We use another method to provide dry nitrogen for venting, valves etc.. Rather than use bottled gas, we take the gas from our liquid nitrogen vessel. It is dried and as yet (touch wood) we have had no problems. This is after approximately 18 months operation.
The vessel is large, 2000 litre capacity but the gas is used for a TEM, SEM, quadrupole mass. spec. and a few other instruments. It is also used to vent ALL vacuum systems in the E.M. lab.
It sure beats having to change gas bottles!!
##################################################################### ********************** * Between the idea * 0------* And the reality * } ---|--- { * Between the motion * | * And the act * / \ * Falls the Shadow * _/ \_ * T.S. Eliot * ********************** Colin Veitch Tel + 61 (0)52 47 2611 CSIRO Division of Wool Technology Tel + 61 (0)52 47 2891 (dir.) P.O. Box 21 Fax + 61 (0)52 47 2657 BELMONT Vic 3216 Australia
To biological light microscopists, I want to make hydrophobic rings on glass microscope slides so that tissue-processing and immunocytochemistry can be carried out on tissue on the slide. To date, I have been smearing a thin ring of silicone (auto and caravan sealant!) and allowing it do dry. The difficulty with this is that the silicone has to be shaved off with a razor blade before the tissue is cover-slipped. I have heard of a product called a PAP pen which does the same thing, with the advantage that the hydrophobic layer is very thin. I found one such product made by "Agar" Essex, UK, distributed by "Alltech" (Australia) but the asking price seems outrageous, approx $Aus120 per pen (approx $US80). 1. Is there a similar product available from other companies (addresses, fax. nos appreciated)? 2. Is there a "home-brew" alternative, perhaps using liquid silicone?
Thanks for any advice, Dave Merritt Zoology University of New England Armidale NSW Australia dmerritt-at-metz.une.edu.au
I used to use a Sorval MT2-B. The company that serviced it for me was "RMC" in Arizona. Their phone number is 602-889-7900. They have service people all over the US. You may want to try them.
I've been charged with purchasing a high resolution coating system to use with both scanning probe and electron microscopes. My lab is primarily a scanning force microscope lab where we look at proteins and polysaccharides on polymers and cell membranes. We are interested in starting a substantial correlative microscopy effort using field emission SEM's on the same samples that we've previously probed with AFM or STM. The only coating systems currently available to us are mechanically pumped DC sputtering units. I have ~$25k to spend and have spent some time looking at the options. It looks like a turbo pumped sputtering system with a rotary tilt stage would best suit our needs. However, I'm in the dark concerning what parameters to assess in determining an optimal system. I know that grain size varies with the sputtered material. Does it also vary between different sputtering systems? Is a thermal evaporation source something to consider? Is it important to spend the money on an oiless backing pump for the turbo? What questions am I not asking that I should be asking?
To make hydrophobic rings on glass microscope slides an old homebrew method is to dilute a resin mountant such as DePex or Permount or Canada Balsam in the appropriate solvent and to use this as ringing agent. Try diluting the mountant 10 to 50 times with the recommended solvent (usually xylene or toluene) and apply with the sharpened end of an applicator stick. The advantage is that you do not have to remove the stuff before mounting under a coverslip. It is also a useful method to keep serial monitor sections in the correct order, each on its own separate little water droplet inside its own hydrophobic ring.
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
We are using fluorescent dyes on bleached wood pulp fibres (ie cellulose) for confocal microscopy. We are having problems with "bleeding" or leaching of dye. Are there any fluorescent dyes which are reactive with cellulose or can be fixed similar to textile dyes (RIT, etc.) so that they will not leach out in aqeous environments? Thanks in advance.
James Drummond Pulp and Paper Research Institute of Canada Vancouver, B.C. Canada
We are using fluorescent dyes on bleached wood pulp fibres (ie cellulose) for confocal microscopy. We are having problems with "bleeding" or leaching of dye. Are there any fluorescent dyes that bind to cellulose or can be fixed similar to textile dyes (RIT, etc.) so that they will not leach out in aqueous environments? Thanks in advance.
James Drummond Pulp and Paper Research Institute of Canada Vancouver, B.C. Canada
} There is a potential alternative for bottled N2. There are driers for } compressed air that dry it to a lower dew point than lab grade N2. In } theory bleeding some of this into the tank instead of N2 from a } cylinder should work.
I haven't tried this, but you still would probably have the problem of all the hydrocarbons from a standard air compressor... How do these driers work? If they use some kind of cold trap, you might be okay. Liquid nitrogen will freeze both water and hydrocarbons out of an atmosphere.
Chris Krenn Graduate Student UC Berkeley Dept. of Materials Science
At the risk of swimming upstream (and against the apparent popular opinion), I have used PAP pens for slide mounted tissue processing, and if that is your method of choice, I think the PAP pen works just fine. I say if that is your choice only because i much prefer processing immunocytochemical material as free-floating sections rather than slide-mounted - better penetration, more uniform staining, and (extremely important) better rinsing (at least in my experience). The PAP pen, as best as I can smell, is a mixture of bees wax dissolved in xylene (or some similar concoction). It goes on easily if you have a dry area around your tissue, and it comes clean in a xylene rinse before mounting. It requires a little practice to get the right pressure so you don't get too much liquid out of the pen, but it is not brain surgery!
The PAP pen is available for about $30 US from RPI:
Research Products International 410 North Business Center Drive Mount Prospect, IL 60056
Ph: 1-800-323-9814
Let me know if you have any further questions. Cheers,
David Morilak Dept Pharmacology Univ Texas Health Science Center San Antonio, TX 78284
Some of the problems can be due to using a disposable blade. We cut routinely serial cryosections of young chick brains cyroprotected in 30% sucrose and snap frozen in dry-ice or nitrogen chilled heptane and mounted on the chuck with OCT. Fixation is with just about every imaginable fix possible. Crank the temperature down, I like -25 to -35, use a real blade and keep it clean, especially the back of the blade. Sometimes I swing the anti-roll plate out of the way and use a #2 paintbrush (keep it inside the cryostat) to prevent curl - touch it to the base of the section as it gbegins to come off the blade and coordinate your arm so that you can move the brush in rythmn with the section as it moves with the cutting stroke. I've tried disposable blades for paraffin, to cut serial paraffin sections, without success. But the standard blades will allow me to cut ribbons as long as my arm can stretch holding the free end of the ribbon.
