM:
-A 1 mm hole is punched in the centre of a 3 mm TEM disk made out of stainless steel (e.g. SS 316) about 300 micron thick, -A 1 mm disk is punched from our ferritic specimen (about 300 micron thick), -This 1mm disk is inserted - slightly forced - in the hole of the 3 mm disk, -A drop of glue (epoxy) is applied (to make sure there is no leak during the electropolish that follows). The 3 mm disk holding the 1 mm disk can then be used as a normal TEM disk: -Mechanical polish down to about 50-80 microns, -Electropolishing with a 10 % vol. perchloric/methanol solution, 18V, 0C.
The final specimen is strong and the remaining magnetism is minimal in the TEM: we can now move and tilt around without too much realignment. Note that a puncher of high quality is essential. The one from 'Eckert' doesn't introduce visible deformation in the centre of the specimen; we tested that in annealed pure Fe.
Dear David: We have 15 year old Reichert Polyvar which, so far, has given us little trouble. We use Osram HLX Xenophot bulbs, 12V, 100 W. Oddly enough, we also have a Reichert MeF3 metallograph, about 10 years old.The light on this instrument flickers erraticly. We replaced the lamp control board with no success, switched lamps, etc. To no avail.Our service man thinks the trouble lies in the transformer, which is no longer available. We are buying a new separate light source but have not received it yet. We are not happy with Reichert/Leica concerning assistancein this problem. They have been less than helpful. On the other hand the service man(Dave Olney) from W. Nuhsbaum, McHenry IL, has been very helpful. I would be interesrted in whatever solution to this problem that you may come up with. How old is your instument?
I have financial interest in either Reichert/Leica or W. Nuhsbaum. } ---------- } From: O'Neil, David } Sent: March 2000 1:56 PM } To: Microscopy Listserver } Subject: LM: Fluorescence lamp instability } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We've been having stability problems with our mercury vapor lamps on } our Reichert Polyvar microscope. We have been told by one source that the } power supply is likely the problem and another source tells us that the } bulbs are the problem. The source illumination, when imaged, can be seen } to } flicker at random intervals and causes the illumination to vary. Does } anyone have experience with this microscope, and if so which brand of } lamps } do you use? This microscope uses a 220W/4 L1 lamp. Thanks. } } } David O'Neil } National Research Council of Canada } Institute for Marine Biosciences } 1411 Oxford Street } Halifax, NS B3H 3Z1 } ph: (902)426-8258 } fax: (902)426-9413 } e-mail: david.o'neil-at-nrc.ca }
} } Hi, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University } Leslie -
My method, FWIW, is simplicity itself. I have a scheduling calendar posted on the outer door of the lab, with a pen provided. Clients simply sign themselves up in whatever day/time slot they wish, provided it's still open. They can even sign up for days/times in future months, if they so desire. Haven't caught anyone erasing a previous booking yet....but then that's why I use a pen..... This method certainly favours the folks who have their act together far enough in advance so that they can schedule the instruments at their most convenient times. Clients who show up at the last minute looking for openings at their prefered time may well be out of luck. That's what we call a learning experience. I'm sure there is software available that can be mounted on your local network which would do exactly the same thing, but, you know, I think it makes more of a commitment for the client to actually make a trip down to the lab to physically sign up, and then they're less likely to forget about the scheduled session later. It's worked pretty well for me for about 5 years now, and haven't seen a good screaming match yet.
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
} Hi, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University
***************************** Dear Leslie, I have run a multi-user EM facility for 12 years, a now also manage an optical microscopy facility. Both are core facilities for the medical school. For both facilities, users must be trained in the operation of the instruments before they are given free access (we have coded locks). During training, people set up appointments with me. Once "cleared fro soloing" they are given an idivualized code and are then free to reserve the instruments by means of a weekly sign-up sheet. For now, the sheet for the following week is posted on Thursday or Friday. Our in-house computing people keep promising us an on-line sign up. I can't wait since the 2 facil/lab group may reserve on a given day. This has worked well for us so far. It also seems to satisfy the granting instituions. For more details, you can check out the school's web site (http://www.med.cornell.edu/research/cores/index.html)select either electron microscopy or optical microscopy to get the operating procedures.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
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Hello everyone, We are having to justify using cervical dislocation without anesthesia on a mouse prior to removing the liver for fixation for our EM class. Does anyone have a reference on hand that we can cite to show that direct cervical dislocation is better (or not) for best preservation of tissue under the circumstances? We won't be doing perfusion, which would obviously provide better fix. Please respond directly to me at: jpshield-at-arches.uga.edu
Background: The class is held once a year, and one mouse is used (usually an extra, "too old" mouse from the monoclonal facility). The animal-use officer wants us to hike down to another building with the class and use CO2 anesthesia, prior to euthanasia, so the mouse suffers less (?). Then hike back up the hill with the tissue in fixative. We've never had a problem with our animal-use request until this year and the guy is busting our chops.
Thanks in advance, john ******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu
Hi Leslie We are a multi-user facility, used by the Faculty of Medicine and the Faculty of Science. Last fiscal year we had 653 individual users, the majority frequent users. We have TEM, SEM, confocal, fluorescent/brightfield, darkrooms and digital imaging equipment.
We use the first-come, first-served approach for many years and so far there hasn't been a problem with sign-up conflicts with the instruments. Most people around here are reasonable, thinking people.
We do not give open access after hours (9-5). But if a user is experienced with the instrument, and I know he/she is not going to mess it up for the next user, and he/she will take a responsibility lecture from me, there is a floating key. I used to be more lax with letting users in after hours, but after a week of just trouble shooting, I got tough. Since then, it has worked well. I remember when I did my PhD, how much more work I got done when there were no interuptions or distractions. So I have a sympathy for allowing responsible researchers in when I am not there, especially as this is a busy facility during the week day.
If it ever came to a conflict of access to the instruments, I am in charge. I would think poorly of researchers who didn't play by the rules. The only time, any problem has occured has been when someone has a crutial deadline. Then when the situation has been put to the person who has it booked, they can usually see themselves in that situation and have always been accommodating. Maybe it is the Canadian way, but I should think it is far more universal!
Good luck with your grant. Let me know if you need more information. Elaine
} } Hi, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
At 08:22 AM 3/16/00 -0500, Leslie Eibest wrote: } Hi, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. }
Leslie,
I have been managing a multi-user facility since 1979. First it was sign up sheets. Now it is a web-based calendar. Have a look at out web site. This works fine for us. We use the web calendar because it is easier for people across campus to sign up this way than to trek over to the facility. Also, I don't have to answer the phone to make their appointments.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Dear Jose, One other method that might work for you is the old "window-polishing" technique. This was the preferred method before the jet-polishing machines were in wide use. A sheet of the metal about one inch by two inches and as thin as possible is masked by stopping lacquer on the edges and electropolished in a bath of electrolyte. When it has polished down to the "lacey" point, a small protrusion of the metal is sliced off with a razor blade and sandwiched in a folding grid or between two grids. The edges, except for the sliced part, are thin enough to examine. It is a bit of an art and takes some practice to get right, but the resulting piece is small and securely held. -----Original Message----- } From: Jose Maria Manero [mailto:manero-at-cmem.upc.es] } To: Microscopy-at-sparc5.microscopy.com } Subject: [TEM] Problem with magnetic samples
} Dear all,
} I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order } to study the dislocations substructure. But I have some problems, for } example, I can not focus the image due to the magnetism of the ferrite.
} I am looking for suggestions to avoid this problem. What could I do?
} Thank you in advance.
} Dr. J. M. Manero } Universidad PolitŽcnica de Catalunya. } E.T.S.I. Industriales de Barcelona. Spain. } e-mail: manero-at-cmem.upc.es
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Leslie, You can post a calendar on your internal web page for users to log onto, or if you do not have a web page post a calendar at each microscope. Let your users sign up as far in advance as they can, this eliminates most of the conflicts and the habitual "last minute guy" learns to get him/herself more organized. It also gives you a record of use, I also log in all down time due to maintenance service etc. The best part is this system is really easy once everyone buys into it. Good luck! } ---------- } From: Leslie Eibest } Sent: Thursday, March 16, 2000 5:22 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Multi-user facility managers } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University } }
We have modified the basic sign up sheet due to very heavy demand on our microscope. Except for special cases (eg., time lapse studies, emergencies) we allow users to sign up for no more than two hour blocks of time and they cannot sign up for additional time until that block is completed. In addition, if they are more than 15 minutes late for their time the entire block of time is forfeited. (This has rarely happened but emphasizes the need for responsible behaviour.) This system was adapted from the one that Mei Li Wong uses for electron microscopes. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
Hi Leslie, Yikes! this reeks of infectious administratium but since you need to deal with the request... I have quite a few systems that are considered multiple user facilities. Over the last 9 years I can say I have never had a conflict to resolve or even heard about one... meaning the users work well together. Users make reservations on a calendar. That is their time. If the machine is down, that time is lost. There is not a ripple effect or compression of the schedule to squeeze them in Frankly if any user asked another for time due to there state of desperation I have no doubt that they would work together. I have a rule for my EMs that only one 4 hour period may be reserved at a time. I think this keeps the level of respect for someone else's time high. My good fortune aside, you could probably get an outline for conflict resolution from your HR dept. & append it to your proposal.
Bruce Brinson, Rice U.
Leslie Eibest 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, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University
We are working on a research project and we need an electron gun with 5-10 kV (wehneldt assembly) and a lens system (for electrons, not photons) that is typically found in SEM systems. Does anyone have a decommissioned or un-needed SEM or TEM equipment from which we could get these components at low or no cost (we are on a very tight budget)? We will even move out a complete system if it will help someone who needs the space.
I recently responded to a thread about Amray FE maintenance contracts. My response contained two errors: one numerical, one in perception. I wish to correct them at this time.
First the numerical correction: I stated that the JEOL FE gun exchange cost was $17,000.00: it is actually $7,000.00. The $17,000.00 figure was obtained from two JEOL customers which I presumed to be true as I am not in the habit to ask other competitors their pricing schedules.
Second the perception error: I stated that " Hitachi is doing well and doesn't need to worry about "locking" third party maintenance or anyone else out." I did not mean to imply that JEOL or any other manufacturer's business was poor -- just that Hitachi was doing well. I am sure that JEOL's business is excellent as they have an excellent product. Moreover, their service organization is one of the best. As their competitor we have a difficult time competing with JEOL.
Keep in mind that I do not have a "cozy" relationship with Hitachi. I just respect the technology & the professional co-operation I get from them & other manufacturer's like them. I can buy schematics, parts lists, & even service manuals from Hitachi that other manufacturers don't offer because they consider it proprietary. It makes my job easier.
I was using Hitachi as one example because in a pure free enterprise system the customer is the ultimate winner.
I apologize for the errors. I wish to thank JEOL for pointing out the error and their gracious attitude in dealing with this error.
We have a calendar that people fill in. As a courtesy to others, we ask that they not block more than 4 hours at a time. So far this has worked well. On the few occasions when another user was in a bind, scheduled users have been accommodating because they know that they could be in the same position.
Ron
-----Original Message----- } From: Leslie Eibest [SMTP:leibest-at-duke.edu] [} ] [} ] I would greatly appreciate insights from managers of multi-user facilities of any sort as to your approaches to allocation of access to instrumentation, and methods of resolving conflicts among researchers.
Place: IBM Microelectronics, E. Fishkill, NY, West Complex, Bldg 600.
The IBM Corporation has graciously agreed to be our host for this meeting at their facility in East Fishkill, NY (West Complex, Bldg 600). Attendees will be able to purchase lunch in the adjacent IBM cafeteria. Due to size and security issues IT IS ESSENTIAL THAT MEMBERS PRE-REGISTER so that an attendee list can be delivered to the IBM security folks to prepare guest badges and escorts. DUE TO THE TIGHT SECURITY CLEARANCE REQUIREMENTS, WALK-INS CANNOT BE ACCOMMODATED.
THE REGISTRATION DEADLINE IS MARCH 24th AND CAN BE ACCOMPLISHED ELECTRONICALLY. Please respond via email (or fax) to Evan Slow directly. A simple email note or a completed fax of the registration form is all that s required to register. You can then bring the required fee with you to the meeting. The meeting fee, which does not include lunch, is $15.00. ON-SITE REGISTRANTS WILL BE CHARGED $25.00 BUT, AGAIN, UNLESS YOU HAVE BEEN PREVIOUSLY CLEARED THROUGH IBM SECURITY, WE CANNOT ACCOMMODATE YOU.
9:45 - 10:45 : CHARACTERIZATION OF MICROSTRUCTURES IN MICROELECTRONIC INTERCONNECTS, Lynne Gignac, IBM T.J. Watson Research Center.
10:45 - 11:45 : IMAGING, DIFFRACTION AND SPECTROSCOPY WITH FIELD EMISSION GUN SEM (FEG SEM) AT LOW VOLTAGE, Vinayak Dravid, Northwestern U.
11:45 - 12:45 : DEEP-UV CONFOCAL MICROSCOPY OF SUB-MICRON FEATURES, Mike Torres, Metron Technology.
12:45 - 1:30 : LUNCH -- available for purchase in the IBM cafeteria.
1:30 - 2:30 : THE USE OF MONTE CARLO CALCULATIONS FOR QUANTITATIVE X-RAY MICROANALYSIS, Eric Lifshin, GE Corporate Research & Development.
2:30 3:30 : STUDIES OF SAMPLES HAVING SHALLOW SURFACE TOPOGRAPHY BY THE LOW-LOSS ELECTRON (LLE) METHOD IN THE SCANNING ELECTRON MICROSCOPE (SEM), Oliver Wells, IBM T. J. Watson Research Center.
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id RAA11115 for dist-Microscopy; Thu, 16 Mar 2000 17:58:04 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id RAA11112 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 16 Mar 2000 17:57:33 -0600 (CST) Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id RAA11099 for {microscopy-at-sparc5.microscopy.com} ; Thu, 16 Mar 2000 17:57:19 -0600 (CST) Received: from mel ([129.94.150.25]) by sam.comms.unsw.EDU.AU (8.8.8/8.8.8 Kenso-Central-NO-SPAM) with SMTP id KAA22727 for {microscopy-at-sparc5.microscopy.com} ; Fri, 17 Mar 2000 10:52:09 +1100 (EST) Message-Id: {3.0.1.32.20000317105504.00986dc0-at-pop3.unsw.edu.au} X-Sender: s7001031-at-pop3.unsw.edu.au X-Mailer: Windows Eudora Pro Version 3.0.1 (32)
At 08:22 16/03/00 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have run a first come-first served policy for 30 years. Its the way to go.
It now is self-administering through our equipment booking software.
VERY seldom we need to reason with excessive users. We can almost always sort problems out with time trading.
Use Internet explorer - Go to http://srv.emunit.unsw.edu.au/ - instrument booking and use login {guest} password {guest} to get its flavour.
} } Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400
It helps to have a good management plan in place whether the facility is extremely busy or not. It avoids most conflicts and gives manager/coordinator a set of rules to use that can be applied to any situation that might pop up. You might not that busy currently on conflicts are occuring but later on the situation can quickly change. Granting agencies want to know if the equipment they are funding are accessible and not misused. We are a fairly busy facility and have been using calendars, lined up together, on a wall for each piece of equipment including work stations. We have one for the current week and below that one for the next week users can sign up. Depending on the instrument from two to three hour slots at a time during peak hours. After 6PM and before 8:30AM longer durations are possible. One can only cancel a day in advance on an instrument. No shows are charged. We cross out time slots for instrument maintenance as needed and as much inadvance as possible. Potential users first fill out an investigator profile form before they can cycle into the facility. This helps us target the instrument/technique they need. Based on several factors we prioritize users. All users must take a tutorial and demonstrate competency before they can solo. We do tutorials only one day a week unless there is an important reason that they cannot do it on that day. In a few weeks, I hope, we are going to our web based scheduler in which users have the a couple months to schedule in advance. It will automatically restrict a users' time usage to only peak hours or to peak and after hours depending on their competency, and other criteria. It will prevent users from making cancellations less then the day before. Down times, etc can be easily entered on the calender. The web based schedular will make it allot more convenient for users and core staff. And best of all, please god let it work smoothly, data from the scheduler will flow into our billing data base for semi automatic billing. So far, with the written calendar syst conflicts, however, usage is so high on some instruments not every user can get the slot they need and so are unhappy campers. In this case, we encourage users, if it is possible, to time their experiments based on when they can get on the instrument. I hope this helps.
Hank Adams Lab Manager Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, TX
At 08:22 16/03/00 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have run a first come-first served policy for 30 years. Its the way to go.
It now is self-administering through our equipment booking software.
VERY seldom we need to reason with excessive users. We can almost always sort problems out with time trading.
Use Internet explorer - Go to http://srv.emunit.unsw.edu.au/ - instrument booking and use login {guest} password {guest} to get its flavour.
} } Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400
Christian, I have a few approaches you might consider: a. Where possible, I usually obtain a culture of the bacteria thought to adhere to a substrate and prepare it exactly the same way:fix, dehydrate, cpd and sputter-coat. b. Prepare two samples but after full prep, apply a cellulose acetate film (or duco cement) to one surface moistened with a drop of acetone and try to strip the bacteria from the bone cement -- I don't know about damage to the methacrylate. This technique produces a replica of the surface. I have used it to remove bacteria colonizing a planchet of hornblende. We were able to locate the pits made by the bacteria and image the bacteria removed, i.e., stripped from the mineral. c. If the above approach fails to produce the results you need, try making a replica with double sided C tabs, the type sold by EMS for X-ray microanalysis. d. If staph epidermidis has a glycocalyx you might try adding ruthenium red to the GA in the primary fix and prep as usual but instead of coating with Au or Au/Pd, evap C and use a backscatter detector to obtain a contrast BE image where the brighter areas should be the Ru stained bacteria and/or collect a spectrum to check for the presence of Ru. If there is a characteristic peak, you could map the Ru and localize the bacteria with an x-ray dot map. I would be interested in knowing how you resolved this problem. Rosemary Walsh The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu {"http://www.lsc.psu.edu/stf/em/home.html" eudora="autourl"} http://www.lsc.psu.edu/stf/em/home.html
I sell the DruAedge blades. Same quality as the accu-edge, but at a better price. Please contact me for pricing and free samples. I carry both the High Profile and Low Profile Teflon coated blades.
Ford M. Royer Analytical Instruments, LLC 9921 13th Ave. N. Minneapolis, MN 55441 phone: (800) 565-1895, ext. 17 fax: (612) 929-1895 froyer-at-bitstream.net
Diana Papoulias 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 } -----------------------------------------------------------------------. } } Could someone please tell me where I can purchase accu-edge microtome } blades? } } Thank you. } } Diana Papoulias } USGS } 4200 New Haven Rd } Columbia, MO 65201 } } T:573 876 1902 } F:573 876 1876 } E:Diana_Papoulias-at-usgs.gov
I am trying to localise a beta-1,3-glucanase gene after Russian wheat aphid infestation. However I am getting quite heavy labeling in the chloroplasts and from the literature this haven't been previously documented. My control serum shows a small amount of labeling in cell walls but these seems to be random. The controls (before infestation) shows very little labeling. Work done with the same antibody on rust infested wheat plants have shown no, to very little labeling in the chloroplasts. My question is how can I make sure this is not artifacts and that the glucanase are for sure in the chloroplasts. Martin Wilding Department Botany & Genetics University of the Orange Free State P.O. Box 339 Bloemfontein 9300 South Africa
Tel +2751 4012818 Fax +2751 4488772 Email paam-at-rs.uovs.ac.za
E.A. Fischione Instruments, Inc. is seeking an individual for the position of a Sales Engineer. The successful candidate will be responsible for one or more states from both inside the office in Export, PA. and in the field. The candidate's responsibilities will include defining contacts, maintaining information in the sales database, qualifying leads and determining need, providing information and quotations for standard products, handling service/spare parts requests and coordinating the same with the Service Department. Must develop a thorough technical knowledge of standard products. Extensive travel required.
The candidate should have an Associates Degree in either Electron Microscopy technology, Material Science, Engineering, or Physics as a minimum; a B.S. or M.S. would be preferred.
Salary is commensurate with experience.
E.A. Fischione Instruments, Inc. is an equal opportunity employer. Please send your resume and salary requirements to:
Human Resources Director, MSA LS E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone (724) 325-5444 FAX (724) 325-5443 E-mail: info-at-fischione.com www.fischione.com
This does not directly address the problem with your current lamp but you might want to consider looking at this new lamp assembly. We recently evaluated the system and there were no visible stability problems as well as several advantages over the standard mercury lamp housing.
The company is called EFOS. This unit replaces the standard mercury source for fluorescence and possibly transmitted light techniques. It uses an external 50W miniature arc lamp incorporated into an elliptical reflector which is connected to the microscope by a liquid light guide. Our unit was mounted on a Zeiss Axioscope.
You can visit the EFOS web site at http://www.efos.com/products/x-cite.htm
Thanks, Louie
At 2:56 PM -0400 3/15/00, O'Neil, David wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
} I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. }
Dear Leslie, Our facility is funded by NIH as a Biotechnological Resource, so we deal with both in-house and outside researchers. Like the other responders, we have not had any conflicts between users since we started. Because the outside users must spend con- siderable time arranging to travel here to use the instruments, we have had a policy which maximizes the utility of their stay here. We have a staff person whose function is outside user liason; she ascertains what the user wants, suggests ways to prepare the specimen to achieve that, examines trial grids, schedules the user, and stays with the user during the run. For the inexperienced user, she does almost everything except select the area of the grid to be photographed; for users who are familiar with our facility, she changes specimens, develops film, and is available for any other user needs. For the occasional user who can operate the HVEM on his own, she still changes the film and is available. The rest of the staff is responsible for starting the HVEM up in the mode the user wants, taking care of any problems which arise, and setting up for non-routine use--EDS, diffraction, etc. In-house users can be bumped to accomodate outside users and can only sign up a few weeks in advance. Outside users can sign up for as far in advance as they wish, and if any problems arise with the scope, our liason person calls to let them know so they can change plans if necessary. In periods of very high use, the staff arranges to keep the scope up for as long as 16 hours a day and will also arrange to be here after hours and/or on week- ends to accomodate users' schedules. Yours, Bill Tivol
How fast must the sequential image capture be to perform image ratios for dtermining something like calcium ion levels? If you have a fast interline or frame transfer camera, does a single filter wheel like a Ludl change filters fast enough?
SmithKline Beecham, a world class leader in Research and Development, continues to pioneer innovative pharmaceutical and healthcare products and services. We have the following opportunity available at our state-of-the-art suburban Philadelphia facility. Working in our Safety Assessment department you will provide technical support and scientific input into the design and execution of studies to elucidate the cellular mechanisms of drug-induced toxicities. You will prepare biological specimens for transmission and scanning electron microscopy, confocal microscopy, X-ray microanalysis and immunohistochemistry. You will also perform qualitative and quantitative data analysis using computer assisted image analysis systems. We require a BS/MS in a biological science, and 3-5 years of microscopy experience in cell biology, physiology, toxicology. SmithKline Beecham is dedicated to an innovative workplace and supports you with career long opportunities and learning. We offer a competitive benefits and compensation package. For confidential consideration, please forward your scannable resume to: SmithKline Beecham Attention: Human Resources AD CODE: 2K0325W c/o National Resume Processing P.O. Box 1070 Burlington, MA 01803 USA
Indicating ad code is essential. Principals only, no agencies, please. For a full listing of current opportunities, or to submit a resume online, visit our website at www.sb.com/careers.
Leslie, Just to add my two cents: I supervise a private Photopathology Lab for the Wellman Labs of Photomedicine at MGH. We have an Axiophot (Zeiss) photomicroscope that has an image analysis system attached. We have used a monthly sign-up calendar successfully for 10 years. The room is secured-- anyone wishing to use the system, must be checked out first by one of us. Once we are con- fident they know what they are doing, they then sign up ahead for a day and time. We allow after- hour use; many researchers work weekends and nights. I sign out a lab key to the individual and the room key is in a desk drawer. People are instructed to make sure the microscope is off, doors are locked, lights are off and room is secure. Other than the occasional light left on, the system has worked. The other advantage to having people sign-up is that if there is a problem with the micro- scope and/or room, we can go back to the last person who used it and advise them of the problem. We also use a sign-up system for a multi-headed teaching scope.
Whatever system you end up using--sign-up sheets or on-line, you have a record of use, which granting agencies are most interested in: making sure you get the most for your buck!
Good luck!
Peggy Sherwood Wellman Labs of Photomedicine-MGH 50 Blossom Street Boston, MA 02114 617-726-6983 (Photopathology Lab) 617-724-4839 (voice mail) 617-726-3192 (fax) e-mail: sherwood-at-helix.mgh.harvard.edu
Thanks so much to all of you who took the time to respond to my question. My management techniques are basically the same as most of yours, but my presentation was woefully inadequate. Thanks to you, I'm well-armed now!
We have had several instruments and many instrument users during each the last twenty five years. Originally, we used calendar sign-up and log book entries to compile data for accounting and annual reports. This became too time-consuming so we developed an in-house automated (computer-based) sign-up (registration) system in 1990. The system serves as an instrument reservation unit and an automated accounting system. We recently developed (continue to develop) a web-based system which allows researchers to reserve time from their offices or anywhere they can access the internet. You may wish to view it at the following URL: http://signu.la.asu.edu:8001/. (Login: guest Password: testit) You will not be able to view the accounting data. The sign-up program allows only qualified users to operate an instrument. Also, rules for successive time reservation are integrated into the program. Optional holders and spectrometers can be reserved at the time the instrument is signed for.
We presently have 90 research users. In addition, there are approximately sixty student users (grad and undergraduates) who are taking semester classes in microscopy or they have one or two sessions on individual instruments to better understand the role of electron microscopy for solving problems in Geology, solid state chemistry etc.
Each instrument has a keypad timer interfaced to either the filament or high voltage on/off switch, depending on ease of accessibility. The timer is interfaced to a 486 computer (obtained from surplus-there are lots of these available). The computer stores user time for each account as well as the number of films exposed for the instrument used. Because we have to pay for approximately 90% of our operations costs, we have to charge for use of the instruments. We also charge different hourly rates depending on user status (inside or outside user).
At the end of each month, the user time/film number data is dumped into another computer which compares the reserved time from the web-based reservation system to the actual use time and charges for the greater of the two. If the instrument develops problems during the use time and can't be used any more, the individual user is able to send a DOWN message to the reservation computer and the user is charged only for actual time used. The monthly use data is formatted into a spreadsheet which contains microscope use times and film numbers used. A principal investigator (PI) may have ten grad students who use five different instruments and three different grant account numbers during the month. He/she receives a statement at the end of the month which identifies the microscope user by name, the account number used, the hours used (rounded to the nearest minute)for each instrument and the total number of films exposed for each use time. The total costs for instrument use is given on the last line of the statement.
An advantage of this system is that within thirty minutes, one can sort data files and determine how many hours each microscope is used over any period of time; how many hours each user utilizes an instrument(s) over time; number of hours used for each account and total number of hours for all instruments over time. This info is useful for reports to administrators as well as for funding agency applications.
If you have questions about our system, please e-mail me directly or call at the number listed below.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
A short time ago there was a discussion about the different EMs RCA produced and when they were produced. The other day I was digging through my sfiles and came across a copy of the August 1967 issue of the RCA Scientific Instruments News which has pictures of all the models produced and the year of introduction:
EMA 1939 I don't think this model was ever sold commercially. I believe it was built in the RCA Labs for developmental uses
EMB 1940 This was the first commercial model
EMC 1944 This model had a horizontal column
EMT 1950 This was an inexpensive table model that was designed with use in high schools and small colleges in mind.. As I recall it had lenses made of permenant magnets to eliminate the need for lens power supplies and to hold the cost down.
EMU 1944 This was probably RCA's most popular model. It was on their leading EM for at least 10 years.
EMD 1948 This was an electron diffraction instrument, designed specifically for obtaining ED patterns by the reflection from solid surfaces at a grazing incidence.
EML 1954 This was the forerunner of the popular EM-U# and EM-U4 models, but provided an accelerating voltage of only 50kV. It was the model in which RCA pioneered the modern ergonomic layout of controls, with a desk-like design.
EMU-3 1955 Similar in appearance to the EML, but providing 100 kV.
EMU-4 1966 A refinement of the EMU-3, providing a double condenser lens, a specimen stage airlock system, etc.
RCA ceased marketing electron microscopes in 1969. I was President of the EMSA that year and discussed the matter with James Hillier, one of the pioneers in the field of electron microscopy, and a Vice President of RCA at that time. He said that it was determined that RCA could make more money selling records and related electronic devices than EMs, and so was phasing out of the EM business.
The issue of RCA Scientific Instruments News in question also has a picture of each model on the cover. I have had a copy run off in jpg format. I understand that it is not appropriate to transmit such info via this list server; however, if anyone would like a copy, let me know and I'll send it to you individually.
Happy St. Patrick's Day,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
} Dear Listservers, } } I recently responded to a thread about Amray FE maintenance contracts. } My response contained two errors: one numerical, one in perception. } I wish to correct them at this time.
[snipped for brevity sake]
Thanks Earl for the update and correction. This is good info. And I see some room for additional discussion and inquiry to wrap up this topic.
The prospects for third party maintenance contracts are rather variable--depending mostly, I think, on what is being maintained under contract. This would be based on the availability of schematics, software, special parts, etc., etc. If some FESEM makers do not "guard" their FE guns relative to others, then of course, that will weigh heavily on third party options. For the record, here are some figures for Amray maintenance contracts. An 1830 LaB6 system runs $13,500 annually. This excludes "consumables" such as apertures, LaB6 cathode, pump oil, etc. But it does include on-site service, troubleshooting, travel, lodging, meals, rental car, things that break down, like Balzers 240 turbo pump or Edwards E2M8 mechanical pump, or the invariable broken wire. Actually, I have found the 1830 to be very reliable and a good performer.
The 1910FE runs $15,000 annually. Same terms as the 1830 but includes the computer control system. What I don't know is whether Amray will sell the FE gun assemblies to third party maintainers. I do know that for Amray users who are not under an Amray contract, the 305FE gun costs about $14,000 total to replace as a per-diem item. Again, I have found that this SEM also is very reliable. But what about performance?
Performance wise, I suppose we could have an "image shoot off" contest based on a standard specimen and different SEM instruments of different models and from different makers. But there are some basic differences among SEMs that would otherwise seem to be the same--but are not. Notable among these differences, for example, is the design and construction of the final lens pole piece. This is due, I think, to two or three driving forces. One is the need to image physically large specimens at various WDs. This means that one needs a large chamber with a stage that offers accommodation of large samples and wide WD range. The second force is the requirement to image whole semiconductor wafers ranging in diameters from 4" to 8" and to have a wide travel distance across the wafer and to operate well at low KV (photo resist examination, for example). The third force is the impact on the SEM when needing to do X-ray analysis.
The flat lens design is quite useable for large and small specimens when high tilt angles do not increase the WD to an unsatisfactory figure. In the case of IC wafers, the conical lens is required. These are available as 45 or 60 degree designs. Since most semiconductor qualification imaging is done at either zero or 45 degrees, the 60 degree lens is probably optimal. What Amray has done in regards to conical lenses is an evolutionary path. The 1830 has a conical lens to accommodate high tilt angles when working with wafers. It also can be configured with a large chamber. However, the design of the lens is such that high resolution is obtained at 10KV-15KV and above. It is not a high resolution SEM at low KV. Alternatively, with a 25KV or 30KV power supply, it has ample voltage to support the beam currents needed for x-ray work. I have no personal information on what the other makers have done and currently offer in regards to IC wafer inspection and analysis.
The Amray 3600LEAP was a major design departure. This system also has a 60 degree conical lens, a huge chamber but a specially designed lens and other features key to semiconductor work. The LEAP lens was designed for low KV imaging. At 2-3KV, the LEAP is virtually identical in resolution to the other lens types at 15KV and at the same WD. This is a key feature and requirement for imaging fine pitch and small feature size wafers and photoresist. The other important feature of the 3600LEAP SEM is the 5-place heated final aperture holder. Since the 3600 runs with 35u and 70u apertures (or less) as routine, these Pt apertures are continuously heated to keep the effects of contamination (photo resist again) at a minimum. Not only does this keep the apertures clean, but it also prevents accumulation of PR in the scan coil liner. This FESEM is a very capable instrument from my experience with it. It can easily image 0.15u wide gate poly at high resolution and 2.5KV at WD=6mm.
If one considers where I have wound up with this discussion, it is cause for pause to ask a fundamental question. That is, "What is a research SEM?" Different applications must surely lead one to select one type of instrument over another. Does it also direct a path to one manufacturer over another? Since I have not seen the myriad number of SEM models from all of the vendors, I cannot answer this question. But while there might be some disdain for one maker versus others, it seems tough for me to make any sort of quantitative decision when there are quite a few variables involved--solely based on application, much less vendors and maintenance options.
What do you think about this, based on your experience?
TOUR SPEAKER (Society for Applied Spectroscopy) - 4/12/2000 Meeting in Mountain View
"RAMAN SPECTROSCOPY AS A PROBE OF STRUCTURE/ORIENTATION IN POLYMERIC FIBERS"
by DR. BRUCE CHASE (DuPont Central Research, Wilmington DE)
at Michaels at Shoreline, Mountain View (Directions to follow)
Register 6:45 PM Dinner 7:30 PM Talk 8:30 PM
Cost: $30 for dinner ($10 students and teachers), free for talk.
Dinner Choices: (Indicate Choice when RSVP'ing)
Breast of Chicken, Piccata Chilean Sea Bass, Ginger Shallot Sauce Grilled Vegetable Brochette with Wild Rice
Reserve by April 10 to Steve Rabin, 650-564-5315 or steve.rabin-at-alza.com
(Please let us know if you're coming for dinner, or just the talk for headcount purposes)
Also, see our web site at www.sas-ncss.org
ABSTRACT Polymeric macroscopic properties such as tenacity, modulus, elasticity, etc. are determined by molecular level effects such as conformational state populations, orientation, and intermolecular interactions. Significant changes occur at the molecular level as a molten or solution phase polymer is spun into fiber form. The polymer goes from an unoriented, amorphous state to an oriented, semi-crystalline material via the deformations imposed by spinning and drawing. Raman spectroscopy can potentially provide information on both structure and orientation. Changes in band intensities can be related to conformational populations and to formation of crystalline regions. Changes in relative intensities as a function of incident and scattered polarization yields information on chain orientation. The use of polarized Raman scattering done in both 90 and 180 degree scattering geometries has been used to determine the second and fourth order orientation functions for both polyethylene and PET fibers. Raman measurements have been made in both the laboratory frame and on-line for 66 nylon and PET fibers. For both PET and 66 nylon, crystallinity levels and a qualitative measure of orientation can be obtained.
DIRECTIONS to Michaels (in Shoreline Park, Mountain View):
} From Anywhere in the Bay Area:
Get to Highway 101 toward Mountain View (from SF and the Peninsula, southbound; from San Jose and the East Bay via 237, northbound)
Go to the Shoreline Rd. exit and turn left at the end of the exit road. Follow Shoreline Rd. into (approx 1 mile) through Mountain View Park gate. Continue on the single lane road (golf course on your left) for approx 1 mile, turn left at Michaels restaurant sign into the parking lot.
Following up on the Hg burner problems, does anyone have a schematic of the "transformer" for these? There is assumed to be a true transformer plus a rectifier and filter. Without a circuit diagram, there is too much speculation for me about what is actually going on. It seems that there are some common problems out there. These ought to be readily solvable.
It really cannot be that difficult, can it? Maybe so.
Diana, These blades are available from Allegiance Scientific (used to be Baxter, used to be Scientific Products, used to be....). As far as I know, this is the only place that carries this brand. I have no financial interest in Allegiance, just passing along a fact. Wanda Shotsberger (HT ASCP)
-----Original Message----- } From: Diana Papoulias [mailto:Diana_Papoulias-at-usgs.gov] Sent: Thursday, March 16, 2000 2:53 PM To: microscopy-at-sparc5.microscopy.com
Could someone please tell me where I can purchase accu-edge microtome blades?
Thank you.
Diana Papoulias USGS 4200 New Haven Rd Columbia, MO 65201
I would like to get hold of an RCA model EMT for display for our museum collection. Ed Sharpe archivist for SMECC
{ { Subj: RCA EMs Date: 3/17/00 2:47:16 PM Pacific Standard Time From: bigelow-at-engin.umich.edu (Wil Bigelow) To: AAmy-at-dtsc.ca.gov, JRowe6427-at-aol.com, cgarber-at-2spi.com (Chuck Garber), john.mardinly-at-intel.com, microscopy-at-sparc5.microscopy.com (Microscopy Listserver), oshel-at-shout.net
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Hi all:
A short time ago there was a discussion about the different EMs RCA produced and when they were produced. The other day I was digging through my sfiles and came across a copy of the August 1967 issue of the RCA Scientific Instruments News which has pictures of all the models produced and the year of introduction:
EMA 1939 I don't think this model was ever sold commercially. I believe it was built in the RCA Labs for developmental uses
EMB 1940 This was the first commercial model
EMC 1944 This model had a horizontal column
EMT 1950 This was an inexpensive table model that was designed with use in high schools and small colleges in mind.. As I recall it had lenses made of permenant magnets to eliminate the need for lens power supplies and to hold the cost down.
EMU 1944 This was probably RCA's most popular model. It was on their leading EM for at least 10 years.
EMD 1948 This was an electron diffraction instrument, designed specifically for obtaining ED patterns by the reflection from solid surfaces at a grazing incidence.
EML 1954 This was the forerunner of the popular EM-U# and EM-U4 models, but provided an accelerating voltage of only 50kV. It was the model in which RCA pioneered the modern ergonomic layout of controls, with a desk-like design.
EMU-3 1955 Similar in appearance to the EML, but providing 100 kV.
EMU-4 1966 A refinement of the EMU-3, providing a double condenser lens, a specimen stage airlock system, etc.
RCA ceased marketing electron microscopes in 1969. I was President of the EMSA that year and discussed the matter with James Hillier, one of the pioneers in the field of electron microscopy, and a Vice President of RCA at that time. He said that it was determined that RCA could make more money selling records and related electronic devices than EMs, and so was phasing out of the EM business.
The issue of RCA Scientific Instruments News in question also has a picture of each model on the cover. I have had a copy run off in jpg format. I understand that it is not appropriate to transmit such info via this list server; however, if anyone would like a copy, let me know and I'll send it to you individually.
Happy St. Patrick's Day,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
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Dear microscopist, Kindly give me information and websites available on adsorption, desorption and diffusion study of Chromium on Tungsten by field emission and field ion microscopy. Also nucleation and growth of Chromium on Tungsten. Thanking you,
Mangesh Bagade Govt College Of Engineering Pune Maharashtra, India.
____________________________________________________________________ Get free email and a permanent address at http://www.netaddress.com/?N=1
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Dear microscopist, Kindly give me information and websites available on adsorption, desorption and diffusion study of Chromium on Tungsten by field emission and field ion microscopy. Also nucleation and growth of Chromium on Tungsten. Thanking you,
Mangesh Bagade Govt College Of Engineering Pune Maharashtra, India.
____________________________________________________________________ Get free email and a permanent address at http://www.netaddress.com/?N=1
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H i Gary,
Your e-mail poses some interesting question & perspectives.