On Thu, 28 Apr 1994, Greg Erdos ICBR EM Core Lab Univers wrote:
} From: TOWER::GWERDOS "Greg Erdos ICBR EM Core Lab Univers" } To: IN%"pataky-at-bcu.ubc.ca" } CC: GWERDOS } Subj: Re: Cryostat help! } } } From: IN%"pataky-at-bcu.ubc.ca" } } Subj: Cryostat help! } } } } Return-path: {BIOSCI-REQUEST-at-net.bio.net} } } Received: from net.bio.net by gnv.ifas.ufl.edu (PMDF V4.2-11 #3240) id } } {01HBOOXBZ5DS8X4B47-at-gnv.ifas.ufl.edu} ; Wed, 27 Apr 1994 22:18:06 EST } } Received: (from daemon-at-localhost) by net.bio.net (8.6.8.1/8.6.6) id TAA23358 } } for cytonet-list; Wed, 27 Apr 1994 19:04:06 -0700 } } Received: (from news-at-localhost) by net.bio.net (8.6.8.1/8.6.6) id TAA23346 for } } cytonet-arpanet; Wed, 27 Apr 1994 19:04:05 -0700 } } Date: Wed, 27 Apr 1994 23:50:46 +0000 (GMT) } } From: pataky-at-bcu.ubc.ca (Dave Pataky) } } Subject: Cryostat help! } } To: cytonet-at-net.bio.net } } Message-id: {pataky-270494154951-at-steevlab.generes.ca} } } Content-transfer-encoding: 7BIT } } Followup-To: bionet.cellbiol.cytonet } } NNTP-Posting-Host: steevlab.generes.ca } } } } Anybody out there know why Tissue-Tek, presumably designed for embedding } } samples for cryostat cutting, is so $##-at-***%* annoying to work with? The } } problem: I'm cutting chick embryonic brain tissue, anywhere from 10-40 } } microns, at -20 and no matter what I try the sections won't stop curling } } up, rolling into tubes as soon as I lift the antiroll plate. I've tried } } adjusting the blade angle, the "anti-roll" (that's a joke) plate, the } } temperature, speed of cutting, new blade (disposable), nothing works. } } Ideally I'd like to see nice flat sections that stay that way, preferably } } "ribboning" off the blade so I can mount several at once. Know any tricks } } or alternatives to Tissue Tek which may work better? Any ideas how to deal } } with static electricity? (sometimes the sections stick to the "anti-roll" } } plate). Feels like I'm battling the antichrist (and losing!), } } } } } } -- } } Dave Pataky } } Dept of Zoology, UBC } } pataky-at-bdc.ubc.ca } } } Brain can be particularly difficult. I suggest cryoprotecting in 20% sucrose } with 3% PEG MW 400. Cut at -25. Don't expect to get a ribbon. We are usually } satisfied with one section at a time. Rarely do we get two. I have a nice } handout on cryosectioning that I picked up at a meeting. Give me your FAX # } or mailing address and I will send a copy. } } ********************************************************** } * Greg Erdos ** * } * Director, ICBR EMCL ** Phone 904-392-1295 * } * 218 Carr Hall ** FAX 904-392-8598 * } * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * } * Gainesville, FL 32611 ** * } ********************************************************** } ********************************************************** } * Greg Erdos ** * } * Director, ICBR EMCL ** Phone 904-392-1295 * } * 218 Carr Hall ** FAX 904-392-8598 * } * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * } * Gainesville, FL 32611 ** * } ********************************************************** }
FOR THOSE OF YOU INVOLVED IN EDITING JOURNALS, WRITING BOOKS, ETC. IN THE FIELD, AND ANYONE ELSE WITH AN OPINION, WHAT DO YOU THINK ARE THE MOST ACCEPTABLE ACRONYMS TODAY FOR: HIGH RESOLUTION (TRANSMISSION) ELECTRON MICROSCOPY " " (SCANNING) " " LOW VOLTAGE, FIELD EMISSION, SEM HIGH PRESSURE OR ENVIRONMENTAL SEM
WOULD APPRECIATE SHORT RESPONSES WITH SUGGESTED ACRONYMS, TKS, LINDA SAWYER
We have a JEOL JSM-35C SEM in outstanding condition for sale. It was purchased in 1980 and was on service contract until April 21, 1994. It comes with a Tracor Northern (Noran) EDS detector and TN-2000 analyzer. Asking $8,000.00. Contact Dr. Stanley L. Flegler Center for Electron Optics Michigan State University Flegler-at-pilot.msu.edu
FYI, using FETCH, I reached your ftp volume at pub/ncsu/jruss, not pub/jruss as indicated in your note to the NIH Image list. Would this be a better place to exchange things rather than mailing?
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
Microscopy list: My apologies for that last note. It was intended for John Russ in response to one of his helpful explanations on the NIH Image mail list.
Again, my apologies.
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
If there are any groups in the preperation of pulp that bind feulgen, the material will give a brilliant red signal appropriate for most confocal instruments. It is very "fast" if it binds.
Bill Tivol asked: Does anyone know a system with the sensitivity to quantitate small num- bers (1 or 2) of electrons, with the ability to produce a signal proportional to electron number for the more intense reflections (i.e. no saturation)? Since each electron hitting a film grain exposes that grain, it's hard to beat that quantum efficiency. Positional accuracy is less important for us than accurate quantitation, including background subtraction.
I don't have any inherent bias towards Hammamatsu; it is merely the only vendor whose literature I've gotten so far, and whose name I was familiar with because of their image processing boxes. They have a series of CCD based photon counters which also have a wide dynamic range (10^6)
Chris Krenn Graduate Student UC Berkeley Dept. of Materials Science
Thanks for your reply. Word of your good results came through the grapevine, and was one of the reasons we are looking at CCD instead of SIT. Hopefully we will have a somewhat functional system by the end of the year.
Received: from MAILQUEUE by MICROBIO1 (Mercury 1.11); Tue, 3 May 94 11:36:44 EST Return-path: {Stanley_Hayes-at-rml.niaid.pc.niaid.nih.gov} Received: from emoryu1.cc.emory.edu by transporter.microbio.emory.edu (Mercury 1.11); Tue, 3 May 94 11:36:37 EST Received: from anlemc.msd.anl.gov by emoryu1.cc.emory.edu (5.65/Emory_cc.3.4.21) via SMTP id AA14758 ; Tue, 3 May 94 12:34:03 -0400 Return-Path: Stanley_Hayes-at-rml.niaid.pc.niaid.nih.gov
Dear Alfred,
We have a Pall air dryer, not to air the HVEM, but to pump dry air through our high-voltage tanks in the event that they have been opened. We find that using the dried air makes a big difference in residual water in the tanks after the air is pumped out and replaced by the SF6 insulating gas which is the normal fill gas for the tanks. These tanks are 2-3 meters in diameter and about 4 meters tall with a connecting piece about 1 meter**3. The air dryer has a pre-filter to remove oil, the dessicant, an oil vapor filter, and a particle afterfilter. The specified dewpoint reached is -60o F or below. It's also noisy enough that I'd hate to have to turn it on each time I wanted to air the microscope, but it could be mounted in a sound-absorbant cabinet.
Message-Id: {199405040807010548-at-qmgate.secrc.abb.se} X-Mailer: InterCon Dispatcher/SMTP for QuickMail X-Priority: 4
Looking for someone that has been working with oxidation, hydriding and wear properties of surface coated or surface modified zirconium base alloys (like Zircaloy 2 or 4). The environment is water or steam at ]300!C. I am interested in all topics related to the subject; coating or surface modification methods ( how to get a coating without defects going through the coating?), corrosion tests, investigation of tested samples (LOM, SEM, TEM, SAM, XRD, electrochemical impedance spectroscopy etc.).
Bengt Stridh ABB Corporate Research S-721 78 Vasteras Sweden E-mail: best-at-secrc.abb.se Fax: +46-21-13 41 00 Phone: +46-21-32 30 67
================================================ An Update Message from the Microscopy Listserver ================================================
G'day All Microscopy Listserver subscribers....
In sorting out a mailserver problem, today, I just noticed that about 2 weeks ago we received our 1000th subscription request. As in previous milestones, I hearby offer to by a beer (or any other appropriate concoction) for
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at the next microscopy meeting that we both attend. I'll definitely be at the MSA New Orleans meeting so Mikey whoever you are, make sure you look me up if your there. I hear tell there may be a pub/bar or two in town. Hmm.. at this rate (1000+ in 6 months of operation), I may be buying more than 1 or 2 beers in New Orleans, hope I don't exceed my limit being the quiet, unassuming person that I am. ;-)
We now have subscribers on every continent except Antarctica anyone know a frozen microscopist/microanalyst down there that's on Email??
As promised earlier in the year a new computer system has arrived, and I'm slowing getting around to shifting software to that machine. I'm having some trouble linking the MSA BBS into the new machine so things are taking longer than expected (as usual). The transition should be seemless at your end as there will be no changes in addresses or functionality just new hardware.
P.S. Another reminder: if you want to unsubscribe from the listserver you MUST supply the original username-at-host address , which you originally subscribed to the listserver using. If you used an alias then you must unsubscribe using that alias name. I still get frustrated messages from individuals who try to unsubscribe, but do so using an unregistered address which the software just simply ignores, and hence they are not unsubscribed and continue to receive Email.
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Message-Id: {9405051128.AA17490-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: NOMENCLATURE: HREM, HRSEM, LVSEM, LVHRSEM, ETC. Orig-Author: {lcs-at-rlmtc.DNET.hcc.com}:ddn:wpafb -----------------------------------------------------------
FOR THOSE OF YOU INVOLVED IN EDITING JOURNALS, WRITING BOOKS, ETC. IN THE FIELD, AND ANYONE ELSE WITH AN OPINION, WHAT DO YOU THINK ARE THE MOST ACCEPTABLE ACRONYMS TODAY FOR: HIGH RESOLUTION (TRANSMISSION) ELECTRON MICROSCOPY " " (SCANNING) " " LOW VOLTAGE, FIELD EMISSION, SEM HIGH PRESSURE OR ENVIRONMENTAL SEM
WOULD APPRECIATE SHORT RESPONSES WITH SUGGESTED ACRONYMS, TKS, LINDA SAWY
Does anyone know of a low to moderately priced sharpener for carbon coater rods? The motorized ones I've seen in microscopy supply houses are going for about $1000 which seems a bit over-priced to me. I saw one once that was based on an ordinary manual type pencil sharpener, have not been able to find one. Any help on this subject would be greatly appreciated.
********************************************************************** * Glenn Poirier glenn_p-at-geosci.lan.mcgill.ca * * Electron Microprobe Lab Phone: (514) 398 6774 * * Earth and Planetary Sciences Fax: (514) 398 4680 * * McGill University THERE ARE THREE SIDES TO EVERY STORY; * * Montreal, Quebec YOUR SIDE MY SIDE AND THE TRUTH * **********************************************************************
The X-ray lab at the US Bureau of Mines has a carbon coater rod sharpener that is like a pencil sharpener. Where they aquired it i am not sure. You should contact Gary Van Landyut at PO Box 280, Rolla, Mo 65401 or call 314-364-3169. C
This may sound a little crazy but... Try a theatrical lighting company-one that specializes in high intensity spotlights. these apparently use carbon rods which are sharpened in a similar manner.
Mark Elliott UBC-Pulmonary Research Laboratory, St. Paul's Hospital, Vancouver, BC
I'm interested in any recommendations for cooled CCD cameras and associated MAC software for light-level immunofluorescent studies. We currently use a Hamamatsu SIT camera and NIH Image. The sensitivity is OK for our studies, but I'd like better resolution. Our cells are very small and we want to image several monoclonals against antigens in the same cell. What are the advantages of the various CCD brands? Has anyone used IPLabs software with a Power Mac? Will we see a big improvement in resolution over the SIT camera (an NTSC-based video camera) we now use? Thanks for any replies.
Does anybody have a proven method (recipe and conditions) for the electropolishing of Ta single crystal samples? Thanks. Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
Date 5/6/94 Subject Carbon Rod Sharpeners From Chris Bowser To Magnavox Internet
Subject: Time:7:03 AM OFFICE MEMO Carbon Rod Sharpeners Date:5/6/94 smtp%"microscopy-at-anlemc.msd.anl.gov"
I have a carbon rod sharpener that I purchased about 10 yr ago from the Ernest F Fullam company. You might try them. Also the Ted Pella company has a small hand sharpener for about $40.00.