As far as the variation of Third Party maintenance contracts, Scanservice generally offers contracts that are exactly the same as the given manufacturer. The customer can then compare "apples against apples". Whenever there is a question regarding coverage we always look to see what the manufacturer's contract covers. The variation in contracts is only between the different manufacturer's. Cambridge (LEO) did not cover scan coils, ETEC did not cover rotary pumps or CRTs, etc. This eliminates the confusion for our customers. We can and do offer modified contracts where the customer is responsible for parts or a limited number of service calls but only at the customer's request.
All manufacturer's , SEM & otherwise, are required by Federal Law to sell parts to third party organizations. To withhold parts is considered a "restriction of trade" and has some hefty consequences. I have had accounts set-up for just about all the manufacturers. It is relatively easy process & most manufacturer's treat us like any other customer in need of parts.
The only exception is Amray that has a multi-step process for obtaining parts which is as follows:
1. Request part number. 2. Wait two to three days for part number. 3. Send credit references. 4. Wait two to three days for response. 5. Send banking references & information. 6.Wait two to three days for response.
The above process has occurred at least three times in the last three years. Finally, I requested that they send the parts COD or I could prepay. The response I got was that I "could have done that all along."
Bottom line: manufacturer's must sell to Third party organizations, most are very professional & co-operative.
As far as an "image shoot-off", I really don' think that would be very practical as there are significant variations even between the same model from a given manufacturer. I would expect the best performing SEM from each manufacturer involved, rather than an average of the industry.
When I worked at ETEC, we noticed differences between the different SEM columns. The demo unit always got the best column. Even today I recommend that customers purchase the "demo unit" as it probably has the best of everything & is tuned up. Moreover the "demo unit" can be sold at a lesser price as it is technically used. I also recommend that customers considering an SEM purchase use the model they are considering at another customer's site. In this way, the potential customer can evaluate the SEM model on their own terms using their own standards and without the applications engineer's influence. Most manucfacturer's want to use the standard "gold on carbon" to highlight their given SEM. I generally do not see many of my customers imaging "gold on carbon" on a daily basis. Customers has such a variety of samples: photo resist, uncoated teflon, etc.
This reminds me of a customer at Hughes who I recommended that she follow the above procedure of evaluating the SEM at a customer site & requesting that the demo unit be sold to her Company. She did neither. When she requested that the demo unit be sold to her Company, the manufacturer declined. No reason was given. She bought the FESEM anyway. After about three months I inquired about the status of her new FESEM. Instead of excitement she responded, "Oh it's OK. I guess I will just have to get used to it". She continued to use her standard tungsten SEM over the FESEM.
As far as your analysis of the SEM chambers & polepieces, historically the SEM industry has seen the large chamber sizes and conical polepieces you decribe for at least last twenty years. When I entered this industry in 1973, I remember the JEOL JSM-2 with a semi-conical lens. The lens was not a true conical lens as it did have a "flat tip" of about 4 cm.dia. but it did allow one to tilt samples at a higher angle. I was surprised to see JEOL had a JSM-1 (circa 1969) that had the same column as the JSM-2.
ISI had a model DS-130 that could be ordered with different final lenses: the standard semi-conical lens, wide bore (WB) for 0 working distances, or conical lens. The ISI conical lens had multiple angles on their design. It started with a 30 degree cone, the mid-section was 45 degree, and ended with a 70 degree cone. For it's day the DS-130 had pretty good optics. The electronics were junk in my opinion. The DS-130 had some extremly large chambers. One was known by the ISI engineers as a "turkey baster" chamber. It measured about 24 in. x 36 in. x 18 in high. Hitachi has had the S-806 and S-808 for semiconductor work. An full 8-inch wafer could be viewed as well as tilted 45 degrees. Hitachi also offerred an S-806C which is a conical lens. I believe the vintage for these machines was in the early eighties.
The JEOL JSM-U3 (circa 1970) had a heated final aperture strip that allowed the user to clean the aperture without removing it. This aperture design was replaced in the JEOL 35U and JEOL 35C (circa 1974?). The JEOL 35 series had a continuously heated final aperture. A smaller current (about 2 amps) was run through the aperture therby cleaning the aperture while using it.
I recently (last year) had the opportunity to attend a friend's/customer retirement party. Attending were many older & retired SEM engineers. They were reminiscing about the SEM industry and how the machine had evolved from their days at Westinghouse (yes Westinghouse) in the 1960's. The same questions and problems that we have today they had in the 1960's. Chamber sizes were initially large to accomodate their sample and stage sizes but vacuum was poor (10-4 torr). Chamber sizes were reduced to improve vacuum and lens designs were changed to accomodate tilt angles at the expense of resolution. And everyone could only dream of a stable FE gun. SEM nerds.
The upshot of all of this is that all of these issues has been historically addressed. Large & small chambers, different lenses, etc. will be with us as long as the application permits. When a manufacturer claims to have a "breakthough", it only remind me of what Steve Jobs (Apple Computer) said about Windows 95: " Windows 95: MacIntosh 1986." I think the next real breakthrough will be the lenses made from super-conductive wire or, better yet, the electrostatic columns being developed.
It is my understanding that the electrostatic columns are Shotky FE sources with backscatter dectectors fully integrated onto the "final lens". Future features include an integrated EDS detector. The columns are small & cheap enough to be "thrown away".
I really don't have a disdain for any given Company. I would only like to comment that every Company has a different "flavor" depending upon whom is running it. Most are a pleasure to work with, while others leave a "bad taste in your mouth".
Please don't take these comments personally as I respect your opinion & have throughly enjoyed the subjects you have brought up on this listserve & look forward to future your comments.
Regards,
Earl Weltmer
At 01:54 PM 3/16/00 , you wrote:
} Dear Listservers, } } I recently responded to a thread about Amray FE maintenance contracts. } My response contained two errors: one numerical, one in perception. } I wish to correct them at this time.
[snipped for brevity sake]
Thanks Earl for the update and correction. This is good info. And I see some room for additional discussion and inquiry to wrap up this topic.
The prospects for third party maintenance contracts are rather variable--depending mostly, I think, on what is being maintained under contract. This would be based on the availability of schematics, software, special parts, etc., etc. If some FESEM makers do not "guard" their FE guns relative to others, then of course, that will weigh heavily on third party options. For the record, here are some figures for Amray maintenance contracts. An 1830 LaB6 system runs $13,500 annually. This excludes "consumables" such as apertures, LaB6 cathode, pump oil, etc. But it does include on-site service, troubleshooting, travel, lodging, meals, rental car, things that break down, like Balzers 240 turbo pump or Edwards E2M8 mechanical pump, or the invariable broken wire. Actually, I have found the 1830 to be very reliable and a good performer.
The 1910FE runs $15,000 annually. Same terms as the 1830 but includes the computer control system. What I don't know is whether Amray will sell the FE gun assemblies to third party maintainers. I do know that for Amray users who are not under an Amray contract, the 305FE gun costs about $14,000 total to replace as a per-diem item. Again, I have found that this SEM also is very reliable. But what about performance?
Performance wise, I suppose we could have an "image shoot off" contest based on a standard specimen and different SEM instruments of different models and from different makers. But there are some basic differences among SEMs that would otherwise seem to be the same--but are not. Notable among these differences, for example, is the design and construction of the final lens pole piece. This is due, I think, to two or three driving forces. One is the need to image physically large specimens at various WDs. This means that one needs a large chamber with a stage that offers accommodation of large samples and wide WD range. The second force is the requirement to image whole semiconductor
wafers ranging in diameters from 4" to 8" and to have a wide travel distance across the wafer and to operate well at low KV (photo resist examination, for example). The third force is the impact on the SEM when needing to do X-ray analysis.
The flat lens design is quite useable for large and small specimens when
high tilt angles do not increase the WD to an unsatisfactory figure. In the case of IC wafers, the conical lens is required. These are available as 45 or 60 degree designs. Since most semiconductor qualification imaging is done at either zero or 45 degrees, the 60 degree lens is probably optimal. What Amray has done in regards to conical lenses is an evolutionary path. The 1830 has a conical lens to accommodate high tilt angles when working with wafers. It also can be configured with a large chamber. However, the design of the lens is such that high resolution is obtained at 10KV-15KV and above. It is not a high resolution SEM at low KV. Alternatively, with a 25KV or 30KV power supply, it has ample voltage to support the beam currents needed for x-ray work. I have no personal information on what the other makers have done and currently offer in regards to IC wafer inspection and analysis.
The Amray 3600LEAP was a major design departure. This system also has a 60 degree conical lens, a huge chamber but a specially designed lens and other features key to semiconductor work. The LEAP lens was designed for low KV imaging. At 2-3KV, the LEAP is virtually identical in resolution to the other lens types at 15KV and at the same WD. This is a key feature and requirement for imaging fine pitch and small feature size wafers and photoresist. The other important feature of the 3600LEAP SEM is the 5-place heated final aperture holder. Since the 3600 runs with 35u and 70u apertures (or less) as routine, these Pt apertures are continuously heated to keep the effects of contamination (photo resist again) at a minimum. Not only does this keep the apertures clean, but it also prevents accumulation of PR in the scan coil liner. This FESEM is a very capable instrument from my experience with it. It can easily image 0.15u wide gate poly at high resolution and 2.5KV at WD=6mm.
If one considers where I have wound up with this discussion, it is cause for pause to ask a fundamental question. That is, "What is a research SEM?" Different applications must surely lead one to select one type of instrument over another. Does it also direct a path to one manufacturer over another? Since I have not seen the myriad number of SEM models from all of the vendors, I cannot answer this question. But while there might be some disdain for one maker versus others, it seems tough for me to make any sort of quantitative decision when there are quite a few variables involved--solely based on application, much less vendors and maintenance options.
What do you think about this, based on your experience?
If you are looking for the traces of the tools which were used to make these surfaces, sounds like a job for an interferometer. There are several small ones which can be used for low to medium power (10x-50x objective mag) work. Hach used to carry a small Michelson; also, check with Nikon for both Michelson and Tolansky varieties. While the Tolansky is a contact method system, it is non-destructive and, since it is a multiple beam system, gives very high precision results. I would recommend photographing the interferograms then correlating with the "tooth marks" left by various tools. The old Polyvar also had an interferometer module.
For a more upscale, automated version, I'd suggest that you find a lab with either a Zygo New View or a Wyko RST. These devices are Scanning White Light Interferometers (SWLIs); both are automated, 3D imaging systems which might provide interesting insight but may also be overkill for your type of project. Zygo is just south of us, in central Connecticut, while Wyko is in Tucson. I believe that both have contract labs. (I have done some work in the Zygo facility)
Please contact me if you are interested in pursuing this approach.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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}
At 07:29 PM 12/12/99 +0100, S¿ren Albek wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A client of ours makes a cut in a polymer block (I can check on the exact material, if you need) then looks at two parameters: 1) the angle made by the sides 2) the radius of curvature of the bottom
He uses a conventional image analysis system for both, using just the simple angle measurement function for the first and the diameter of a circle of best fit for the second. He had been using a stereo microscope but we took a look at the cut under a 10x objective/compound microscope on a recent visit and found that there was a lot to be learned about the quality of the cut from the shards and shredding left on the surface (I can't remember whether it was the upper or lower, so take a look at both).
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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}
At 07:04 PM 3/13/00 -0400, Rosemary Walsh wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think you must be referring to the famous Bill Miller, who was my "partner in crime" at Sarastro. You can reach him at Bill Miller {microbill-at-mohawk.net} .
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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}
At 11:12 AM 3/14/00 -0500, Ric Felten wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Raman is going to become as available as regular confocal. As a matter of fact, it is on the same price scale but can do both confocal and chemical imaging. It is an analytical technique which works well alongside fluorescence, but you need to collect the Raman signal separately. I just came back from PITTCON (THE major analytical chemical meeting) and some companies are collecting the signal in the near UV, others in the near IR.
A good starting point is Jack Koenig's book "Spectroscopy of Polymers" (Am. Chem. Soc.. Washington DC, 1992). I just saw him at PITTCON and he said that the new edition is either just coming out or about to come out.
The move to Raman is part of the merging of microscopy and spectroscopy. There are now several interesting true hybrid instruments which permit chemical as well as microscopy imaging. One is the Continuum from Spectra-Tech (Shelton, CT); the other is a series built on the DuraScope from SensIR (Danbury, CT). I had a chance to teach a microscopy class to the IR specialists from SpectraTech in January, so really had a chance to have my hands on the system. In addition to running full FT-IR chemical spectra, I was able to do low power darkfield. They also have regular phase and DIC objectives available (the Phase was a bit tricky to implement) and have just introduced a fluorescence module.
I will be doing a review for American Lab (Watch for Am Lab: "Focus on Microscopy") and will post pertinent excerpts for you within the next few weeks.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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}
At 03:00 AM 3/2/00 +0000, Jose Feijo wrote: } Since the subject was raised, could anyone point me out a good paper or internet source to understand Raman microscopy, and how to make it work? On the side, from the gurus, how much should we expect from it in the future? Is it like, it's going to substitute something already available, or otherwise it will complement specific aspects of visualisation of some special biological structure? What does it involve, special lasers, special optics, special computers? } } Thanks in advance } Jose } }
A propos of our earlier discussion, this just came across the wire from the Society for Appl. Spectroscopy:
TOUR SPEAKER (Society for Applied Spectroscopy) - 4/12/2000 Meeting in Mountain View
"RAMAN SPECTROSCOPY AS A PROBE OF STRUCTURE/ORIENTATION IN POLYMERIC FIBERS"
by DR. BRUCE CHASE (DuPont Central Research, Wilmington DE)
at Michaels at Shoreline, Mountain View (Directions to follow)
Register 6:45 PM Dinner 7:30 PM Talk 8:30 PM
Cost: $30 for dinner ($10 students and teachers), free for talk.
Dinner Choices: (Indicate Choice when RSVP'ing)
Breast of Chicken, Piccata Chilean Sea Bass, Ginger Shallot Sauce Grilled Vegetable Brochette with Wild Rice
Reserve by April 10 to Steve Rabin, 650-564-5315 or steve.rabin-at-alza.com
Also, see our web site at www.sas-ncss.org
ABSTRACT Polymeric macroscopic properties such as tenacity, modulus, elasticity, etc. are determined by molecular level effects such as conformational state populations, orientation, and intermolecular interactions. Significant changes occur at the molecular level as a molten or solution phase polymer is spun into fiber form. The polymer goes from an unoriented, amorphous state to an oriented, semi-crystalline material via the deformations imposed by spinning and drawing. Raman spectroscopy can potentially provide information on both structure and orientation. Changes in band intensities can be related to conformational populations and to formation of crystalline regions. Changes in relative intensities as a function of incident and scattered polarization yields information on chain orientation. The use of polarized Raman scattering done in both 90 and 180 degree scattering geometries has been used to determine the second and fourth order orientation functions for both polyethylene and PET fibers. Raman measurements have been made in both the laboratory frame and on-line for 66 nylon and PET fibers. For both PET and 66 nylon, crystallinity levels and a qualitative measure of orientation can be obtained.
I've been asked whether it is possible to look at a frozen powder using Cryo-SEM. The powder will be frozen and stored at -80 C and must be mounted whilst frozen (it may be possible to raise the temperature to -20 C) any ideas of an adhesive I could use at these low temperatures??
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hort.cri.nz
} As far as your analysis of the SEM chambers & polepieces, historically } the SEM industry has seen the large chamber sizes and conical polepieces } you decribe for at least last twenty years.
For those, like me, who were interested in your recollections about SEM chamber and lens design, and who may be considering attending ICEM-15 in South Africa in 2002, make a note that there will be an exhibition at the congress retracing the history of electron microscopy in South Africa. This will include a variety of old EM equipment, including some of the SEM's to which you referred.
Regards
Rob
Robin H Cross Chairman : 15th International Congress on Electron Microscopy (ICEM-15) tel: +27 46 603 8168/9 - fax: +27 46 622 4377 email: r.cross-at-ru.ac.za http://www.ru.ac.za/emu/icem-15.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
Ian, I've used Duro-Tak¨ 80-1061 to hold particles during analysis at liquid nitrogen temperatures . For more information see: http://www.fbi.gov/programs/lab/fsc/backissu/july1999/ward.htm
Dennis ________________________________________________________________________ Dennis C . Ward voice: 202-324-2982 FBI Laboratory fax: 202-324-4018 Microanalysis Laboratory email: DCWard-at-concentric.net
----- Original Message ----- } From: "IAN HALLETT" {ihallett-at-hort.cri.nz} To: {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, March 19, 2000 7:45 PM
Hello Ian
} I've been asked whether it is possible to look at a frozen powder } using Cryo-SEM.
That shouldn't pose any problems. Choice of adhesive would depend on the characteristics of the powder.
Rob
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
I would like to thank those who took the time to respond to my e-mail. I received a lot of valuable information and advice on different techniques to be used (and NOT to be used) when studying carbides in tool steels. I appreciate your help.
Kind regards, Jesper
---------------------------------------------------- Jesper Vejloe Carstensen Research Scientist, M.Sc., Ph.D. Materials Research Department Risoe National Laboratory P.O. Box 49 DK-4000 Roskilde, Denmark Phone: +45 4677 5776 Fax: +45 4677 5758 E-mail: jesper.v.carstensen-at-risoe.dk Web: http://www.risoe.dk/AFM/Personal/Jeca/jeca.htm ----------------------------------------------------
We are planning to buy a new diamond knive for sectioning biological samples. My question is if anybody has experience with using 35 degree angle diamond knives and if they have any disantvantages compared to 45 degree angle diamond knives. We are mainly sectioning unicellular algae embedded in LR-gold and hope to reduce compression problems by using a 35 degree angle knive. Our problem is that, especially algae with thick cell walls, tend to fall out of the sections. This problem seems to be caused by compressions, because using a brand-new 45 degree angle diamond knive I had much less algae falling out of the sections. Because we can effort only one new knive we have to use it as an all-round knive. Is a 35 degree knive suitable to replace a 45 degree knive or is it much more fragile and/or less durable?
Thanks in advance for your time and help.
Sincerely,
Stefan Geimer
*********************** Stefan Geimer University of Cologne Botanical Institute Gyrhofstr. 15 D-50931 Cologne Germany phone: +49-(0)221-470-3795 fax: +49-(0)221-470-5181 ************************
Apparently there is more equipment lying around that needs a good home. If anyone is interested, please respond directly to Mark Chambers via mark-at-semiconductor.com or at (613) 599-6500 ext 4269.
Marisa
} -----Original Message----- } From: Mark Chambers } Sent: Friday, March 17, 2000 4:18 PM } To: Marisa Ahmad } Subject: RE: Liquidation of assets } } Hi Marisa, } } Is the listserver you used for the PGT EDS announcement a suitable venue } for the plasma asher and the Mosaid tester? } } The barrel etcher is a Plasmaline model 411 and whoever wants it will need } to get a vacuum pump for it. The etcher was working but it was not an } efficient way to decap packages. } } The Mosaid tester is a model MS2300 and that's basically all I know about } it. } } Thanks } Mark
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You can buy them from Electron Microscopy Sciences. 1-800-523-5874
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
I have been asked to find a new home for a fully working JEOL 100CX tem which is currently in use in the south of England. It has the ASID stem attachment, and it could also be available with a Link AN10 analysis system.
Probably the only cost will be the dismantling & removal charges. If anyone is interested, please reply to me directly. My only commercial interest is that I might be involved with the dismantling etc.
Bob Phillips MicroServiS bob.phillips-at-microservis.co.uk
Please check out the web-site for ICHC 2000 (11th International Congress for Histochemistry and Cytochemistry) 3-8th Sept. York, UK.
We have put together a fabulous speaker list covering a range of the most important areas of development and application of cutting edge techniques in Cell Biology and Imaging today. Lead speakers include Roger Tsien, Stefan Hell, Alan Fine, Alan Bode, Hans Tanke, Jennifer Lippincott-Schwartz, Angus Lamond, Jim Smith, Richard Haugland, Paul Monaghan, Mike Ormerod, Simon Gilroy Nick White etc. etc. etc.
www.med.ic.ac.uk/external/ichc_2000
ABSTRACT DEADLINE 15TH MAY 2000.
Best wishes
Gary Coulton Dr. Gary Coulton Molecular Pathology Division of Biomedical Sciences Imperial College School of Medicine The Sir Alexander Fleming Building South Kensington London SW7 2AZ
tel 0044 (0)171 594 3190 fax 0044 (0)171 594 3022
PLEASE NOTE AS FROM APRIL 1ST 2000 THE FOLLOWING NUMBERS WILL REPLACE THOSE ABOVE AND SHOULD BE USED.
------------------------------------- Announcing the 11th International Congress of Histochemistry and Cytochemistry (ICHC 2000)
"Understanding Biocomplexity: The Post-Genome Challenge"
September 3-8, 2000, York, United Kingdom
ICHC 2000 will comprise 27 symposia addressing the latest developments and applications of histochemistry and cytochemistry in the life sciences including medicine.
Many leading experts to speak
On-line registration now open!!!!!!!!!!
For further details of the meeting and how to pre-register please visit our web-site at http://www.med.ic.ac.uk/external/ichc_2000
The Department of Materials Science and Engineering at the University of Pennsylvania is seeking an engineer or scientist for a technical staff position in its advanced characterization facility. This facility contains a number of state-of-the-art microscopes and scattering facilities as well as ancillary detectors, specimen preparation equipment, and data processing and computational hardware and software. The facility serves Penn research programs, as well as academic, industry and government users from the Delaware Valley region and beyond. The University of Pennsylvania is located in Philadelphia, one of the nation's most vibrant cities. The university, a member of the Ivy League, was founded by Ben Franklin and is the fourth oldest and first secular university in the US. The Laboratory for Research on the Structure of Matter was constructed to house one of the original three Materials Research Laboratories (MRLs) in the US. The university has a continuing tradition of leading materials research and has housed the MRL (now MRSEC) continuously for 40 years.
Under the direction of the facility manager, the successful candidate will be responsible for conducting experiments with users on facility equipment, training new users and maintenance of facility equipment not covered under service contract. The successful candidate must have the communication skills and self-confidence to interact with technical users with a wide range of expertise and background. Job performance will be assessed based on the success in experimental interactions with users, the operational state of equipment for which the staff member is responsible, progress in the acquisition of new skills, and facility appearance.
A Bachelors degree in physical science or engineering is required, although an advanced degree is preferred. The successful candidate must have experience with electronics, ultra-high vacuum technology and computer interfacing of equipment. Experience in the operation and use of electron microscopes and ion beam lines and associated end-stations is desirable. Experience in the use of Auger electron spectroscopy or scanned probe imaging is also desirable.
The text of the official job listing from the University of Pennsylvania web site follows. Please refer to the Penn Human Resources web site for the official hiring policy of the university at www.hr.upenn.edu. For information about the specific position, please contact the facility manager, Dr. Douglas Yates, at dmyates-at-lrsm.upenn.edu.
Text of web site listing:
Reference Number: 00034808DL Job Title: RESEARCH COORDINATOR SR School/Center: ENGINEERING & APPLIED SCIENCE Department: MATERIALS CHARACTERIZATION Date Posted: 3/9/00
Duties: Train or coordinate training of facility users; assist users performing experiments; compose monthly MCF users billing summary; organize purchasing of consumable laboratory materials; maintain or coordinate maintenance of equipment; maintain working environment & appearance of laboratory.
Qualifications: BA/BS in Physical Science or Engineering required, advanced degree preferred; 3 to 5 years experience with electronics, ultra-high vacuum technology & computer interfacing of equipment & in operation & use of electron microscopes, ion beam lines and/or scanned probe imaging.
******************************************************* Douglas M. Yates, Ph.D. Manager, Materials Characterization Facility
(215) 573-6123 dmyates-at-lrsm.upenn.edu
University of Pennsylvania Department of Materials Science & Engineering 3231 Walnut Street Philadelphia, PA 19104-6272 *******************************************************
One of your best sources of information is going to be Diatome USA. They have an actual lab set up to test cutting problems of a large variety. They also have access to the information of the company who makes these knives.
Please consider that your problem may not be with the knife or the angle at which the knife is sharpened. Most "breaking out" problems are related to the production of the block, especially 1) the infiltration 2) the match between the specimen and the formulation used. Contact Diatome, at EMS, and ask to speak with Stacy Kirsch, who is an expert with these problems. (I have no business interest in Diatome - I have just received the most excellent advice from them over the years)
Hildy Crowley Sr. Electron Microscopist University of Denver Denver, CO
P.S. Should processing be the basic problem, please contact me. I may be able to help you.
My specialists tell me, that Boron shows a plasma loss at 22.7 eV and a K-edge at 188 eV. They also tell me, that it is hard to see B in samples as it usually disappears rapidly. To see B, they suggest focusing on a different sample area and then only expose the area in question for the PEELS acquisition.
In case you are using our software on a LEO microscope for the acquisition, you should select the MDF option (Micro Dose Focusing).
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Exchange Administrator Sent: Saturday, March 11, 2000 12:35 PM To: Michael Bode
Need help on PEELS } } To all, } } I am working on grain boundary chemistry measurement } with Digipeels. Try to find out the segregation } behavior of boron, which is about 0.5at% averagely. } However, I cannot find any edge for boron in the } spectrum. As I don't know what's the optimum setting } for PEELS to check the grain boundary segregation, if } you have any idea on this, please let me know. } } Thanks, } Lung __________________________________________________ Do You Yahoo!? Talk to your friends online with Yahoo! Messenger. http://im.yahoo.com
We bought a JEOL 100SX for about $130,000 in 1987. The scope was for student use in an undergraduate course. It was used for 4 to 8 weeks per year for 7 years. It was used very little. About 1994, the water chiller failed and cooked the objective mini-lens and some other electronics. It was one year off the service contract. JEOL could not guarantee that they could fix it for less than $11,000 so it was moth balled. It was functional at 8000X and higher. We have to get rid of this in a real hurry. It someone wants some or all of this machine please contact me ASAP. Within about a week it will be in the dump. I recall that at the time it failed there was a user at Stanford (I think) who had the same model. If anyone knows who that was, please have him contact me.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
It seems that nowadays every SEM vendor is offering variable pressure models, which are conventional SEMs with a plumbing modification that allows the sample to be at pressures of up to about 4 torr (500 Pa) while the electron column operates at the conventional high vacuum. I am very interested in buying a variable pressure SEM with an EDS, but was recently warned that EDS has very poor spatial resolution in the variable pressure mode because of the large beam spread due to electrons scattering off the gas molecules in the sample chamber. Is this really a significant issue? Do any of you have experience using EDS with variable pressure SEMs? Thanks,
In short, it is an issue, but not intractable. For careful quantitative work we reduce the vacuum as much as the sample permits. We can usually get by with about 40 Pa of helium atmosphere on concrete samples. Even at 40 Pa we can pick up percent levels of elements away from the beam. For example, a Co chip in a standard mount can easily show 1% Fe from the stainless steel carrier. For qualitative work, that is not much of a problem, but if you are looking for trends in that element (Fe in Co), it would be impossible in low vacuum mode.
BTW, Helium greatly reduces the scattering compared to air at the same pressure.
I would encourage you to go ahead and get the VP SEM, but work through a few exercises with it to get a feel for its limits.
At 05:22 PM 3/20/2000 -0500, you wrote: } It seems that nowadays every SEM vendor is offering variable pressure } models, which are conventional SEMs with a plumbing modification that } allows the sample to be at pressures of up to about 4 torr (500 Pa) while } the electron column operates at the conventional high vacuum. I am very } interested in buying a variable pressure SEM with an EDS, but was recently } warned that EDS has very poor spatial resolution in the variable pressure } mode because of the large beam spread due to electrons scattering off the } gas molecules in the sample chamber. Is this really a significant issue? } Do any of you have experience using EDS with variable pressure SEMs? } Thanks,
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
It is a good point to contact DIATOME. It reminds to me, also, that they (DIATOME) may cut your samples on site and sent back the grids (if I understand them correctly) for free. So, you may try 45 and 35o knife and compare the results before you make final decision. As for 35o. 45o is more universal and durable. If you rich enough only for one diamond knife, it should be 45o, I believe. 35o supposed to be a "second knife" for "special occasions". I have no financial interest in DIATOME, but happy user of DIATOME knife
Sergey
} Date: Mon, 20 Mar 2000 12:00:52 -0700 (MST) } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } Subject: Re: 35 Degree Angle Diamond Knive } X-Sender: hcrowley-at-odin.cair.du.edu } To: "Stefan.Geimer" {stefan.geimer-at-uni-koeln.de} } Cc: Microscopy-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Diatome, for one, can supply you with clear ev idence for significantly reduced compression with a 35 degree knife over a 45 degree knife. No surprise there, since compression should be reduced with knife angle in materials which compress significantly. We are a 'materials science' lab, where we section all manner of metals and alloys, composites, powders, fibres, coatings, thin films, etc, etc. For us reduced compression is not a big deal, but durability obviously is. I can report that, for the last 5 years or so since their acquisition, our two Diatome 35 degree knives are used about 90% of the time, with no catastrophic failures or large increase in sharpening frequency.
I agree with Hildegard Crowley that breaking out is far more likely to be due to embedding/bonding problems. another possibility is simply a dull edge. Your increased success with a fresh (presumably demo) knife could indicate the latter. If the edge is quite rounded from use, the effective angle at the point of sectioning will shoot up incredibly, which will then increase the compressive forces that expose any embedding/bonding deficiencies.
That said, if you can only afford one knife, look in the mirror and ask yourself how careful are the people who might use the knife. A simple consideration of the forces on a knife edge during sectioning will tell you that a lesser angle should be slightly more susceptible to side forces. However, just the thought of a reduced angle might make an inexperienced operator sufficiently nervous to increase that risk greatly!
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: Stefan.Geimer } Sent: Monday, March 20, 2000 9:59 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: 35 Degree Angle Diamond Knive } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are planning to buy a new diamond knive for sectioning biological } samples. My question is if anybody has experience with using 35 degree } angle diamond knives and if they have any disantvantages compared to 45 } degree angle diamond knives. We are mainly sectioning unicellular algae } embedded in LR-gold and hope to reduce compression problems by using a } 35 degree angle knive. Our problem is that, especially algae with thick } cell walls, tend to fall out of the sections. This problem seems to be } caused by compressions, because using a brand-new 45 degree angle } diamond knive I had much less algae falling out of the sections. Because } we can effort only one new knive we have to use it as an all-round } knive. Is a 35 degree knive suitable to replace a 45 degree knive or is } it much more fragile and/or less durable? } } } Thanks in advance for your time and help. } } Sincerely, } } Stefan Geimer } } } } *********************** } Stefan Geimer } University of Cologne } Botanical Institute } Gyrhofstr. 15 } D-50931 Cologne } Germany } phone: +49-(0)221-470-3795 } fax: +49-(0)221-470-5181 } ************************ } } } }
Can anyone help Karen Maxwell?, reply to her as well as the list as she is not a subscriber.
Nestor Your Friendly Neighborhood SysOp
Below is the result of your feedback form. It was submitted by (maxwell-at-lec.med.utoronto.ca) on Wednesday, March 15, 2000 at 17:03:53 ---------------------------------------------------------------------------
Question: I have been looking at a paper from 1984 (Kochan et al) where they used EM to study the preconnector from bacteriophage lambda. They discovered that the axial hole in the ring shaped structure was 2.2-2.5 nm in diameter. They used negative staining with ammonium molybdate, uranyl acetate, and uranyl formate. In more recent studies (Valpuesta et al, 1999), the connector of phi29 was examined using a three-dimensional cryo-reconstruction, and the axial hole was found to be 3.3 nm. These are two different phage, but my question is, could the old techniques used in the case of the bacteriophage lambda preconnector (negative staining) have under-estimated the diameter of the axial hole? And if so, do you have an idea of the percent error that may be inherent in the technique? Is the cryo-reconstuction likely to give a value that is more accurate? Could you recommend any references that would address these questions?
At 04:22 PM 3/20/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have not had an opportunity to try these systems but from what I can glean, they are not totally similar to conventional SEMs. In particular, the VP SEMs do not use the E-T detector, but rather a gas discharge detector. There are some good examples of these images around but I do wonder what the actual difference is. Perhaps owners can tell us. Or warn us???
At 04:22 PM 3/20/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have not had an opportunity to try these systems but from what I can glean, they are not totally similar to conventional SEMs. In particular, the VP SEMs do not use the E-T detector, but rather a gas discharge detector. There are some good examples of these images around but I do wonder what the actual difference is. Perhaps owners can tell us. Or warn us???
Everett Ramer wrote: } It seems that nowadays every SEM vendor is offering variable pressure } models, which are conventional SEMs with a plumbing } modification that allows the sample to be at pressures of up to } about 4 torr (500 Pa) while the electron column operates at the } conventional high vacuum. I am very interested in buying a } variable pressure SEM with an EDS, but was recently warned that } EDS has very poor spatial resolution in the variable pressure } mode because of the large beam spread due to electrons } scattering off the gas molecules in the sample chamber. Is this } really a significant issue? Do any of you have experience using } EDS with variable pressure SEMs? } Thanks,
The fraction of primary electrons that are scattered by the gas in the specimen chamber is approximately given by:
1-I/Io = 1 - exp(-psL/kT)
where p is the pressure s the total scattering cross section for scattering of x keV electrons on the gas used L the distance between the last pressure limiting aperture and the sample k the Boltzmann constant and T the absolute temperature.
You can therefore reduce the contribution of X-rays excited by scattered primary electrons by reducing p (lowest possible pressure), s (use gas with low scattering cross section and the highest possible acceleration voltage balanced against overvoltage considerations) and L.
As Warren Straszheim wrote this will overcome the beam skirt problem to a large degree. In the situations where this is not sufficient, you can use an extrapolation method that has been developed here at Risoe National Laboratory. In short, you perform the analysis at various different pressures and use the above equation to extrapolate to the result you would get under zero pressure. A more detailed description can be found at the web-address
***************************** J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I was wondering if anybody was using a GIF filter. We have one mounted on a 120 kV TEM and are trying to image small particles (5-10 nm) and doing some energy filtering to image different elements in the particles (for example gold, silicon, copper, cerium...) . This is not really what I would call trivial work and we are still working out the set up (window size and positionning for example) in a rather empiric way. The main problem is often to get a decent signal in the interesting energy region and still keep the windows fairly narrow. I would add that we often have to work at low dose because of the beam sensitivity of our samples, but I guess our GIF setups can be improved. Any suggestion ?
I am staining virus infected cell DNA (in cell culture) with Hoechst 33258 fluorescent stain. I need to make correlation of the fluorescent spots with cellular structure.
Do you know of any counter-stain that can be used without destroying the fluorescent Hoechst staining?
Thanks Dr. A.P. Alves de Matos biologist apmatos-at-ip.pt
Diana: TAAB Laboratories Equipment, Ltd, 3 Minerva Court House,Calleva Park Aldermaston, Berks,RG7 8NA, UK
They list single edge razor blades, with and without a bac k in stainless and carbon steels. They also list double edge razor blades, 0.004 in. thick, in stainless steel.
Best regards,
Sam Purdy } ---------- } From: Jeff & Wanda Gray } Sent: March 2000 10:12 PM } To: Diana Papoulias; microscopy-at-sparc5.microscopy.com } Subject: RE: accu-edge blades } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Diana, } These blades are available from Allegiance Scientific (used to be Baxter, } used to be Scientific Products, used to be....). As far as I know, this is } the only place that carries this brand. } I have no financial interest in Allegiance, just passing along a fact. } Wanda Shotsberger } (HT ASCP) } } -----Original Message----- } } From: Diana Papoulias [mailto:Diana_Papoulias-at-usgs.gov] } Sent: Thursday, March 16, 2000 2:53 PM } To: microscopy-at-sparc5.microscopy.com } Subject: accu-edge blades } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Could someone please tell me where I can purchase accu-edge microtome } blades? } } Thank you. } } Diana Papoulias } USGS } 4200 New Haven Rd } Columbia, MO 65201 } } T:573 876 1902 } F:573 876 1876 } E:Diana_Papoulias-at-usgs.gov } } }
The extrapolation method (aka Variable Pressure Method) described by Dr. Bilde-Sorenson has been implemented by EDAX in a software feature called ViP-Quant. With this method the EDS analysis under Bad-Vacuum conditions can be done with nearly the same accuracy as under High-Vacuum conditions. Although this method was developed with the ESEM microscope in mind, it of course can be applied to all Bad-Vacuum scanning electron microscopes.
Please contact your local EDAX representative for more information and a copy of the new ViP-Quant brochure, or request one through www.edax.com.
} -----Original Message----- } From: Joergen Bilde-Soerensen 5802 [mailto:j.bilde-at-risoe.dk] } Sent: Tuesday, March 21, 2000 10:04 AM } To: Everett Ramer } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: EDS in Variable Pressure SEM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Everett Ramer wrote: } } It seems that nowadays every SEM vendor is offering variable pressure } } models, which are conventional SEMs with a plumbing } } modification that allows the sample to be at pressures of up to } } about 4 torr (500 Pa) while the electron column operates at the } } conventional high vacuum. I am very interested in buying a } } variable pressure SEM with an EDS, but was recently warned that } } EDS has very poor spatial resolution in the variable pressure } } mode because of the large beam spread due to electrons } } scattering off the gas molecules in the sample chamber. Is this } } really a significant issue? Do any of you have experience using } } EDS with variable pressure SEMs? } } Thanks, } } The fraction of primary electrons that are scattered by the gas in } the specimen chamber is approximately given by: } } 1-I/Io = 1 - exp(-psL/kT) } } where p is the pressure } s the total scattering cross section for scattering of x keV } electrons on the gas used } L the distance between the last pressure limiting aperture and } the sample } k the Boltzmann constant } and T the absolute temperature. } } You can therefore reduce the contribution of X-rays excited by } scattered primary electrons by reducing p (lowest possible } pressure), s (use gas with low scattering cross section and the } highest possible acceleration voltage balanced against } overvoltage considerations) and L. } } As Warren Straszheim wrote this will overcome the beam skirt } problem to a large degree. In the situations where this is not } sufficient, you can use an extrapolation method that has been } developed here at Risoe National Laboratory. In short, you } perform the analysis at various different pressures and use the } above equation to extrapolate to the result you would get under } zero pressure. A more detailed description can be found at the } web-address } } { HYPERLINK http://www.risoe.dk/afm/news1new.htm }http://www.risoe.dk/afm/news1new.htm
Best regards, Joergen Bilde-Soerensen
***************************** J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
Dear Colleagues, I want to fix insects at particular moments of muscle activity. I plan to freeze the bugs in liquid nitrogen and section their heads, after including them in hard Durcupan. I have no experience in fixating and including frozen pieces. Note that hard resin should be used, provided the hardness of the insect cuticle. Does anybody know a simple method for the transference from liquid nitrogen to plastic? Thank your very much in advance, Claudio Dr. Claudio R.Lazzari lazzari-at-bg.fcen.uba.ar
------------------------------------------ Laboratorio de Fisiolog’a de Insectos Dpto. Cs. Biol—gicas, Univ. Buenos Aires Ciudad Universitaria, 1428 Buenos Aires Argentina Tel.(54 11) 4576 3300, ext. 332, FAX (54 11) 4576 3384
Gabriel Adriano Rosa Centro de Imagenes y Microscopia Departamento de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA TE -(54-11)-4576-3349 e-mail cimic-at-bg.fcen.uba.ar FAX (54-11)-4576-3384
I don't think 7-AAD will work - it is used as a live/dead stain, and I know that it works much better on detergent-permeabilized cells than on merely PFA-fixed ones.
If you are contemplating microinjection (ugh!), maybe try the fluorescent dextrans instead of UTPs...they won't mess up your nucleic acids. We use them as injection markers and they don't seem to harm the cells.
Tamara Howard CSHL
On Tue, 21 Mar 2000, Donald O'Malley wrote:
} Hi Folks, } } Thanks for the flurry of info about DNA/RNA staining. I've summarized some of this info below in case anyone else has needs related to ours. } What I neglected to mention is that we are trying to count nuclei inside of living mouse embryos. That's why we're searching for a non-invasive, membrane crossing, nuclear-selective dye. } } Summary of recent listserv messages: } } } Possible Dyes for live TISSUE, visible wavelength, } selective NUCLEAR staining: } } 7 aminoactinomycin. But does it cross membranes? } microinjected Alexa-UTPs
I just got back from PITTCON. When I was visiting the EDAX booth they showed me a new program through which they have resolved this problem. I would suggest at least inquiring.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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 is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 05:22 PM 3/20/00 -0500, Everett Ramer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Alves de Matos, Regarding your question about counter staining for cellular structure--have you considered scanning your Hoechst image and overlaying a Nomarski image? If this doesn't yield enough detail there are fluorescent dyes specifically for organelles such as mitochondria and golgi. Molecular Probes sells them.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
Karl E. Garsha, Doctoral Student AOP Fellow Chief Coordinator-Graduate Student Association Department of Biological Sciences University of Wisconsin-Milwaukee P.O. Box 413 Milwaukee, WI 53201 Office: 459 Lapham Hall Phone: (414) 229-4316 Mobile: (414) 617-4295 E-Mail: keg-at-csd.uwm.edu
---------- } From: "Karl E. Garsha" {keg-at-csd.uwm.edu} To: maxwell-at-lec.med.utoronto.ca
Firewire Camera Anyone? Does anyone know of, or have information on current or upcoming digital cameras for light microscopy that utilize the Firewire or USB interface for quick and easy importing of light microscope images through a color digital camera, for example to a Mac G4 (or even the iMac G3)? Does this technology exist in a microscope camera? (yet)
In a message dated 03/21/2000 5:15:50 PM US Mountain Standard Time, HUGGINBJ-at-BP.com writes:
{ { Firewire Camera Anyone? Does anyone know of, or have information on current or upcoming digital cameras for light microscopy that utilize the Firewire or USB interface for quick and easy importing of light microscope images through a color digital camera, for example to a Mac G4 (or even the iMac G3)? Does this technology exist in a microscope camera? (yet) } }
The only one I am aware of is the Optronics MagnaFire, which is a cooled camera with the IEEE-1394 FireWire interface.
Previous versions of the Optronics cameras used a parallel (printer) port interface so they had to be used on PC's only. It would be worth inquiring about Mac compatibility. Sorry I don't have the answer to this part...I only know the MagnaFire has the FireWire interface.
Visit {http://www.optronics.com} and click on "MagnaFire" for specs.
We are in the midst of writing a proposal for a new film scanner and I am hoping that someone out there can help with a recommendation for a top of the line film scanner. The major application would be TEM negatives. A model we are currently looking at is the Nikon LS-4500AF Multi Format Film Scanner.
If anyone has archived recommendations from previous discussions and could send them on to me, that would be wonderful.
Thank you, Valerie Leppert
Dept. of Chem. Eng. and Mat. Sci. U. of California, Davis
I do not work for Optronics, but know a Mac version of Drivers is due out very shortly for the Magnafire Camera. For more information, check out....
http://www.optronics.com
Also, Nikon has an SLR based 2.74 megapixel Digital Camera suitable for most Microscopy (including bright emission Fluorescence with higher ISO) called the D1, based on a N90s Body with a F-mount and Firewire connection. Approx. $5300
Nikon also has a much less expensive digital camera Coolpix 990 coming in the next few weeks. This is a 3.34 million pixel, true resolution camera with live NTSC out (for simultaneous real-time video) and USB connectivity with an Electronic Shutter Release built-in for around $1000. It too, can handle most Microscopy techniques, but is limited in fluorescence by its 100 ISO rating. It is a C-mount type camera.
**I have no corporate affiliation with/nor any financial agreement with Optronics, Inc. I do however have strong ties with Nikon (as you can see), but make very little money from either product ;)
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} } In a message dated 03/21/2000 5:15:50 PM US Mountain Standard Time, } HUGGINBJ-at-BP.com writes: } } { { Firewire Camera Anyone? } Does anyone know of, or have information on current or upcoming digital } cameras for light microscopy that utilize the Firewire or USB interface for } quick and easy importing of light microscope images through a color digital } camera, for example to a Mac G4 (or even the iMac G3)? Does this } technology exist in a microscope camera? (yet) } } } } } The only one I am aware of is the Optronics MagnaFire, which is a cooled } camera with the IEEE-1394 FireWire interface. } } Previous versions of the Optronics cameras used a parallel (printer) port } interface so they had to be used on PC's only. It would be worth inquiring } about Mac compatibility. Sorry I don't have the answer to this part...I only } know the MagnaFire has the FireWire interface. } } Visit {http://www.optronics.com} and click on "MagnaFire" for specs. } } Cheers, }
Dear Karl I disagree with you, that "UA in particular, will positively stain proteins". Let start from definition: positively stain object is a dark object on the light background (therefore stain penetrate/interact with object making it "stained", dark in the terms of EM); "negative staining" (NS) is opposite. We have light (stain do not interfere with the sample) object on the dark background of stain (or dark ring of stain around the object, depends from stain and technique). The beauty of the NS is that stain (UA in particular) easily penetrates cavities in the object making visible fine details (for instance, I was able to recognize variable and constant domains of the Fab of the IgG molecules as well as domains of the Fc using UA staining). May be this effect you call "positively staining". But it is not.
Yes, UA may stain RNA and DNA positively. For this reason UA gives us "mixed stain" for ribosomes (negative for proteins and positive for RNA components). The disadvantage UA is its low pH.
As for subject of the current discussion. I think Cryo in general will give us more correct information than others EM techniques. But, we have to keep in mind, that in Cryo-technique, to make image relatively contrast, people should go to very high overfocusing (from half to couple microns, I believe). For this reason, the best Cryo-results I know, yielded only 2-3 nm resolution (and this is a huge problem in Cryo - to get better resolution): the same or even worse as for "negative staining" (I expect 1.5 nm resolution for UA). We have to count resolution when compare the data. I lost the original message from which this discussion was started, but it seems to me, that difference between NS and Cryo was not so dramatic and may be comparable if we will count the resolution. In general, I don't believe any EM data if resolution is not shown. Talking about resolution is complicated because of the difference between "resolution of the instrument" (0.14 nm for TEM currently), "physical resolution of the method" (resolution limit for overfocusing, for instance, radiation damage, drift, noise/signal ratio etc) and "resolution of the sample" (for biological samples they are not the same: preparation of the sample may affect sample's intactness/structure). Cryo is the best for sample preservation, but it is low in contrast. UA may affect sample's structure (may not) but the resolution limited only by granularity of stain (in theory it is a couple angstroms).
As for "averaging" of the images, I am a little bit skeptical about that. It is great deal if your sample is uniform in shape (symmetry is a great help too). If your biological sample is functionally flexible (IgG for instance), averaging will "hide" some interesting details even if you will distribute images in classes.
In my point of view, it is difficult to extract absolute numbers from EM. Inside one methodology you could easily compare the data, but it becomes difficult when you want to compare the data obtained by different techniques. This is reality of EM. I think Scanning Probe Microscopy is very promising to obtain the direct measurements on the samples under physiological conditions (EM is so far from "physiological conditions", unfortunately). I am so sorry, EM!
} Date: Mon, 20 Mar 2000 16:46:25 -0600 } From: "Karl E. Garsha" {keg-at-csd.uwm.edu} } Subject: FW: neg stain vs cryo EM plus image averaging } To: microscopy-at-sparc5.microscopy.com } X-Mailer: Microsoft Outlook Express for Macintosh - 4.01 (295) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I posted a previous message asking for a counterstain for Hoechst 33258, however I fail to mention that I want to use the counterstain in brignt field ilumination, switching to fluorescence as needed. Something that reveals celular structure like Giemsa would be OK. Giemsa however does not alow simultaneous staining with Hoechst as far as I can see.
Regarding EFTEM on small particles, this should actually work quite well. That is, elemental mapping works reasonably if you choose the right objective apperture. You should simulate the spatial resolution to see what can be attained under your conditions (HT, Cc, Cs etc). However it will be very difficult to get real concentration information out of these images. I'm trying EELS with nanoprobe but so far no succes (everything gets destroyed before I take a spectrum, allignment is very very difficult) Still, focussing remains a problem. Comon practice is to focus at 100eV and then suppose everything is OK for higher losses. I do not understand the theory of this technique (blurring by finite slitwidth & Cc?) nor thus it work well. Any comments on this are welcome.
Jo
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
On Tue, 21 Mar 2000, GuessWho wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } I was wondering if anybody was using a GIF filter. We have one mounted } on a 120 kV TEM and are trying to image small particles (5-10 nm) and } doing some energy filtering to image different elements in the particles } (for example gold, silicon, copper, cerium...) . This is not really what } I would call trivial work and we are still working out the set up } (window size and positionning for example) in a rather empiric way. The } main problem is often to get a decent signal in the interesting energy } region and still keep the windows fairly narrow. } I would add that we often have to work at low dose because of the beam } sensitivity of our samples, but I guess our GIF setups can be improved. } Any suggestion ? } } Any general comment on the GIF warmly welcome ! } } Thanks } } Olivier } } }
A few weeks ago, when our server was working correctly (*??!!*), there were e-mails talking about the purchase of a new SEM. Now we appear to be back on line I felt that our ideas may be of use to the mailers?
As part of the consultancy side of our business we purchase SEM for our clients. This means we are buying instruments regularly and are therefore probably one of the few professional SEM purchasing organisations?
In our team we have staff who have worked for most of the SEM manufacturers as demonstrators and application specialists, so we have seen the purchase game from both sides and of course try to use this to our clients advantage. Unlike the average microscopist, who probably only buys two instruments in their career, we have been able over the years to formulate a purchase plan, a purchasing criteria that we apply to each purchasing project. In our position we are unable to buy from the nicest salesman or the normal "gut" feeling, we must be absolutely certain that we are purchasing the best instrument for our clients applications.
Set out below are the headings from a lecture that I give on instrument purchase. Only the basic ideas are presented ( I guess if you need more I shall be attacked by Microscopy Today to give just that) but you will get the theme. It is important to realise that the specimen chamber of a SEM, the specimen-detector geometry, determines what we will see. For this reason you will find that one instrument will do a particular task better than any other in the price range, you need test specimens that will sort this out for you. If you have a multi instrument laboratory I always feel that it is a good idea to have instruments from different manufacturers optimising the laboratory for optimising a wider range of applications.
"Buying a New Microscope"
How Do You Start?
Collect ALL the brochures and prices Form a view of the new facilities being made available If you do not understand any features ASK the salesman Check through all the users desired facilities? Determine who will use the instrument now? Would there be others if you purchased particular accessories?
Formulate a Purchase Specification
Price range - we always look one level higher Set essential instrument specifications Set desired instrument specifications give them points and produce a "desirability assessment"
Why use a consultant?
Only he will without prejudice talk to all interested parties Interdepartmental feuds could spoil the case A different questioner provides different answers Their wide knowledge base may develop additional features They will have experience producing a "desirability assessment" which often brings surprises. They will be experienced with ways of dealing with the sales staff, they will keep them off your back
How to handle the demonstration? DO NOT LET THEM
Show you how wonderful the alignment procedures are - you will not spend all day aligning the instrument! Dominate YOUR demonstration Take you into totally uninteresting areas of the instrument - you should be buying because of the image quality Show their favourite gimmick Use their own favourite specimen Take you out of the room when about to load one of your specimens Take you to a two hour lunch with lots of drink
How to handle the demonstration before you set off
Select a maximum of three specimens that you know really well and that are important to your laboratory The specimens must be capable of meaningful low and high magnification imaging Develop a demonstration criteria for each specimen Have a tie break specimen available as suggested by the consultant Provide each company with an exact demonstration programme
What a consultant would do during a demo
Time the demonstrator to produce images at specific magnifications under very specific conditions Encourage the demonstrator (this is not a customer v demonstrator competition) allow them to try their own ideas with each specimen also, give them information that you have found to be of importance with your specimens Stop the demonstrator taking you away from your standard path - you do wish to compare each instrument under exactly the same conditions there is no time for other deviations
After the demo
Layout the results Compare performance instrument to instrument and time taken to obtain the results Produce a short list Compare the short list with the "desirability assessment" Research local and national service performance of the best two instrument manufacturers, and the manufacturers reputations
Finally Decide upon the instrument that you want, the specification and then haggle
Hope this helps?
Steve Chapman Senior Consultant Protrain for Training and Consultancy in EM World Wide Tel & Fax 44+ 1280 814774 e-mail protrain -at-emcourses.com web site www.emcourses.com
The ESEM with a cryostage has been used to observe such things as ice cream and asphalt/solvent mixtures. The ice cream work was published out of the Cavendish Labs [THiel, Donald]. The other was a lab experiment.
Hi! I have problem obtaining a good replica using a rotary shadow coating method. Samples are proteins suspended in a Tris, NaCl and CaCl2 buffer. They are absorbed onto a mica and dried in vacuum. I am using platinum wire (0.1 mm in diameter, 8 cm long) wrapped around tungsten filament followed by carbon coating. This replica is then transferred onto the grids. Coating is done in Hitachi HUS 5GB vacuum evaporator: vacuum 10-6 torr, 20 mA current through the electrode for 12 sec. The distance from the electrode to the plate with the samples is 10 cm. The problem is that I get very little coating (so little that there is no replica) and if I go higher then 20 mA tungsten brakes. I have no problem when I use palladium/gold wire with same parameters but that gives coarse granulation. This is the first time I used this method and I run out of ideas how to improve it. Thanks in advance for your replies. Dorota
I have just caught up on some of the questions on this server and I think I can contribute to the blade discussion. As far as I remember Accu-Edge Blades were re-branded Feather blades sold by Anglia Scientific, a UK microtome manufacturer in the 1970's and 80's. Anglia were taken over by Shandon Scientific and in turn they all disappeared into Life Sciences International. Phil Parker of Anglia I understand is still designing microtomes for LSI.
In short for Accu-Edge read Feather. We supply these Feather blades as do other microtome suppliers,
Best regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, England
Call for Papers Michigan Microscopy & Microanalysis Society Spring Meeting
The Michigan Microscopy & Microanalysis Society (Local Affiliate of MSA) will hold its Spring Meeting May 12th, 2000 at the Genoe Woods Conference Center in Brighton, Michigan. If you are interest in presenting a microscopy related paper or attending, please contact Deborah Rothe for more information, email: drrothe1-at-dow.com .
The society encourages student participation by providing free meeting registration for students whom present a paper. A cash award is sponsor by the society vendors for the best student paper.
Robert C. Cieslinski Michigan Microscopy & Microanalysis
Robert C. Cieslinski The Dow Chemical Company Microscopy & Microanalysis (517) 636-6875 email: rccieslinski-at-dow.com
} The other Firewire camera I know if currently is the MicroImager. } Check out http://www.qimaging.com/ Dave
} } The only one I am aware of is the Optronics MagnaFire, which is a cooled } camera with the IEEE-1394 FireWire interface. } } Previous versions of the Optronics cameras used a parallel (printer) port } interface so they had to be used on PC's only. It would be worth inquiring } about Mac compatibility. Sorry I don't have the answer to this part...I only } know the MagnaFire has the FireWire interface. } } Visit {http://www.optronics.com} and click on "MagnaFire" for specs. } } Cheers, } } Bob Chiovetti
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
I would suggest that you look at UMAX Powerlook III. It is a flatbed scanner with built in transparency adapter, so you can scan reflective and transmissive media. Its resolution is 1200x2400 pixels. This should handle most anything you might have. I use this scanner all the time. They are about $1100 these days. For super high quality 35mm scanning, I use the Polaroid SprintScan 35+. It does 2700 dpi, D=3.4. It runs about $1500. For medium format and 4x5" transparent media, I use the Polaroid SprintScan 45. It runs about $4500 but will handle 35mm and 6x6cm media.
I would not recommend the Nikon.
gary g.
At 07:48 PM 3/21/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Valerie, I can't comment on the Nikon scanner but I have plenty to say about the Polaroid Sprintscan 45, most of which is probably best left unsaid! Let me summarize. We purchased the Polaroid Sprintscan 45 in late 1998 and planned to operate it on a PC with Windows 98 operating system. Our first unit was defective and was replaced, however, the trouble shooting process was very time consuming. Our second unit functioned intermittently. Again, after much trouble shooting we returned the unit to Polaroid Canada. After having it tested we were told that there were no problems with the Sprintscan 45. The unit was returned to us but continued to cause us grief with its erratic behavior. We spent more time trying to diagnose the problem and even called in a computer specialist who spoke directly with Polaroid's tech support personnel in an effort to resolve the problem. In the end we found that the Sprintscan appears not to like Windows 98. The scanner has been running sucessfully on an older PC with Windows 95. Why? Has there been an identified problem associated with Windows 98? If so why aren't customers alerted? Having this knowledge would have saved us an enormous amount of time and money. I have been waiting for a comment from Polaroid for well over a month.
I would be interested in hearing from other Sprintscan users who experienced similar problems.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
} -----Original Message----- } From: Valerie Leppert [SMTP:vjleppert-at-ucdavis.edu] } Sent: Tuesday, March 21, 2000 8:48 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Film Scanner Recommendations? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all- } } We are in the midst of writing a proposal for a new film scanner and I } am hoping that someone out there can help with a recommendation for a top } of the line film scanner. The major application would be TEM negatives. A } model we are currently looking at is the Nikon LS-4500AF Multi Format Film } Scanner. } } If anyone has archived recommendations from previous discussions and could } send them on to me, that would be wonderful. } } Thank you, } Valerie Leppert } } Dept. of Chem. Eng. and Mat. Sci. } U. of California, Davis } } vjleppert-at-ucdavis.edu }
Yes, there is a community of researchers involved in the use of EDS in ESEM (mostly with the Electroscan now Phillips microscope but the work is not exclusive of other low vacuum SEMs). See the Microscopy and Microanalysis Meeting Proceedings from the past several years (in 1999-see Wight pg 290, Carlton pg 292). EDS spatial resolution can be an issue depending on your conditions. Some general rules (to reduce the scattering and therefore improve the EDS spatial resolution) are to reduce the beam-gas-path length (shorten the distance the primary electrons need to travel thru the gas molecules), lower the chamber pressure (reduce the number of gas molecules in the path), use a less scattering gas (Helium has already been suggested), and use as high an accelerating voltage as reasonable. Several corrections have been suggested for removing the unwanted contributions to the EDS spectrum: the extrapolation method (already mentioned), beam stop method, spectral subtraction method, and continuum method. There are also simulation models that predict scattering by the gas, I am not aware of any that predict EDS spectra based on a multiphase or multicomponent system. I highly recommend that you take a typical sample and ask each of the microscope manufacturers demonstrate imaging and EDS in their scope.
I have no interest in any microscope or EDS system, {fontfamily} {param} Times {/param} Certain commercial software, equipment, instruments, or materials are identified in this report to specify adequately the experimental procedure. Such identification does not imply recommendation or endorsement by the National Institute of Standards and Technology, nor does it imply that the materials or equipment identified are necessarily the best available for the purpose. {/fontfamily}
} It seems that nowadays every SEM vendor is offering variable pressure models, which are conventional SEMs with a plumbing modification that allows the sample to be at pressures of up to about 4 torr (500 Pa) while the electron column operates at the conventional high vacuum. I am very interested in buying a variable pressure SEM with an EDS, but was recently warned that EDS has very poor spatial resolution in the variable pressure mode because of the large beam spread due to electrons scattering off the gas molecules in the sample chamber. Is this really a significant issue? Do any of you have experience using EDS with variable pressure SEMs?
} Thanks,
-------------------note: new mailing address----------------------
Scott Wight e-mail: scott.wight-at-nist.gov
NIST 222/A113 W voice: 301-975-3949
100 Bureau Dr STOP 8371 | fax: 301-417-1321
Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
} We are in the midst of writing a proposal for a new film } scanner and I am hoping that someone out there can help } with a recommendation for a top of the line film scanner. } The major application would be TEM negatives. ...
I'll not knock the Nikon scanner, but I do believe your ultimate choice should be based on trial. Most film scanners anticipate normal films and normal exposures ... which addresses the film's optical density (... defined as Dmax minus Dmin ... approximately 3 f/stops for every OD unit ...). For normal negatives this generally doesn't exceed '3', and for normal positives, rarely exceeds '3.4'. However, the OD for x-ray films and electron induced exposures are reported to approach '4'. You should take one of your most problematic and representative films and test the scanners, paying particular attention to the detail/noise in the densest areas of the negative. I'll also mention ... you need not be limited to dedicated film scanners. I don't recall the PPI resolution of the Nikon scanner, but some of the newest flatbeds, together with their transparency options, I'm sure can approach the the resolution of the Nikon (e.g., 1600ppi for the new Epson) ... although I would tend to believe the film scanners are the most likely to deliver the best recognition for higher Dmax.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ 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/
Hans, Could you please explain the main idea of the Vip-Quant algorithm. I cannot even imagine how quantitative analysis could have nearly the same accuracy as in high-vacuum mode without a priori knowledge of the composition of surrounding areas. Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } Sent: Tuesday, March 21, 2000 7:54 AM } To: Everett Ramer } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } Subject: RE: EDS in Variable Pressure SEM } } } Dear Everett, } } The extrapolation method (aka Variable Pressure Method) } described by Dr. } Bilde-Sorenson has been implemented by EDAX in a software } feature called } ViP-Quant. With this method the EDS analysis under Bad-Vacuum } conditions can } be done with nearly the same accuracy as under High-Vacuum conditions. } Although this method was developed with the ESEM microscope } in mind, it of } course can be applied to all Bad-Vacuum scanning electron microscopes. } } Please contact your local EDAX representative for more } information and a } copy of the new ViP-Quant brochure, or request one through } www.edax.com. } } With best regards, } } Hans Dijkstra }
We are planning to upgrade our water supply from a reverse-osmosis, deionization system by adding a "polishing" system. Since our volume requirements are low, we're considering the Millipore Simplicity Ultrapure Water System. Specs for this countertop unit are a resistivity of 18.2 megaohms-cm, with total organic content of {15 ppb for the product water. The water would be used for routine EM use, i.e., making up reagants, staining, immunolabeling, etc.
My question is: does anyone have any experience with these systems and would you be willing to share it with us? Please feel free to respond off-line.
Thanks very much. Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
We have a SprintScan 45, and while we don't work it to the bone, it has been relatively reliable(We've had it serviced once since it was purchased, and that was to have the internal motherboard replaced). I'm sorry to hear that you've had so many problems Paul. We've had ours since late 97, and it's been scanning consistently on a Win95 machine. It IS very fussy with SCSI--let this be a warning to everyone. It won't work with the new fast cards, but won't work with the really old cards either. We've currently got it on a Adaptec AHA 15XX series SCSI2 card with UW negotiation. We haven't been able to try it on a Win98 machine because all of our Win98 machines have SCSI cards which are too fast. I've heard good things about the Nikon(especially about Digital ICE). Minolta also has Digital ICE. The SprintScan isn't perfect, but it gets our job done. I would NOT recommend a flatbed with transparency adapter for dedicated film scanning tasks--in general graphic arts professionals agree that if you want to maximize scan quality from film, it's best to get the dedicated film scanner rather than the multi-option flatbed. Dedicated film scanners tend to have better bit depth too.
Of course, if money is no object, a drum scanner or an Imacon flextight scanner will bring you the best images.
Disclaimer--I have no financial stake in any of the aforementioned corporate entities blah blah blah
Sony makes 2 firewire cameras, the DFW-V300 and DFW-V500. Both are c-mount cameras and can be used on a light microscope. They are priced in a range of $2000 or so.
Seth Grotelueschen MIS, Inc. www.paxit.com sethg-at-paxit.com
Hans, Could you please explain the main idea of the Vip-Quant algorithm. I cannot even imagine how quantitative analysis could have nearly the same accuracy as in high-vacuum mode without a priori knowledge of the composition of surrounding areas. Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } Sent: Tuesday, March 21, 2000 7:54 AM } To: Everett Ramer } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } Subject: RE: EDS in Variable Pressure SEM } } } Dear Everett, } } The extrapolation method (aka Variable Pressure Method) } described by Dr. } Bilde-Sorenson has been implemented by EDAX in a software } feature called } ViP-Quant. With this method the EDS analysis under Bad-Vacuum } conditions can } be done with nearly the same accuracy as under High-Vacuum conditions. } Although this method was developed with the ESEM microscope } in mind, it of } course can be applied to all Bad-Vacuum scanning electron microscopes. } } Please contact your local EDAX representative for more } information and a } copy of the new ViP-Quant brochure, or request one through } www.edax.com. } } With best regards, } } Hans Dijkstra }
Hi Ya'll: We are looking for a CCD camera for our Philips 430 TEM. We would like to get the best camera for the best price, e.g., either buying a used camera or a new non-Gatan camera (Gatan seems to be twice as expensive as the others). We would be using the camera for bright field and high resolution TEM of materials (semiconductors) rather than for biological specimens. Does anyone have a camera they would like to sell/donate or does anyone have recommendations as to a less-expensive camera that they know can be used for materials applications. Thanks, Mike Coviello Lab Manager University of Texas -at- Arlington
I probably have rotary pump oil in my oil vapor diffusion pump (long story, situation remedied, I hope). My question is, how well do I have to clean out the diffusion pump? Solvent clean and bake out, or only wipe down? What are the probable effects of contamination? Will it affect the vacuum quite a bit?
After overhauling the vacuum system on our Zeiss 10/A TEM because a faulty anti-suction device allowed (a LOT) of r.p. oil to migrate throughout the vacuum lines, I am only getting a vacuum of 6 X 10-4 when I should get 5 X 10-5. Either I've got a leak or a contaminated d.p. When I cleaned out the d.p. I only wiped it out well, but did not wash it with solvent and bake it out. I am trying to clean it in place rather than desolder the 13 wires (again) to get it all the way out of the scope, and then have to resolder (again) while lying on my stomach in the dark recesses of the instrument with glasses that are the wrong focal length... Call me lazy, if you wish, but I'm trying to find the easiest but effective way to do this!
Any tips and tricks appreciated!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
The Sprintscan 45; some people swear by them, others swear at them.
I swear by mine. But only after much trouble, and what sounds identical to your experience. Here's what I found:
It really makes no difference as I see it whether one uses Win95 or Win98. The crux of the problem is the SCSI controller and how it talks to the scanner. The Sprintscan 35+ and the 45, the Agfa Arcus II, and several other scanners were totally flaky and crash prone on my system. I too sent back the Sprintscan 45 only to be told that it was fine. The SCSI adapter is a key item in this whole picture. Which adapter were/are you using?
At the time, I was running a FW SCSI system with an Adaptec AHA-2940UW. The rear plate connector on this host adapter is a 68-pin FW connector. The scanners are SCSI-I/II and are narrow, not wide....and definitely not fast. So, in order to run a scanner on a FW connector, one needs a FW--} SCSI-I cable and (this is the kicker) a high byte terminator on the host adapter external connector and a standard narrow SCSI terminator at the scanner. Good luck trying to get this high byte terminator. Adaptec has a part number for it but I gave up trying to get one from them. Maybe you will be luckier than I was.
The alternative is to connect to the narrow SCSI bus at the host adapter and snake that out the rear of the PC using a blank panel interface that accepts the 50-pin ribbon cable connector and presents a SCSI-II high density connector at the rear of the computer. This can work and it did with the Agfa scanner. But it did not work with the Sprintscans. Even when set for 10MB/s and no negotiation, it still would hang the system. The solution? I got a Mac G3/266. I still have it and I still use it for all scanning and photo input.
Very interesting, and totally successful. All scanners work flawlessly on the Mac's legacy external SCSI bus. I still am running FW SCSI inside for the disks, but using the external SCSI for scanners, CD-R, CD-RW and 8mm tape. The easiest way to run the scanners is via a common TWAIN interface plug-in in Photoshop. My current main flatbed is a UMAX Powerlook III. I have not tried it on my PC. My current PC is ATA-66 EIDE with an Adaptec AHA-2930 narrow SCSI adapter. It might run the UMAX and the Polaroid but everything is hooked up to the Mac. With DAVE on the Mac, all PCs, printers and the Mac talk to one another via TCP/IP on a 10BaseT LAN. Any node that needs external access gets it via an ISDN router. So there is no problem transporting files across platforms. Worst case is a Zip disk.
Any Mac system before the G4 should work fine. The pre-G3 systems are really cheap now. G3 systems can also be found rather inexpensively. A simple system is all that one needs. Heavy duty computing is done on the PC but photo input and scanning is done on the Mac. This solved all of my problems--and from your discussion, they were the same as you experienced.
Hope this helps.
gary g.
At 09:27 AM 3/22/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sergey, I appreciate your feedback. It is obvious to me that you are pretty good at this. First, I was unclear about the message to which I was responding. I was responding to the subject "bacteriophage question" by way of Nestor. To my limited knowledge, UA will positively stain proteins-UA is used as a positive stain in thin section TEM. I understand the concept(s) of negative staining well, and I appreciate that UA is one of the best negative stains for high resolution work (small granularity+nice density). Totally asymmetrical molecules may be 3-D reconstructed. It just takes a brute force approach involving thousands of micrographs of the molecule of interest randomly orientated in vitreous ice. Although your point about symmetry makes good sense-a more symmetrical molecule should yield a higher resolution reconstruction (require fewer micrographs for the same level of confidence in a structure). Your point about intrument limitations is also well recieved. A quick concept: both TEM negative staining and Scanning Probe (AFM) studies of protein macromolecules usually involve the use of charged surfaces to facilitate adherance and/or orientation of the molecules (i.e. carbon+glow discharge). Some feel that this may distort the shape of the molecule (and indeed it will depending on the charge). In the case of Scanning Probe, there are those who would say that the "quasi-physical" interaction of the probe (presumably in tapping mode for molecules under physiological conditions) distorts the reconstructed image of the protein. I'ld like to say "sorry probe", but I don't have enough confidence in these statements to do so. Cheers, Karl G.
----- Original Message ----- } From: Sergey Ryazantsev {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 22, 2000 1:09 AM
Dear Dorota
You have to use such amount of platinum that it will create a small drop of the melted metal at the end of the "V"-shape tungsten filament. If the drop will be large, it is very likely that filament will broke. It seems to me, 8 cm is a huge amount of platinum. Again, what is diameter of the filament? 20 mA is a tiny current, seems to me, for thermal evaporation. But, I never work on HUS 5GB, may be they have special setup. As for granulation, I don't think that platinum will dramatically improve the quality of your shadowing. Try platinum-palladium at least. May be, you have to try buffers, which are able to vaporize in vacuum: ammonium acetate or bicarbonate adjusted by CO2, for instance. For such applications I am using for many years 50-150 mM ammonium acetate buffer (with some additives if necessary, magnesium acetate, for instance) and tungsten shadowing by electron gun. I also use thickness monitor to control shadowing. I also use one or bi-directional shadowing: it resolves details better. I am using rotary shadowing only for DNA and not in all cases. If I have any problems with shadowing, I am using "test-object" - latex spheres to determine the problem. Sometimes the problem is just wrong angle of shadowing. If you have further questions, you may contact me off-listServer.
Sergey
} Date: Wed, 22 Mar 2000 10:13:39 -0400 } From: Dorota Wadowska {wadowska-at-upei.ca} } Subject: TEM-shadow coating } To: microscopy-at-sparc5.microscopy.com } Organization: University of P.E.I. } Priority: normal } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Our specimen was sprayed on to freshly cleaved mica, rotary shadowed with platinum, and carbon coated. We are having difficulty removing the carbon replica. Scoring around the edge did not help. Would anyone have a suggestion? Thankyou.
Donald Gantz Boston Univ School of Medicine gantz-at-med-biophd.bu.edu
Dear Listservers, Does anyone have a good method for skeletal muscle fixation for Transmission Electron Microscopy? We will not be able to use a perfusion method. A fixative method would also be greatly appreciated.
Many Thanks
Margery Stark Abbott Laboratories Department of Microscopy and Microanalysis D-45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202
Steve Chapman writes: } If you have a multi instrument laboratory I always feel } that it is a good idea to have instruments from different manufacturers } optimising the laboratory for optimising a wider range of applications. } Steve, You make some excellent points. However, I believe that, (just as you state) potential benefits can be had by choosing different manufacturers - at the same time you are missing some very significant opportunities if you create a facility with many different manufacturers. Two important, potential opportunities are: reduced training time and learning curves for multiple users, and, discounted service contracts from the manufacturer.
For example, we have in the recent past had the benefit of significantly reduced service contract costs by having multiple instruments from JEOL and Philips. At one time our research facility here, happened to have four JEOL EMs. These four SEMs covered three vastly different SEM instrumentation ranges (and eras) and serviced different applications. They were all very capable of handling the tasks for which they were purchased, and we were able to save big bucks each year over the cost of single instrument service contract prices. In addition we had multiple users who could go from one instrument to another without any significant additional training (or retraining) because of the relatively familiar user interface that accompanied these microscopes.
Just another perspective. Have Fun, Brad Huggins
} ---------- } From: Steve Chapman[SMTP:PROTRAIN-at-CompuServe.COM] } Sent: Wednesday, March 22, 2000 5:49 AM } To: American EM Soc } Subject: Buying a SEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi } } A few weeks ago, when our server was working correctly (*??!!*), there } were } e-mails talking about the purchase of a new SEM. Now we appear to be back } on line I felt that our ideas may be of use to the mailers? } } As part of the consultancy side of our business we purchase SEM for our } clients. This means we are buying instruments regularly and are therefore } probably one of the few professional SEM purchasing organisations? } } In our team we have staff who have worked for most of the SEM } manufacturers } as demonstrators and application specialists, so we have seen the purchase } game from both sides and of course try to use this to our clients } advantage. Unlike the average microscopist, who probably only buys two } instruments in their career, we have been able over the years to formulate } a purchase plan, a purchasing criteria that we apply to each purchasing } project. In our position we are unable to buy from the nicest salesman or } the normal "gut" feeling, we must be absolutely certain that we are } purchasing the best instrument for our clients applications. } } Set out below are the headings from a lecture that I give on instrument } purchase. Only the basic ideas are presented ( I guess if you need more I } shall be attacked by Microscopy Today to give just that) but you will get } the theme. It is important to realise that the specimen chamber of a SEM, } the specimen-detector geometry, determines what we will see. For this } reason you will find that one instrument will do a particular task better } than any other in the price range, you need test specimens that will sort } this out for you. If you have a multi instrument laboratory I always feel } that it is a good idea to have instruments from different manufacturers } optimising the laboratory for optimising a wider range of applications. } } "Buying a New Microscope" } } How Do You Start? } } Collect ALL the brochures and prices } Form a view of the new facilities being made available } If you do not understand any features ASK the salesman } Check through all the users desired facilities? } Determine who will use the instrument now? } Would there be others if you purchased particular accessories? } } Formulate a Purchase Specification } } Price range - we always look one level higher } Set essential instrument specifications } Set desired instrument specifications give them points and produce a } "desirability assessment" } } Why use a consultant? } } Only he will without prejudice talk to all interested parties } Interdepartmental feuds could spoil the case } A different questioner provides different answers } Their wide knowledge base may develop additional features } They will have experience producing a "desirability assessment" which } often } brings surprises. } They will be experienced with ways of dealing with the sales staff, they } will keep them off your back } } How to handle the demonstration? DO NOT LET THEM } } Show you how wonderful the alignment procedures are - you will not spend } all day aligning the instrument! } Dominate YOUR demonstration } Take you into totally uninteresting areas of the instrument - you should } be } buying because of the image quality } Show their favourite gimmick } Use their own favourite specimen } Take you out of the room when about to load one of your specimens } Take you to a two hour lunch with lots of drink } } How to handle the demonstration before you set off } } Select a maximum of three specimens that you know really well and that are } important to your laboratory } The specimens must be capable of meaningful low and high magnification } imaging } Develop a demonstration criteria for each specimen } Have a tie break specimen available as suggested by the consultant } Provide each company with an exact demonstration programme } } What a consultant would do during a demo } } Time the demonstrator to produce images at specific magnifications under } very specific conditions } Encourage the demonstrator (this is not a customer v demonstrator } competition) allow them to try their own ideas with each specimen also, } give them information that you have found to be of importance with your } specimens } Stop the demonstrator taking you away from your standard path - you do } wish } to compare each instrument under exactly the same conditions there is no } time for other deviations } } After the demo } } Layout the results } Compare performance instrument to instrument and time taken to obtain the } results } Produce a short list } Compare the short list with the "desirability assessment" } Research local and national service performance of the best two instrument } manufacturers, and the manufacturers reputations } } Finally } Decide upon the instrument that you want, the specification and then } haggle } } Hope this helps? } } Steve Chapman } Senior Consultant } Protrain for Training and Consultancy in EM World Wide } Tel & Fax 44+ 1280 814774 } e-mail protrain -at-emcourses.com } web site www.emcourses.com }
Dear Karl, As my knowledge on thin sections, UA stains DNA and RNA, lead citrate - some proteins and OSO4 - some lipids. Sometimes UA (as well as any others stains) may just penetrate hydrophilic areas in the plastic sections (which correspondent, usually, with biological material areas). In this case, I agree, it may be named "positive staining". But in this case UA stain all areas, which accessible to water, therefore it is not specific "positive" stain for proteins.
As for 3D reconstruction. This is a subject for long and very interesting discussion. I know just a few examples of the complete 3D reconstruction of asymmetrical particles. Mostly, these works comes from Frank's (Spider program) and Marin van Heel (Imagic program) groups. The resolution on it is about 2 nm - not so impressive, if you count, that they did 3D on 26 nm ribosome. Frank's algorithm with random tilted particles (on the support film) has "dead angle" (between 60 and 90), therefore, the reconstruction generally speaking is not "complete". Van Hell's algorithm supposed to be free from such limitation, but it is not enough reconstructions performed to prove this technique. But, I am very optimistic about that in general. Both algorithms used "classification" of particles by type. Criteria for such classification determined by operator, therefore it is possible that in one class we will count slightly different particles (different shape or different orientation or both), averaging of these particles will enhance major features and lose fine details. Combination X-ray analysis and 3D reconstruction data (for phasing of the X-ray data set at the beginning) may help in this case. Marat Yusypov's grop did it for ribosome last year with great success. But, I have to point, using X-ray crystallography you studied some particular conformation under conditions of crystallization, which is not necessary correctly reflect the real conformation in solution.