We are planning on using freeze-substitution as our standard protocol for specimen preparation for thin sections. I would like to get an idea of how many subscribers to this newsgroup use freeze-substitution and what general comments you may have. I have several questions, some of which cannot be answered by reading published methods. We will be freezing our specimens (bacteria) in liquid nitrogen cooled liquid propane. Should a special grade of propane be used for this? What safety precautions should be used with liquid propane? What types of substitution media are preferred? I have seen that both methanol and acetone are commonly used as solvents. Most substitution media contain glutaraldehyde, osmium tetroxide and uranyl acetate. Are there any tricks to making this mixture up? Once the substitution medium has been made up, should it be stored frozen in liquid nitrogen? Thanks in advance for any comments.
J. W. Austin Microbiology Research Division Food Directorate Health Protection Branch Health Canada Ottawa, Ont.
The Microscopy Listserver has been down due to hardware problems for the last 2-3 days. Things should be back to normal shortly. Please report any major problems to: Zaluzec-at-anlemc.msd.anl.gov
Sometimes ago I posted a questions about difference between digital and other type confocal. I got an address for a server group. I tried subcribing, but could not. Does anyone has that server address, and can someone give me information about confocals.
S. D. Walck's contribution to the backfill question had a lot of good things to say. I'd like to add two cents worth: Using teflon tubing, available, e.g. from Cole-Parmer works better than tygon in LN2.
i'm looking for a U.S. distributor, if there is one, for vertical slide staining dishes and racks. like standard dishes + racks, but the slides go in vertically and use less solution, and can hold more slides (25x75)than a coplin jar - 20 or 30 i think. they're evidently standard in Japan, and we have a quote from a Matsunami Trading Co. in Osaka for them at $10.70 for racks and $11.36 for dishes, which is reasonable, but Air Parcel Post at 2/3 of the list price isn't. so i was wondering if anyone might know of a U.S. source for these things. haven't come across any in the obvious places.
------------------------------------------------------------------ |Glenn Holm Internet:karuzis-at-wccf.mit.edu| |M.I.T Dept. of Brain + Cog. Sci. Bitnet:karuzis-at-mitwccf | |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | ------------------------------------------------------------------
Try Fisher Scientific - the staining sets are plastic, hold from 100-250 ml depending on how much of the slide you need immersed, and the racks hold 25 slides. The only problem I have found using these setups is that evaporation of solution is serious if you leave them sit. I keep my solutions in bottles and pour them back when I'm finished staining (the ones that aren't made up fresh of course). The sets are made by Tissue-Tek.
Cheers
David Morilak Dept Pharmacology UT Health Science Center San Antonio morilak-at-uthscsa
I was wondering if anyone could recommend a couple of good books on TEM and SEM of plant materials. I've always worked with animal and human tissue, and microorganisms so all of my books are related to these topics.
Thanks,
Phil Rutledge p.s. If you know the publisher and ISBN that would help also.
Message-Id: {MAILQUEUE-101.940511084750.751-at-prl.pulmonary.ubc.ca} To: microscopy-at-anlemc.msd.anl.gov
Does anyone know of an easy way to fix a scratch on a 40X dry objective lens for an Olympus scope, other than returning it to Olympus??? Someone here scratched their's and are on limited budget and need it fast. Any suggestion greatly appreciated.
Thanks, Mark Elliott UBC-Pulmonary Research Lab, Vancouver
Message-Id: {MAILQUEUE-101.940511095225.756-at-prl.pulmonary.ubc.ca} To: MICROSCOPY-at-anlemc.msd.anl.gov
a FEW SUGGESTIONS; Introduction to Biological Electron Microscopy: Theory and Techniques, By Clinton J. Dawes Ladd Research Industries, Inc, Burlington Vermont, Library of Congress # 86-082930-not sure of ISBN #
Also Books by Robards, not sure of title-if find will let you know. Can also check for books by Hyatt, not sure which one but one has plant material in it.
Depends on what type of plant material you are dealing with-Fungi, marine algae or vascular plants-they are all different and their methods of preparation are different.
We have been using Denka LaB6 {100} 60 deg filaments in our JEOL 4000 with less than great results. Brightness is poor and after a week of use the center of the filament is small and dim and most of the illumination is from the lobes. Has anyone tried the LaB6 filaments from Kimble Physics or FEI in a JEOL 4000? Do they seem better or worse? Do they last?
Thanks Roseann Csencsits Electron Microscopy Center Argonne National Laboratory Argonne, IL
If anyone has a Noran 5500 no longer in service with good display boards and wants to make a deal. Please contact me directly by e-mail or voice.
*************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
} We have been using Denka LaB6 {100} 60 deg filaments in our JEOL 4000 } with less than great results. Brightness is poor and after a week of } use the center of the filament is small and dim and most of the } illumination is from the lobes. Has anyone tried the LaB6 filaments } from Kimble Physics or FEI in a JEOL 4000? Do they seem better or } worse? Do they last? } } Thanks } Roseann Csencsits } Electron Microscopy Center } Argonne National Laboratory } Argonne, IL
We were trying the Kimball filaments when I first got here two years ago. After blowing a few, about which they were VERY understanding and generous, we decided to go back to tungsten, until everyone could sit down and figure out what was happening.
Peter Sewell, who makes the filaments, has been very helpful and accessible. At the time, he told me that they were doing some more testing on why they were getting runaway heating in JEOL scopes, but not Philips. It has to do with the control circuitry of the gun and what is being regulated. (I'm neither a physics nor an electronics whiz, so please don't ask for more specifics from me!)
The bottom line is that conditioning of the filament to be used in a JEOL, as well as conditioning of the gun, is essential if you're going to use their filaments. We have a JEOL 2000 EXII and have adopted their procedure at installation of tungsten filaments. We haven't gone back to trying the Kimball ones yet.
Filament conditioning: after installation, set gun bias to zero, turn up filament heating SLOWLY, when the vacuum starts to degrade stop, when it stabilizes continue turning up till you get to near your normal operating conditions. Only after heating the filament and getting beyond the outgassing should you turn up the bias to get emission. After conditioning this way, you shouldn't get any more outgassing. From what I can tell, you should be able to do this at any accelerating voltage. This conditioning has taken me one and a half hours. Be patient. I'm expecting very long filament life after this treatment.
I'd be interested in others experiences with this kind of procedure.