As for Scanning Probe Microscopy, I agree that probe may distort the sample. To eliminate such problem, people scan the same area in different directions. If the images are the same, it means, that distortion from probe at least smaller than observed creatures. SPM may be very successful for membrane proteins (which are difficult for EM), pore complexes, etc. It is not necessary to use "modified" surfaces (glow-discharge, etc) for SPM or "negative staining" (I never use it for NS, the secret is a quality of the support carbon film). But, I agree with you, Karl that, as any "microscopy", SPM is limited in some way in their abilities. Sorry, STM.
Karl, thank you so much for such nice discussion.
Sergey.
} Date: Wed, 22 Mar 2000 15:39:32 -0600 } From: Karl Garsha {keg-at-csd.uwm.edu} } Subject: Re: neg stain vs cryo EM plus image averaging } To: Microscopy-at-sparc5.microscopy.com, Sergey Ryazantsev {sryazant-at-ucla.edu} } X-Mailer: Microsoft Outlook Express 5.00.2615.200 } X-MSMail-Priority: Normal } X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 } } Dear Sergey, } I appreciate your feedback. It is obvious to me that you are pretty good at } this. First, I was unclear about the message to which I was responding. I } was responding to the subject "bacteriophage question" by way of Nestor. } To my limited knowledge, UA will positively stain proteins-UA is used as } a positive stain in thin section TEM. I understand the concept(s) of } negative staining well, and I appreciate that UA is one of the best negative } stains for high resolution work (small granularity+nice density). } Totally asymmetrical molecules may be 3-D reconstructed. It just takes } a brute force approach involving thousands of micrographs of the molecule of } interest randomly orientated in vitreous ice. Although your point about } symmetry makes good sense-a more symmetrical molecule should yield a higher } resolution reconstruction (require fewer micrographs for the same level of } confidence in a structure). } Your point about intrument limitations is also well recieved. } A quick concept: both TEM negative staining and Scanning Probe (AFM) } studies of protein macromolecules usually involve the use of charged } surfaces to facilitate adherance and/or orientation of the molecules (i.e. } carbon+glow discharge). Some feel that this may distort the shape of the } molecule (and indeed it will depending on the charge). } In the case of Scanning Probe, there are those who would say that the } "quasi-physical" interaction of the probe (presumably in tapping mode for } molecules under physiological conditions) distorts the reconstructed image } of the protein. I'ld like to say "sorry probe", but I don't have enough } confidence in these statements to do so. } Cheers, } Karl G. } } ----- Original Message ----- } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, March 22, 2000 1:09 AM } Subject: FW: neg stain vs cryo EM plus image averaging } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Karl } } I disagree with you, that "UA in particular, will positively stain } } proteins". Let start from definition: positively stain object is a dark } } object on the light background (therefore stain penetrate/interact with } } object making it "stained", dark in the terms of EM); "negative staining" } } (NS) is opposite. We have light (stain do not interfere with the sample) } } object on the dark background of stain (or dark ring of stain around the } } object, depends from stain and technique). The beauty of the NS is that } } stain (UA in particular) easily penetrates cavities in the object making } } visible fine details (for instance, I was able to recognize variable and } } constant domains of the Fab of the IgG molecules as well as domains of the } } Fc using UA staining). May be this effect you call "positively staining". } } But it is not. } } } } Yes, UA may stain RNA and DNA positively. For this reason UA gives us } } "mixed stain" for ribosomes (negative for proteins and positive for RNA } } components). The disadvantage UA is its low pH. } } } } As for subject of the current discussion. I think Cryo in general will } } give us more correct information than others EM techniques. But, we have } } to keep in mind, that in Cryo-technique, to make image relatively } contrast, } } people should go to very high overfocusing (from half to couple microns, I } } believe). For this reason, the best Cryo-results I know, yielded only 2-3 } } nm resolution (and this is a huge problem in Cryo - to get better } } resolution): the same or even worse as for "negative staining" (I expect } } 1.5 nm resolution for UA). We have to count resolution when compare the } } data. I lost the original message from which this discussion was started, } } but it seems to me, that difference between NS and Cryo was not so } dramatic } } and may be comparable if we will count the resolution. In general, I } don't } } believe any EM data if resolution is not shown. Talking about resolution } } is complicated because of the difference between "resolution of the } } instrument" (0.14 nm for TEM currently), "physical resolution of the } } method" (resolution limit for overfocusing, for instance, radiation } damage, } } drift, noise/signal ratio etc) and "resolution of the sample" (for } } biological samples they are not the same: preparation of the sample may } } affect sample's intactness/structure). Cryo is the best for sample } } preservation, but it is low in contrast. UA may affect sample's structure } } (may not) but the resolution limited only by granularity of stain (in } } theory it is a couple angstroms). } } } } As for "averaging" of the images, I am a little bit skeptical about that. } } It is great deal if your sample is uniform in shape (symmetry is a great } } help too). If your biological sample is functionally flexible (IgG for } } instance), averaging will "hide" some interesting details even if you will } } distribute images in classes. } } } } In my point of view, it is difficult to extract absolute numbers from EM. } } Inside one methodology you could easily compare the data, but it becomes } } difficult when you want to compare the data obtained by different } } techniques. This is reality of EM. I think Scanning Probe Microscopy is } } very promising to obtain the direct measurements on the samples under } } physiological conditions (EM is so far from "physiological conditions", } } unfortunately). I am so sorry, EM! } } } } } Date: Mon, 20 Mar 2000 16:46:25 -0600 } } } From: "Karl E. Garsha" {keg-at-csd.uwm.edu} } } } Subject: FW: neg stain vs cryo EM plus image averaging } } } To: microscopy-at-sparc5.microscopy.com } } } X-Mailer: Microsoft Outlook Express for Macintosh - 4.01 (295) } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } _____________________________________________________ } } } } } } Karl E. Garsha, Doctoral Student } } } AOP Fellow } } } Chief Coordinator-Graduate Student Association } } } Department of Biological Sciences } } } University of Wisconsin-Milwaukee } } } P.O. Box 413 } } } Milwaukee, WI 53201 } } } Office: 459 Lapham Hall } } } Phone: (414) 229-4316 } } } Mobile: (414) 617-4295 } } } E-Mail: keg-at-csd.uwm.edu } } } } } } ---------- } } } } From: "Karl E. Garsha" {keg-at-csd.uwm.edu} } } } To: maxwell-at-lec.med.utoronto.ca } } } Subject: neg stain vs cryo EM plus image averaging } } } Date: Mon, Mar 20, 2000, 4:45 PM } } } } } } } } } Dear Karen, } } } My gut reaction: cryo-EM/3-D reconstruction will yield a much more } accurate } } } value with regard to measurement of macromolecular features. } } } Some "negative" stains, UA in particular, will positively stain } } } proteins, and so it would be expected that pore size might, in theory, be } } } over-estimated in neg. stain images. This is a shortcoming of neg. } staining } } } as opposed to cryo-EM (if the protein isn't glycosylated). I have no } doubt } } } that investigators more experienced than I would argue this point } however. } } } The real power of 3-D reconstruction is that it averages many images } and } } } determines a structure that is statistically probable...the derived } } } structure has a certain level of confidence. 3-D reconstruction, as well } as } } } 2-D averaging, may be performed on either neg. stained specimens } (Wilkens, } } } et. al., Journal of Biological Chemistry, 1999) or cryo-prepared } specimens } } } (many important citations...one of my favorites is Boisset, et. al., } Journal } } } of Structural Biology 109, 39-45 (1992). } } } Either way you need the computer software/hardware and theory } (Journal } } } of Structural Biology Vol. 116, Number 1, 1996). } } } My main point is that a (responsibly) digitally enhanced image should } } } provide more accurate measurement of macromolecular features than a raw } } } negative stain photo. } } } Best Regards, } } } Karl G. } } } } } } } } } _____________________________________________________ } } } } } } Karl E. Garsha, Doctoral Student } } } AOP Fellow } } } Chief Coordinator-Graduate Student Association } } } Department of Biological Sciences } } } University of Wisconsin-Milwaukee } } } P.O. Box 413 } } } Milwaukee, WI 53201 } } } Office: 459 Lapham Hall } } } Phone: (414) 229-4316 } } } Mobile: (414) 617-4295 } } } E-Mail: keg-at-csd.uwm.edu } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } http://www.bol.ucla.edu/~sryazant } } } } } } } } } } _____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I thank you as well for your knowledge and insight, Dr. Ryazanstev. I've enjoyed this discussion. -Karl ----- Original Message ----- } From: Sergey Ryazantsev {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 22, 2000 5:26 PM
At 05:59 PM 3/21/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It exists. But...and this is a big but....what performance do you seek? Most of these, at least from what I have seen, are not industrial strength. They are more consumer- than research-grade units. But maybe this will satisfy your needs. The higher quality units tend to use SCSI from what I have seen and used.
Donald Gantz wrote: =============================================== Our specimen was sprayed on to freshly cleaved mica, rotary shadowed with platinum, and carbon coated. We are having difficulty removing the carbon replica. Scoring around the edge did not help. Would anyone have a suggestion? Thankyou. =============================================== There are probably as many different ways to approach this kind of problem as there are people doing platinum replicas! The most reliable method we have ever discovered is to use a 1% aqueous polyacrylic acid solution, deposited as one drop on the surface with the stubborn replica. Twenty four hours later, the drop has spread, the water has evaporated leaving a thin but very tough PAA film. Then with something sharp (e. g. scalpel blade, but wear eye protection for this!), slide the blade edge underneath the edge of the PAA film, and if you have the "art", it will literally just "pop off" . Now you have the tough PAA layer and the carbon coating, and where it once rested on the mica, should now be a corresponding area free of replica.
Next the PAA is floated on a surface of water, carbon side up, and again, do other things and come back 24 hours later. The PAA will have all dissolved into the water, leaving the carbon/Pt film floating on the surface of the water. The film is then picked up on grids as you would any other floating film.
Note: Patience of clearly a virtue, be sure to give it the full 24 hours or your grids will be PAA contaminated, with a major loss of contrast. Use a deep petri dish for this so that there is sufficient volume of water to efficiently dissolve the PAA.
If this sounds too complicated and you don't have patience, there is an alternative we also use, it involves the use of Victawet® for EM (see our URL http://www.2spi.com/catalog/spec_prep/victawet.html
The Victawet is evaporated from a tungsten basket (see website instructions) and a very thin layer of the release agent is deposited onto the mica (or glass slide). Then apply your samples, shadow with Pt/C and the replica now is almost guaranteed to float off on the first try.
One note: The "better" the grade of mica, I am told, the easier is the release of such films. Grade V1 mica supposedly releases easier and that might be because there are fewer cleavage steps. We have not tested that theory ourselves so on that there can be no guarantees.....but it does make sense. If you are not familiar with the different mica "grades", see URL http://www.2spi.com/catalog/submat/chart.html
Disclaimer: SPI Supplies is a supplier of Victawet for Electron Microscopy and mica substrate materials.
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.spi.cc ######################## ============================================
I have one of these for sale. I may have two. These are specifically for microscopy. At highest resolution, they take a 28MB TIF file. This is as high of resolution I have ever seen. Interface is SCSI-II small sized narrow-SCSI connector.
For platinum: try 1-10% aqueous HF. Use it instead water to float replica. Insert mica slowly. Pickup replica on the grid and rinse with water. Use corrosion-resistant tweezers. Do not immerse grids into HF solution if it is copper. You have to do it fast to avoid corrosion of the grid. I was using copper grids with carbon perforated (holey) film. Perhaps, carbon protected grid form HF. This is easiest way for platinum shadowing.
You may use even 50% HF - it did not affect platinum shadowing as well as carbon. Be careful, HF is extremely corrosive, avoid direct contact and vapors.
If you have questions, you may contact to me off-line.
Good luck!
Sergey
} Date: Wed, 22 Mar 2000 17:31:48 -0400 (EDT) } From: "GANTZ-at-med-biophd.bu.edu"-at-sparc5.microscopy.com } Subject: Removal of Carbon Replica from Platinum-shadowed sample } To: MICROSCOPY-at-sparc5.microscopy.com } X-VMS-To: MICROSCOPY-at-MSA.MICROSCOPY.COM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I would like to thank those who have replied to my query about diff pump contamination. Most have asked for more details...
How long have I let the scope pump after cleaning it out and changing diff pump oil? Just shy of two weeks. The most improvement in vacuum came within 4 days, then leveled off.
Do I think I got oil into the viewing chamber or column? Yeah, lots. Lots. Most was on the anode plate, and quite a few droplets on the liquid nitrogen anticontaminator plate. And even a drop hanging off the filament! I can see all of you squirming... I cleaned the entire column, including a couple of parts I had never been into before, and all the vac lines. I was as thourough as possible, under the circumstances.
I presume I will have a fair amount of vapor around for years to come. However, the scope operation and resolution aren't bad, considering. The only thing that keeps me from slitting my wrist is that we have this great, new, fancy LEO 912 EFTEM that I have been trying to get all my users trained on, anyway. This means that the old Zeiss 10 can be my hobby scope. It reminds me of the Volkswagens I used to work on. Great German engineering!
So one friend thinks that small amounts of rotary pump oil in the diffusion pump will "burn off" and cease to be a problem, one thinks that it will only migrate up into the column, and a couple of third party service people don't have a clue what to expect. No "official" word from Zeiss/LEO yet. Hoping some of you clever microscopists will have either direct knowledge or a grand theory backed by thoughtful physics!
Aloha, Tina
Life's not all fun and Photoshop. **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
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With our super clean e-mail address lists you'll send less...and get better results...
* Y O U G E T W H A T Y O U P A Y F O R *
Our NEW 10 Million, Volume 9, address list CD will result in:
* Higher Response Rates * Higher Sales Conversion Ratios * More Receptive prospects; Less Flames & Non-Buyers. * Less Contact With Anti-Commerce Radicals & Extremists.
Remember that potential income chart at the beginning of this message? Can you imagine the kind of money you could make if you mailed one million pieces and sold only one tenth (.01%) of one percent? You do the math, you'll be amazed!
We've been in the list brokerage business for over 5 years and we've never compromised on quality. We won't release any address list until it passes our "high standards" test.
This is not a rental list that is restricted to a one-time mailing. You are purchasing an e-mail address list for your own personal mailings and may use it over-and-over.
DON'T HESITATE on this offer or you will miss out on the least expensive, legal and most effective way to market... PERIOD!
Order within 72 hours and we'll include the following FREE Bonuses... we call this our "BUSINESS ON A CD" bonus.
1. To help you get started we include basic proven Professional Mailing Software. This software has sold for as high as $499.00 in the past. No demo, but a full working version (SORRY, SINCE THE SOFTWARE IS FREE WE CANNOT OFFER ANY TECHNICAL SUPPORT, however set-up instructions are included).
2. Every survey has always indicated that the most profitable product to sell on the Internet is INFORMATION!
Our "BUSINESS ON A CD" gives you 650 reports/manuals/books that are yours to use and sell. With these "Special Reports" you may instantly start your "Information Product" business... plus a sample SALES LETTER is included to help you GET STARTED FAST!
3. "THE BULK E-MAIL SURVIVAL GUIDE" A manual/guide that addresses the Bulk E-Mail business. Especially useful for beginners. "THE BULK E-MAIL SURVIVAL GUIDE" will answer most of your questions and concerns about Bulk E-Mail. An exclusive for our customers... INCLUDED FREE.
4. "LISTMATE" - This is the software the Pro's use to manage their mailing lists. We've included two versions, both are fully functional demo's, the only limit is the file size.
5. "SCIENTIFIC ADVERTISING"! This book is responsible for untold millions of dollars in sales and profits. Many of today's internet "gurus" have used this powerful book as the foundation for marketing courses that they have written & sold for as much as $495. Marketeer's that have studied this book have been so deeply inspired, that it has changed their entire way of doing business, and they've gone on to make fortunes -- it's yours FREE with your order!
This "BUSINESS ON A CD" bonus is yours absolutely FREE if you order within the next 72 hours --- After that... . it's gone!
***SPECIAL BONUS*** Order within the next 72 hours and receive an additional 972,565 e-mail addresses as a prompt ordering bonus. Order Now!
D O N ' T H E S I T A T E E-Marketing is the most effective and fastest way to market anywhere... PERIOD!
O R D E R N O W . . . SAME DAY SERVICE (M-F) if your order is received before 2pm Pacific. 24hour fax service, just fax to: 1-435-806-3011
To order, via credit card simply cut/paste and print out the EZ ORDER FORM below and fax to our office today.
**** MILLIONS CD - Volume 9 ****
**** NOW ONLY $247! ****
This "Special Price" is in effect for the next 72 hours, after that we go back to our regular price of $299.00 ... Don't delay... you can be in business tomorrow!
We accept Visa, Mastercard, Amex and Checks by Fax. Fax your order to: 1-435-806-3011
----------------------Cut & Paste---------------------- ---------------------EZ Order Form---------------------
_____Yes! I want everything! I am ordering within 72 hours. Include my FREE "Business On A CD" bonus along with your 10 Million Vol. 9 E-Mail address CD (plus 972,565 bonus addresses) for the special price of only $247.00 + shipping as indicated below.
_____Oop's I missed the 72 hour "special". I am ordering Vol. 9 at the regular price of $299.00 + shipping.
***PLEASE SELECT YOUR SHIPPING OPTION***
____I would like to receive my package FedEx OVERNIGHT. I am including $15 for shipping. (Hawaii & Alaska $20 - Canada $25, all other International add an *additional* $25 [$40 total] for shipping)
____I would like to receive my package FedEx 2 DAY delivery. I'm including $10 for shipping. (Sorry FedEx 2nd Day is not available for shipments to Alaska, Hawaii, Canada or any International destination - Continental U.S. shipping addresses only).
***Please Print Carefully***
NOTE: Orders cannot be shipped without complete information including your signature. No exceptions!
COMPANY NAME____________________________________________
ADDRESS_________________________________________________ (FedEx can only ship to street addresses - no P.O. boxes)
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PHONE NUMBER____________________________________________ (required for shipping & tracking)
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NAME ON CARD___________________________________________
TOTAL AMOUNT (Including Shipping): $___________________
DATE:x__________________
(Required) SIGNATURE:x_________________________________ I understand that I am purchasing the Millions Vol. 9 e-mail address CD, the addresses are not rented, but are mine to use for my own mailing, over-and-over. Free bonuses are included, but cannot be considered part of the financial transaction. I understand that it is my responsibility to comply with any laws applicable to my local area. As with all software, once opened the CD may not be returned, however, if found defective it will be replaced with like product at no charge.
You may fax your order to us at: 1-435-806-3011
CHECK BY FAX SERVICES!
Please Note: Sorry, we can only accept checks drawn on U.S. banks.
If you would like to fax a check, tape your check below and fax it to our office along with the EZ Order Form to: 1-435-806-3011
If You fax a check, there is no need for you to mail the original. We will prepare a one-time draft, with the exact information on your original check. All checks will be held for bank clearance. (7-10 days) Make payable to: "CD-Marketing"
******************************************************** Do not reply to this message - ******************************************************** To register your e-mail address for removal from unsolicited mailings, visit: http://www.OptList.com ********************************************************
As they say, "It's the simple ideas that make money." And this is SIMPLE & GREAT INCOME!
Our research has found that many people have tried one or more of the following...
Free Classifieds? They just don't work anymore Web Site? Takes thousands of surfers Banners? Expensive and losing their punch E-Zine? Hope they have a -huge- subscriber list Search Engines? Forget it, unless you're in the top 20
S O W H A T D O E S W O R K ?
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2. Next, we use a filter list of 400+ words/phrases to clean even more. No address with inappropriate or profane wording survive!
3. Then we used our private database of thousands of known "extremists", opposed to commercial e-mail, and kicked off every one we could find.
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5. All domains have been verified as valid.
WHAT DID WE END UP WITH?
Volume 9... 10 Million Addresses Strong!
An address list so clean you'll want to start mailing today!
N O B R A G - J U S T F A C T
With our super clean e-mail address lists you'll send less...and get better results...
* Y O U G E T W H A T Y O U P A Y F O R *
Our NEW 10 Million, Volume 9, address list CD will result in:
* Higher Response Rates * Higher Sales Conversion Ratios * More Receptive prospects; Less Flames & Non-Buyers. * Less Contact With Anti-Commerce Radicals & Extremists.
Remember that potential income chart at the beginning of this message? Can you imagine the kind of money you could make if you mailed one million pieces and sold only one tenth (.01%) of one percent? You do the math, you'll be amazed!
We've been in the list brokerage business for over 5 years and we've never compromised on quality. We won't release any address list until it passes our "high standards" test.
This is not a rental list that is restricted to a one-time mailing. You are purchasing an e-mail address list for your own personal mailings and may use it over-and-over.
DON'T HESITATE on this offer or you will miss out on the least expensive, legal and most effective way to market... PERIOD!
Order within 72 hours and we'll include the following FREE Bonuses... we call this our "BUSINESS ON A CD" bonus.
1. To help you get started we include basic proven Professional Mailing Software. This software has sold for as high as $499.00 in the past. No demo, but a full working version (SORRY, SINCE THE SOFTWARE IS FREE WE CANNOT OFFER ANY TECHNICAL SUPPORT, however set-up instructions are included).
2. Every survey has always indicated that the most profitable product to sell on the Internet is INFORMATION!
Our "BUSINESS ON A CD" gives you 650 reports/manuals/books that are yours to use and sell. With these "Special Reports" you may instantly start your "Information Product" business... plus a sample SALES LETTER is included to help you GET STARTED FAST!
3. "THE BULK E-MAIL SURVIVAL GUIDE" A manual/guide that addresses the Bulk E-Mail business. Especially useful for beginners. "THE BULK E-MAIL SURVIVAL GUIDE" will answer most of your questions and concerns about Bulk E-Mail. An exclusive for our customers... INCLUDED FREE.
4. "LISTMATE" - This is the software the Pro's use to manage their mailing lists. We've included two versions, both are fully functional demo's, the only limit is the file size.
5. "SCIENTIFIC ADVERTISING"! This book is responsible for untold millions of dollars in sales and profits. Many of today's internet "gurus" have used this powerful book as the foundation for marketing courses that they have written & sold for as much as $495. Marketeer's that have studied this book have been so deeply inspired, that it has changed their entire way of doing business, and they've gone on to make fortunes -- it's yours FREE with your order!
This "BUSINESS ON A CD" bonus is yours absolutely FREE if you order within the next 72 hours --- After that... . it's gone!
***SPECIAL BONUS*** Order within the next 72 hours and receive an additional 972,565 e-mail addresses as a prompt ordering bonus. Order Now!
D O N ' T H E S I T A T E E-Marketing is the most effective and fastest way to market anywhere... PERIOD!
O R D E R N O W . . . SAME DAY SERVICE (M-F) if your order is received before 2pm Pacific. 24hour fax service, just fax to: 1-435-806-3011
To order, via credit card simply cut/paste and print out the EZ ORDER FORM below and fax to our office today.
**** MILLIONS CD - Volume 9 ****
**** NOW ONLY $247! ****
This "Special Price" is in effect for the next 72 hours, after that we go back to our regular price of $299.00 ... Don't delay... you can be in business tomorrow!
We accept Visa, Mastercard, Amex and Checks by Fax. Fax your order to: 1-435-806-3011
----------------------Cut & Paste---------------------- ---------------------EZ Order Form---------------------
_____Yes! I want everything! I am ordering within 72 hours. Include my FREE "Business On A CD" bonus along with your 10 Million Vol. 9 E-Mail address CD (plus 972,565 bonus addresses) for the special price of only $247.00 + shipping as indicated below.
_____Oop's I missed the 72 hour "special". I am ordering Vol. 9 at the regular price of $299.00 + shipping.
***PLEASE SELECT YOUR SHIPPING OPTION***
____I would like to receive my package FedEx OVERNIGHT. I am including $15 for shipping. (Hawaii & Alaska $20 - Canada $25, all other International add an *additional* $25 [$40 total] for shipping)
____I would like to receive my package FedEx 2 DAY delivery. I'm including $10 for shipping. (Sorry FedEx 2nd Day is not available for shipments to Alaska, Hawaii, Canada or any International destination - Continental U.S. shipping addresses only).
***Please Print Carefully***
NOTE: Orders cannot be shipped without complete information including your signature. No exceptions!
COMPANY NAME____________________________________________
ADDRESS_________________________________________________ (FedEx can only ship to street addresses - no P.O. boxes)
CITY, STATE, ZIP________________________________________
PHONE NUMBER____________________________________________ (required for shipping & tracking)
EMAIL ADDRESS___________________________________________ (Print Carefully - required in case we have a question and to send you a confirmation that your order has been shipped)
NAME ON CARD___________________________________________
TOTAL AMOUNT (Including Shipping): $___________________
DATE:x__________________
(Required) SIGNATURE:x_________________________________ I understand that I am purchasing the Millions Vol. 9 e-mail address CD, the addresses are not rented, but are mine to use for my own mailing, over-and-over. Free bonuses are included, but cannot be considered part of the financial transaction. I understand that it is my responsibility to comply with any laws applicable to my local area. As with all software, once opened the CD may not be returned, however, if found defective it will be replaced with like product at no charge.
You may fax your order to us at: 1-435-806-3011
CHECK BY FAX SERVICES!
Please Note: Sorry, we can only accept checks drawn on U.S. banks.
If you would like to fax a check, tape your check below and fax it to our office along with the EZ Order Form to: 1-435-806-3011
If You fax a check, there is no need for you to mail the original. We will prepare a one-time draft, with the exact information on your original check. All checks will be held for bank clearance. (7-10 days) Make payable to: "CD-Marketing"
******************************************************** Do not reply to this message - ******************************************************** To register your e-mail address for removal from unsolicited mailings, visit: http://www.OptList.com ********************************************************
This sounds very like the problem we had removing polymer single crystals deposited on mica. The heavy metal tends to "key" to the mica, as it also does if one is shadowing a bulk polymer specimen directly. What we do in both cases is to put on a tiny amount of carbon first, about one-tenth of the thickness of the main carbon film.
If the chemistry of your specimen allows, one might spray onto a polystyrene surface, and then dissolve the PS with butanone.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
On Wed, 22 Mar 2000 GANTZ-at-med-biophd.bu.edu-at-sparc5.microscopy.com wrote:
} Our specimen was sprayed on to freshly cleaved mica, rotary } shadowed with platinum, and carbon coated. We are having difficulty } removing the carbon replica. Scoring around the edge did not help. } Would anyone have a suggestion? Thankyou. } } Donald Gantz } Boston Univ School of Medicine } gantz-at-med-biophd.bu.edu }
More chemical can access the replica if the replica is scored into 3mm squares - which is required for TEM anyway. A backing of wax or plastic is useful if the replica tends to fall apart - I don't expect that this would happen on a mica surface. For separating and cleaning use a spotting plate or a small watchglass within a Petrie dish. I suggest to immerse the mica (or replicas) alternating in a solution of chromic acid (made up from Pot dichromate) and then concentrated household bleach - with a water rinse in between. Leave the mica and later the specimen in those solutions for several hours or over night. 2 periods in both solutions would be minimal for most tissues, but in your case, once the replica has separated, the actual cleaning should be minimal. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, March 23, 2000 7:32 AM, "GANTZ-at-med-biophd.bu.edu"-at-sparc5.microscopy.com [SMTP:"GANTZ-at-med-biophd.bu.edu"-at-sparc5.microscopy.com] wrote: } } Our specimen was sprayed on to freshly cleaved mica, rotary } shadowed with platinum, and carbon coated. We are having difficulty } removing the carbon replica. Scoring around the edge did not help. } Would anyone have a suggestion? Thankyou. } } Donald Gantz } Boston Univ School of Medicine } gantz-at-med-biophd.bu.edu
Doehne, E., A New Correction Method for High-Resolution Energy-Dispersive X-ray Analyses in the Environmental Scanniing Electron Microscope. Scanning 1997, Vol. 19, 75-78.
Carlton, R. A., Energy Dispersive X-ray Spectrometry in the Environmental Scanning Microscope. Microscope 1999, Vol. 47:1, 5-11.
Cheers,
Stephen M. Harmon Electron Microscopist United States Environmental Protection Agency M.S. 681 26 W. Martin Luther King Blvd. Cincinnati, OH 45268 513.569.7184 Harmon.Stephen-at-epa.gov
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It seems that nowadays every SEM vendor is offering variable pressure models, which are conventional SEMs with a plumbing modification that allows the sample to be at pressures of up to about 4 torr (500 Pa) while the electron column operates at the conventional high vacuum. I am very interested in buying a variable pressure SEM with an EDS, but was recently warned that EDS has very poor spatial resolution in the variable pressure mode because of the large beam spread due to electrons scattering off the gas molecules in the sample chamber. Is this really a significant issue? Do any of you have experience using EDS with variable pressure SEMs? Thanks,
Hi, I am interested in buying an ultramicrotome which is not necessary be a new one. Does anybody have idea where I should contact? Thank's in advances. Sincerely,
Young Ho Koh NSB program UMass Amherst, MA 01003 kohyh-at-bio.umass.edu
At 4:45 PM -0600 3/22/0, Stark,Margery wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
**************************
I've worked with muscle (skeletal and cardiac) for years. If you can't perfuse, you need to keep the muscles from contracting down on themselves. Depending on the size of the muscle to be fixed, it can be handled as a whole or blunt dissected into small (1-2mm diam) bundles and tied to applicator sticks before immersing it in fix. It helps if you've boiled the sticks in water for 10 minutes or so to extract any resins, etc. The smaller the bundle, the better your initial fixation. You may want to wash the bundles in bufer with "high" potassium (100mM KCl) to relax it. I fix in 2.5% glut, 4% paraform. + approx. 0.02% picric acid (2 ml of a sat'd sol'n in 40 ml of fix) in 0.1 M Na-cacodylate buffer(Ito & Karnovsky J Cell Bio 89(abstr. 418) 1968). You can keep the 100mM KCl in this too. I would suggest that you fix for 30 min or so at RT them overnight in the 'fridge. Wash well, then cut into smaller pieces before osmium. dehydrate, etc as usual. I like Spurr's resin, but feel free to use your favorite.
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
University of Central Florida Advanced Materials Processing and Analysis Center (AMPAC)
The Advanced Materials Processing and Analysis Center (AMPAC) is seeking candidates for a position as Assistant in Research to support the Materials Characterization Facility (MCF). The individual must have a Master degree or a Bachelor degree from an accredited institution in an appropriate area of study related to surface science and materials characterization, and have at least two years of experience in materials characterization or related areas. The person will be responsible for establishing and maintaining laboratory infrastructure, laboratory maintenance, equipment installation, operation and maintenance, and be an investigator in contracted research. The person must be able to conduct independent research and development in materials characterization with an emphasis in ion beam technology and have the ability to interact with students and provide instruction in the use and interpretation of data. The individual must have extensive knowledge and experience in the operation and maintenance of materials characterization equipment. AMPAC is particularly interested in individuals desiring an academic setting to further their professional goals. The applications will be reviewed beginning April 17, 2000 and will continue to be reviewed until the position is filled.
Applicants should send vitae and three references to Dr. Vimal Desai, Director, 12424 Research Parkway, Suite 408, Orlando, FL 32826. UCF is an equal opportunity/affirmative action employer. As an agency of the State of Florida, all application materials and selection procedures are available for public review.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Hi Ya''ll: In reference to my previous message: I'm sorry I might have given the impression that I feel the Gatan digital camera is overpriced...We have lots of Gatan equipment and are extremely happy with it. Maybe I should have said: we would like to know what are the lower-priced options for a CCD camera for a Philips 430 (without singling out Gatan). I am sorry for any offense this may have caused. Regards, Mike Coviello University of Texas -at- Arlington
The main idea of the ViP-Quant procedure is rather straight-forward and elegant. It is based on the phenomenon that as the pressure increases the size of the skirt effect remains fairly constant, although the number of skirt electrons increases, and thus the contribution of the skirt-effect to the EDS signal increases. This intensity increase is linear with the pressure.
This is valid for low- to medium pressures, or at a higher (ESEM) pressure with a very short free-path length of the skirt electrons, since in both cases you can assume the skirt electrons have scattered only once. At higher pressures with longer working distances these assumptions are no longer fully valid.
So to do an EDS analysis you take 2 measurements at different pressures, the low-pressure one at a pressure where you can just avaid charging, and a high-pressure one at at least twice that pressure. The measured intensities of all elements are then plotted as a function of pressure, and extrapolated to pressure zero, i.e. high vacuum. The extrapolated results will be very close to the results hat would have been obtained directly if the sample could have been analyzed at high vacuum conditions.
Of course anyone can do this plotting and extrapolation manually, that is: if your EDS software allows you to enter intensities manually! The only thing the EDAX ViP-Quant is offering as an extra is that the software does it automatically for you. There are no proprietary miracles involved, EDAX has just listened carefully to what science has to offer....
Best regards,
Hans Dijkstra
Disclaimer: The above is my personal opinion, and not necessarily EDAX's. ------------------------------------------------------------- EDAX Europe www.edax.com Ringbaan Noord 103 Tel.: +31-13-5364000 P.O. Box 4144 Fax.: +31-13-5356279 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl the Netherlands -------------------------------------------------------------
} -----Original Message----- } } From: "Dusevich, Vladimir", INTERNET:DusevichV-at-umkc.edu } To: Everett Ramer, INTERNET:Everett.Ramer-at-netl.doe.gov } "'Hans Dijkstra'", INTERNET:hans.dijkstra-at-edax.nl } } CC: Joergen Bilde-Soerensen 5802, INTERNET:j.bilde-at-risoe.dk } [unknown], INTERNET:Microscopy-at-sparc5.microscopy.com } } Date: 3/22/00 4:35 PM } } RE: RE: EDS in Variable Pressure SEM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hans, } Could you please explain the main idea of the } Vip-Quant algorithm. I cannot even imagine } how quantitative analysis could have nearly the } same accuracy as in high-vacuum mode without } a priori knowledge of the composition of } surrounding areas. } Thank you, } } Vladimir } } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } -----Original Message----- } } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } } Sent: Tuesday, March 21, 2000 7:54 AM } } To: Everett Ramer } } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } } Subject: RE: EDS in Variable Pressure SEM } } } } } } Dear Everett, } } } } The extrapolation method (aka Variable Pressure Method) } } described by Dr. } } Bilde-Sorenson has been implemented by EDAX in a software } } feature called } } ViP-Quant. With this method the EDS analysis under Bad-Vacuum } } conditions can } } be done with nearly the same accuracy as under High-Vacuum conditions. } } Although this method was developed with the ESEM microscope } } in mind, it of } } course can be applied to all Bad-Vacuum scanning electron microscopes. } } } } Please contact your local EDAX representative for more } } information and a } } copy of the new ViP-Quant brochure, or request one through } } www.edax.com. } } } } With best regards, } } } } Hans Dijkstra } } } } } } } } } ----------------------- Internet Header -------------------------------- } Sender: Microscopy-request-at-sparc5.microscopy.com } Received: from ultra5.microscopy.com ([206.69.208.10]) } by spdmgaac.compuserve.com (8.9.3/8.9.3/SUN-1.9) with ESMTP } id LAA16645; } Wed, 22 Mar 2000 11:34:33 -0500 (EST) } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id KAA01363 } for dist-Microscopy; Wed, 22 Mar 2000 10:27:23 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id KAA01359 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 22 } Mar 2000 10:26:53 -0600 (CST) } Received: from UMKC-MAIL01.umkc.edu (email.exchange.umkc.edu } [134.193.71.1]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id KAA01352 } for {Microscopy-at-sparc5.microscopy.com} ; Wed, 22 Mar 2000 } 10:26:41 -0600 (CST) } Received: by umkc-mail01 with Internet Mail Service (5.5.2650.21) } id {HJ6P423S} ; Wed, 22 Mar 2000 10:21:30 -0600 } Message-ID: {95A711A70065D111B58C00609451555C01CA7552-at-UMKC-MAIL02} } From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} } To: "'Hans Dijkstra'" {hans.dijkstra-at-edax.nl} , } Everett Ramer } {Everett.Ramer-at-netl.doe.gov} } Cc: Microscopy-at-sparc5.microscopy.com, } Joergen Bilde-Soerensen 5802 } {j.bilde-at-risoe.dk} } Subject: RE: EDS in Variable Pressure SEM } Date: Wed, 22 Mar 2000 10:21:28 -0600 } MIME-Version: 1.0 } X-Mailer: Internet Mail Service (5.5.2650.21) } Content-Type: text/plain; } charset="iso-8859-1" } Errors-to: Microscopy-request-at-sparc5.microscopy.com } }
I sometimes found this and never had a great explanation of why it happened. I would make sure that your bell jar is clean, i.e.free from oil. I did find that the thickness of the carbon sometimes mattered. If it was too thick it shattered. Too thin you cannot see it or it won't stay together. Sometimes in between it would not float off.
Good luck.
ML } } } Our specimen was sprayed on to freshly cleaved mica, rotary } shadowed with platinum, and carbon coated. We are having difficulty } removing the carbon replica. Scoring around the edge did not help. } Would anyone have a suggestion? Thankyou. } } Donald Gantz } Boston Univ School of Medicine } gantz-at-med-biophd.bu.edu
Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
At 11:25 AM -0500 3/23/0, koh young ho wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
***************** for used instruments in the NY Metropoitan are you can try: M.O.C. at (914)268-6450 and Marcus Meyerhoff (I'lm not sure I've spelled that correctly) at (732) 747-6228
My name is George Laing from National Graphic Supply. NGS is a vendor of electronic imaging and traditional photographic products to scientific markets. Our customers have had excellent results scanning TEM negatives with the Agfa Duoscan T2500 scanner. The T2500 looks like a traditional "flatbed" scanner but utilizes a unique "Twin Plate" design similar to a negative carrier, that eliminates the use of glass in the film holder. This eliminates Newton rings,dust on the scanner glass, etc. Optical resolution is 2500x2500ppi(maximum resolution 5000x5000), Dmax is 3.5. The T2500 has Apochromatic optics and also includes glassless film holders for 35mm, 120/220 and 4x5 films. Also included is a glass drawer for transparent originals up to 8"x10". Agfa's Fotolook software is included for Mac and Windows, interface is SCSI. In addition, the T2500 will also scan reflective originals up to 8.5" x 14". There are three models in the Duoscan family ranging from $750 to $5175 list price. I can provide sample output and literature for anyone who wishes to contact me directly or visit http://www.agfahome.com/product/CatProd_DisplayPublic.html?id=3115
George Laing National Graphic Supply E-mail: scisales-at-ngscorp.com Phone: (800) 223-7130 X3109 USA
Valerie, I can't comment on the Nikon scanner but I have plenty to say about the Polaroid Sprintscan 45, most of which is probably best left unsaid! Let me summarize. We purchased the Polaroid Sprintscan 45 in late 1998 and planned to operate it on a PC with Windows 98 operating system. Our first unit was defective and was replaced, however, the trouble shooting process was very time consuming. Our second unit functioned intermittently. Again, after much trouble shooting we returned the unit to Polaroid Canada. After having it tested we were told that there were no problems with the Sprintscan 45. The unit was returned to us but continued to cause us grief with its erratic behavior. We spent more time trying to diagnose the problem and even called in a computer specialist who spoke directly with Polaroid's tech support personnel in an effort to resolve the problem. In the end we found that the Sprintscan appears not to like Windows 98. The scanner has been running sucessfully on an older PC with Windows 95. Why? Has there been an identified problem associated with Windows 98? If so why aren't customers alerted? Having this knowledge would have saved us an enormous amount of time and money. I have been waiting for a comment from Polaroid for well over a month.
I would be interested in hearing from other Sprintscan users who experienced similar problems.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
} -----Original Message----- } From: Valerie Leppert [SMTP:vjleppert-at-ucdavis.edu] } Sent: Tuesday, March 21, 2000 8:48 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Film Scanner Recommendations? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all- } } We are in the midst of writing a proposal for a new film scanner and I } am hoping that someone out there can help with a recommendation for a top } of the line film scanner. The major application would be TEM negatives. A } model we are currently looking at is the Nikon LS-4500AF Multi Format Film } Scanner. } } If anyone has archived recommendations from previous discussions and could } send them on to me, that would be wonderful. } } Thank you, } Valerie Leppert } } Dept. of Chem. Eng. and Mat. Sci. } U. of California, Davis } } vjleppert-at-ucdavis.edu }
hello, I was wondering if I could get a sampling of opinions on the different attachments to light microscopes, dissecting microscopes to display video to an external monitor. I've seen a few different products available, and would like to know any experiences people may have had with different systems. Any comments are greatly appreciated, and I will happily make a summary of the information.
Sincerely, Gordon Vrdoljak.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
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The ongoing discussion about replica techniques continues to be very educational. I wish to suggest another approach to this analysis problem. Occasionally I jet polish foils of metals which contain precipitates. Certain acid mixtures seem more "agressive" and preferentially remove the precipitates. This seems prone to happen when using nitric or perchloric acid mixtures. However, a few years ago when I began to use "non-acid" electrolytes, precipitates were sometimes thinned beautifully and were still retained in the thinned foil. I once got a hole in the center of a large precipitate! For more information, see Ultramicroscopy, Vol.19,(1986). Perhaps more research should be funded along this "thread". A second approach has been to "design" less agressive electrolytes that rely on hydrochloric acid, for example, which sometimes works for some materials but is not in The general literature. Within safty considerations,worthwhile electrolyte improvements may be achieved with a bit of work! Good luck.
Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Avenue Argonne, Il., 60439
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id PAA05355 for dist-Microscopy; Thu, 23 Mar 2000 15:14:34 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id PAA05350 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 23 Mar 2000 15:14:04 -0600 (CST) Received: from UMKC-MAIL01.umkc.edu (email.exchange.umkc.edu [134.193.71.1]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id PAA05343 for {microscopy-at-sparc5.microscopy.com} ; Thu, 23 Mar 2000 15:13:52 -0600 (CST) Received: by umkc-mail01 with Internet Mail Service (5.5.2650.21) id {HJ6P40N0} ; Thu, 23 Mar 2000 15:08:43 -0600 Message-ID: {95A711A70065D111B58C00609451555C01CA7555-at-UMKC-MAIL02} "Dusevich, Vladimir" {DusevichV-at-umkc.edu} Cc: Microscopy Listserver {microscopy-at-sparc5.microscopy.com}
Hans, Thank you very much for explanations. But for me this method will not work - quite often I analyze wet specimens at pretty high pressure (5-6 Torr). Thank you again,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } Sent: Thursday, March 23, 2000 12:19 PM } To: DusevichV-at-umkc.edu } Cc: Microscopy Listserver } Subject: RE: EDS in Variable Pressure SEM } } } Dear Vladimir, } } The main idea of the ViP-Quant procedure is rather } straight-forward and } elegant. It is based on the phenomenon that as the pressure } increases the } size of the skirt effect remains fairly constant, although } the number of } skirt electrons increases, and thus the contribution of the } skirt-effect to } the EDS signal increases. This intensity increase is linear with the } pressure. } } This is valid for low- to medium pressures, or at a higher } (ESEM) pressure } with a very short free-path length of the skirt electrons, } since in both } cases you can assume the skirt electrons have scattered only } once. At higher } pressures with longer working distances these assumptions are } no longer } fully valid. } } So to do an EDS analysis you take 2 measurements at different } pressures, the } low-pressure one at a pressure where you can just avaid } charging, and a } high-pressure one at at least twice that pressure. The } measured intensities } of all elements are then plotted as a function of pressure, } and extrapolated } to pressure zero, i.e. high vacuum. The extrapolated results } will be very } close to the results hat would have been obtained directly if } the sample } could have been analyzed at high vacuum conditions. } } Of course anyone can do this plotting and extrapolation } manually, that is: } if your EDS software allows you to enter intensities } manually! The only } thing the EDAX ViP-Quant is offering as an extra is that the } software does } it automatically for you. There are no proprietary miracles } involved, EDAX } has just listened carefully to what science has to offer.... } } Best regards, } } Hans Dijkstra } } Disclaimer: The above is my personal opinion, and not } necessarily EDAX's. } ------------------------------------------------------------- } EDAX Europe www.edax.com } Ringbaan Noord 103 Tel.: +31-13-5364000 } P.O. Box 4144 Fax.: +31-13-5356279 } 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl } the Netherlands } ------------------------------------------------------------- } } } -----Original Message----- } } } } From: "Dusevich, Vladimir", INTERNET:DusevichV-at-umkc.edu } } To: Everett Ramer, INTERNET:Everett.Ramer-at-netl.doe.gov } } "'Hans Dijkstra'", INTERNET:hans.dijkstra-at-edax.nl } } } } CC: Joergen Bilde-Soerensen 5802, INTERNET:j.bilde-at-risoe.dk } } [unknown], INTERNET:Microscopy-at-sparc5.microscopy.com } } } } Date: 3/22/00 4:35 PM } } } } RE: RE: EDS in Variable Pressure SEM } } } } } } } -------------------------------------------------------------- } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } } } } Hans, } } Could you please explain the main idea of the } } Vip-Quant algorithm. I cannot even imagine } } how quantitative analysis could have nearly the } } same accuracy as in high-vacuum mode without } } a priori knowledge of the composition of } } surrounding areas. } } Thank you, } } } } Vladimir } } } } } } Vladimir M. Dusevich, Ph.D. } } Electron Microscope Lab Manager } } 3127 School of Dentistry } } 650 E. 25th Street } } Kansas City, MO 64108-2784 } } } } Phone: (816) 235-2072 } } Fax: (816) 235-5524 } } Web: http://www.umkc.edu/dentistry/microscopy } } } } } -----Original Message----- } } } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } } } Sent: Tuesday, March 21, 2000 7:54 AM } } } To: Everett Ramer } } } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } } } Subject: RE: EDS in Variable Pressure SEM } } } } } } } } } Dear Everett, } } } } } } The extrapolation method (aka Variable Pressure Method) } } } described by Dr. } } } Bilde-Sorenson has been implemented by EDAX in a software } } } feature called } } } ViP-Quant. With this method the EDS analysis under Bad-Vacuum } } } conditions can } } } be done with nearly the same accuracy as under } High-Vacuum conditions. } } } Although this method was developed with the ESEM microscope } } } in mind, it of } } } course can be applied to all Bad-Vacuum scanning electron } microscopes. } } } } } } Please contact your local EDAX representative for more } } } information and a } } } copy of the new ViP-Quant brochure, or request one through } } } www.edax.com. } } } } } } With best regards, } } } } } } Hans Dijkstra } } } } } } } } } } } } } } } } } ----------------------- Internet Header } -------------------------------- } } Sender: Microscopy-request-at-sparc5.microscopy.com } } Received: from ultra5.microscopy.com ([206.69.208.10]) } } by spdmgaac.compuserve.com (8.9.3/8.9.3/SUN-1.9) with ESMTP } } id LAA16645; } } Wed, 22 Mar 2000 11:34:33 -0500 (EST) } } Received: (from daemon-at-localhost) } } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id KAA01363 } } for dist-Microscopy; Wed, 22 Mar 2000 10:27:23 -0600 (CST) } } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP } id KAA01359 } } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 22 } } Mar 2000 10:26:53 -0600 (CST) } } Received: from UMKC-MAIL01.umkc.edu (email.exchange.umkc.edu } } [134.193.71.1]) } } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP } id KAA01352 } } for {Microscopy-at-sparc5.microscopy.com} ; Wed, 22 Mar 2000 } } 10:26:41 -0600 (CST) } } Received: by umkc-mail01 with Internet Mail Service (5.5.2650.21) } } id {HJ6P423S} ; Wed, 22 Mar 2000 10:21:30 -0600 } } Message-ID: {95A711A70065D111B58C00609451555C01CA7552-at-UMKC-MAIL02} } } From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} } } To: "'Hans Dijkstra'" {hans.dijkstra-at-edax.nl} , } } Everett Ramer } } {Everett.Ramer-at-netl.doe.gov} } } Cc: Microscopy-at-sparc5.microscopy.com, } } Joergen Bilde-Soerensen 5802 } } {j.bilde-at-risoe.dk} } } Subject: RE: EDS in Variable Pressure SEM } } Date: Wed, 22 Mar 2000 10:21:28 -0600 } } MIME-Version: 1.0 } } X-Mailer: Internet Mail Service (5.5.2650.21) } } Content-Type: text/plain; } } charset="iso-8859-1" } } Errors-to: Microscopy-request-at-sparc5.microscopy.com } } } } }
I've heard from several SEM/TEM independent maintenance companies that some of the manufacturer's have engaged in "price dumping" when negotiating a service contract.
In other words, if the manufacturer is aware that they have competition
for a service contract they will reduce the contract by as much as forty
percent.
I am all for the free enterprise system, but I thought that when a contract price is offered it is the same for all customers especially government contracts that must abide by the GSA rules. GSA rules states
that the price offered is the lowest price for this service. It would be illegal for the manufacturer (or anyone) to offer a contract at less than the cost offered to GSA customers.
I am surprised by this move as our contracts are the same as we abide by the GSA rules.
EDS in ESEM is a pretty similar thing to EDS in SEM if you do not need trace/minor (~1%) element analysis and use some standard precautions. Some vendors even sell as an option gaseous detectors which could be placed close (1mm) to a sample and greatly reduce a beam scattering. I routinely use EDS at water vapor pressure up to 6 Torr. I've put an X-ray map taken in environmental conditions at our web site http://www.umkc.edu/dentistry/microscopy If low concentrations are of no interest to you then you will not see real difference in performance of an EDS in VP and high vacuum modes, especially if you do not work with wet specimens which need really high pressure.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} } } It seems that nowadays every SEM vendor is offering variable } pressure models, which are conventional SEMs with a plumbing } modification that allows the sample to be at pressures of up } to about 4 torr (500 Pa) while the electron column operates } at the conventional high vacuum. I am very interested in } buying a variable pressure SEM with an EDS, but was recently } warned that EDS has very poor spatial resolution in the } variable pressure mode because of the large beam spread due } to electrons scattering off the gas molecules in the sample } chamber. Is this really a significant issue? Do any of you } have experience using EDS with variable pressure SEMs? } Thanks, } }
Some of the K-12 classrooms we serve in our Bugscope (http://bugscope.beckman.uiuc.edu/) program have expressed interest in sending us aquatic or marine invertebrates to prepare for their sessions. What I'd like to know is how I should ask them to prepare the samples for me (e.g., should I ask them to overnight pondwater or seawater samples in plastic bottles?), and after that, what should I do with them? I can't really be sending glutaraldehyde and cacodylate to gradeschool kids, but I'm assuming I'll want to do some sort of standard fixation/dehydration/HMDS treatment, presumably on Millipore filters, once I get samples. Any tips?
} We are in the midst of writing a proposal for a new film scanner and I } am hoping that someone out there can help with a recommendation for a top } of the line film scanner. The major application would be TEM negatives. A } model we are currently looking at is the Nikon LS-4500AF Multi Format Film } Scanner. } } Valerie Leppert } Dept. of Chem. Eng. and Mat. Sci. } U. of California, Davis
We use a Polaroid SprintScan 45 for SEM, TEM and other large film but find that the negatives need to be somewhat tailored to the scanner's needs. We have great difficulty with very dense or very high contrast negatives, especially from the TEM. Staining and microscope settings need to be adjusted to produce negatives that are within the range of the scanner. It then produces very good results.
_____________________________________ Tom Gore, Advanced Imaging Laboratory Biology Department, University of Victoria Box 3020, Station CSC, Victoria, BC, V8W 3N5 Canada voice (250) 721-7134 fax (250) 721-7120 web: http://web.uvic.ca/ail/
We are looking for an independant service provider for our JEOL-880 SEM. They would need to be very familiar with JEOL equipment since the 880 is a rare bird (the only one in the States, I believe). I believe it's a 1200 TEM column married to 840 electronics. We are located in central Oklahoma. TIA.
Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
I am looking for a vendor to purchase The Image Processing Tool Kit by John Russ. I placed an order with Amazon nearly a month ago but have yet to receive anything. Hmmmm.
Thank you,
John B.
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
-----Original Message----- From: Michael Coviello [SMTP:coviello-at-mae.uta.edu] Sent: Thursday, March 23, 2000 4:44 AM To: Microscopy-at-sparc5.microscopy.com Subject: TEM-Digital camera recommendations
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Hi Ya'll: We are looking for a CCD camera for our Philips 430 TEM. We would like to get the best camera for the best price, e.g., either buying a used camera or a new non-Gatan camera (Gatan seems to be twice as expensive as the others). We would be using the camera for bright field and high resolution TEM of materials (semiconductors) rather than for biological specimens. Does anyone have a camera they would like to sell/donate or does anyone have recommendations as to a less-expensive camera that they know can be used for materials applications. Thanks, Mike Coviello Lab Manager University of Texas -at- Arlington
MARCH 30TH MEETING AT GENENTECH TOPIC: Microscopy and Public Health.
Our two speakers come from the USDA, Agricultural Research Service in Albany, CA. Robert Mandrell's presentation is titled "Analysis of Human Pathogens on Food Surfaces by Stereo- and Confocal Micros-copy". Robert is the Research Leader of the Food Safety and Health Unit at the USDA facility. Also speaking is De Wood on "Immunolocalization using FESEM and a Backscatter Detector". De heads up the Microscopy & Imaging Lab at the USDA.
Please call Greg Lum at 415/338-1339 or Email: "glum-at-sfsu.edu" for dinner reservations and entree choice by Monday March 27th. The meeting starts at 5:00 p.m. with a beverage reception. The dinner will start at 6:00 pm.
DINNER RESERVATIONS Entree choice (select one) $20 nonmember, $15 regular members; $8 student members Menu choices for Genentech meeting on March 30th [ ] Chicken Parmesan [ ] Flank Steak with Mushroom Sauce [ ] Pasta Primavera
Please call Greg Lum at 415/338-1339 or Email: "glum-at-sfsu.edu" for dinner reservations and entree choice by Monday March 27th. The meeting starts at 5:00 p.m. with a beverage reception. The dinner will start at 6:00 pm.
There are two labs on campus with the Agfa Duoscan T2500 scanners. As stated in the message from National Graphics Supply, they have both a flat bed and a transparency drawer. They will scan at 16-bit gray scale as well as in 8-bit and RGB. Weve been very happy with ours, primarily using it for TEM and SEM negatives, and for 35 mm. One caveat, the 2500 dpi capability is limited to a 4 inch x 14 inch area (1/2 width of the scan area). Set-up was simple and no crashes (so far). The software is easy to learn with identical interfaces for both the standalone application and the Photoshop compatible plug-in.
We purchased ours locally from a professional photography retailer.
Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
Hum.... This is very interesting. Government customers are one thing, non-government customers are another. GSA rules and principles do not apply to non-government customers. If you would please cite the FAR that is the basis for your assertion, this could help a lot to clarify your statement.
I have never heard of "price dumping." But I have heard of competition. One must keep U.S. government business separate from others. Even then, how does one mingle them together? On what basis is this done?
How much of this "competition" is based on multiple unit discounts versus actual price reduction? And as a consumer, what real difference does it make? If the consumer can negotiate a good deal, all the better for the consumer, right?
Mixing GSA into free enterprise is like apples and oranges, so to speak.
What do you think?
gary g.
At 03:19 PM 3/23/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You are right. The 880 is an "immersion lens" SEM, a very rare animal. There are only two that I know of: One at OU, the second was at IBM in France but is now in the back of my office sadly used for parts.
Earl Weltmer
Bill Chissoe wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are looking for an independant service provider for our JEOL-880 SEM. } They would need to be very familiar with JEOL equipment since the 880 is } a rare bird (the only one in the States, I believe). I believe it's a } 1200 TEM column married to 840 electronics. We are located in central } Oklahoma. TIA. } } Bill } } -- } ============================================================= } Bill Chissoe III } Electron Microscopist } University of Oklahoma } 770 Van Vleet Oval } Norman, Ok. 73019 } E-mail: wchiss-at-ou.edu Ph. (405)325-4391 } =============================================================
The Company providing the service does certify to the GSA customer that the prices given are the same or lower than the prices given to any other entity: public, government or private.
Earl Weltmer
"Dr. Gary Gaugler" wrote:
} Hum.... This is very interesting. Government customers are one } thing, non-government customers are another. GSA rules and } principles do not apply to non-government customers. If you } would please cite the FAR that is the basis for your assertion, } this could help a lot to clarify your statement. } } I have never heard of "price dumping." But I have heard of } competition. One must keep U.S. government business separate } from others. Even then, how does one mingle them together? } On what basis is this done? } } How much of this "competition" is based on multiple unit discounts versus } actual price reduction? And as a consumer, what real difference } does it make? If the consumer can negotiate a good deal, all } the better for the consumer, right? } } Mixing GSA into free enterprise is like apples and oranges, so } to speak. } } What do you think? } } gary g. } } At 03:19 PM 3/23/00 , you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi All, } } } } I've heard from several SEM/TEM independent maintenance companies that } } some of the manufacturer's have engaged in "price dumping" when } } negotiating a service contract. } } } } In other words, if the manufacturer is aware that they have competition } } } } for a service contract they will reduce the contract by as much as forty } } } } percent. } } } } I am all for the free enterprise system, but I thought that when a } } contract price is offered it is the same for all customers especially } } government contracts that must abide by the GSA rules. GSA rules states } } } } that the price offered is the lowest price for this service. It would } } be } } illegal for the manufacturer (or anyone) to offer a contract at less } } than the cost offered to GSA customers. } } } } I am surprised by this move as our contracts are the same as we abide } } by } } the GSA rules. } } } } Regards, } } } } Earl Weltmer
OK....to a GSA customer. What about to other customers? And what precedence does one GSA contract or schedule have for other instantiations? They are, in my experience, single point events. They are not precedence events.
Where is the original thread that spawned this message?
Note, that in my experience, GSA contracts are distinct from other Federal government contracts. i.e., GSA is one thing, other gov contracts are another. GSA negotiates rates....they do not establish them. Therefore, for a particular GSA contract or rate structure, the rates are pre-negotiated for fed users to adopt as per their own contracting department. Or, they can use the GSA schedule and go with that vehicle and its added surcharge. (Yes, GSA contracts cost more than face value). GSA has two flavors: negotiated rates (the user does their own contracting) and negotiated contracts (the user buys into the GSA contract and pays a surcharge for doing so).
Which flavor are you talking about?
gg
At 09:19 PM 3/23/00 , you wrote: } The Company providing the service does certify to the GSA customer that the } prices given are the same or lower than the prices given to any other entity: } public, government or private. } } Earl Weltmer } } "Dr. Gary Gaugler" wrote: } } } Hum.... This is very interesting. Government customers are one } } thing, non-government customers are another. GSA rules and } } principles do not apply to non-government customers. If you } } would please cite the FAR that is the basis for your assertion, } } this could help a lot to clarify your statement. } } } } I have never heard of "price dumping." But I have heard of } } competition. One must keep U.S. government business separate } } from others. Even then, how does one mingle them together? } } On what basis is this done? } } } } How much of this "competition" is based on multiple unit discounts versus } } actual price reduction? And as a consumer, what real difference } } does it make? If the consumer can negotiate a good deal, all } } the better for the consumer, right? } } } } Mixing GSA into free enterprise is like apples and oranges, so } } to speak. } } } } What do you think? } } } } gary g. } } } } At 03:19 PM 3/23/00 , you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hi All, } } } } } } I've heard from several SEM/TEM independent maintenance companies that } } } some of the manufacturer's have engaged in "price dumping" when } } } negotiating a service contract. } } } } } } In other words, if the manufacturer is aware that they have competition } } } } } } for a service contract they will reduce the contract by as much as forty } } } } } } percent. } } } } } } I am all for the free enterprise system, but I thought that when a } } } contract price is offered it is the same for all customers especially } } } government contracts that must abide by the GSA rules. GSA rules states } } } } } } that the price offered is the lowest price for this service. It would } } } be } } } illegal for the manufacturer (or anyone) to offer a contract at less } } } than the cost offered to GSA customers. } } } } } } I am surprised by this move as our contracts are the same as we abide } } } by } } } the GSA rules. } } } } } } Regards, } } } } } } Earl Weltmer
On the very low end A Snappy digitizer and a surveillance video camera produce what I would consider the minimal acceptable image. It is good enough for some work I would say it is comparable to a Polaroid image or slightly worse.
} From a quality per dollar point of view it can't be beat. I would say it is good enough for collaboration, web publishing and some publishing.
This set up and 35mm camera should meet all quality needs as long as you don't need real-time images. It does not compare to a top of the line dedicated system. But it does better than anything but a dedicated system.
My best effort to date is: http://www.couger.com/microscope/onion.jpg This is the raw image with no enhancement and the enhanced version is: http://www.couger.com/microscope/onionE.jpg This version was contrast enhanced and mapped to false color. The subject is a partially dehydrated onion cell. There are depth of focus problems and possibly some vibration problems. There is only 7.5 bits of information in the raw image. I attribute this to loss of dynamic range in the camera due to age.
This was taken with a Leitz darkfield condenser and a Leitz 63x 0.85 objective with a variable aperture for darkfield work. The camera was a monochrome RCA surveillance videocron camera and a Snappy digitizer.
I don't think the microscope end can be improved on much in the onion image. I still had dynamic range problems. The only solution I can think of is to take multiple pictures at different light levels and combine them. A high resolution CCD camera will give better resolution but I don't think the dynamic range problem will go away using video cameras. Maybe I will get around to exposing a 4X5 negative and see what is really there. But after using digital capture it is an awful lot of trouble.
I am strictly an amateur with a microscope but I do have professional experience in digital images.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
----- Original Message ----- } From: "Gordon Vrololjak" {gvrdolja-at-nature.Berkeley.EDU} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 23, 2000 1:56 PM
Try going direct:
Reindeer Games, Inc. 235 S. Main St. #201 Gainesville, FL 32601 352.384.1850
http://members.aol.com/foveapro
jcr6-at-aol.com
gg
At 06:21 PM 3/23/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am looking for a vendor to purchase The Image Processing Tool Kit by John Russ. I placed an order with Amazon nearly a month ago but have yet to receive anything. Hmmmm. } } Thank you, } } John B. } } #################################################################### } John J. Bozzola, Ph.D., Director } Micro-Imaging and Analysis Center } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
} I am looking for a vendor to purchase The Image Processing Tool Kit by } John Russ.
I believe you can purchase it directly from John Russ.
Rob
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
Scott Some aquatic invertebrates such as gastrotrichs are difficult to fix for SEM in a life-like relaxed state because aldehyde fixatives induce violent contraction. A paper by May+ advocates relaxing the organisms (rotifers in this instance) first by narcosis with procaine, or similar anaesthetics. However, in our experience gastrotrichs can sense these anaesthetics, and considerable distortion results. There is a considerable literature on this topic covering the properties of EDTA, menthol, osmium tetroxide, etc. etc. for this purpose, but mostly these solutions are ineffective at least on gastrotrichs.
We have found sodium azide at ~0.1% to be very effective at preventing distortion. Gastrotrichs seem not to be irritated by it, but simply go to sleep. We have usually mounted them for LM in glycerol, in which they can be preserved and collected for transit or for subsequent processing and examination. The fixation/ dehydration/ HMDS procedure you suggest would probably be very effective, but you might also try Ensikat & Barthlott's* approach of simply examining them in the SEM in the glycerol-wet state, or after drying them from glycerol under vacuum.
+May, L. (1985) The use of procaine hydrochloride in the preparation of rotifer samples for counting. Verh. Internat. Verein. Limnol. 22, 2897-2990 *Ensikat & Barthlott (1993) Liquid substitution - a versatile procedure for sem specimen preparation of biological-materials without drying or coating Journal of Microscopy 172, 195-203
Sodium azide is obviously a serious poison, and schoolkids cannot handle the powder, but would it be acceptable for them to use 0.1% solution under supervision by a teacher? I guess this depends on local legislation and attitudes, and also on the age and level of experience and responsibility of the students.
Hope this helps Chris
Date sent: Thu, 23 Mar 2000 16:16:36 -0600 To: Microscopy-at-sparc5.microscopy.com } From: Scott Robinson {sjrobin-at-itg.uiuc.edu}
Dear Vladimir,
Under the circumstances you describe you are probably on the edge of the condition where the virtual composition changes linearly with the pressure, but this depends on the working distance and the gas that you use. However, if you still have some pressure range to work with, then making several measurements at a range of pressures may still allow you to use a non-linear extrapolation.
The main problem might be that wet specimens often have a thin water film on the sample, which may adversely affect EDS analysis. Keeping exact control over sample temperature, pressure and humidity is required to get suitable EDS conditions.
} -----Original Message----- } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] } Sent: Thursday, March 23, 2000 10:09 PM } To: 'Hans Dijkstra'; Dusevich, Vladimir } Cc: Microscopy Listserver } Subject: RE: EDS in Variable Pressure SEM } } } Hans, } Thank you very much for explanations. } But for me this method will not work - } quite often I analyze wet specimens at } pretty high pressure (5-6 Torr). } Thank you again, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } -----Original Message----- } } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } } Sent: Thursday, March 23, 2000 12:19 PM } } To: DusevichV-at-umkc.edu } } Cc: Microscopy Listserver } } Subject: RE: EDS in Variable Pressure SEM } } } } } } Dear Vladimir, } } } } The main idea of the ViP-Quant procedure is rather straight-forward and } } elegant. It is based on the phenomenon that as the pressure increases the } } size of the skirt effect remains fairly constant, although the number of } } skirt electrons increases, and thus the contribution of the skirt-effect to } } the EDS signal increases. This intensity increase is linear with the pressure. } } } } This is valid for low- to medium pressures, or at a higher (ESEM) pressure } } with a very short free-path length of the skirt electrons, since in both } } cases you can assume the skirt electrons have scattered only once. At higher } } pressures with longer working distances these assumptions are no longer } } fully valid. } } } } So to do an EDS analysis you take 2 measurements at different pressures, the } } low-pressure one at a pressure where you can just avoid charging, and a } } high-pressure one at at least twice that pressure. The measured intensities } } of all elements are then plotted as a function of pressure, and extrapolated } } to pressure zero, i.e. high vacuum. The extrapolated results will be very } } close to the results hat would have been obtained directly if the sample } } could have been analyzed at high vacuum conditions. } } } } Of course anyone can do this plotting and extrapolation manually, that is: } } if your EDS software allows you to enter intensities manually! The only } } thing the EDAX ViP-Quant is offering as an extra is that the software does } } it automatically for you. There are no proprietary miracles involved, EDAX } } has just listened carefully to what science has to offer.... } } } } Best regards, } } } } Hans Dijkstra } } } } Disclaimer: The above is my personal opinion, and not necessarily EDAX's. } } ------------------------------------------------------------- } } EDAX Europe www.edax.com } } Ringbaan Noord 103 Tel.: +31-13-5364000 } } P.O. Box 4144 Fax.: +31-13-5356279 } } 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl } } the Netherlands } } ------------------------------------------------------------- } } } } } -----Original Message----- } } } } } } From: "Dusevich, Vladimir", INTERNET:DusevichV-at-umkc.edu } } } To: Everett Ramer, INTERNET:Everett.Ramer-at-netl.doe.gov } } } "'Hans Dijkstra'", INTERNET:hans.dijkstra-at-edax.nl } } } } } } CC: Joergen Bilde-Soerensen 5802, INTERNET:j.bilde-at-risoe.dk } } } [unknown], INTERNET:Microscopy-at-sparc5.microscopy.com } } } } } } Date: 3/22/00 4:35 PM } } } } } } RE: RE: EDS in Variable Pressure SEM } } } } } } } } } } } -------------------------------------------------------------- } } ---------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------- } } ---------. } } } } } } } } } Hans, } } } Could you please explain the main idea of the } } } Vip-Quant algorithm. I cannot even imagine } } } how quantitative analysis could have nearly the } } } same accuracy as in high-vacuum mode without } } } a priori knowledge of the composition of } } } surrounding areas. } } } Thank you, } } } } } } Vladimir } } } } } } } } } Vladimir M. Dusevich, Ph.D. } } } Electron Microscope Lab Manager } } } 3127 School of Dentistry } } } 650 E. 25th Street } } } Kansas City, MO 64108-2784 } } } } } } Phone: (816) 235-2072 } } } Fax: (816) 235-5524 } } } Web: http://www.umkc.edu/dentistry/microscopy } } } } } } } -----Original Message----- } } } } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } } } } Sent: Tuesday, March 21, 2000 7:54 AM } } } } To: Everett Ramer } } } } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } } } } Subject: RE: EDS in Variable Pressure SEM } } } } } } } } } } } } Dear Everett, } } } } } } } } The extrapolation method (aka Variable Pressure Method)described by Dr. } } } } Bilde-Sorenson has been implemented by EDAX in a software feature called } } } } ViP-Quant. With this method the EDS analysis under Bad-Vacuum conditions can } } } } be done with nearly the same accuracy as under High-Vacuum conditions. } } } } Although this method was developed with the ESEM microscope in mind, it of } } } } course can be applied to all Bad-Vacuum scanning electron microscopes. } } } } } } } } Please contact your local EDAX representative for more information and a } } } } copy of the new ViP-Quant brochure, or request one through www.edax.com. } } } } } } } } With best regards, } } } } } } } } Hans Dijkstra
Thanks for the info. I really didn't want to go through & look up the FAR again.
The GSA wasn't my real inquiry anyway. I just wanted to find out how widespread this practice extends. The GSA issue seems to get in the way.
OK to rephase: How many have experienced an extreme reduction in service contract price (20% or more) when confronted with competition?
Thank You,
Earl Weltmer
"Allen R. Sampson" wrote:
} Actually, both are right here. Government stipulations have apparently } eased recently, but a part of the government contract procedure has in the } past been a certification by the service provider that the rates being } quoted are normal and appropriate. This has essentially required details } of a few other comparable contracts that were similar in price and } requirements. } } I have not had to go through that particular body cavity search for some } time. I assume the government learned that certifications of this sort are } basically worthless, as there are companies large and small who are willing } to do a little spin control in order to secure contracts. Government } agencies are probably unlikely to bother verifying compliance with their } regulations - not a problem in this field alone. } } My memory goes only so far, as does my desire to look up old records. } However, if you're really interested, you may want to start with the old } DAR (Defense Acquisition Regulation) 7-1903.41 - Service Contract Act of } 1965, also updated and known as FAR 52.222-40. In the late 80's, it seems } to have been a part of the small business set-aside program (FAR 52.219-04) } to require a price list or a verifiable listing of 3 similar items sold to } establish a market price. You might also look into the Walsh-Healey Public } Contract Act, FAR 52.222-20. } } DOD FAR Supplement (46 CFR Chapter 2) clause 11.111-10 states - "...The } Contractor agrees that the prices for the supplies or services furnished } under this contract are as low or lower than those charged the supplier's } most favored customer for comparable quantities under similar terms and } conditions, in addition to any discounts for prompt payment." } } Frankly each government contract I have generates about 1/2" of paper work } per year. I give little concern to pricing matters, either I win a } contract or not. The same goes for commercial customers. I try to price } my services to all customers based on the expenses I expect to incur and a } modest income for myself (perhaps way too modest, I reflect, as the time to } finish the tax year has come). There are many other regulations and laws } that I have to pay more attention to. } } Whether these, and other, regulations, laws and requirements have any } actual practical effect is open to debate. I can't say that I have noticed } any recent trend for manufacturers to discount their service contracts - } although I don't normally inquire about other bids submitted. I wouldn't } be surprised, though, if manufacturers are finding it advantageous to } discount service prices now to capture customers while the capital } expenditures are running high in the current economy. However, when those } capital budgets dry out in the next economic cycle, they will find that the } service end will have a larger effect on their profitability and they will } increase their profit margins as best they can. } } Consider this a normal result of the economic cycles that we are all slaves } to. As a third party service provider, you can generally think of me as an } independent car repair shop. In good times, people will go to } manufacturer's service, or buy a new car - in bad times they will make } every effort to stretch their capital investment. In good economic times, } I catch a good deal of lower end business from those who have instruments } orphaned by their manufacturer or are very price conscious. In bad } economic times, I'm the guy who can help you avoid laying off personnel by } reducing your costs. } } In either case, just be glad that there are independent service sources } available for these instruments and consider that they will only be there } as long as it is economically feasible for them to survive all economic } scenarios. That is even more true for manufacturers who would rather sell } you a new instrument than service a 3 year old instrument. That } short-sighted corporate position has proven to be a death knell in this } industry. In many cases it has only been the availability of third party } service that has allowed a company to survive in a given market for a } little longer. Public relations is a two-sided blade - it can cut real } deep when times are bad. } } Government laws, rules and regulations can have a significant effect on } manufacturers serving both government and commercial customers. As has } been stated before, manufacturers wishing to do business with the } government are often required to provide service for a stated period of } time and, as stated here, are required to provide service at a market rate. } A first year law student would recognize that, in this country, commercial } concerns benefit from the standards set by the government. It would } behoove any company to provide an even pricing structure across the board, } as any other position opens them up to legal action from either side. } } Since the government rules require comparison under similar "terms and } conditions", the question of multiple instrument discounts is appropriately } accounted for. We are not talking '"apples and oranges" here. Rather, we } are talking about a closed feedback loop where any change in either side } must affect the other. The only wiggle-room here is in the quality of } service provided. Yes, the consumers, both government and commercial, can } bid the service price down. But in the end, only the service quality will } be compromised. } } Allen R. Sampson, Owner } Advanced Research Systems, St. Charles, Illinois 60174 } voice 630.513.7093 fax 630.513.7092 } } -----Original Message----- } From: Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] } Sent: Thursday, March 23, 2000 6:18 PM } To: Earl Weltmer } Cc: MSA listserver } Subject: Re: Dumping of Service Contracts costs } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hum.... This is very interesting. Government customers are one } thing, non-government customers are another. GSA rules and } principles do not apply to non-government customers. If you } would please cite the FAR that is the basis for your assertion, } this could help a lot to clarify your statement. } } I have never heard of "price dumping." But I have heard of } competition. One must keep U.S. government business separate } from others. Even then, how does one mingle them together? } On what basis is this done? } } How much of this "competition" is based on multiple unit discounts versus } actual price reduction? And as a consumer, what real difference } does it make? If the consumer can negotiate a good deal, all } the better for the consumer, right? } } Mixing GSA into free enterprise is like apples and oranges, so } to speak. } } What do you think? } } gary g. } } At 03:19 PM 3/23/00 , you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi All, } } } } I've heard from several SEM/TEM independent maintenance companies that } } some of the manufacturer's have engaged in "price dumping" when } } negotiating a service contract. } } } } In other words, if the manufacturer is aware that they have competition } } } } for a service contract they will reduce the contract by as much as forty } } } } percent. } } } } I am all for the free enterprise system, but I thought that when a } } contract price is offered it is the same for all customers especially } } government contracts that must abide by the GSA rules. GSA rules states } } } } that the price offered is the lowest price for this service. It would } } be } } illegal for the manufacturer (or anyone) to offer a contract at less } } than the cost offered to GSA customers. } } } } I am surprised by this move as our contracts are the same as we abide } } by } } the GSA rules. } } } } Regards, } } } } Earl Weltmer
We find that we get better silver staining of cross-sections of hair viewed bt TEM when embedded in LR White than Araldite resin.
We are looking for a way to allow us to orientate a bundle of hair fibres (about 5mm long) and keep the bundle intact whilst heat-curing the LR White anaerobically. We do not have the means to lower the temperature of the resin, nor use UV light for curing.
Can anyone suggest a means of keeping the bundle of hairs intact within the LR White whilst embedding and curing? Any support must be able to withstand the effects of acetone and LR White resin in liquid form.
Thanks for your attention
Jeremy Sanderson Please reply to jb_sanderson-at-yahoo.com
********************************************************************** Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT Director, Medical Informatics Unit * CELLULAR SCIENCE University Department of Cellular Science * Room 5501, John Radcliffe Hospital, * OXFORD Headington, Oxford, OX3 9DU * Tel 44 (1865) 222039 * website: FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk **********************************************************************
Here is a question for those of you who use sucrose to provide cryoprotection for tissues:
We are preparing perfusion fixed brain tissue by freeze substitution for post-embedding immuno gold. Following fixation we prepare 500 micron vibratome sections, which we then wish to infiltrate with 2 M sucrose in buffer prior to freezing. We have been transferring the brain sections to 1 M sucrose, then when they sink, transferring again to 2 M. The problem is that the sections never sink in the 2 M sucrose.
Do others have this problem? Do we have to cut the sections into small pieces first, or is there some other way to get them to go under? Or should we not worry about making them sink?
Thanks for your advice.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-1936
I have done similar things in a manner such as this: Take the cap off a BEEM capsule and lay it down open side up. Lay your hair fibers flat in the cap and add LRW. Place a glass slide on top of the cap to seal it off during polymerization. Heat polymerize. Use a dremel moto-tool or jewelers hacksaw to trim out a rectangle of the LRW. Good luck, Tom
} } } We find that we get better silver staining of cross-sections of hair } viewed bt TEM when embedded in LR White than Araldite resin. } } We are looking for a way to allow us to orientate a bundle of hair fibres } (about 5mm long) and keep the bundle intact whilst heat-curing the LR } White anaerobically. We do not have the means to lower the temperature of } the resin, nor use UV light for curing. } } Can anyone suggest a means of keeping the bundle of hairs intact within } the LR White whilst embedding and curing? Any support must be able to } withstand the effects of acetone and LR White resin in liquid form. } } Thanks for your attention } } Jeremy Sanderson } Please reply to jb_sanderson-at-yahoo.com } } ********************************************************************** } Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT } Director, Medical Informatics Unit * CELLULAR SCIENCE } University Department of Cellular Science * } Room 5501, John Radcliffe Hospital, * OXFORD } Headington, Oxford, OX3 9DU * } Tel 44 (1865) 222039 * website: } FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk } **********************************************************************
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Jeremy, We have had good luck keeping roots straight by sanwiching them between films of formvar on wire loops. We make a loop with a 3mm diameter and a long stem out of very thin copper wire (36 gauge in usa). We then cast rectangles of formvar about 4 mm x 8 mm and line the loop up over the floating rectangle so that the plane of the loop is parallel to the short side of the rectangle and right in the middle. Then we quickly plunge the loop straight down onto the formvar and into the water and pull it back out again. This gives us a nice film on the loop. Give this at least a few hours to dry (but they will keep for ages--we plant the stem of the loop in some wax and cover with a beaker). Then when ready to go, place your sample, either before or after fixation, on the loop and repeat with a second formvar rectangle.Now you have the sample sandwiched. It is very nice for small samples because solution exchange becomes a snap. The formvar definitely stands up to actetone, butyl methyl methacrylate, Spurs, and I am 95% sure LR white. It is easy to dehydrate through and get other things through even antibodies go through (although they are slowed down a bit).