John chandler-at-lamar.ColoState.EDU Fort Collins CO
} From: IN%"jgoodhouse-at-molecular.Princeton.EDU" "Goodhouse, Joseph" } Subj: Silver enhancement, ultra structure } } Return-path: {jgoodhouse-at-molecular.Princeton.EDU} } Received: from anlemc.msd.anl.gov by gnv.ifas.ufl.edu (PMDF V4.2-11 #3240) id } {01HC7XIRCF4G8X5IJZ-at-gnv.ifas.ufl.edu} ; Wed, 11 May 1994 16:49:27 EST } Date: Wed, 11 May 1994 09:48:00 -0400 (EDT) } From: "Goodhouse, Joseph" {jgoodhouse-at-molecular.Princeton.EDU} } Subject: Silver enhancement, ultra structure } To: Micrscopy {Microscopy-at-anlemc.msd.anl.gov} } Message-id: {2DD0E2AB-at-molecular.princeton.edu} } X-Mailer: Microsoft Mail V3.0 } Content-transfer-encoding: 7BIT } Encoding: 10 TEXT } } } I am trying to do silver enhancent on 3nm gold probes in Drosophila embryos } for ultra structure morpholgy. The problem I am experiencing is that the } reaction is occurring too rapidly and is blowing the embryos apart. Can } some one direct me to a few good references on this technique for cells and } tissues, or provide me with a working protocol. Much appreciation } Joe Goodhouse } Dept. of Molec. Bio. } Princeton University } jgoodhouse-at-molecular.princeton.edu #################%%%%%%%%%%%%%%%%%%%%%%%%%%%##################%%%%%%%%%%%%%% If you are using one of the daylight kits like the one from BioCell you might try diluting the reagents with water to slow the reaction. Their rep. told me this at a meeting one time. Maybe lowering the temp would work too ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * ********************************************************** ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
} From: IN%"KARUZIS-at-wccf.mit.edu" "GLENN HOLM" } Subj: RE: U.S. supplier for vertical staining dishes? } } Return-path: {KARUZIS-at-wccf.mit.edu} } Received: from WCCF.MIT.EDU by gnv.ifas.ufl.edu (PMDF V4.2-11 #3240) id } {01HC7V1A7RVK8X8BBU-at-gnv.ifas.ufl.edu} ; Wed, 11 May 1994 15:38:11 EST } Received: from wccf.mit.edu by wccf.mit.edu (PMDF V4.2-14 #2603) id } {01HC7UYC8CK48WXF5V-at-wccf.mit.edu} ; Wed, 11 May 1994 15:37:26 EST } Date: Wed, 11 May 1994 15:37:26 -0500 (EST) } From: GLENN HOLM {KARUZIS-at-wccf.mit.edu} } Subject: RE: U.S. supplier for vertical staining dishes? } To: GWERDOS-at-gnv.ifas.ufl.edu } Message-id: {01HC7UYC8CK68WXF5V-at-wccf.mit.edu} } Organization: Mass. Inst. Tech. - Whitaker College } X-VMS-To: IN%"GWERDOS-at-gnv.ifas.ufl.edu" } MIME-version: 1.0 } Content-transfer-encoding: 7BIT } } greg- } } thanks for the reply. i looked at the Fisher staining dish, but what we } want is a bit larger and holds the slides vertically in a rack, not in } grooves in the side of the dish like a coplin jar. will keep hunting. } ------------------------------------------------------------------ } |Glenn Holm Internet:karuzis-at-wccf.mit.edu| } |M.I.T Dept. of Brain + Cog. Sci. This VAX doesn't do NeXTmail | } |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | } ------------------------------------------------------------------
############%%%%%%%%%%%%%%%%##############%%%%%%%%%%%%%%########## I just remembered where I saw it. Shandon & Lipshaw Catalogue. Purveyors of goods for pathology, cytology and mortuary. Call 800-547-7429 and they will send a catalogue that smells like a morgue. Check out page 184 for a rack that holds 38 slides vertically on end.
Or if you are close to a hospital pathology lab or autopsy lab they may have the catalogue.
A few years ago, I used a very nice silver enhancement kit from Janssen called IntenseII. I used it only for light microscopy, but I know that people have used it successfully for EM as well. It was simple and quite forgiving with regard to development time. I also believe the reaction can be slowed by diluting the reagents. I don't remember who the US distributor for Janssen is now, but for some reason I'm thinking it might be Vector.
Cheers
David Morilak Dept Pharmacology UT Health Science Center San Antonio morilak-at-uthscsa.edu
Dr. Avalos, I lost your E-mail address. Please send it. Thank you. My apologies to the others on the list. Dr. Stanley L. Flegler flegler-at-pilot.msu.edu
I can't afford a new scanner but the thought of upgrading my Hitachi S450 with the Semicaps imaging system has excited me. My questions are...does anyone have the system on an analog scope? How does it grab the image off the scope? Can I use it with my TEM also? Is it as good as the other systems out there?
John Grazul Rutgers University Electron Microscope Facility
I saw a request recently from Michael Winton concerning TEM sample preparation of silicon on sapphire, but I saw no responses posted to the listserver.
Did anyone out there have a good (or reasonably good!) answer for him?
Thanks!
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
Now I remember. The comercial daylight kits for silver enhancement require the addition of gum arabic in order to slow the reaction. I can't immediately recall the deatails but maybe it will ring a bell in a younger brain ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
} Does anyone have proven sample preparation techniques for thin film } silicon on sapphire substrates?
Hi, Michael:
I tried to post this mail last weekend but the listserver was down. I hope this can be any help to you.
I made such a sample once. It came out beautifully. I assume that you are talking about cross-sectional TEM sample because it is easy to make plane-view sample. The recipe is as following:
1. Stack two pieces face to face together with a very thin layer of M-bond. Curing samples at 170 degree C for two hours. 2. Grind and polish one side. (with SiC paper first then finish with diamond paste). The thickness of the sample is 100 um now. 3. Dimple the other side to less than 25 um. 4. Ion mill the sample.