Hope this helps.
You got more questions, let me know.
Tobias Baskin
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We are facing a similar problem here and have just had some preliminary success with polymerizing LR White in a vacuum dessicator placed in an oven. We were using gelatin capsules full of LR White placed upside down over coverslips upon which cells have been grown. Standard polymerization did not adhere the gel caps to the coverslips, supposedly due the presence of oxygen (it was worth a try). However, the trial run in the dessicator seems to have worked well, and the cover slip detached cleanly when placed in liquid nitrogen.
You might try this with flat embedding molds to preserve the orientation of your bundles. If you do, please let us know the results.
Another approach might be to embed the hair bundles and later cut them out of the LR White in a piece of resin, orient them the way you want, and glue the resin piece onto the tip of a blank block for ultramicrotomy.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
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We find that we get better silver staining of cross-sections of hair viewed bt TEM when embedded in LR White than Araldite resin.
We are looking for a way to allow us to orientate a bundle of hair fibres (about 5mm long) and keep the bundle intact whilst heat-curing the LR White anaerobically. We do not have the means to lower the temperature of the resin, nor use UV light for curing.
Can anyone suggest a means of keeping the bundle of hairs intact within the LR White whilst embedding and curing? Any support must be able to withstand the effects of acetone and LR White resin in liquid form.
Thanks for your attention
Jeremy Sanderson Please reply to jb_sanderson-at-yahoo.com
********************************************************************** Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT Director, Medical Informatics Unit * CELLULAR SCIENCE University Department of Cellular Science * Room 5501, John Radcliffe Hospital, * OXFORD Headington, Oxford, OX3 9DU * Tel 44 (1865) 222039 * website: FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk **********************************************************************
off the top of my head: pack hairs into a small tube made out of thin paper - velin tissue or Rizla cigarette paper wrapped round a cocktail stick - the gum on the edge of a cigarette paper should survive the resin
Date sent: Fri, 24 Mar 2000 08:50:13 -0600 To: Microscopy-at-sparc5.microscopy.com } From: Kingsley Micklem {kingsley.micklem-at-cellular-science.oxford.ac.uk}
Hi, all,
Many suggestions have been given to me after my posting the Personal TEM Website http://syli.homepage.com in this list sever. Thanks a lot to these contributers!
Now the site has been updated according to these good suggestions. More journals are added into the TEM-related Journals. and an additional page for JOB LIST and RESUMES was included for head-huntings in TEM region. Now you may send me your head-hunting ads or your resume to me and I will place it in this part. Thanks again.
Shu-You Li ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Please visit my new homepage - a personal website on transmission electron microsocpy (TEM) - at http://syli.homepage.com at your convenience! It contains not only my current research work, but also LINKS to useful journal informations, instruction for authors, on-line EELS database, periodic table, physical constants, online TEM course and etc. It also contains JOB list and RESUMEs for head-hunting in TEM region. I am looking forward to your invaluable suggestions! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ************************************************** Shu-You Li, Dr. Institut fuer Physikalische Chemie Johannes Guttenberg Universitaet Jakob-Welder-Weg 11 D-55099 Mainz, Germany
OK - this is extremely low-tech, but it works (if none of the other suggestions you receive do the trick). Cotton thread! I've made bundles of fibres by tying them up with fine sewing thread. You can tie several places along the length of the bundle so it will be held more firmly if you need to. You can process, dehydrate, and embed. Just make certain that you have cotton thread - some of the polymer ones might melt in the solvents or resins.
I've also heard of using fishing monofilament for stuff that doesn't require nasty solvents - this is probably just the "guy version" of the thread technique :)
Tamara Howard CSHL
On Fri, 24 Mar 2000, Kingsley Micklem wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We find that we get better silver staining of cross-sections of hair } viewed bt TEM when embedded in LR White than Araldite resin. } } We are looking for a way to allow us to orientate a bundle of hair fibres } (about 5mm long) and keep the bundle intact whilst heat-curing the LR } White anaerobically. We do not have the means to lower the temperature of } the resin, nor use UV light for curing. } } Can anyone suggest a means of keeping the bundle of hairs intact within } the LR White whilst embedding and curing? Any support must be able to } withstand the effects of acetone and LR White resin in liquid form. } } Thanks for your attention } } Jeremy Sanderson } Please reply to jb_sanderson-at-yahoo.com } } ********************************************************************** } Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT } Director, Medical Informatics Unit * CELLULAR SCIENCE } University Department of Cellular Science * } Room 5501, John Radcliffe Hospital, * OXFORD } Headington, Oxford, OX3 9DU * } Tel 44 (1865) 222039 * website: } FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk } ********************************************************************** } } } }
Of course you should all consider that cutting or grinding ANY embedding resin should be undertaken with the knowledge that resin dust can cause serious health effects. I don't know if any of my allergies or other health quirks have been 'encouraged' from exposure to resin dust or solvents, but judging from the discussions on the listserver about some people's experiences with resins, I recommend that you treat that dust with the same respect as paraformaldehyde powder or asbestos!
Just thought I'd share that safety concern.
Gregg Sobocinski Parke Davis Pharmaceutical Research Ann Arbor, Michigan, USA Gregg.Sobocinski-at-wl.com
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Friday, March 24, 2000 10:28 AM To: Kingsley Micklem Cc: Microscopy-at-sparc5.microscopy.com
I have done similar things in a manner such as this: Take the cap off a BEEM capsule and lay it down open side up. Lay your hair fibers flat in the cap and add LRW. Place a glass slide on top of the cap to seal it off during polymerization. Heat polymerize. Use a dremel moto-tool or jewelers hacksaw to trim out a rectangle of the LRW. Good luck, Tom
} } } We find that we get better silver staining of cross-sections of hair } viewed bt TEM when embedded in LR White than Araldite resin. } } We are looking for a way to allow us to orientate a bundle of hair fibres } (about 5mm long) and keep the bundle intact whilst heat-curing the LR } White anaerobically. We do not have the means to lower the temperature of } the resin, nor use UV light for curing. } } Can anyone suggest a means of keeping the bundle of hairs intact within } the LR White whilst embedding and curing? Any support must be able to } withstand the effects of acetone and LR White resin in liquid form. } } Thanks for your attention } } Jeremy Sanderson } Please reply to jb_sanderson-at-yahoo.com } } ********************************************************************** } Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT } Director, Medical Informatics Unit * CELLULAR SCIENCE } University Department of Cellular Science * } Room 5501, John Radcliffe Hospital, * OXFORD } Headington, Oxford, OX3 9DU * } Tel 44 (1865) 222039 * website: } FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk } **********************************************************************
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
hi earl- i've been considering the whole service contract routine lately since i have an older sem, a newer fesem, and a vanilla tem under contract. the sum of my contracts (from the vendors) is about $40K. i have a hard time justifying this level of expenditure, but have been burned in the past with replacement part costs under "limited" arrangements. in my role i have to recover all lab costs, and contracts are a substantial portion of them.
my main vendor (SEMs) has offered me a 5% "discount". this seems a bit light (to me) because i am limited in terms of my cost recovery mechanisms. a commercial or industrial lab would seem to have more flexibility than a shared facility at a small university. currently i'm negotiating with them to extend this discount. i'm not sure what will happen if i can't keep these costs under control...perhaps this is an opportunity for a crafty entrepreneur. at the very least it may be a signal to look for 3-rd party service, in-house methods, and/or go bare on older equipment.
brian ******************** } OK to rephase: How many have experienced an extreme reduction in service } contract price (20% or more) when confronted with competition? } } Thank You, } } Earl Weltmer
---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
Does anyone have for sale, or know of a supplier of Au/Pd target for the older model Balzers-Union Sputtering Device Type 07120-A, serial number 235? It is an early version of that coater series and uses targets for an SCD010 type coater with serial numbers 101-317. The target is ~5 cm diameter with a threaded mounting hole. Thanks!
Hello Listers, We are trying to do some electron microprobe work on the Yucca Mountain Project for the DOE. However, we may need external verification of our standards by a laboratory that is on the Qualified Supplier List, specifically for the YMP. Therefore, unless a lab has done work directly for Yucca Mountain specifically, they don't count (even DOE run
national labs). This is all according to our QA people, and I am hoping
that someone out there may have direct knowledge of the YMP QA and/or know someone who does. Please respond offline to me. Many thanks in advance, Sarah
Sarah A.W. Lundberg Lab (EPMA) (702) 895-2660 or Electron Microanalysis and (SEM) (702) 895-2462 Imaging Laboratory Office (702) 895-1134 Department of Geoscience, UNLV Fax (702) 895-4064 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010 Dept Office (702) 895-3262
I am thinking of trying to put together a laser tweezer setup on my microscope. It doesn't seem too difficult for a simple system, but I am having trouble finding the appropriate laser so I don't have to build it myself from the module. Has anyone out there found a good source for a near IR laser appropriate for this application? Any other words of wisdom for a laser/laser tweezer novice appreciated. Dave Knecht --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Micklem, I embed a variety of fibers for cross-sectioning using the following. Collect little round pieces of paper from your office paper hole punch. Using a sharp needle punch an appropriately sized hole in the center of several of them. Thread your fibers to be embedded through the center hole of the paper circles. The number required will depend on the length, diameter and mass of the fibers to be sectioned. I find that a fine pair of tweezers and a disectin microscope make this job easier. Gently place this assembly into your embedding capsule (I use BEEM 00). With a pipette, slowly add resin down the sides to fill the capsule (I use Spurrs with Z-6040 adhesion promoter). The fiber/paper assembly will self-center in the capsule and maintain this orientation through curing. Be careful with your choice of paper, it must be dry, and some porous paper will outgas bubbles. Test paper samples ahead of time to identify a suitable type. I'm sure you can adapt this to work with LR White.
Good luck,
Phil Swab Engineering Development Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com
-----Original Message----- } From: Kingsley Micklem [SMTP:kingsley.micklem-at-cellular-science.oxford.ac.uk] Sent: Friday, March 24, 2000 6:50 AM To: Microscopy-at-sparc5.microscopy.com
We find that we get better silver staining of cross-sections of hair viewed bt TEM when embedded in LR White than Araldite resin.
We are looking for a way to allow us to orientate a bundle of hair fibres (about 5mm long) and keep the bundle intact whilst heat-curing the LR White anaerobically. We do not have the means to lower the temperature of the resin, nor use UV light for curing.
Can anyone suggest a means of keeping the bundle of hairs intact within the LR White whilst embedding and curing? Any support must be able to withstand the effects of acetone and LR White resin in liquid form.
Thanks for your attention
Jeremy Sanderson Please reply to jb_sanderson-at-yahoo.com
********************************************************************** Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT Director, Medical Informatics Unit * CELLULAR SCIENCE University Department of Cellular Science * Room 5501, John Radcliffe Hospital, * OXFORD Headington, Oxford, OX3 9DU * Tel 44 (1865) 222039 * website: FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk **********************************************************************
A client asked me to photograph bacilli and endospores on a cotton thread (he supplied the sample already sputtered with au/pd). I took one really nice photo, then began experiencing movement of the thread fibers due to electrostatic charges. It seems the cells & endospores are concentrated at the unsupported ends of the fibers; when I look at thte interior, bundled fibers, which are better restrained, they are practically devoid of cells. I can only guess that the cells/spores were drawn to the outer edges of the thread as the buffer evaporated.
Any advice, insights or ideas will be rewarded with a big Thank You, Thank You, Thank You.
Cheers,
Paul Grover :o) Chief Microscopist and Bottle Washer Microvista Laboratory 1220 Cincinnati St. Lafayette, IN 47904
} From: "Sobocinski, Gregg" {Gregg.Sobocinski-at-WL.com} } } } Of course you should all consider that cutting or grinding ANY embedding } resin should be undertaken with the knowledge that resin dust can cause } serious health effects. I don't know if any of my allergies or other health } quirks have been 'encouraged' from exposure to resin dust or solvents, but } judging from the discussions on the listserver about some people's } experiences with resins, I recommend that you treat that dust with the same } respect as paraformaldehyde powder or asbestos! } } Just thought I'd share that safety concern.
The unrecalled products cause more reaction problems than the cured product.
But as one that has been too careless with air quality don't risk you health breathing dusts, chemicals or solvents any more than absolutely necessary.
I think the reaction products of an automobile my be worse than most things in the lab so you may need to wear you respirator on your drive to work. About the time the catalytic converter came along my asthma really flared. some how I don't thing H2SO3 is good for me.
Take good care of you health if the average age at death continues it present course you may have to live to 120 or more.
Take care Gordon
G. C,. Couger 624 Cheyenne Stillwater, OK 74075-1411 405 624 2855 between 3:00 pm to 1:00 am CST 405 742 2758 cell
We solved the resin dust/ Dremel tool problem by rigging a small vacuum cleaner with the intake very close to where we would be grinding, which was usually under a disecting scope. I got the idea while watching a cast being cut from a broken foot. The cast cutter had a vacuum attached. Our rig takes care of the fine particles, while larger ones generally fall to the earth and can be cleaned up later. This more of an issue with epoxies, I suspect. LR White is a dental resin, I believe, and the dentist will grind in your mouth or over you using your chest as a table. I don't know if that means it is safe.
Greg Erdos University of Florida Greg Erdos 5410 SE 185th AVe Micanopy, FL 32667 352-466-0843
You might be able to buy a metal foil of a suitable size and fix it to what you already have (or a custom made holder) using silver-loaded epoxy?
Goodfellow Metals (England) used to have such foils. I could check this further if you wish, plus send you their contact details when I am back in the lab.
Regards - Keith
_______________________________ Dr. Keith Ryan Marine Biological Association of the UK The Laboratory Citadel Hill Plymouth Devon PL1 2PB England
Tel. ++44 (0)1752 633249 Tel. ++44 (0)1752 633279 The 279 number has an answering machine
Hi, I had done an evapouration of cobalt on a quartz sample. Now I want to etch the quartz sample to remove the substrate so that I can do Transmission Microscopy for charactrization. My problem is that If I etch the substrate with HF will it be harmful to carbon? If it is so what are the other materials that can be used to etch quartz. I will be very pleased if you can answer me. Thanking you Ramu, Senior student in M.Sc, Dept of Physics, IIT Bombay.
meet me at ramu8sp-at-ccs.iitb.ernet.in suvee-at-wowmail.com, srisuvee-at-yahoomail.com *******************************************************************************
Does anyone know of some sort of vacuum vessel that would hold a good number of boxed specimens under vacuum? I have several mechanical pumps (dual stage) but have not seen any sizable, sturdy containers. I would like to store my specimens under a vacuum at all times unless loading into the SEM. The purpose is to keep any fungi, moisture and dust from contaminating the specimens.
The problem with large vacuum containers is strength and therefore cost. Its those 14.7 pounds for every sq inch the atmosphere exerts on vacuum vessels, a fact EMists know and understand. Some don't realize that another order of magnitude in high vacuum makes no real difference to that pressure and a very poor vacuum still exerts over 14 lbs. on every sq inch of surface area.
The practical and economic solution is the use airtight cabinets with drying agents or a trickle of dry nitrogen gas. Afterall its mostly moisture that impedes pumping in the SEM or causes specimens and coatings to fall apart with time. Disclaimer: Proscitech ships world-wide a large of desiccating cabinets and vacuum desiccators. See Online } Contents } Page E7 Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Sunday, March 26, 2000 11:05 AM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote:} } Does anyone know of some sort of vacuum vessel that } would hold a good number of boxed specimens under } vacuum? I have several mechanical pumps (dual stage) } but have not seen any sizable, sturdy containers. I would } like to store my specimens under a vacuum at all times } unless loading into the SEM. The purpose is to keep any } fungi, moisture and dust from contaminating the specimens. } } Vendor responses are welcome. } } gary g. }
I am trying to help a new SEM user w/ a tough resolution shot. For some reason he was not able to find the quoted resolution for the SEM (JEOL JSM 6400) he was using. If anybody out there happens to know its exact or approximate quoted resolution, that would be very helpful. His electron source is W.
thanks to the many responders about this post. Let me add some more info to help zero in on a solution--if there is one.
The reason I am seeking a vacuum container is two-fold. First, I have very limited N2 availability. I use industrial grade bottled N2 and dry it with a molecular sieve for venting my SEM chamber during specimen exchange. I vent at about 5psi. The bottles are at 1900 psi and hold 130cu ft of N2. One bottle lasts about two months. I have two bottles--on-line and standby. A "trickle" of N2 through a dessicator would work but for how long per bottle? I don't know.
The other factor for a vacuum container is that if a specimen is under a roughing pump vacuum, if it is at all defective, the body of the specimen will implode. Thus saving the time of fooling with it in the SEM. If it does not implode, then it will be dry and not require very much pump down time before opening the column isolation valve. Thus ensuring minimal effect on the column ion pump.
Steve D'Angelo showed a very nice metal dessicator unit, which would be great if I had a lot of N2....I presume a lot of N2. If I take one of my 130 cu ft cylinders and leak N2 into the dessicator, about how long will it last? And what is the minimum leak pressure needed to make the dessicator function as a drying unit? I could get another cylinder of N2 and another sieve for this unit if the gas volume was sufficient to run the box for two to three months.
University of Central Florida Advanced Materials Processing and Analysis Center (AMPAC)
The Advanced Materials Processing and Analysis Center (AMPAC) is seeking candidates for a position as Assistant in Research to support the Materials Characterization Facility (MCF). The individual must have a Master's degree or a Bachelor's degree from an accredited institution in an appropriate area of study related to surface science and materials characterization, and have at least three years of experience in materials characterization or related areas. The person will be responsible for establishing and maintaining laboratory infrastructure, laboratory maintenance, equipment installation, operation and maintenance, and be an investigator in contracted research. The person must be able to conduct independent research and development in materials characterization with an emphasis in ion beam technology and have the ability to interact with students and provide instruction in the use and interpretation of data. The individual must have extensive knowledge and experience in the operation and maintenance of materials characterization equipment. AMPAC is particularly interested in individuals desiring an academic setting to further their professional goals. The applications will be reviewed beginning April 17, 2000 and will continue to be reviewed until the position is filled.
Applicants should send vitae and three references to Dr. Vimal Desai, Director, 12424 Research Parkway, Suite 408, Orlando, FL 32826. UCF is an equal opportunity/affirmative action employer. As an agency of the State of Florida, all application materials and selection procedures are available for public review.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Gary, Are you concerned about rmp backstreaming, thereby contaminating your specimens with hydrocarbon. I assume you are using a filter with a baking element? -Ken
Hi Gary, How about a TEM film dessicator? Available from most EM suppliers, vacuum companies and some scrapped TEMs.
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. ============================================================================
you mention using a small vacuum cleaner to clean up resin dust. You don't specify whether it has an exhaust filter so I think I should just mention that the typical domestic vacuum cleaner exhausts fine particulates and may well make the problem worse because it may be pumping the very size of resin particles out that you want to avoid. We once used a small portable vacuum cleaner for cleaning up resin dust but stopped when I realized that there was a potential risk.
I know that there are now domestic and industrial vacuum cleaners which have very fine exhaust filters and there are also the small photocopier/toner vacuum cleaners. Has anyone investigated their use for resin dust?
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
greg erdos wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We solved the resin dust/ Dremel tool problem by rigging a small vacuum } cleaner with the intake very close to where we would be grinding, which was } usually under a disecting scope. I got the idea while watching a cast } being cut from a broken foot. The cast cutter had a vacuum attached. } Our rig takes care of the fine particles, while larger ones generally fall } to the earth and can be cleaned up later. } This more of an issue with epoxies, I suspect. LR White is a dental resin, } I believe, and the dentist will grind in your mouth or over you using your } chest as a table. I don't know if that means it is safe. } } Greg Erdos } University of Florida } Greg Erdos } 5410 SE 185th AVe } Micanopy, FL 32667 } 352-466-0843
In Australia we can readily purchase industrial dry N2, this obviates the drying agent and filtering of the gas. In WDS, gas proportional spectrometers are fed with about one bubble a second (holding the outlet into water). At that rate a cylinder routinely last for over a year and although an expensive gas is used for that purpose, cylinder rental invariably is the greater cost. A tight drying cabinet only requires a similar amount of gas to maintain a positive pressure, however slight. Specimens added to the cabinet should be dry and I think that a tray of desiccant is a good idea.
No additional cylinder would be required since Gary already has two. I used to deplete the N2 cylinder after use on the EMs, to nitrogen burst during film development. Since the EM duty cylinder does not go empty unexpectedly, the cylinder feeding the drying cabinet can be removed in time obtain a full cylinder. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Monday, March 27, 2000 1:59 AM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } thanks to the many responders about this post. Let me add some } more info to help zero in on a solution--if there is one. } } The reason I am seeking a vacuum container is two-fold. First, } I have very limited N2 availability. I use industrial grade bottled N2 } and dry it with a molecular sieve for venting my SEM chamber } during specimen exchange. I vent at about 5psi. The bottles } are at 1900 psi and hold 130cu ft of N2. One bottle lasts about } two months. I have two bottles--on-line and standby. A "trickle" } of N2 through a dessicator would work but for how long per } bottle? I don't know. } } The other factor for a vacuum container is that if a specimen is } under a roughing pump vacuum, if it is at all defective, the body } of the specimen will implode. Thus saving the time of fooling with } it in the SEM. If it does not implode, then it will be dry and not } require very much pump down time before opening the column } isolation valve. Thus ensuring minimal effect on the column ion } pump. } } Steve D'Angelo showed a very nice metal dessicator unit, which } would be great if I had a lot of N2....I presume a lot of N2. If } I take one of my 130 cu ft cylinders and leak N2 into the dessicator, } about how long will it last? And what is the minimum leak pressure } needed to make the dessicator function as a drying unit? I could } get another cylinder of N2 and another sieve for this unit if the } gas volume was sufficient to run the box for two to three months. } } Appreciate your feedback. } } gary g. } }
Does anyone no of a good web source for info on light microscopy for undergraduates. Ideally I'm after reference material on brightfield, phase contrast and fluorescent microscopies. Some info on new areas would also be useful. Hope someone out there can help !
Thanks in Advance,
Barry Shaw
Barry Shaw Senior Imaging Technician Molecular & Cell Biology E Floor School Of Biomedical Sciences University of Nottingham Medical School NG7 2UH
I am looking to purchase an Zeiss INKO condenser with 4 Wollaston prisms. The turret position/Wollaston prism marked IIII was used in conjunction with the Plan Achromat 6.3x objective.
What was the difference between the INKO condenser (46 52 79) and the INKO slider III (47 44 44) and INKO slider II (47 44 31)? According to the literature, the type II INKO slider was for research microscopes and the type III for the the smaller STANDARD microscope.
Using a Photom I, I have tried the INKO condenser in position IIII with both type III and type II sliders and am not able to obtain DIC with a PLAN achromat x6.3 objective.
I am very familiar with the 3 position Wollaston prism condenser and am able to obtain great interference DIC with position I and the PLAN x16 objective. However, using the 4 position condenser on prism IIII yields no DIC using the x6.3 objective. As a side note, the prisms are in great shape and show no sign of separation.
I have noticed the dark fringe interference figures for IIII, III, and II are oriented at 11:00 and 5:00 when viewed through an inverted condenser, while in position I, the dark fringe is oriented at 1:00 and 7:00. (hand held with the condenser inverted, the polarizer is placed on top of the dove-tailed retainer ring to position and lock the condenser into the condenser holder, and the INKO slider is held at 45 degrees to the east -west oriented polarizer under the top condenser lens element, i.e., the entire system is inverted.) If this makes sense, great!
Any suggestions as to why the number IIII position does not yield DIC using the 6.3 objective?
Thanks Ken --------------- Ken Tiekotter, Adjunct Professor The University of Portland Dept. of Biology 5000 N Willamette Bldv. Portland, OR 97303
Hi, I am looking for carbon rods with spectrographic quality for Edwards vacuum evaporator. The rods should be about 1.5mm in diameter and 30mm in length. I have tried several suppliers without success. If any of you have information regarding who supplies this kind of carbon rods could you please let me know? Your help will be greatly appreciated. Thank you.
} Chesapeake Society for Microscopy } } Is pleased to announce } } The PhotoShop Workshop } } April 11th,2000 } } Morning Session } 9:00am-12: 00 noon } Introduction and Intermediate level } } Afternoon Session } 1:00pm- 4:00pm } More Advanced Issues and Discussion } } Location: NIH, Building 4 Room 433 } } } Seating is limited to 50 people } Preregistration for CSM members by March 31st –No Charge } After March 31st –Open Registration } CSM members –No charge } Non-members - $20.00 per session } } To register Contact Andrea Weisberg } Phone: (301) 435-1977 } e-mail: aweisberg-at-nih.gov } } } Please register early, we can only seat 50 people. } Ask me about how to become a CSM member } Membership $10.00 per year } } Andrea S. Weisberg } NIH/NIAID/LVD } Bldg. 4/Room 210 } 4 Center Drive } Bethesda, MD 20892-0445 } phone: (301) 435-1977 } fax: (301) 480-1147 } e-mail: aweisberg-at-nih.gov }
I would be very concerned about vacuum pump oil contamination of your specimens if you us oil sealed roughing pumps on such a vacuum chamber. Oil backstreaming will occur unless you use a trap and are meticulous about trap maintenance.
Ronald Vane XEI Scientific
-----Original Message----- } From: Dr. Gary Gaugler {gary-at-gaugler.com} To: MSA listserver {Microscopy-at-sparc5.microscopy.com}
Hi Brian,
Service contract can be likened to "insurance policies'. We are in the service business & offer service contracts for less mostly because we can work on several manufacturers and save on travel time & costs. Still we allot about 20% of the contract costs to parts.
When you "go bare" (self insure) the equipment will probably be OK for about two years if the maintenance has been properly done. After that it is anyone's guess.
When we offer reduced service contract prices for reduced risk (customer buys the parts or limited number of service calls) the discount is about 20-30% to cover parts costs. The largest expenditure is labor and parts in that order.
Competition generally helps the equation but the question that I had is when "competition" is only done when faced with a start-up entrepreneurs. I would ask for a further discount of at least 20%.
Regards,
Earl Weltmer (714) 573-9158
Brian McIntyre 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 earl- } i've been considering the whole service contract routine lately since i } have an older sem, a newer fesem, and a vanilla tem under contract. the } sum of my contracts (from the vendors) is about $40K. i have a hard time } justifying this level of expenditure, but have been burned in the past with } replacement part costs under "limited" arrangements. in my role i have to } recover all lab costs, and contracts are a substantial portion of them. } } my main vendor (SEMs) has offered me a 5% "discount". this seems a bit } light (to me) because i am limited in terms of my cost recovery mechanisms. } a commercial or industrial lab would seem to have more flexibility than a } shared facility at a small university. currently i'm negotiating with them } to extend this discount. i'm not sure what will happen if i can't keep } these costs under control...perhaps this is an opportunity for a crafty } entrepreneur. at the very least it may be a signal to look for 3-rd party } service, in-house methods, and/or go bare on older equipment. } } brian } ******************** } } OK to rephase: How many have experienced an extreme reduction in service } } contract price (20% or more) when confronted with competition? } } } } Thank You, } } } } Earl Weltmer } } ---------------------------------------------- } Brian McIntyre } Electron Microscopy Lab- River Campus } Univ of Rochester } Rochester, NY 14627 } 716-275-3058/4875
A Cambridge S240 scanning electron microscope is available to any interested party for moving expenses and best offer. Please contact me directly. Thank you and kind regards, Diana
Diana L. Kittleson Pillsbury Technology East 737 Pelham Blvd. St. Paul, MN 55114 651-917-5859 651-917-5850 fax www.tpclabs.com
______________________________________________________________________ This e-mail and any attachment contains information which is private and confidential and is intended for the addressee only. If you are not an addressee, you are not authorized to read, copy or use the e-mail or any attachment. If you have received this e-mail in error, please notify the sender by return e-mail and then destroy it.
I have used a Snappy digitizer for the last 6 years with good results. I use it with a Javelin color camera. I,m curious, how did you get it to work with a monochrome camera?? The Snappy syncs on the color burst portion of the video waveform. I have never been able to get a oicture with a monochrome camera or electron microscope.
Dear Sir: I am looking for a PAM interface board/card which would plug into a slot in a Personal Computer. I have a Link Analytical EDX connected to a Hitachi S520LB SEM. The pulse processor for the EDX is a stand alone unit which connects via a 25 pin ribbon cable to a personal computer of older vintage i.e.. 80286 or 80386 technology. The connection is into a PAM Interface Card. The person I purchased this unit from was unable to locate the PAM Interface Board. I am looking for this board. Does anyone know where I can locate one? Thank You, David Browning dbrownng-at-stargate.net
Gary: Why not store samples in a standard vacuum desiccator (Fisher Scientific, VWR, etc). They come in glass (with ground glass or o-ring seals) and plastic (with o-ring seals). However, don't leave samples in a desiccator under active pumping. All pumps backstream. It's just a matter of how much contamination your samples can tolerate. Pump the desiccator to some nominal reduced pressure (a few seconds is fine) and close the valve. A silica gel or similar desiccant can be used concurrently in the dessicator and will more effectively pump residual water out of your samples at this reduced pressure.
Phil Swab Engineering Development Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com
-----Original Message----- } From: Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] Sent: Sunday, March 26, 2000 7:59 AM To: MSA listserver Cc: Steve D'Angelo
thanks to the many responders about this post. Let me add some more info to help zero in on a solution--if there is one.
The reason I am seeking a vacuum container is two-fold. First, I have very limited N2 availability. I use industrial grade bottled N2 and dry it with a molecular sieve for venting my SEM chamber during specimen exchange. I vent at about 5psi. The bottles are at 1900 psi and hold 130cu ft of N2. One bottle lasts about two months. I have two bottles--on-line and standby. A "trickle" of N2 through a dessicator would work but for how long per bottle? I don't know.
The other factor for a vacuum container is that if a specimen is under a roughing pump vacuum, if it is at all defective, the body of the specimen will implode. Thus saving the time of fooling with it in the SEM. If it does not implode, then it will be dry and not require very much pump down time before opening the column isolation valve. Thus ensuring minimal effect on the column ion pump.
Steve D'Angelo showed a very nice metal dessicator unit, which would be great if I had a lot of N2....I presume a lot of N2. If I take one of my 130 cu ft cylinders and leak N2 into the dessicator, about how long will it last? And what is the minimum leak pressure needed to make the dessicator function as a drying unit? I could get another cylinder of N2 and another sieve for this unit if the gas volume was sufficient to run the box for two to three months.
Balzers Union renamed to BAL-TEC AG in 1992. But the product line was maintained and new developments are carried out. There are several coating systems of this older type in use. So of course we can deliver these targets furthermore. Article No. of this target: BU 007 129 -T
Our reprepresentative in USA / Canada:
Techotrade International 7 Perimeter Road Manchester, NH 03103-3343 Mr. Johnny Hagen T: +1 603 622-5011
Our representative will contact you directly to assist with further information.
Ulrike Zeile
BAL-TEC AG EM-Technology and Application FL-9496 Balzers Tel. +423 388 12 36 Fax +423 388 12 60
-----Ursprüngliche Nachricht----- Von: Mandayam V. Parthasarathy [mailto:mvp2-at-cornell.edu] Gesendet: Freitag, 24. März 2000 21:22 An: Microscopy-at-sparc5.microscopy.com Betreff: Balzers-Union putter coater
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Does anyone have for sale, or know of a supplier of Au/Pd target for the older model Balzers-Union Sputtering Device Type 07120-A, serial number 235? It is an early version of that coater series and uses targets for an SCD010 type coater with serial numbers 101-317. The target is ~5 cm diameter with a threaded mounting hole. Thanks!
Balzers Union renamed to BAL-TEC AG in 1992. But the product line was maintained and new developments are carried out. For additional information have a look at our webside www.bal-tec.com. There are several coating systems of this older type in use. So of course we can deliver these targets furthermore. Article No. of this target: BU 007 129 -T
Our reprepresentative in USA / Canada:
Techotrade International 7 Perimeter Road Manchester, NH 03103-3343 Mr. Johnny Hagen T: +1 603 622-5011
Our representative will contact you directly to assist with further information.
Ulrike Zeile
BAL-TEC AG EM-Technology and Application FL-9496 Balzers Tel. +423 388 12 36 Fax +423 388 12 60
-----Ursprüngliche Nachricht----- Von: Mandayam V. Parthasarathy [mailto:mvp2-at-cornell.edu] Gesendet: Freitag, 24. März 2000 21:22 An: Microscopy-at-sparc5.microscopy.com Betreff: Balzers-Union putter coater
of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html ------------------------------------------------------------- ----------. ++ ++ ++ Does anyone have for sale, or know of a supplier of Au/Pd ++ target for the ++ older model Balzers-Union Sputtering Device Type 07120-A, ++ serial number ++ 235? It is an early version of that coater series and uses ++ targets for an ++ SCD010 type coater with serial numbers 101-317. The target is ~5 cm ++ diameter with a threaded mounting hole. Thanks! ++ ++ ++ ******************************************************************* ++ ++ M.V. Parthasarathy ++ Prof. of Plant Biology, Adjunct Prof. of Biomedical Sciences (Vet), & ++ Director, Cornell Integrated Microscopy Center (CIMC) ++ Section of Plant Biology ++ 228 Plant Science Building ++ Cornell University, Ithaca, NY 14853 ++ E-Mail: mvp2-at-cornell.edu ++ Plant Biology Office: 268 Emerson; Telephone: 607-255-1734 ++ Plant Biology Fax: 607-255-5407 ++ CIMC Office: C1 054 Vet. Med. Center; Telephone: 607-253-3803 ++ CIMC Office Fax: 607-253-3803 ++ CIMC web site: http://www.cimc.cornell.edu ++ ++ ++
Like Malcolm I gave up on a hand held vacuum cleaner when sawing resin as I suspected it leaked fine particles. Now I saw up resin in a large bag in a fume cupboard. The bag has lasted 5 years. Anyone got a neater/safer method?
Dave
On Mon, 27 Mar 2000 10:35:33 +0100 Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greg } } you mention using a small vacuum cleaner to clean up resin dust. You } don't specify whether it has an exhaust filter so I think I should just } mention that the typical domestic vacuum cleaner exhausts fine } particulates and may well make the problem worse because it may be } pumping the very size of resin particles out that you want to avoid. We } once used a small portable vacuum cleaner for cleaning up resin dust but } stopped when I realized that there was a potential risk. } } I know that there are now domestic and industrial vacuum cleaners which } have very fine exhaust filters and there are also the small } photocopier/toner vacuum cleaners. Has anyone investigated their use for } resin dust? } } Malcolm } } Malcolm Haswell } Electron Microscopy } School of Sciences } Fleming Building } University of Sunderland } SUNDERLAND SR1 3SD } Tyne and Wear } UK } } Tel (0191) 515 2872 } e-mail: malcolm.haswell-at-sunderland.ac.uk } } greg erdos wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } We solved the resin dust/ Dremel tool problem by rigging a small vacuum } } cleaner with the intake very close to where we would be grinding, which was } } usually under a disecting scope. I got the idea while watching a cast } } being cut from a broken foot. The cast cutter had a vacuum attached. } } Our rig takes care of the fine particles, while larger ones generally fall } } to the earth and can be cleaned up later. } } This more of an issue with epoxies, I suspect. LR White is a dental resin, } } I believe, and the dentist will grind in your mouth or over you using your } } chest as a table. I don't know if that means it is safe. } } } } Greg Erdos } } University of Florida } } Greg Erdos } } 5410 SE 185th AVe } } Micanopy, FL 32667 } } 352-466-0843 }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I have a Hummer VII sputter coater, which has an automatic processing cycle. As the vacuum drops from 60 millitorr to 40 millitorr (during which time the high voltage should switch on), the entire process shuts down. I have tried running the system with the argon valve open and shut.
Any suggestions from users out there?
Thanks,
Don ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
} Does anyone no of a good web source for info on light microscopy for } undergraduates. Ideally I'm after reference material on brightfield, } phase contrast and fluorescent microscopies. Some info on new areas } would also be useful.
Barry, Try looking at these web sites. The virtual microscopy is fun. You might want to filter out the stuff of interest to materials and geological sciences... or leave it in in the name of interdisciplinary microscopy !
A number of universities are now using "Optimizing Light Microscopy for Biological and Clinical Laboratories". It covers all the areas you mentioned and more. It also includes a number of small experiments which can be done as a group or on independent study. See our website for further info.
Caveat: MME does have financial interest in this book.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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} ******************************************************
At 01:46 PM 3/27/00 GMT0BST, BARRY SHAW wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Ping, The last time I went looking for spec-pure carbon rods, our Stores man found them at: ultra carbon, 900 Harrison St., Bay City, MI48708-8244. They weren't that exact size, but they are a good place to try. At 09:38 AM 3/27/00 -0400, you wrote:
} } Hi, I am looking for carbon rods with spectrographic quality for Edwards } vacuum evaporator. The rods should be about 1.5mm in diameter and 30mm } in length. I have tried several suppliers without success. If any of you } have information regarding who supplies this kind of carbon rods could } you please let me know? Your help will be greatly appreciated. Thank } you. } } Ping } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} Does anyone no of a good web source for info on light microscopy for } undergraduates. Ideally I'm after reference material on brightfield, } phase contrast and fluorescent microscopies. Some info on new areas } would also be useful. Hope someone out there can help ! } } Barry -
I can suggest a good CD-ROM: Pagliaro, l., et al., 1997 Microscopy-Tutor. You'll find a description and ordering information in the Project MICRO bibliography (URL below).
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
} Here is a question for those of you who use sucrose to provide } cryoprotection for tissues: } } We are preparing perfusion fixed brain tissue by freeze substitution for } post-embedding immuno gold. Following fixation we prepare 500 micron } vibratome sections, which we then wish to infiltrate with 2 M sucrose in } buffer prior to freezing. We have been transferring the brain sections to } 1 M sucrose, then when they sink, transferring again to 2 M. The problem } is that the sections never sink in the 2 M sucrose. }
Dear Marie, The Handbook of Chemistry and Physics gives a specific gravety for 2 M sucrose of about 1.25. Is the sp. gr. of your tissue (exclusive of the exchangable fluids, of course) greater than this? If not, the sections will never sink. If it is marginally greater, then the sections will sink when hell freezes over, and you can do cryo easily. If you can determine the weight of a tissue section of known volume first with fixation fluid, then after infiltration with 1 M sucrose (and also measure the volume change), you could extrapolate to the situation for 2 M sucrose (or some- one with too much time on his/her hands could do this for many sucrose concentrations). Good luck. Yours, Bill Tivol
We need to localize zinc in prostate tissue for TEM. Has anyone used Sulfide-silver method or Zinc-dithizonate method? If so, is there a method better than the other or new methods that might work on such tissue?
I am also trying to locate a reference by Pihl E. Falkmer S: Trials to modify the sulfide-silver method for ultrastructural tissue localization of heavy metals. Acta Histochem 27:34-41, 1967. If you have a copy of this method please email me.