The experiences of grinding, polishing and using dimpler are necessary. Since sapphire is brittle and hard, you need patience. Good luck. -- Tan-Chen Lee Cornell University tanchen-at-msc.cornell.edu
Dear Clayton Smith: Thank you for your letter of May 4, 1994 and the catalog. Dr. Curzon will make a decision. My apologies to the others on the list. Sandy Burany
Subject: Time:1:49 PM OFFICE MEMO N2 from LN2 Date:5/12/94 In the recent discussions about using dry nitrogen to backfill vacuum systems, several of us have mentioned collecting the gas that boils off liquid nitrogen storage vessels and using it for this purpose. There is a very inexpensive way to do this that I devised some 20 years ago and used for many years with good success. What you do is to run a flexible, non-collapsible plastic tube (I prefer polyethylene tubing to Tygon, because Tygon seems to exude so much plasticizer - which might find its way into the vacuum system) from the liquid nitrogen container to the gas inlet of the vacuum system. At a convenient site, insert a tee joint into this tube. A large, flexible, inflatable, plastic container (a beach ball or childs toy animal) is attached to this tee joint by means of a length of soft, highly-flexible surgical rubber tubing. Before installing this tubing, however, take a sharp scalpel or razor blade and make a clean slit about 100 mm long in it. This slit will ordinarily close tightly enough so that the nitrogen from the storage vessel will flow into the plastic ball. When the ball becomes full, however, the slit will serve as a primitive pressure release valve by opening slightly to allow the gas to escape, thereby preventing the ball from rupturing. When the gas inlet is opened, the dry nitrogen in the ball will flow into the system under atmospheric pressure, and so there is no danger of over-pressurization. A small weight can be placed on the ball to sustain flow after the system reaches atmospheric pressure, if desired. A ball that is a half meter in diameter will store enough gas to fill most laboratory systems several times. This is a very inexpensive arrangement, but one that works quite well, and an inflated dinosauer that is hooked to your SEM will provide a topic of conversation for everyone who visits your lab. Again I mention that this technique, and many others, are described in my recent book on Vacuum Methods in Electron Microscopy, which can be ordered from Portland Press Ltd., c/o Ashgate Publishing Co, Old Post Road, Brookfield, VT 05036-9704, USA. (Fax: 802-276-3837); or Portland Press Ltd., Commerce Way, Colchester, CO2 8HP, UK (Fax: 0206-799331)
check Y_D Stierhof, et al J of electron microsc tech 17:336 for a nice comparison of different silver enhancement techniques
------- Forwarded Message Follows - - - - - - -
Now I remember. The comercial daylight kits for silver enhancement require the addition of gum arabic in order to slow the reaction. I can't immediately recall the deatails but maybe it will ring a bell in a younger brain ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
Hello We're looking at living cultured cells in a perfusion bath maintained at 37 deg C. To minimise heat loss through the objective lens, we heat the lens during the experiments. I'm looking for a source of an appropriate immersion oil that does not change refractive index over the range 20-40 deg C to reduce any abberations in images of the cells. Any suggestions would be appreciated. Thanks Malea
************************************************************************ Malea Kneen Department of Physiology The University of Melbourne Grattan Street, Parkville, 3052 Victoria, AUSTRALIA
Re: Donald Grimes request for microscopy software details:
There are four PC packages published by the UK Institute of Materials,
titled
Scanning Electron Microscopy by John Humphreys Transmission Electron Microscopy by Peter Goodhew Electron Diffraction by Peter Goodhew Analysis in the EM by Peter Goodhew, John Humphreys and Graham Cliff
There is also a nice package on Stereographic Projection by John Humphreys, which is useful for those working with crystals.
They are all intended for introductory use (for people new to EM) and employ quite a lot of graphics. They are NOT research tools for competent microscopists. We use them with 2nd and 3rd year undergrads and people attending short courses on EM.
Each package costs about US$145. They are distributed by
Ashgate Publishing Co Old Post Road Brookfield VT 05036 USA Tel 802 276 3162 Fax 802 276 3837 (for the world except Europe)
and
Institute of Materials 1 Carlton House Terrace London SW1Y 5DB Tel 071 976 1338 Fax 071 839 2078 (for UK and Europe)
Work is in progress to upgrade these programs from DOS to Windows versions but there are no immediate plans for Mac versions (unless someone would like to volunteer to translate/re-emgineer them).
Peter Goodhew (series editor) University of Liverpool goodhew-at-liv.ac.uk (44) 51 794 4665 Fax (44) 51 794 4675
How are publication quality trace analysis results generated? I can get a stereogram with all the poles etc plotted on a computer, but how do you go about drawing the great circles (which are best done using a Wulff net} ? Some Bezier curves can be drawn to get the right curvature etc. but this would be pretty inaccurate, generally. Drawing by hand, unless done by a professional is not publication quality. Any suggestions? Thanks, Sharath dakshs-at-rpi.edu
Message-ID: {MAILQUEUE-101.940513102236.352-at-bmg.bhs.uab.edu} To: microscopy-at-anlemc.msd.anl.gov
Don, Signal Analytics Corporation makes several exellent digital imaging packages sold under the name IP Lab Spectrum for the Macintosh. Their forte is that the software is easy to install, configure, and use and the price is relatively (?) inexpensive, their base package starting around $1500. We have such a system up and running in our digital imaging facility at the University of Alabama at Birmingham and have had great success with it. Their address is:
Signal Analytics Corporation 440 Maple Avenue East Suite 201 Vienna, Virginia 22180
Phone 703-281-3277 Fax 703-281-2509 They are not on Internet yet but anticipate entering the network in the near future. I am currently working with the company to set up a user's group archive that will be accessable via Internet using Gopher. We hope to have this up an working in the next month or two. Any input from any interested parties would be greatly appreciated
Kevin McCarthy Kevin McCarthy Assistant Professor Department of Cell Biology University of Alabama at Birmingham Birmingham, Alabama 35294 Phone 205-934-9923/9924 Fax 205-934-7029
Message-Id: {MAILQUEUE-101.940513084352.998-at-prl.pulmonary.ubc.ca} To: microscopy-at-anlemc.msd.anl.gov
To Malea Kneen could you please repeat the last part of your message about an appropriate immersion oil. For some reason I did not recieve the complete message so am not sure what type of oil you are looking for. Thanks Mark Elliott, UBC-Pulmonary Research Lab Vancouver, Canada
As many of the readers of this list have funding grants from DoE(Energy & Water Budget), NSF, and NASA (VA-HUD-IA Budget) I thought that you might want to know about this gem which is going through the US Congress.. It's not microscopy, however, it will likely impact our research funding. Apologizes in advance if you've already seen this information, or if you are one of our International Subscribers who isn't interested in the US research budget.
Nestor J. Zaluzec ANL EMCenter
---------- WHAT'S NEW by Robert L. Park Friday, 13 May 94 Washington, DC
1. BUDGET: CONGRESS TURNS OFF THE LIGHT AT THE END OF THE TUNNEL. Yesterday, the House Appropriations Committee divvied up the FY 95 budget among the 13 Subcommittees. Only Military Construction was up from last year. Energy and Water came out $81M below the President's request. VA-HUD-IA, which includes both NASA and NSF, came out $361M below the President's request; Subcommittee chair Louis Stokes (D-OH) said he had "grave concerns about whether this allocation can fund the space station." The specter of the SSC hangs over the space station debate; supporters of the space station like to point out that the money saved from the SSC last year did not go to science. Actually, no one in their right mind expects savings from the space station to go to science either-- the important thing is stop the money from going the other way.
Alexey Sidorenko phone: (7-095) 932-88-61 Moscow State University E-mail: avs-at-srdlan.npi.msu.su Institute of Nuclear Physics http://www.npi.msu.su/people/avs/avs.html
I just got my hands on the softcover book "The internet Companion-A beginner's guide to global Networking", and recommend it highly to anyone who is not a computer expert. Even though, I used the internet now for over two years, not until browsing through the text, did I finally understand some of the things I was doing without knowing why. If everyone in the userlist were to read the book, we would probably not get certain message-types that are explicitly discouraged in chapter three of the book.
***** ************ ************** ************ *Cesar D. Fermin, Ph.D |Fax (504) 587-7389 *Tulane Medical School |Answ. Mach.(504) 584-2618 *Pathology/SL79 |Secretary (504) 584-2436 *New Orleans, La 70112 | Lab (504) 584 2521 ***** ***************** ***********************
Message-Id: {MAILQUEUE-101.940516144653.198-at-prl.pulmonary.ubc.ca} To: microscopy-at-anlemc.msd.anl.gov
Does anyone know of a similar network to the microscopy one that deals with tissue culture problems???? We are having problems with guinea pig lymphocyte stimulation with phytohemaglutinin and need advice. Any help would be appreciated. Thanks Mark Elliott Pulmonary Research LAb Vancouver Canada
Have you checked out the "Big Dummies Guide to the Internet"? It is a Hypercard stack which covers most aspects of using the Internet. It is available via ftp. I think I got my copy from sumex-aim.stanford.edu.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On 16 May 1994, Fermin, Cesar wrote:
} } Hello: } } I just got my hands on the softcover book "The internet Companion-A beginner's } guide to global Networking", and recommend it highly to anyone who is not a } computer expert. Even though, I used the internet now for over two years, not } until browsing through the text, did I finally understand some of the things I } was doing without knowing why. If everyone in the userlist were to read the } book, we would probably not get certain message-types that are explicitly } discouraged in chapter three of the book. } } ***** ************ ************** ************ } *Cesar D. Fermin, Ph.D |Fax (504) 587-7389 } *Tulane Medical School |Answ. Mach.(504) 584-2618 } *Pathology/SL79 |Secretary (504) 584-2436 } *New Orleans, La 70112 | Lab (504) 584 2521 } ***** ***************** *********************** }
Mark Elliott queries about a similiar discussion group about tissue culture. In looking through a list of numerous listservers/discussion groups out their I can up with this: MEDCC-L-at-UAFSYSB Issues related to the study of media, culture, etc. Hope this helps.
I'm sorry for my annoying message to this mailing list. I wanted to subscribe and expected to get an automatic response with information.
Thanks to everyone who helped me.
Alexey
Alexey Sidorenko phone: (7-095) 932-88-61 Moscow State University E-mail: avs-at-srdlan.npi.msu.su Institute of Nuclear Physics http://www.npi.msu.su/people/avs/avs.html
you might try any of the bio. or sci. news groups. Reading "news" can be a real chore, and you have to wade through a lot of noise, but you do find the occasional gem. There are a handful of bionet news groups that function more or less like our mail lists, ie posting of questions and answers that all interested parties get to "eavesdrop" in on. I believe you can ftp a utility called TheNews from sumex-aim which holds your hand as you custom,ize your newsgroup subscriptions and set up your sessions. It also gives a listing of the zillion or so groups that are available. Good luck.
David Morilak UT Health Science Center Dept Pharmacology San Antonio morilak-at-uthscsa.edu
I have just tried to access sumex-aim.stanford.edu to obtain a copy of "Big Dummies Guide to the Internet", but the server is not available. Does anyone know of another ftp site?
_____________________________________________________________________ | James V. Jester | Dept Ophthalmology | | jester-at-crnjjsgi.swmed.utexas.edu | UT Southwestern Medical Center | | (214)648-7215 | 5323 Harry Hines Blvd | | | Dallas, TX 75235-9057 | |__________________________________|__________________________________|
Hi there, I am sorry about not sending the complete reference for the book. I hope that the following is of help.
RE:Internet Companion-Laquey
Source for Internet comp-Laquey The publisher is Addison-Wesley Publishing Co. ISBN 0-201-62224-6 Fourth Printing March 1993 ($11.00)
However, the text can be downloaded by anonymous FTP from FTP.STD.COM. If you have a Word Wide Web (WWW) browser like Mosaic you can get it from many sources, including our here at Tulane, by looking in the category internet information and is free, I like the book because is a nice ready reference. Good luck.
P.D. Other commonly recommended texts are also available from the same source.
Just received a demo disk for the Electron Flight Simulator program from Small World. It looks interesting. Does anyone have any experience with, or comments about this product? They're asking $449. Is it worth it?
.............................................................................. John Hudak hudakjm-at-mcmaster.ca I.M.R. - Electron Optics (905) 525-9140 Ext.24890 ABB Rm. 331 FAX: (905) 521-2773 McMaster University 1280 Main St. West Hamilton, Ont. L8S 4M1 Canada
Does anyone know of a commercial source for various thin ceramic (150 - 300 angstrom thick) films on salt flats (1" x 1" minimum area)? We need these for some "non-microscopy" experiments we are considering. The compositions required are SiO, SiO2, and/or SiN. Non-stoichiometric films are OK, we only need the average molecular weight to be correct.
Of course, we need these yesterday! Thanks to all in advance.
Regards, Bill Lamberti.