Thank you
Karen Kelley Senior Electron Microscopist UF Biotechnology Electron Microscopy Core Lab Box 118525 Gainesville Florida lab:352-392-1184 fax: 352-846-0251 email: klv-at-biotech.ufl.edu
} I had done an evapouration of cobalt on a quartz sample. Now I want to } etch the quartz sample to remove the substrate so that I can do } Transmission Microscopy for charactrization. My problem is that If I etch } the substrate with HF will it be harmful to carbon? If it is so what are } the other materials that can be used to etch quartz.
Dear Ramu, Quartz can be etched--slowly--in strong base. A 5-to-10 M solution of NaOH should do the trick, but if the quartz thickness is in the mm range, it will take a long time. I don't know whether the base will be harmful to either carbon or cobalt, but I don't think it will. Good luck. Yours, Bill Tivol
} Hi, I am looking for carbon rods with spectrographic quality for Edwards } vacuum evaporator. The rods should be about 1.5mm in diameter and 30mm } in length. I have tried several suppliers without success. If any of you } have information regarding who supplies this kind of carbon rods could } you please let me know? Your help will be greatly appreciated. Thank } you. }
Dear Ping, I got mine from Ted Pella, but I'm surprized you didn't find them in other suppliers' catalogues. Pella lists both carbon and graphite, both
spectroscopically pure and technical grade, in several diameters and lengths. I have no connection to Ted Pella, Inc. except that of customer. Yours, Bill Tivol
Hello everyone, I have a student who wants to prepare tissue culture cells for TEM in Agar blocks. I shall appreciate any suggestions. Thanks in advance. C.L.Singla
We are contemplating replacing our analytical TEM. A basic spec we envisage is STEM, EDS + GIF/PEELS. We are happy to choose the EDS, but we run into trouble with e-guns, i.e. to go LaB6 or FEG. In particular as we would like to bolt a GIF/PEELS on to the system.
We have several concerns: The safety of the FEG gun, considering we are a multi-user facility with users who have a frequent tendency to bend holders and press the wrong buttons. How problematic are FEGs, do you have much down time / running cost due to careless users?
GIF/PEELS: has anybody carried out a systematic comparison between a system with LaB6 or FEG. Is the extra cost of a FEG (over twice the price) really justified for GIF/PEELS or will it quite happily run on a LaB6.
The thoughts of fellow microscopy in the same quandary much a appreciated.
Keith.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Sucrose infiltration is an essential step in the preparation of cryosections for immunocytochemistry. Aldehyde-fixed biological tissue is infiltrated in 2.0M to 2.3M sucrose to protect the sample from freezing damage when it is subseuently frozen by immersion in liquid nitrogen. If the sample is fixed, the membranes become permiable to sucrose and the sample can be frozen into a vitreous state by immersion in liquid nitrogen. Although it is advised to infiltrate for 24 hr on a rotator, many years of experience has demonstrated that even after 15 min of exposure to sucrose, small pieces of aldehyde-fixed material are sufficiently cryoprotected to allow for successful vitrification. Take no heed of whether the sample has sunk to the bottom of the tube or not, check to see if it has been fixed and has spent sufficient time in the sucrose. If it has, it will be cryoprotected.
Useful reference: Griffiths, G, McDowall, A, Back, R, Dubochet, J. 1984. On the preparation of cryosections for immunocytochemistry. Ultrastruct. Res. 89:65-78 They showed by quantitative mass measurements that fixed cells are freely permeable to sucrose.
Best regards,
Paul Webster
Paul Webster, Ph.D. House Ear Institute 2100 West Third St. Los Angeles, CA 90057
Bill Tivoli wrote: The Handbook of Chemistry and Physics gives a specific gravety for 2 M sucrose of about 1.25. Is the sp. gr. of your tissue (exclusive of the exchangable fluids, of course) greater than this? If not, the sections will never sink. If it is marginally greater, then the sections will sink when hell freezes over, and you can do cryo easily. If you can determine the weight of a tissue section of known volume first with fixation fluid, then after infiltration with 1 M sucrose (and also measure the volume change), you could extrapolate to the situation for 2 M sucrose (or some- one with too much time on his/her hands could do this for many sucrose concentrations). Good luck.
Marie E. Cantino wrote: We are preparing perfusion fixed brain tissue by freeze substitution for post-embedding immuno gold. Following fixation we prepare 500 micron vibratome sections, which we then wish to infiltrate with 2 M sucrose in buffer prior to freezing. We have been transferring the brain sections to 1 M sucrose, then when they sink, transferring again to 2 M. The problem is that the sections never sink in the 2 M sucrose.
Hi, Our safety people are concerned about having our Polaron c.p.d pressure tested at regular intervals. Does any one out there know of anyone who does this sort of thing, preferably on site? Also I would be interested to hear how other users safety check this sort of equipment. thanks, Christine.
Dear Keith, If you are seriously worried about the experience of some of the users then you should consider the Philips/FEI Tecnai series of instruments. The person in charge of the microscope can determine the level of access to all the features for each user i.e. basic, medium and expert user profiles. The microcope, in a similar way to fly-by-wire aircraft, will simply prevent users from blowing the tip up or anything else detrimental to the 'scope. This should take out the worry about user/FEG problems.
The LaB6 and FEG work identically with the GIF/PEELS, they're only electrons after all. The FEG will give you a smaller energy width (good for EELS:ELNES .etc) and more current (good for Energy Filtered TEM, STEM:HAADF, EELS & EDS). Then there is the option of adding a biprism with the FEG if you want to try off-axis holography. The FEG machine will simply give you a wider tolerance range in terms of specimen thickness and experimental conditions i.e greater signal to noise ratios than with the LaB6, something that is a problem with STEM based methods. FEG alignment procedures will be slightly different too.
We have taken delivery of a 300kV Tecnai and I have to say that the machine is very impressive. The machine is very stable and the high brightness FEG is a major improvement over the LaB6. The GIF is also quite a God-send for zero-loss filtering and for mapping.
I hope this helps, Jon
P.S. I am not affiliated or have any commercial connection to Philips/FEI. ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
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Dear Friends,
The Pirani Gauge of the Oxford Hexland - CT1000 cryo unit mounted on SEM S120 is not working. Can anyone guide me in purchase of this? Could this be manufactured in-house? Please help me out.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road Pune - 411 004, India Phone 91-20-5653680/5654357 e-mail : rajdeep-at-aripune.ernet.in
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id GAA03992 for dist-Microscopy; Wed, 29 Mar 2000 06:33:01 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id GAA03989 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 29 Mar 2000 06:32:31 -0600 (CST) Received: from ultra3-4.rz.Uni-Hohenheim.DE (ultra3-4.rz.uni-hohenheim.de [144.41.4.28]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id GAA03980 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 29 Mar 2000 06:32:19 -0600 (CST) Received: from uni-hohenheim.de (eiche.botanik.uni-hohenheim.de [144.41.33.45]) by ultra3-4.rz.Uni-Hohenheim.DE (8.8.3/8.8.3) with ESMTP id OAA29921 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 29 Mar 2000 14:27:06 +0200 (MET DST) Message-ID: {38E1F5A4.96CAA8EC-at-uni-hohenheim.de}
We work with plant material and after ultra thin sectioning our sections show lines of compressions in cell wall areas. Of course, we try to stretch the Epon sections with a heat pen or with chloroform, but very often we are not successful. The sections just move on the water surface and nothing is happening. Any advice would be gratefully appreciated. Regards, Anne Heller -- Dr. Anne Heller, AG Elektronenmikroskopie, Institut fŸr Botanik (210), UniversitŠt Hohenheim, Garbenstra§e 30 D-70593 Stuttgart
Hi Jo, Thanks for your answer. We use our GIF in a similar way, focusing around the energy loss we want to take the image at with a slit width covering the 3 energy windows if possible. This works best for rather thick samples (compared to our 5 nm particles)But as you say, maybe it's just the spatial resolution which is not good enough, our microscope was designed to enhance contrast for biological studies, unfortunately loosing some resolution in the process. We are not yet trying to get concentrations out of the pictures...rather get a qualitative mapping.
I tried to do EELS measurements a few times and got very few succes. The best results are obtained on beam resistant samples simply because I can then move the zero loss peak off the CCD and increase intensity (or acq time) at will. I start with a very low intensity and optimise it in turboview mode to get a good signal out. However I could seldom see well defined peaks. One point which I find strange is that we can often get reasonnably good elemental maps even when we don't see any peak in the EELS spectrum. The question is then if the map is really trustworthy...
Any comments welcome !
Olivier
Jo Verbeeck wrote:
} Hello, } } Regarding EFTEM on small particles, this should actually work quite well. } That is, elemental mapping works reasonably if you choose the right } objective apperture. You should simulate the spatial resolution to see } what can be attained under your conditions (HT, Cc, Cs etc). } However it will be very difficult to get real concentration information } out of these images. I'm trying EELS with nanoprobe but so far no succes } (everything gets destroyed before I take a spectrum, allignment is very } very difficult) } Still, focussing remains a problem. Comon practice is to focus at 100eV } and then suppose everything is OK for higher losses. I do not understand } the theory of this technique (blurring by finite slitwidth & Cc?) nor thus } it work well. Any comments on this are welcome. } } Jo } } ************************************************************* } * Jo Verbeeck * } * University of Antwerp * } * Dept. EMAT (Electron Microscopy for Materials Research) * } * e-mail: joverbee-at-ruca.ua.ac.be * } * tel: +32(0)3 218 02 49 * } * fax: +32(0)3 218 02 57 * } ************************************************************* } } On Tue, 21 Mar 2000, GuessWho wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear all, } } } } I was wondering if anybody was using a GIF filter. We have one mounted } } on a 120 kV TEM and are trying to image small particles (5-10 nm) and } } doing some energy filtering to image different elements in the particles } } (for example gold, silicon, copper, cerium...) . This is not really what } } I would call trivial work and we are still working out the set up } } (window size and positionning for example) in a rather empiric way. The } } main problem is often to get a decent signal in the interesting energy } } region and still keep the windows fairly narrow. } } I would add that we often have to work at low dose because of the beam } } sensitivity of our samples, but I guess our GIF setups can be improved. } } Any suggestion ? } } } } Any general comment on the GIF warmly welcome ! } } } } Thanks } } } } Olivier } } } } } }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We work with plant material and after ultra thin sectioning our sections } show lines of compressions in cell wall areas. Of course, we try to } stretch the Epon sections with a heat pen or with chloroform, but very } often we are not successful. The sections just move on the water } surface and nothing is happening. Any advice would be gratefully } appreciated. } Regards, } Anne Heller } -- } Dr. Anne Heller, AG Elektronenmikroskopie, } Institut fŸr Botanik (210), UniversitŠt Hohenheim, } Garbenstra§e 30 } D-70593 Stuttgart } } Tel.: (0049)-711-459-2180 } Fax.: (0049)-711-459-3355
Hi Anne,
My first guess is that the mechanical properties of your Epon mix differs too much from that of the fixed walls. However, when I worked with plant material, I generally used Xylene to stretch floating sections (rarely in Epon). I'm not sure if was the aromatic character or the boiling point difference that made it better at swelling and relaxing the sections than CHCl3.
Other suggestions would be using a harder resin (my favorite was VCD and Quetol 651 (equiequivalent) hardened with NSA (or HSA), which I called Spurtol ;-)). I found it stains easier than Spurr's yet is low viscosity and about as hard as a medium Spurr's. Reducing the knife angle might help. Post curing the existing blocks at elevated temperature might harden them some more.
Cheers,
John
John Heckman MSM Department Michigan State University
I don't know if you have interest in this but it is good info. Wallace
-----Original Message----- } From: Parker, Jo Sent: Wednesday, March 29, 2000 8:45 AM To: All SOD Faculty & Staff
Dear Jonathan and Keith,
I just want to add a few comments to what Keith said about FEG's. One of the major advantages of a FEG over conventional souces for AEM is the spot size. The new FEG scopes have STEM resolution of 2 angstroms with great brightness. (I have only seen the Philips/FEI scope, but I think the other companies have similar specs.) When one is deciding whether or not to go for the FEG, I think it will depend on your applications. If a small intense beam and good energy resolution are necessary, go for the FEG. Of course, there is also the consideration of the cost of service contract ($$$$).
Christine Richardson wrote: =================================================== Hi, Our safety people are concerned about having our Polaron c.p.d pressure tested at regular intervals. Does any one out there know of anyone who does this sort of thing, preferably on site? Also I would be interested to hear how other users safety check this sort of equipment. =================================================== I might not be the last word on this, but I had described to me the kind of large liquid filled tank (I think it is water) that pressure vessels like this are tested in, and to me at least it did not sound like it would be the kind of thing that one would "carry around" in a portable kind of unit to do this kind of testing, on site. This description came, some years ago, from Dr. Wilf Gee (now deceased) and who was very much involved in the original manufacturing of this product and who had involvement for many years with the safety testing of the unit.
When your CPD unit was first delivered, it would have come with a copy of a report from the testing agency showing the results and conditions of the pressure testing. I might be off by a bit on this, but it was my recollection that the vessel is tested to a pressure level that is three or four times higher than what would be required to blow out the rupture disc. In other words the unit is tested to a level that, if the rupture disc did not blow exactly where it was supposed to blow, there is some very large margin of error so that the disc still would blow long before the vessel.
There are probably more CPD units of this design installed in the world than any other design. I have never heard of anyone having any kind of a problem with it (e.g. an explosion), except an occasional blowing of a rupture disc, but certainly not the pressure vessel itself. Just don't ever try to run the unit by replacing the rupture disc, if one is not handy, with a cut metal disc. That is very dangerous and should never be done. Believe it or not, we do uncover once in a while, people who do do that sort of thing, out of naivety of course and that is why I do not waste an opportunity to point out the danger in doing that sort of thing. That is why I always have recommended, for multi-user environments in particular, to always have a small supply of replacement rupture discs near by the unit just to avoid even the slightest temptation (such as by a student) to by-pass this important safety feature.
So I for one would be very interested in hearing the logic and rationale of your safety committee that is of the opinion that you should have your pressure vessel tested from time to time.
However, there is one kind of situation where one should have the vessel pressure tested and that is if one has attempted any mechanical modifications to the chamber itself. Then the system should be pressure tested. And since the manufacturer of this particular CPD unit (e.g. Polaron) has had on the order of thirty years of experience doing this kind of testing, on this particular design of vessel, I would recommend having them do the testing for you in the UK. Today the testing of the Polaron unit is done to the CE standard, without doubt the most stringent in the world.
If you do not have the original test certificate, if you send me your S/N I could try to get a copy of it for you. That alone might satisfy the questions being asked of your safety people.
Disclaimer: SPI Supplies has offered this particular CPD unit, especially to customers outside the USA, for nearly twenty five years and we are unaware of any requirement to have the vessel retested at periodic intervals for safety reasons.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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I am passing this on for a fellow colleague. Thank you in advance.
Ginger (Baker) Hendricks EM Lab Manager Oklahoma State University College of Osteopathic Medicine
"I am looking for an immunogold-silver enhancement method for labelling a cytoplasmic antigen in cultured cells and viewing with Light Microscopy. We will be using Protein Aand a monoclonal antibody. Once we have determined that we have specific labelling, we will proceed to visualizing the gold with TEM."
} Hello everyone, } I have a student who wants to prepare tissue culture cells for TEM in Agar } blocks. I shall appreciate any suggestions. } Thanks in advance. } C.L.Singla } It would be helpful to know how the tissue culture cells are being grown...in plastic wells where they will be scraped off or on plastic or glass coverslips where the entire surface will be embedded. What is the ultimate goal of the TEM examination...morphology, interactions? Will the samples be ultimately embedded in plastic or expoxy?
Dr. Tina Schwach Microscopy Consulting Services, Inc.
I would appreciate it very much if someone can refer me to a good book or protocol for preparation of fungal spores for SEM and TEM. This is very foreign to me since I have been working with mammalian tissues exclusively for so many years but now that the lab is a core facility I am getting somel requests for other types of samples. Thank you,
Cora Bucana ******************************************************* Corazon D. Bucana, Ph.D. Department of Cancer Biology U.T. M.D. Anderson Cancer Center 1515 Holcombe Blvd. Box 173 Houston, Texas 77030 Phone: (713) 792-8106 FAX: (713) 792-8747 Email:bucana-at-audumla.mdacc.tmc.edu FAX: (713) 792-8747
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } Our safety people are concerned about having our Polaron c.p.d } pressure tested at regular intervals. } Does any one out there know of anyone who does this sort of thing, } preferably on site? } Also I would be interested to hear how other users safety check this } sort of equipment. } thanks, } Christine.
Hi Christine,
Over here in the US we have an Interstate Commerce Commission that checks pressure vessels via hydrostatic testing. This is required every 5 years and the tops of gas cylinders, at least, are stamped with the last test date. It's always amazed me just how long N2 cylinders last (reading hydrostatic test dates is something to do while your plates are processing). I've seen bottles with dates that were before W.W.I. All that age and 2250 psi. I've never heard of them requiring the testing of lab equipment, though. Might check with your bottled gas supplier to see if they'd do it.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We work with plant material and after ultra thin sectioning our sections } show lines of compressions in cell wall areas. Of course, we try to } stretch the Epon sections with a heat pen or with chloroform, but very } often we are not successful. The sections just move on the water } surface and nothing is happening. Any advice would be gratefully } appreciated. } Regards, } Anne Heller } -- } Dr. Anne Heller, AG Elektronenmikroskopie, } Institut für Botanik (210), Universität Hohenheim, } Garbenstraße 30 } D-70593 Stuttgart } } Tel.: (0049)-711-459-2180 } Fax.: (0049)-711-459-3355 } } } } Hi,
If I get any reaction in a section floating in a boat from xylene, heat pen, chloroform, etc., I know that something is wrong. My personal test of a good embedment is nonreactivity of floating sections. Why?
In industry, if it is important that there are no free monomers in the section, the material is exposed to the above solvents to soak them out.
If you have enough unpolymerized monomers in your section, solvents will affect that section. (Note: Embedments that are too soft in formulations are excluded from this discussion at this time).
So - keep your formulations well mixed at all times - do not allow it to sit in the hood. It must be in motion at all times. Push infiltration! More changes, longer times. If using epoxy of any sort, heat the infiltration medium to 37 deg C for 1 hour after every new change of resin while the sections are on the rotator. A plain 60W light bulb is ideal for this. Polymerize well. At least for 48 hours. If needed, repolymerize at 95 deg C for an hour especially if your formulation contains NMA.
Your formulation may be wrong for your material. Try going harder. Also with existing blocks, try reheating, then try a lower knife angle and a slower speed of cutting.
Most important: Keep records. Know what you are doing and why. Do not haphazardly try this, try that. You will get crazy! Embedments are very complex systems that belong into the realm of material science - that is why biologists have so much trouble with them. (I have had mega fights lasting years with epoxies)
Keith If you have a new FEG machine with a Schottky emitter, then these are relatively easy to operate and fairly robust, but they do, of course, cost a lot more and so you may not want to spend that money and have ham-fisted users operating it. For high-resolution analytical TEM/STEM there is no doubt that FEG is the preferred source. The small source size and small energy spread combined with high gun brightness gives great performance. A word of warning,though. When using a GIF with any TEM it is often necessary to operate at low mag because of the additional mag (18 - 20 times) introduced by the GIF. In this situation (depending on the mag), the LaB6 source may be preferable in certain situations because the total current is higher than the FEG source. I have used both types of machine-Schottky FEG plus GIF and LaB6 plus GIF and there do not appear to be any severe problems with the low mag mode operating with a Schottky source. The cold FEG source may not be so good from this point of view, since the source size is smaller than the Schottky FEG and so the cross-over probe size above which the current in the cold FEG probe is less than the LaB6 is small and probably in the range 10-100 nm. I have not used a cold FEG TEM with a GIF such as the Hitachi and so I can't really make an informed comment, but Profs Dravid and Marks at Northwestern University are very skilled users of the Hitachi machine and they can surely advise you. Best of luck.
Alan Fox Center for Materials Science and Engineering Naval Postgraduate School Monterey California 93940
Keith Moulding wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Greetings, } } We are contemplating replacing our analytical TEM. A basic spec we envisage } is STEM, EDS + GIF/PEELS. We are happy to choose the EDS, but we run into } trouble with e-guns, i.e. to go LaB6 or FEG. In particular as we would like } to bolt a GIF/PEELS on to the system. } } We have several concerns: } The safety of the FEG gun, considering we are a multi-user facility with } users who have a frequent tendency to bend holders and press the wrong } buttons. How problematic are FEGs, do you have much down time / running } cost due to careless users? } } GIF/PEELS: has anybody carried out a systematic comparison between a system } with LaB6 or FEG. Is the extra cost of a FEG (over twice the price) really } justified for GIF/PEELS or will it quite happily run on a LaB6. } } The thoughts of fellow microscopy in the same quandary much a appreciated. } } Keith. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Dr. K. Moulding. } } Materials Characterisation and Preparation Facility } Hong Kong University of Science and Technology, } Clear Water Bay, } Kowloon, } Hong Kong. } } FAX: (852) 2358 2451 } TEL: (852) 2358 8724 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Phone a divers shop and ask who is testing their cylinders. Those people can also test the CPD. It would be a hydrostatic test and they would need to have a fitting to connect to the back of the CPD. To have any meaning the applied pressure would need to be higher than normally applied, I suggest by 50%.
I think its a bad idea to do such a test. The metal housing of those units is vastly over-designed and the window is quartz which ultimately would crack but not shatter. All the test would achieve is to burn up some of your funds and time. Stress the system and if your unlucky crack the window - would the "safety people" pay for that?
It would be interesting to learn how many systems have failed and if there were any special circumstances. I don't know of any instance of a CPD failure. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
Hi, Our safety people are concerned about having our Polaron c.p.d pressure tested at regular intervals. Does any one out there know of anyone who does this sort of thing, preferably on site? Also I would be interested to hear how other users safety check this sort of equipment. thanks, Christine.
ASSOCIATE DEAN, COLLEGE OF ENGINEERING AND APPLIED SCIENCES
Western Michigan University seeks applications and nominations for the position of Associate Dean for Undergraduate Programs and Assessment of the College of Engineering and Applied Sciences.
Western Michigan University is a Carnegie Doctoral I university with 750 FTE tenure-track faculty and an enrollment of about 27,000 students, 25% at the graduate level. It is advancing to Carnegie Research II classification and plans a substantial investment in the College of Engineering and Applied Sciences, including a new physical facility, to accomplish this goal. In addition to its Graduate College and Lee Honors College, Western supports seven degree-granting colleges: Arts and Sciences, Haworth College of Business, Education, Engineering and Applied Sciences, Aviation, Health and Human Services, and Fine Arts. These colleges offer 242 academic programs, including 60 at the masterâs level and 25 at the doctoral level.
The Associate Dean for Undergraduate Programs and Assessment reports to the Dean. This person provides leadership and oversees the development of college policy for undergraduate programs, accreditation, assessment, advising, recruitment, retention, student orientation, co-op, and intern programs. An earned terminal degree in one of the disciplines in the college or a related science is desirable. A doctoral degree, with at least one degree in an engineering or closely related science/engineering discipline, is required. University experience in academic programs and demonstrated excellence in teaching are required. Candidates should have excellent administrative and interpersonal skills and experience utilizing instructional and administrative technology. An excellent record of teaching and research and/or creative achievement that would warrant an appointment as associate or full professor with tenure in one of the academic units in the university is required.
The individual selected will assume academic and administrative responsibilities for undergraduate programs in a dynamic and growing college offering 16 baccalaureate, 11 masterâs and three doctoral programs. The collegeâs staff includes 70 full-time faculty, 10 administrators (8 with faculty rank), 31 funded staff positions, and dozens of contract staff and graduate student assistants, and teaches approximately 2300 students. The college has a strong commitment to education and research, which spans a broad spectrum of engineering and engineering technologies. The college offers courses and programs primarily on Western Michigan Universityâs main campus in Kalamazoo, but also serves as a major resource for off-campus instruction and economic growth at fives sites across West Michigan.
Candidates should submit 1) a curriculum vita, 2) three letters of recommendation, 3) an application letter stating their qualifications for the position, and 4) a statement outlining a personal vision for engineering education to:
Parviz Merati, Chair Associate Dean Search Committee Department of Mechanical and Aeronautical Engineering Western Michigan University Kalamazoo, MI 49008
Review of applications will begin on or about May 1, 2000. Applications will be accepted until the position is filled. Western Michigan University is an equal opportunity /affirmative action employer.
For additional information about WMU and the College of Engineering and Applied Sciences, refer to our Website: http://www.wmich.edu/. ===================================== Pnina Ari-Gur, D.Sc., Professor Materials Science and Engineering CMD Department Western Michigan University Kalamazoo, MI 49008 (616) 387-3372 FAX: (616) 387-6517 email: pnina.ari-gur-at-wmich.edu http://www.wmich.edu/cmd/arigur.htm =====================================
Not announced on ebay, and limited to MSA, I am offering an IBM Thinkpad 355 which is optionally available to make this camera truly portable. $350 extra gets the Thinkpad. You will still need a Slim SCSI PCMCIA card, but these are very low cost. I have one in a Sony VAIO and it works great with the Leaf.
I need information related to EMS software. There was a related posting (enclosed below) recently, replies to which I seem to have missed. My mail to the author of that posting also bounced back. Could somebody please help me get this info? Thanks very much. ---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
-----Original Message----- } From: Divakar R [SMTP:divakar-at-igcar.ernet.in] Sent: Wednesday, March 29, 2000 8:23 PM To: 'Chengge Jiao'
Hi,
Does anybody know the price for following softwares for EM simulation.
1. Diffraction 2.0 (or the most latest version),
2. EMS.
Chengge Jiao
H.H.Wills Physics Laboratory University of Bristol, UK
In the matter of inexperienced users and FEG versus Lab6 I have no fears on our FEGTEM.
We have been running our JEOL3000F for over a year now without any trouble, it has a Schottky emitter so there is no flashing to be carried out. The emitter is the original one used for factory tests, shipped over, used for installation and still in use. We have experience of several LaB6 instruments over the past 15-20 years and are a materials science multi-user facility.
The FE gun runs 24 hours a day, the user just opens the valve to see the beam. The gun and emitter are run up by an experienced person whenever it needs it (every 2-3 months). We have a `Quick Emission Selector' that we have set up to give 3 more emission currents (higher for GIF, lower for less energy spread etc.) and the user cannot readjust these only select between them. There is no room for abuse (even for some of our users!).
The LaB6 guns have to be run up at the start of every session and every time a user changes specimen or films. After a period of time the LaB6 tips slowly deposit insulating LaB6 on the Wenhelt, this charges and the bias has to be adjusted to reset the emission correctly. When a user finishes the session this charge dissipates and the next user gets a high emission current which has to be reset using the bias. The bias has to be adjusted during the first part of the session as the charge builds up. This gives the user lots of room for abusing the tip, it is not possible to prevent access to the bias control as they do need to use it.
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. ============================================================================
Two days ago, the window in our CPD cracked. The run was in every respect normal, but the window cracked at 35 degrees/80 bar. This was a big surprise to us, as we have belived that this could never happen as long as temperature and pressure are within their normal ranges. We would be very interested to receive comments on this.
The week before, the equipment, which is more than 20y old, had been disassembled for cleaning and replacement of the window O-ring.
Best Regards, Gunnar Kopstad.
Vennlig Hilsen dr.ing Gunnar Kopstad overingeni¿r Avd f Patologi, Rit
For the record - and maybe this is a record - our Polaron E3000 CPDA was tested in Feb. 1974 to a proof load of 2,500 lbf/in2. Under "Remarks" on the test certificate it says "Proof pressure applied without any visual signs of distortion or leakage. Safety valve set at 1,900 lbf/in2."
I take "lbf/in2" to be "pounds per square inch".
The unit has been in quite regular use since 1974 with no problems other than failure of seals. I did once have to de-"fur" the waterways but that was a local water supply problem.
Keith Ryan Marine Biological Association Plymouth UK
} I tried to do EELS measurements a few times and got very few succes. The best } results are obtained on beam resistant samples simply because I can then move } the zero loss peak off the CCD and increase intensity (or acq time) at will. I } start with a very low intensity and optimise it in turboview mode to get a } good signal out. However I could seldom see well defined peaks. } One point which I find strange is that we can often get reasonnably good } elemental maps even when we don't see any peak in the EELS spectrum. The } question is then if the map is really trustworthy... } } Any comments welcome ! } } Olivier
With my experience I think it is rather normal than surprising that you can achive relatively good elemental maps in ESI mode but see no recognizable features in the EELS spectrum. With an elemental map you get energy loss information for each single image point. If there is a higher concentration of a certain element, the intensity of the corresponding pixel in the energy window on the edge will be higher than in neighbouring pixels not containing this element or only in less quantity. This does not necessarily mean that you would actually see a peak in the corresponding spectrum of this pixel, it can be just a change in the slope of the background. Since you are comparing the changes in intensity of every pixel this small differences become visible in an map. Furthermore, in spot mode or selecting a small area with an aperture, you will average usually over a larger area than a pixel in image mode. If the features of your specimen are smaller the jump ratio of the edge will be reduced due to the contribution of the surrounding material. If you thoroughly compare spectra acquired on a (large enough) area containing the element of interest and a spectrum of the matrix you will find differences. Furthermore you will see certainly more of the edges in your spectra of thick specimens if you go for a deconvolution. It is still surprising (at least for me) how much information you can gain from a spectrum which did not look much more than a steady decrease in intensity.
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann 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-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I would appreciate it very much if someone can refer me to a good book or } protocol for preparation of fungal spores for SEM and TEM. This is very } foreign to me since I have been working with mammalian tissues exclusively } for so many years but now that the lab is a core facility I am getting somel } requests for other types of samples. Thank you, } } Cora Bucana } ******************************************************* } Corazon D. Bucana, Ph.D. } Department of Cancer Biology } U.T. M.D. Anderson Cancer Center } 1515 Holcombe Blvd. Box 173 } Houston, Texas 77030 } Phone: (713) 792-8106 } FAX: (713) 792-8747 } Email:bucana-at-audumla.mdacc.tmc.edu } FAX: (713) 792-8747
Dear Cora Bucana,
it is depending very much on the type of spore, e.g. mildew with high water content and "thin" walls is easy to prepare with standard protokolls for plant material. Dormant, thick walled spores with less water and high lipid content are quite tricky to prepare for TEM, but easy for SEM. I can send you a detailed protocol if you tell more about your fungal spores.
-- Dr. Anne Heller, AG Elektronenmikroskopie, Institut fŸr Botanik (210), UniversitŠt Hohenheim, Garbenstra§e 30 D-70593 Stuttgart
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Christine Richardson wrote: } =================================================== } Hi, } Our safety people are concerned about having our Polaron c.p.d } pressure tested at regular intervals. } Does any one out there know of anyone who does this sort of thing, } preferably on site? Also I would be interested to hear how other users } safety check this sort of equipment. } =================================================== I might not be the } last word on this, but I had described to me the kind of large liquid } filled tank (I think it is water) that pressure vessels like this are } tested in, and to me at least it did not sound like it would be the } kind of thing that one would "carry around" in a portable kind of unit } to do this kind of testing, on site. This description came, some } years ago, from Dr. Wilf Gee (now deceased) and who was very much } involved in the original manufacturing of this product and who had } involvement for many years with the safety testing of the unit. } } When your CPD unit was first delivered, it would have come with a copy } of a report from the testing agency showing the results and conditions } of the pressure testing. I might be off by a bit on this, but it was } my recollection that the vessel is tested to a pressure level that is } three or four times higher than what would be required to blow out the } rupture disc. In other words the unit is tested to a level that, if } the rupture disc did not blow exactly where it was supposed to blow, } there is some very large margin of error so that the disc still would } blow long before the vessel. } } There are probably more CPD units of this design installed in the } world than any other design. I have never heard of anyone having any } kind of a problem with it (e.g. an explosion), except an occasional } blowing of a rupture disc, but certainly not the pressure vessel } itself. Just don't ever try to run the unit by replacing the rupture } disc, if one is not handy, with a cut metal disc. That is very } dangerous and should never be done. Believe it or not, we do uncover } once in a while, people who do do that sort of thing, out of naivety } of course and that is why I do not waste an opportunity to point out } the danger in doing that sort of thing. That is why I always have } recommended, for multi-user environments in particular, to always have } a small supply of replacement rupture discs near by the unit just to } avoid even the slightest temptation (such as by a student) to by-pass } this important safety feature. } } So I for one would be very interested in hearing the logic and } rationale of your safety committee that is of the opinion that you } should have your pressure vessel tested from time to time. } } However, there is one kind of situation where one should have the } vessel pressure tested and that is if one has attempted any mechanical } modifications to the chamber itself. Then the system should be } pressure tested. And since the manufacturer of this particular CPD } unit (e.g. Polaron) has had on the order of thirty years of } experience doing this kind of testing, on this particular design of } vessel, I would recommend having them do the testing for you in the } UK. Today the testing of the Polaron unit is done to the CE } standard, without doubt the most stringent in the world. } } If you do not have the original test certificate, if you send me your } S/N I could try to get a copy of it for you. That alone might satisfy } the questions being asked of your safety people. } } Disclaimer: SPI Supplies has offered this particular CPD unit, } especially to customers outside the USA, for nearly twenty five years } and we are unaware of any requirement to have the vessel retested at } periodic intervals for safety reasons. } } 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.spi.cc } ######################## } ============================================ } } }
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
Firstly, apologies for the earlier reply that was sent before I gave my comments!
} There are probably more CPD units of this design installed in the } world than any other design. I have never heard of anyone having any } kind of a problem with it
We have used two of these units for more than 25 years and the only problems we have experienced, other than seals which have had to be replaced from time to time, have been one ruptured disc and a leak which developed in the supply pipe from the CO2 cylinder. The latter was repaired (new one made up using existing connectors fitted to new high pressure piping) by our local compressed gas supplier.
Many years ago, having been told that pressure chambers should be checked from time to time, I sent one of our units back to be tested by Polaron in the UK. It came back with a certificate issued by Probe Technical Services saying that it had been tested to 2500 psi with the comment " Proof pressure applied for one minute without any visual sign of leakage or failure".
Regards
Rob
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
Hi Christine, We have a Polaron E300 but no demand for routine testing from our 'safety people' presumably because they know that the burst disc will go long before the body starts to creak. After a major re-fit it was sent to testing specialists who gave it a hydrostatic test to 2500 psi for one minute for a safe working pressure of 2000 psi. It must be possible to do this on site, I cant imagine big autoclaves or air compressor reservoirs being posted off for their tests. It is not as exciting as it sounds. Holes are blanked off and then you pump it full of water so faults are revealed as leaks rather than the more entertaining earsplitting bang and shower of shrapnel which could result from the use of a compressible medium. If the safety people twist your arm try the Yellow Pages under Engineers, Inspection & Testing. ttfn, chris.smith-at-bbsrc.ac.uk Plant Path Dept., IACR-Rothamsted, Harpenden, Herts. UK.
Dear Gunnar It is most likely that there was at least one surface scratch or microcrack that was loaded in a way that resulted in major crack growth, whose result you observed. During glass removal and replacement, there were probably several actions that could have contributed to this -- scratching during cleaning or tool use, altered clamping of the glass, or turning a scratch on one side of the glass from inside to outside (or the reverse). Did anyone clamp the glass in a different way on replacing it, compared with before? (so that the applied stresses are less uniform or at least different, from before?) Moisture is well-known (scientifically and by all who specialize in cutting glass) to enhance crack growth when some component stress applied to the crack tip region helps to open the crack. All these could contribute to the failure. The timing of the failure indicates that one or more of these factors most likely resulted in cracking. Best wishes for avoiding the problem for another 20 years! Brian Robertson
------------------ Brian Robertson Assoc. Prof., Mechanical Engineering, University of Nebraska-Lincoln, on sabbatical at Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH, United Kingdom FAX 44+ 1865 273794 brian.robertson-at-materials.ox.ac.uk
* This e-mail message was sent with Execmail V5.0.x *
Olivier, I find it a bit bizarre that you cannot see our mapped features in your EELS. You're not mapping multiply scattered plasmons are you (a problem with lower lying edges)? How are you acquiring the EEL spectrum: Diffraction pattern (spot/CBED)? Or using the image? What sort of count rates are you getting in the pre-edge images and the maps? Are you sure it is not noise?
For EFTEM practice, I would if possible, try getting hold of some Al or Mg particles/powder (not smoke cubes), preferably mixed, and play with imaging the various plasmons i.e. see which energy windows are best and if possible see if you can image the oxide layers. It's fun because the signal is pretty high so easy to see.
Going to core-loss EFTEM the main problems are resolution: For low loss edges its spatial resolution (mainly delocalization) although there is evidence that this is not as restrictive as it looks on paper (see Z.L.Wang Ultramic Vol 67 (1997) pp105-111 for a demonstration in Al).
For higher edges the signal resolution (object resolution function as Berger & Kohl call it) tells you how good the statistics in a map are. A good paper is A.Berger & H.Kohl (Optik Vol:92 (1993), No4 pp175-193) which tells you about both spatial and signal resolution. The equations are quite rigorous, but it will show you how to get started measuring the signal-to-noise ratio (SNR). With a bit of practice you can then produce some SNR plots like those in Hofer, Grogger, Kothleitner & Warbichler (Ultramicroscopy 67 (1997)pp83-103) which tell you where the energy slit should be positioned to optimise SNR for a given slit width.
I suppose the best advice is find an easy material to practice with and work from thereon. Good luck and have fun.
Jonathan ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
Alan Fox, in a response to the Analytical TEM string, cautioned the question-raiser to beware of people who abuse instruments. Perhaps Nestor will forgive a little microscopy related humor and allow us to start a string on "Operator Horror Stories." Here's ours:
The "ham-fisted user" reference made us chuckle/cringe with the memory of a guest "microscopist" in our lab who hauled himself (220 pounds or so) out of his seat by pulling on the half-inserted specimen rod, bending it about 20 degrees or so!
Henceforth, when we saw him in the hall (he never came into the scope room again), we referred to him as "Conan the Microscopist"!
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Marie: You didn't mention the fixative and length of time fixed. Could it be possible that the tissue is "too well fixed" to allow easy exchange solutions? Also, have you tried thinner vibratome sections? We routinely use 40 micron thick brain sections for immuno labeling. Best regards, Henry *********************************************** Marie E. Cantino wrote: We are preparing perfusion fixed brain tissue by freeze substitution for post-embedding immuno gold. Following fixation we prepare 500 micron vibratome sections, which we then wish to infiltrate with 2 M sucrose in buffer prior to freezing. We have been transferring the brain sections to 1 M sucrose, then when they sink, transferring again to 2 M. The problem is that the sections never sink in the 2 M sucrose. ***************************** Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
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Omega is seeking an individual who is excited about working with varied applications of fluorescence detection in the life sciences. The work involves discussing the best match between laboratory experiments and Omegaâs wide range of light filters for this science. Experience in fluorescence, microscopy, life sciences and instrumentation is essential.
Salary: Competitive with full benefit package.
Contact Information: Email response with attached Word file resume and salary history to: acoutermarsh-at-omegafilters.com or snailmail response to:
Andrew Coutermarsh, SPHR, HR Director Omega Optical, Inc. PO Box 573 Brattleboro, VT 05302
I am posting a message for a colleague who does not subscribe to this list.
He is looking for small embedding oven which can polymerize resins in the range of 40¡-70¡C, but which has a very precise temperature control (holds the temperature well, with very little fluctuation during polymerization). He is having problems with the oven he presently has because the temperature is not controlled well enough, and he is getting uneven polymerization.
Another condition is that he is in the middle of a project and needs the oven right away. He had an oven in mind, but it could not be delivered for 6 weeks!!!
Any suggestions from users or vendors are welcome. Please contact me offline, and I'll forward the replies to my colleague.
Thanks again for all your help.
Paula.
Paula Allan-Wojtas Research Scientist, Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Hallo friends, I have a Bomar SPC 1500 critical point dryer and I am looking for the rupture disc size or part number and a vendor. I looked in the Bomar manual but I can't find any information about it. I shall very much appreciate any information about it. Thanking you in advance. C.Singla
I agree with the disagreement that has been raised to my posting of my personal criteria for good embedment. (A section that stretches in the boat on exposure to solvent fumes is suboptimal and the embedment needs changing.)
I have not dealt much with plant material - mycobacterium is the closest I have come to dealing with cell walls which are tough as Michelin tires. I also have not used glass knives for more than a decade. Even my students start out on an old diamond knife. I am no longer familiar with the vagrancies of glass knives (every one of them seems to have a different cutting edge! Never twice the same!).
Certainly epoxies belong to the organic chemists. But they also belong to the material scientists. I have in front of me a thesis done under the mentoring of Dr. Giammara - A Material Science Evaluation of Epoxy Systems for TEM Applications. At one time I got into a huge fight with embedding filters of certain types. I had a computer program which calculated for me using WPE numbers and molecular weights 36 different formulations. I actually investigated 25 of them. Totally desperate (and mad) I started attending the material science symposia on polymers at the MSA meetings. I found this helpful.
My second favorite book is, HANDBOOK OF EPOXY RESINS. I adore it. (This has caused my friends to question my sanity and make peculiar remarks) This book has been of enormous help in my various battles (lots of them won) with epoxies. Certainly it deals mostly with organic chemistry.
On behalf of the many excellent microscopists I have known for many years I disagree violently with the statement that "and most EM biologists (AT LEAST THE GOOD ONES) understand a lot about organic chemistry." (Capitalizations are mine). One look at this most valuable of books makes it clear that most of the good EM biologists have no hope of knowing enough about polymer chemistry to understand or deal with the varied problems which can occur with epoxies.
Sections in our laboratory do not "stretch". Unless, of course, I want them to, for other special reasons. Our standard sections are the same size as the block face they come from. We deal with keratinized skin which is no picnic compared other biological tissues.
As usual, I always reserve the right to be wrong.
Hildy Crowley Sr. Electron Microscopist Dept of Biol Sciences University of Denver Denver, CO
P.S. My book is under lock and key at all times. If it is stolen, I will have to be hospitalized!
} This is on ebay now. Here is the URL: } } http://cgi.ebay.com/aw-cgi/eBayISAPI.dll?ViewItem&item=296173307 } } Not announced on ebay, and limited to MSA, I am offering } an IBM Thinkpad 355 which is optionally available to make this camera truly } portable. $350 extra gets the Thinkpad. You will still need a Slim SCSI } PCMCIA card, but these are very low cost. I have one in a Sony VAIO } and it works great with the Leaf.
It is on ebay and "Now announced on ebay, ..." It is NOT limited to MSA. Sorry about the poor eye-hand coordination.
Has anyone got a protocol for embedding LR white in a low-temperature chamber using UV light? I have a Leica AFS, so setting and maintaining a temp. is not a problem. I just tried it overnight at -20 C with UV (in filled gelatin capsules, but without activator) and did not get polymerization. Any tips would be appreciated.
Mary McKee Renal Unit MGH Charlestown, MA (617)726-5643
Dear UK CPD users, In the UK pressure systems which operate at pressures greater than 0.5 bar are subject to the Pressure Systems and Transportable Gas Container Regulations 1989. These rules require, among other things, regular inspection of the apparatus. You may also find that your institution's insurers impose a similar rule.
Regards, Eric
---------------------- Dr Eric Lachowski Department of Chemistry University of Aberdeen Meston Walk Old Aberdeen AB24 3UE Scotland +44 (0)1224 272934 fax 272921 e.lachowski-at-abdn.ac.uk
Originally Posted by: shens-at-focus.ruca.ua.ac.be
Hi,
How we can experimentally measure the collection angle when working with GIF? In other words, how to measure magnification between the plane of negatives and the plane of GIF entrance? Using a CCD camera? But normally a CCD camera magnifies an image 20 times of that on a negative, so pictures become uncomparible.
Originally posted by: Beverly_E_Maleeff-at-sbphrd.com
The Microscopy & Microanalysis 2000 Local Arrangements Committee is pleased to announce an additional social event at M&M2000---
TAKE ME OUT TO THE BALL GAME !!
Join us on Tuesday, August 15th as the PHILADELPHIA PHILLIES meet the ARIZONA DIAMONDBACKS at Veterans Stadium. Game time is 7:35 PM.
For $32.50, you'll get: - a ticket to the game (300 level, on the third base line) - a $10.00 coupon, good toward purchases at all concession and souvenir stands - round-trip transportation from the Convention Center to the stadium on a luxury coach (buses leave at 6:00 PM; return to the convention center immediately following the game)
A limited number of packages are available. First come, first served. Maximum 2 packages per person.
Checks, in US$ only, payable to "Microscopy & Microanalysis 2000" can be mailed to:
Bev Maleeff Treasurer, M&M2000 LAC c/o SmithKline Beecham Pharmaceuticals Mail Code UE 0462 709 Swedeland Road King of Prussia, PA 19406
Checks only, please. No credit cards accepted. Your cancelled check will serve as your receipt.
Tickets will be distributed at the M&M2000 Hospitality Booth on Monday and Tuesday, August 14th & 15th.
Further information about this and other M&M2000 events can be found on our web site: http://www.msa.microscopy.com/~mm2000/
Originally posted by: ron.doole-at-materials.ox.ac.uk
Hi All,
I enclose details of post doctoral positions currently available at Dept. of Materials here in Oxford. Please respond directly to the address given for each post. Good luck to any applicants,
Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
********************************************************************** UNIVERSITY OF OXFORD
Department of Materials
POSTDOCTORAL RESEARCH POSITIONS - Salary range £16,286-£24,479p.a.
Applications are invited for the following positions:
Manufacture of Metal Matrix Composite Components
A position is available for a period of 20 months for a suitably qualified research assistant to join an inter-disciplinary research team in the Department of Materials and Department of Engineering Science researching into the manufacture of next generation aeroengine components from long fibre reinforced metal matrix composites. This post will study the fundamentals of fabrication mechanisms, and how these influence subsequent mechanical performance, using a combination of numerical modelling and experiments. Candidates should preferably have expertise and ability in one or more of the following areas: experimental solid mechanics, finite element analysis of plasticity or metal forming, microstructural characterisation of materials, composite materials. Please quote DJ00/6
Interface Modification in Organic LED
A position, sponsored by Opsys Limited, is available in the Department of Materials from April 2000 (or as soon as possible thereafter) for 18 months in the first instance. The project will explore routes to modifying the interfaces of the OLEDs with the objective to improve charge transport and injection. It will involve detailed investigation of the structural and physical properties of the modified interfaces between organo-lanthanide phosphors and other organic and inorganic materials. Techniques will include UPS, SIMS, AFM/STM and electro-optical characterisation of OLEDs. Expertise in one or more of these is highly desirable. Further details are available by email from: oleg.salata-at-materials.ox.ac.uk. Please quote DJ00/7.
Compositional inhomogeneities in information storage materials - effect on physical properties (Principal investigators: Amanda Petford-Long and Alfred Cerezo)
A 3-year position funded by the EPSRC, with support from Seagate Technology, is available in the Department of Materials from May 2000 (or as soon as possible thereafter). The aim of this project is to address how local inhomogeneities in the morphology and composition of thin layered films used for spin-valves and media in information storage technology are controlled by processing of the materials, and how this in turn determines their magnetic properties. The project will primarily involve the use of three-dimensional atom probe (3DAP) analysis to study the chemistry of the films at near-atomic resolution. Results from 3DAP analysis will be combined with measurements of magnetic properties to elucidate the role of the observed nanostructural features on the physical film properties. Some electron microscopy will also be required for comparative microstructural analysis. Expertise in materials characterisation is essential, and some knowledge of thin layered films or magnetic materials would also be helpful. Please quote ref. DJ00/8.
Applications including a full CV, list of publications and the names and addresses of three referees should be sent to The Administrator, Department of Materials, University of Oxford, Parks Road, Oxford, OX1 3PH, from whom further particulars are available. The closing date for applications is 20 April 2000.
It is not "horror" story, but a sort of... Many years ago the colleague from friendly Lab visited me with great project. The idea was simply and beautiful. She argues that because the image in the scope (TEM) is green (green fluorescence of the screen), she wants to modify the sample (I forgot what, some protein, I believe) by red-fluorescent dye to be able to see on the screen of the electron microscope the "double-staining": red on the green background. No comments...
Sergey
} Date: Thu, 30 Mar 2000 09:17:56 -0500 } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } Subject: Operator Horror Stories } To: microscopy-at-sparc5.microscopy.com } Importance: Normal } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b (Intl)|18 } January 2000) at 30/03/2000 09:18:01 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
As further comment on the pressure testing of the Polaron ® CPD units, I have been asked to submit the following statement from David Cheshire, a Manufacturing Manager at VG Microtech, the manufacturers of the Polaron brand of critical point dryers: ================================================= We use an independent test facility that has amongst its credentials the following international approvals : NAMAS, UKAS, NATA. The test certificates issued by ERA are for each individual chamber and we have copies of all these. The test itself is a sustained 50% overpressure (2000 psi) for 1 minute. Obviously any leakage would lead to failure and non certification. In the past units have been overpressured to 2500 and 3000 psi.
I think it is highly unlikely that a "local dive shop" would have the facilities and wherewithal to support the standards imposed by the various associations.
Incidentally the window is not quartz but a far denser and tougher material specifically manufactured for high pressure use.
Best regards Dave Cheshire
Polaron Range Development Manager VG Microtech The Birches Industrial Estate, Imberhorne Lane, East Grinstead, West Sussex. RH19 1UB. UK. =================================================== I believe there was some concern about having this kind of testing, if it was to be done, in any kind of facility that was not certified according to the bodies mentioned in David's message.
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.spi.cc ######################## ============================================
In trying to adapt the SPAM filter last night to catch the latest porn site posting I accidently broke the software. Most messages got rejected after ~ 5 pm Chicago time. I will try to repost those messages , however, I may have missed a few. If you dont' see your posting by the end of the day, please resend it and accept my apology.
I would like to know why they use Argon in the metal coating devices. At our laboratory, we don't use Argon, we use air. What is the advantage of using Argon ? De Pauw Bart Faculty of Veterinary Medicine Morphology Salisburylaan 133 9820 Merelbeke Belgium Phone : 0032(0)9 264.77.19 Fax : 0032(0)9 264.77.90
Does anybody know where I can purchase Ni TEM grids with a holey carbon film ? I can find plenty of copper grids, but the Ni grids seem to be hard to find.
Thanks W. Erasmus
Willem Erasmus Snr. Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 9603954 Fax : +27 +16 9602826 E-mail : willem.erasmus-at-sasol.com
Question: Many years ago the museum bought an AO microscope with an Expostar Shutter control. It appears that the control box (model 1190) has been lost. Is there anyway to buy a replacement? Does the company still exist? Is the microscope manufactored under another name?
Originally Posted by: george.bou-gharios-at-csc.mrc.ac.uk
I am writing to you in my capacity as secretary general of the XIth International Congress of Histochemistry and Cytochemistry which is held in YORK, UK 3-8 July 2000. The above mentioned Congress is Hosted by the Royal Microscopical Society (www.rms.org.uk) and n exciting programme has been put together which can be seen on
www.med.ic.ac.uk/external/ichc_2000/
Dr George Bou-Gharios Muscle Cell Biology Group MRC Clinical Sciences Centre Imperial College School of Medicine Hammersmith Hospital Du Cane Road London W12 0NN Tel: 0181-383 8261 Fax: 0181- 383 8264 email: george.bou-gharios-at-csc.mrc.ac.uk
Hi there folks, just a line to let you know that the schedule of presentations for the IUMAS 2000 meeting to be held in Kona, Hawaii, July 9th-14th 2000, is on the Web at:
http://www.microanalysis.org/iumas2000/
Thanks.
Jfm.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42¡ 16' 48" Long. 83¡ 43' 48"
Hi there folks, just a line to let you know that the schedule of presentations for the IUMAS 2000 meeting to be held in Kona, Hawaii, July 9th-14th 2000, is on the Web at:
http://www.microanalysis.org/iumas2000/
Thanks.
Jfm.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42¡ 16' 48" Long. 83¡ 43' 48"
I would like to know whether the ratio of plastidic to extraplastidic membranes changes in roots of Arabidopsis or any other plant in response to phosphate starvation. Does anyone know of a TEM ultrastructural morphometric analysis or any other type of analysis towards this end?
Thanks, Christoph Benning
Christoph Benning
Assistant Professor Dept. of Biochemistry Michigan State University East Lansing, MI 48824-1319 USA
The problem with Ni grids is that Ni is a magnetic metal, and so Ni grids can give all kinds of trouble. We used to use them back in the early days when they were about all that was available, and when resolution of EMs wasn't so great; now-a-days they have lost their popularity.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
He needs a source of plastic slides and/or film which can be machined and which do not fluoresce. Anybody have any ideas? Contact me offline.
Many thanks Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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} ******************************************************
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id KAA09849 for dist-Microscopy; Fri, 31 Mar 2000 10:17:44 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id KAA09845 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 31 Mar 2000 10:17:14 -0600 (CST) Received: from mhro1.mayo.edu (mhro1.mayo.edu [129.176.100.75]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id KAA09837 for {microscopy-at-sparc5.microscopy.com} ; Fri, 31 Mar 2000 10:16:49 -0600 (CST) Received: from excsrv15.mayo.edu by mhro1.mayo.edu with ESMTP for microscopy-at-sparc5.microscopy.com; Fri, 31 Mar 2000 10:10:39 -0600 Received: by excsrv15.mayo.edu with Internet Mail Service (5.5.2650.21) id {HPJ4T1LM} ; Fri, 31 Mar 2000 10:11:24 -0600 Message-Id: {9DF709FEA025D311B5040090274E63A10401A5-at-excsrv09.mayo.edu}
} ---------- } From: Springett, Margaret J. } Sent: Friday, March 31, 2000 11:04 AM } To: 'springett.margaret-at-mayo.edu' } Subject: nickel grids } } I do an extensive amount of immunolabeling, and nickel grids are } economical, } but two factors need to be considered on a daily basis. The fact that } they } are magnetic can be an advantage as well as a detriment. Their magnetic } quality allows one to rinse grids floating on a bubble with a magnetic } stir } plate, this has been demonstrated in Dr. Bendyan's workshops on } goldlabeling } techniques. When trying to retreive a stubborn grid out of a grid box, a } magnetic tweezer is "heaven sent". Now about magnetic grids in the scope, } if you stigmate the scope after fine-focusing, your micrograph will be } focused. I have used these techniques to produce excellent micrographs of } caveolae at better than 100K magnification. For our use gold grids are } too } expensive, so "work around methods" with nickel grids are necessary. } Marge } } Margaret Springett IEM Specialist Electron Microscopy Core Facility Mayo Foundation and Clinic Rochester, MN 55905
There is an entire FTIR imaging industry out there. SpectraTech has been a leader in that industry and is a good place to start. Suggest you call Ed Manke (203-926-8998). Also, John Reffner has been one of the few people who has really bridged microscopy and spectroscopy. He is the real guru in this area. John is currently working with SensIR technologies and can be reached there at 203-207-9700.
The other companies who are in this area come from more of a spectroscopy background (Jobin Yvon/Instruments SA, Perkin Elmer, Hewlitt Packard, etc.) and would know less about the microscopy interface.
I taught a microscopy course to the apps specialists at Spectra Tech this January and had a chance to work with their Continuum, which has a combination of both glass and reflecting optics on its nosepiece. However, I think that the highest magnifications they had availalable were only about 32x objectives. The NAs were really great as I remember, so you could make use of electronics from the CCD to boost the actual mag to the screen.
Let me know how you make out.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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} ******************************************************
At 08:12 AM 3/31/00 -0600, Nestor J. Zaluzec wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id KAA09893 for dist-Microscopy; Fri, 31 Mar 2000 10:24:41 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id KAA09889 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 31 Mar 2000 10:24:11 -0600 (CST) Received: from mail.map.com (mail.map.com [204.71.19.12]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id KAA09880; Fri, 31 Mar 2000 10:23:28 -0600 (CST) Received: from Barbara Foster [208.168.76.208] by mail.map.com (SMTPD32-6.00) id A06E874A00E2; Fri, 31 Mar 2000 11:21:02 -0500 Message-Id: {3.0.3.32.20000331111416.00de1458-at-mail.map.com} X-Sender: mme-at-mail.map.com X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32)
Bill,
AO became part of Cambridge in 1986 then came under the Leica umbrella in 1990. I'd suggest that you start with Leica in Deerfield, IL. Wayne Buttermore is still there from the "old days" and may have a suggestion: 847-405-7044. The alternative would be to ask around in the used equipment market. Here are some of my contacts: MicroOptical Methods Dennis O'Leary Albany, NY 518-482-8200 Spectra Services Mike Specht Rochester, NY 716-654-9500 Vermont Optechs John Oren 802-425-2040 Martin Instruments Bob Martin Easley, SC 864-859-2688
Good luck. Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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} ******************************************************
At 08:13 AM 3/31/00 -0600, Nestor J. Zaluzec wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We once had a student twist the condenser control knob off a TEM. When asked to turn the knob clockwise, he just kept turning, until he handed me the knob.
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Friday, March 31, 2000 7:02 AM To: Microscopy-at-sparc5.microscopy.com
Originally posted by: sryazant-at-ucla.edu
It is not "horror" story, but a sort of... Many years ago the colleague from friendly Lab visited me with great project. The idea was simply and beautiful. She argues that because the image in the scope (TEM) is green (green fluorescence of the screen), she wants to modify the sample (I forgot what, some protein, I believe) by red-fluorescent dye to be able to see on the screen of the electron microscope the "double-staining": red on the green background. No comments...
Sergey
} Date: Thu, 30 Mar 2000 09:17:56 -0500 } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } Subject: Operator Horror Stories } To: microscopy-at-sparc5.microscopy.com } Importance: Normal } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b (Intl)|18 } January 2000) at 30/03/2000 09:18:01 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I had one faculty operator several years ago who after a SINGLE lesson on the Phillips 200, decided to come back late at night and look some more. After taking some images, he couldn't get the door to the camera chamber off, since he had failed to vent it. After rummaging through the lab, he found a large screwdriver and commenced to pry the camera door off, unconcerned about the marks and gouges that he was putting in the brass. Needless to say, when the vacuum was finally broken, the mercury diffusion pump became very unhappy. Eventually we were able to replace the instrument and the user finally left after being denied tenure.
W. L. Steffens, Ph.D Dept. of Pathology University of Georgia
We get our Ni/holey carbon films from SPI, at http://www.2spi.com/. They will prepare excellent holey carbon films on just about any available grid material (for a price of course...).
Larry
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
Nestor J. Zaluzec 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 } -----------------------------------------------------------------------.} } Originally posted by: willem.erasmus-at-sasol.com } } Dear fellow microscopists } } Does anybody know where I can purchase Ni TEM grids with a holey carbon film } ? I can find plenty of copper grids, but the Ni grids seem to be hard to } find. } } Thanks } W. Erasmus } } Willem Erasmus } Snr. Scientist, Basic Catalysis Research } Sasol Technology } Tel : +27 +16 9603954 } Fax : +27 +16 9602826 } E-mail : willem.erasmus-at-sasol.com
We at Ladd Reseach can supply you with these, as would most of the other supply houses that make their own holey film. Please let us know what size mesh you want and a quantity and we can quote you.
Debbie Sicard Sales Manager
Disclaimer: Ladd Research is a vendor that sells EM supplies. --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The respected PhD I once worked with on our old JEOL JSM-U3 scanner, who was having trouble making out the details of the image on the monitor, so grabbed a flashlight and shined it at the screen so he could see the image a little better... :-).
Larry
} } } Originally posted by: sryazant-at-ucla.edu } } } } } It is not "horror" story, but a sort of... Many years ago the colleague } from friendly Lab visited me with great project. The idea was simply and } beautiful. She argues that because the image in the scope (TEM) is green } (green fluorescence of the screen), she wants to modify the sample (I } forgot what, some protein, I believe) by red-fluorescent dye to be able to } see on the screen of the electron microscope the "double-staining": red on } the green background. No comments... } } Sergey } } } Date: Thu, 30 Mar 2000 09:17:56 -0500 } } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } } Subject: Operator Horror Stories } } To: microscopy-at-sparc5.microscopy.com } } Importance: Normal } } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b } (Intl)|18 } } January 2000) at 30/03/2000 09:18:01 } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Alan Fox, in a response to the Analytical TEM string, cautioned the } } question-raiser to beware of people who abuse instruments. Perhaps Nestor } } will forgive a little microscopy related humor and allow us to start a } } string on "Operator Horror Stories." Here's ours: } } } } The "ham-fisted user" reference made us chuckle/cringe with the memory of } } a guest "microscopist" in our lab who hauled himself (220 pounds or so) out } } of his seat by pulling on the half-inserted specimen rod, bending it about } } 20 degrees or so! } } } } Henceforth, when we saw him in the hall (he never came into the scope room } } again), we referred to him as "Conan the Microscopist"! } } } } } } } } Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com } } } } IBM Analytical Services; http://www.chips.ibm.com/services/asg } } } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant
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
W. Erasmus asked: ================================================ Does anybody know where I can purchase Ni TEM grids with a holey carbon film ? I can find plenty of copper grids, but the Ni grids seem to be hard to find. ================================================= Some times the coating of Ni grids is a bit more difficult than the coating of copper grids. This translates to somewhat lower yields and sometimes higher prices. However, SPI Supplies regularly produces custom coated Ni grids on whatever mesh and grid the customer specifies and at only a slight price premium over copper.
Contact me off line for details or visit our webpage URL http://www.2spi.com/catalog/grids/cusctgrd.html
Some of the other major suppliers of consumables also will coat Ni grids, or at least that is my understanding.
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.spi.cc ######################## ============================================
One day a user of our TEM complained that she could see the beam with the specimen out, but as soon as she put in a grid the screen went black. After playing with it awhile and ruling out magnetism or something on the grid holder, I decided that there must be some small magnetic particle in the stage area that was being pushed around by the grid holder. Of course the service technician I got by phone wanted me to begin taking the column apart from the gun on down, but I grew impatient with this and soon went straight for the stage. I figured I would be looking for some small metal sliver. Imagine my surprise when I found an entire plastic pipettortip lying across the bore through the anticontamination plate in the column! It seems the previous user had been using these tips as forceps covers, and mysteriously lost one the day before. It had gone into the column through the airlock with her grid, not impossible with the Zeiss 10 top-loading system. Although not the only strange thing I have found in that microscope's column, at about an inch and a half long it is the largest.
Aloha, Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I am currently using the peroxidase/DAB pre-embedding method for localization of a novel extracellular matrix protein. I am using 40 micron vibratome sections. While my results give some extracellular labelling, most of the labelling is associated with dendritic microtubules. Has anyone experienced non-specific labelling of microtubules?
Microscpy Core Managers, Are there any EM/Confocal Microscope facilities within academia that are running in the black or at least breaking even without a subsidy. If so what are your rates and overhead, are your salaries included? Also are there any companies or facilities that do tele-microscopy? All info will be shared with the listserver, unless confidentiality is requested. Thank You.
Michael Delannoy Basic Science Microscopy Facility JHMI
Many, many years ago... I was asked to evaluate particle size of a powder. The powder was dried sludge from a radioactive waste storage tank. My downfall was permitting a co-worker to prepare the sample. When I received it, rather than a thin layer on carbon tape, it was caked on. Furthermore, the powder turned out to be sub-micron in size and almost pure Sr-90! Static and air currents began to spread contamination, even before I got it in the chamber. I quickly left the area before I became "hot" too.
I did finish the exam... In anti-contamination dress with a full face respirator at the console!
The job took a few hours. It took two weeks for me and my service engineer to disassemble the SEM, clean (and the room) it to reasonable levels and reassemble. The vacuum system was rather "warm" from that time on... Nevermore! Nevermore!
Moral: Always insist on making input and final approval if someone else wants to prepare your samples.
We had a cousin of Conan , who in his addled age, after inserting the injector tip into the stage of our EM400 could not find his grid in the microscope; he had dropped it on the floor. Of course the tip must have fallen off in the stage, so he promptly took a second tip and properly inserted the second tip on top of the first. Still no grid could be found. Ah ha, I will leave a polite note explaining the problem. It read as follows: " Please check the microscope, I had great difficulty inserting my specimens last evening." Philips kindly replaced the bulk of the stage just so they could keep the original for the museum of what not to do.
There once were two eager graduate students at Florida in the Materials Science and Engineering Department (one has the initials MK and now works for JEOL) that decided that they wanted learn how to align the Philips 300 TEM. Armed with the user manual, they proceeded to align the column, but couldn't do it. They had to call in the service engineer. The first thing that the service engineer did was to use a framing square to help to take the visible bow out of the column. It seems that the user manual they were working from was different from one for a microscope with a STEM attachment. (Not witnessed by me, but heard from reliable sources.)
A professor in the same department was escorting a visitor around the EM lab and the two were discussing possible experiments that might be performed in the Philips 300. They asked the EM technician to give them a hand opening up the microscope so they could look inside. The technician was busy and said that he would be with them in about 5 minutes. Well, busy professors can't wait for lowly technicians so they decided to open the chamber themselves. The professor cranked on the handle to open the gun chamber while it was under vacuum and the voltage was on. Bang, pop, whoosh, whistles, bells brought the technician running to see what had happened. After all, they only wanted to look inside. But, there was a happy ending to the story. The vacuum and diffusion pump were OK and the apertures were cleaned by the supersonic air. (I was around when this one happened.)
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } [mailto:"anderron-at-us.ibm.com"-at-sparc5.microscopy.com] } Sent: Thursday, March 30, 2000 9:18 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Operator Horror Stories } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.} html } } } -------------------------------------------------------------- } ---------. } } } } Alan Fox, in a response to the Analytical TEM string, cautioned the } question-raiser to beware of people who abuse instruments. } Perhaps Nestor } will forgive a little microscopy related humor and allow us to start a } string on "Operator Horror Stories." Here's ours: } } The "ham-fisted user" reference made us chuckle/cringe with } the memory of } a guest "microscopist" in our lab who hauled himself (220 } pounds or so) out } of his seat by pulling on the half-inserted specimen rod, } bending it about } 20 degrees or so! } } Henceforth, when we saw him in the hall (he never came into } the scope room } again), we referred to him as "Conan the Microscopist"! } } } } Ron Anderson, IBM, Hopewell Jct., New York, USA. } anderron-at-us.ibm.com } } IBM Analytical Services; http://www.chips.ibm.com/services/asg } } } }
This is my second go around with this, I got the Nestor porno bump this morning.
I had a student using my JEOL 100 CX II; loading and looking around went well. When she went to remove the holder she got half way out and yanked it toward her. This action put a 70 degree bend in the holder. I could never get the Z height to behave after that for some odd reason.
The other disaster was with my Philips 300, complete with Mercury Dif pump at that time. I needed to do some anode cleaning, so without deflating the air table I got on the console finished my work and got off. When the scope lurched to the right it dumped the contents of the mercury pump into the oil pump. This sent a cloud of mercury up the column creating an alloy wherever it went.
Believe it or not the scope is in fine working order with not a trace of mercury left in in the column. How did I do it? If you can figure it out I will send the winner a Lucent/Bell Labs brief case...at my cost of course.
John Grazul Lucent Technologies
"W. L. Steffens" 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 had one faculty operator several years ago who after a SINGLE lesson } on the Phillips 200, decided to come back late at night and look some } more. After taking some images, he couldn't get the door to the camera } chamber off, since he had failed to vent it. After rummaging through } the lab, he found a large screwdriver and commenced to pry the camera } door off, unconcerned about the marks and gouges that he was putting in } the brass. Needless to say, when the vacuum was finally broken, the } mercury diffusion pump became very unhappy. Eventually we were able to } replace the instrument and the user finally left after being denied } tenure. } } W. L. Steffens, Ph.D } Dept. of Pathology } University of Georgia
Once, over a decade back, I was working EM and Med Tech at the same time at a local hospital..
We had a CAP inspection, the officers were from another hospital in the state.
The question I was expected to reason and answer with a straight face was:
* What other protective measures besides standing behind a lead wall do you take when you put the glass slide into the TEM, to protect yourself from the flying electrons???
seriously!
I was rather stunned, and started looking from the wall socket to him and back, but he didn't catch the connection.
I then tried to tactfully explain that vacuum was required, and that we really didn't need that extra protection!
It was explained to me that he did too know all about EM because he had a one day workshop.
Web Pages: Lab Page http://treefrog.cvm.uiuc.edu Centeral States Microscopy Page http://treefrog.cvm.uiuc.edu/csmms Personal Home Page http://treefrog.cvm.uiuc.edu/lam
} We once had an operator on our JEOL 840A SEM complain about her } sample moving. Her sample was samples of vacuum grease she had gold } coated and was attempting to image in the SEM. When the electron } beam heated the grease it cracked up the gold coating and charged. } She was using the SEM to look for silicon in various greases. I } suggested she use a different technique so we did not end up with } vaporized grease inside the SEM.
David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
It seems everyone has one of these. I had a person who one day complained of a burning smell while she was at the scope. I went to investigate and when I arrived in the room there was no smell... burning or otherwise. Several more times that session she came and got me to investigate this burning smell in the TEM room. This continued for a couple of sessions. I did go and investigate because after all, who wants their TEM on fire? I honestly didn't know what to do about it and was beginning to seriously question her sanity. Finally she convinced me to sit with her while she operated the scope and sure enough there was this faint smell of something burning! I jumped up and tried to find it, but it had gone away. So...she sat down again and began to work and the smell, like plastic kitchenware burning in a dishwasher, came back. Again I jump up and look around....to no avail! I was curious now. The burning smell only happens when she is sitting at the scope. Not being a big believer in spontaneous combustion, there had to be an answer! There was. As part of the TEM course I take the kick panel off the Zeiss 10 to show the students the guts of the vacuum system. The kick panel is right below the desk area where you sit up to the microscope. What she was doing was placing her tennis shoe on the heater of the diffusion pump and the plastic/rubber would melt and burn a bit -- just enough to give the room a burning smell. She never did notice her foot getting warm. When I pointed it out she did admit that her shoe was warm and had some ugly scars on it to boot! She never returned after that semester - I guess that she had enough of EM! Now I carefully point out the hazard and replace the cover quickly when we are finished with the class.
-- ================================================================== 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 ==================================================================
And all of these cute blunders that happened way back when were caused by today's scientists/researchers.
Experience, good and bad, is the only way we all really learn.
Enjoying the stories.....
Harry Ekstrom Honeywell Materials Laboratory
-----Original Message----- } From: White, Woody N [mailto:Woody.N.White-at-mcdermott.com] Sent: Friday, March 31, 2000 2:46 PM To: Microscopy-at-sparc5.microscopy.com
Many, many years ago... I was asked to evaluate particle size of a powder. The powder was dried sludge from a radioactive waste storage tank. My downfall was permitting a co-worker to prepare the sample. When I received it, rather than a thin layer on carbon tape, it was caked on. Furthermore, the powder turned out to be sub-micron in size and almost pure Sr-90! Static and air currents began to spread contamination, even before I got it in the chamber. I quickly left the area before I became "hot" too.
I did finish the exam... In anti-contamination dress with a full face respirator at the console!
The job took a few hours. It took two weeks for me and my service engineer to disassemble the SEM, clean (and the room) it to reasonable levels and reassemble. The vacuum system was rather "warm" from that time on... Nevermore! Nevermore!
Moral: Always insist on making input and final approval if someone else wants to prepare your samples.
Purdue University Department of Biological Sciences
An opening is available immediately for a scientist specializing in macromolecular microscopy at Purdue University. The successful candidate would be responsible for overseeing the day-to-day operations of the high-resolution electron microscopy facility, training new users, and assisting outside visitors with data collection and analysis. He/she would also be expected to work with the cryoEM groups in the department at planning and implementing the future directions of the facility, including the development of new technologies for improved specimen preservation, data collection, and analysis. The ideal candidate should complement the existing strengths represented in the department and pursue independent research funding.
The Department of Biological Sciences at Purdue provides a rich intellectual environment as well as outstanding modern scientific facilities including Philips EM420, CM200-FEG, and CM300-FEG electron cryo-microscopes. Purdue University is home to over 50,000 students, faculty, and staff, and is located in West Lafayette, Indiana - approximately 1 hour northwest of Indianapolis and 2.5 hours southeast of Chicago. The area affords the advantages associated with a major university, while providing an affordable and attractive `small town' environment in which to live.
Requirements for this position include a Ph.D. in biophysics or one of the related sciences, a minimum of 5 years post-doctoral work experience in electron cryo-microscopy of biological molecules, and a proven ability to work well with others. Preference will be given to candidates who have successfully developed new technologies and methodologies for high-resolution electron cryo-microscopy. Salary will be commensurate with experience and includes full benefits.
Interested individuals should send their resume, relevant reprints, a statement of research interests and career goals, and provide contact information (name/phone/e-mail) for three individuals who have agreed to serve as references to:
Dr. Timothy S. Baker (tsb-at-bragg.bio.purdue.edu)
and/or
Dr. Amy McGough (amcgough-at-bcm.tmc.edu)
Department of Biological Sciences Lilly Hall of Life Science Purdue University West Lafayette, IN 47907-1392
Purdue is an equal access/equal opportunity university.
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Is there any truth in the story I was told about a lab in London that replaced the leaded glass on the microscope chamber with regular glass? Apparently the mistake was discovered when all the film on the shelf behind the operator become fogged!
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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Hello All, My two horror stories! I was working with an investigator who wanted to see matrix vessicles. Well, we were so happy that we had a nicely fixed sample with "lots" of vessicles. Nice whole round ones. Well, when the negatives were developed, my director and I discovered that what we really had were nicely fixed, beautiful bacteria!!!! No vessicles! Another time, in school, I was walking down the hallway later in the afternoon when I saw a scope room door slightly open. I popped my head in to see who was there and to ask if they wanted the door closed. When I looked inside, I found a student sound asleep, with hands still holding the knobs of the microscope!!!!!!! Funny huh? Jo
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Alan Fox, in a response to the Analytical TEM string, cautioned the } question-raiser to beware of people who abuse instruments. Perhaps Nestor } will forgive a little microscopy related humor and allow us to start a } string on "Operator Horror Stories." Here's ours: } } The "ham-fisted user" reference made us chuckle/cringe with the memory of } a guest "microscopist" in our lab who hauled himself (220 pounds or so) out } of his seat by pulling on the half-inserted specimen rod, bending it about } 20 degrees or so! } } Henceforth, when we saw him in the hall (he never came into the scope room } again), we referred to him as "Conan the Microscopist"! } } Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com } } IBM Analytical Services; http://www.chips.ibm.com/services/asg
Bart, I believe Argon is used because it is a larger molecule and sputters the metal more efficiently. I also use air with no problem. Possibly the amount of metal sputtered is slightly less per unit time. John Hunt
On Fri, 31 Mar 2000, Nestor J. Zaluzec 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 } -----------------------------------------------------------------------. } } } } Originally posted by: bart.depauw-at-rug.ac.be } } } Hello, } } I would like to know why they use Argon in the metal coating devices. At } our laboratory, we don't use Argon, we use air. What is the advantage of } using Argon ? } De Pauw Bart } Faculty of Veterinary Medicine } Morphology } Salisburylaan 133 } 9820 Merelbeke } Belgium } Phone : 0032(0)9 264.77.19 } Fax : 0032(0)9 264.77.90 } } }
Air contains water vapour, oxygen and carbon dioxide all of which can be broken down in the plasma generated in a sputter coater and gve rise to nasty active species which can damage delicate samples. Argon is used because of its relative high atomic number and this ensures there is multiple scattering of the sputtered metal. There is a Zenon sputter coater on the market which should sputter even better but I have no experience with it. Come to the Lehigh Short Course(contact {slc6-at-lehigh.edu) and we will tell you all about it.
Many years ago we had a Siemens EM-101. This was a relatively new microscope and beautifully machined with German precision. The camera chamber was a work of art. Each film plate (we used glass back then) was encased in it's own light-tight cassette. The camera would move a film cassette into position for exposure and pull off the cassette cover. The operator would then expose the film and move the casette into a drop box. The cassette cover would be pushed closed in the process. This camera worked almost flawlessly. On occassion, an operator, when loading the camera, would put the cassettes in on an angle. When you went to take the first image, the cassette would jam and not move into position. All that was necessary to unjam was to break vacuum, giggle the cassette with your hand to straighten it and then repump the camera chamber and get back to work...a 10 minute process as most. We had one operator who thought she knew it all. She was working on the microscope one evening and the camera jammed. Her solution was to take out the camera and take it apart...screw by screw and spring by spring. She ended up with an incredible heap of parts and of course had no clue as to how to put it back together. Neither did the Siemens service engineers. They had never seen one apart! They had to find another camera (not easy in those days with relatively few of these instruments around), take it apart..carefully marking where each piece came from...and then reassembled each camera simultaneously. It took hours!! Moral of the story...NO ONE fixed ANYTHING without permission of the lab personnel!!
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
I am preparing to purchase a sputter coater for Au/Pd SEM specimen preparation. I have narrowed the search to the VG-Scientific Polaron SC7620. My old Polaron E5100 is very tired.
I have located used Hummer VI, Fullam EMS-76M and Denton Desk II coaters for about $1000 less than new. Some of these include the pump. However, I have several dual stage mech pumps, so this is not an issue.
If anyone knows of a source for a good/recent used coater, I would appreciate knowing about it. Otherwise, I will go with a new one.
These are great stories and would make a wonderful collection, especially to hand out to students of EM. Yes, experience is a great way to learn but it's often best, and cheaper, to learn from some else's.
I heard that one years ago. Also one that supposedly happened at Univ. Wisconsin. Someone apparently cleaned a scope's internal parts in acetone and started pumping. Before it was fully pumped down, they turned on the filament. Bang! Nice story but could it possibly happen? I would think that any acetone residue would evaporate very quickly or is there some other phenomenon here relating to ratio of gases?
Damian Neuberger, Ph.D. Baxter Healthcare, Inc. WG3-2S P.O. Box 490 Round Lake, IL 60073 e-mail: damian_neuberger-at-baxter.co, Tel: 847-270-5888 Fax: 847-270-5897
} Is there any truth in the story I was told about a lab in London that } replaced the leaded glass on the microscope chamber with regular } glass? Apparently the mistake was discovered when all the film on the } shelf behind the operator become fogged! } } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } phone:213 273 8026 } fax: 213 413 6739 } e-mail: pwebster-at-hei.org } http://www.hei.org/htm/aemi.htm
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