William A. Lamberti Office LA-196 Exxon Research and Engineering Company Route 22 East Annandale, NJ 08801 (908)730-2144 office (908)730-2262/2104 labs (908)730-3042/3051 fax Email: walambe-at-erenj.com
John I recieved the same package yesterday. The program does indeed look useful, but I have a serious problem with the price. $500 seems like a lot to pay for a nice interface, especially when non-Window versions are available free on the EMMPDL. Just my two cents. ********************************************************************** * Glenn Poirier glenn_p-at-geosci.lan.mcgill.ca * * Electron Microprobe Lab Phone: (514) 398 6774 * * Earth and Planetary Sciences Fax: (514) 398 4680 * * McGill University THERE ARE THREE SIDES TO EVERY STORY; * * Montreal, Quebec YOUR SIDE MY SIDE AND THE TRUTH * **********************************************************************
I am shopping for a new SEM and wanted to know what active users would recommend or not recommend. Ideally, I need an actual resolution of 75 A at 25K magnification. Our expected budget will be $200,000 and we would also like a microprobe. This microscope will be multi-departmental (biological sciences and chemistry). Thank you in advance, Page Owen
Page Owen Dept. of Botany Connecticut College 270 Mohegan Ave., Box 5555 New London, CT 06320 (203)439-2147
SPECIAL ISSUE - ELECTRON SPECTROSCOPIC IMAGING AND ANALYSIS TECHNIQUES. A SELECTION OF PAPERS FROM THE 4TH EUROPEAN WORKSHOP HELD AT THE INSTITUTE OF PHYSICS, UNIVERSITY OF MUNSTER, GERMANY
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 133 - 142.
KOHLER ILLUMINATION IN THE TEM: FUNDAMENTALS AND ADVANTAGES
G. BENNER & W. PROBST Electron Optics Division, Carl Zeiss, 73446 Oberkochen, Germany
Summary
Kohler illumination is the most favourable design for the illumination path of an electron microscope with a condenser objective lens. The new illumination system of the EM 910 and EM 912 OMEGA allows both wide area (Kohler) illumination for TEM operation and spot illumination for analytical investigations. Compared to conventional systems and objective lenses with a condenser mini lens, this system offers many advantages. In addition to the homogeneous, highly coherent and parallel illumination of every point in the specimen, it offers advantages for selected area diffraction and spot scan mode. Combined with the electron optical selection of a condenser aperture, this illumination system provides the flexibility necessary to achieve optimum illumination for the specimen.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 143 - 148.
INFLUENCE OF ZERO-LOSS FILTERING ON ELECTRON OPTICAL PHASE CONTRAST
P. HIRSCH & L. REIMER Physikalisches Institut, Universitat Munster, Wilhelm-Klemm- Strasse 10, 48149 Munster, Germany
Summary
Complex scattering amplitudes are used to calculate the phase contrast of colloidal gold particles.Comparison of measurements of the phase contrast intensity at the centre of the gold particle as a function of defocus for unfiltered and zero-loss filtered images demonstrates the increase in phase contrast achieved by zero-loss filtering even for a thick carbon substrate film. The granulation of amorphous germanium films is measured by the spatial root mean square values of image intensity in a defocus series.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 149 - 159.
IMAGE-EELS: SIMULTANEOUS RECORDING OF MULTIPLE ELECTRON ENERGY- LOSS SPECTRA FROM SERIES OF ELECTRON SPECTROSCOPIC IMAGES
K.-H. KORTJE Institute of Zoology, University Hohenheim, Garbenstrasse 30, D- 70593 Stuttgart, Germany
Summary
A new approach for element microanalysis with energy-filtering transmission electron microscopy (EFTEM) is presented which was accomplished with the CEM 902 electron microscope (Zeiss, Germany). This method is called Image-EELS, because it is a synthesis of electron energy-loss spectroscopy (EELS) and electron spectroscopic imaging (ESI). Series of energy-filtered images at increasing energy losses are recorded from one area with a TV camera. In a second step the intensity of selected regions in the image stack is measured with an image analysis system and plotted as a function of the energy loss. Thus many spectra from different objects can be calculated from one image series and compared with each other. The spatial resolution of EELS is considerably enhanced, the noise is decreased because many pixels from irregular objects are integrated, and the information from ESI can be analysed as a function of the energy loss.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 161 - 169.
OPTIMAL CONDITIONS FOR THE USE OF CORRESPONDENCE ANALYSIS FOR ELEMENT DETERMINATION IN EELS IMAGES
E. S. GELSEMA, * A. L. D. BECKERS* & W. C. DE BRUIJN + * Department of Medical Informatics, and + AEM-unit Pathological Institute, Erasmus University, PO Box 1738, 3000 DR Rotterdam, The Netherlands
Summary
Correspondence analysis is used for the determination of element localization and element concentration in electron energy-loss imaging. The introduction of a new method for the quantification of element concentration embedded in the protocols of correspondence analysis may be regarded as a useful replacement of the power-law estimation technique, especially in regions of the spectrum where the latter method cannot be used. Conditions for the optimum use of the method are established.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 171 - 182.
QUANTITATIVE ELECTRON SPECTROSCOPIC IMAGING IN BIO-MEDICINE: METHODS FOR IMAGE ACQUISITION, CORRECTION AND ANALYSIS.
A. L. D. BECKERS,* W. C. DE BRUIJN,+ E. S. GELSEMA,* M. I. CLETON-SOETEMAN# & H. G. VAN EIJK# *Department of Medical Informatics, +AEM-unit Pathological Institute, #Department of Chemical Pathology, Erasmus University Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands.
Summary
Many questions about the metabolism of specific elements in the human body might be answered if elemental concentrations could be measured in situ in cells. With electron energy-loss spectroscopic imaging (ESI), concentrations can potentially be determined with high spatial resolution. The theory of the quantification procedure has already been derived. Many practical instrument-related problems, however, have to be solved. In the current research an energy-filtering TEM is used and the image-acquisition chain is examined in detail. Quantification requires images to be recorded over a large dynamic range. To solve this problem, the use of optical attenuation filters has been introduced. The use of the combination of a scintillator screen and a TV-camera as a detection system has consequences for the processing of the data. Corrections for the camera photometric sensitivity and , to some extent, for shading are necessary. Further consequences of such a detection system for the correction of the element-aspecific spectral background and element detection are discussed. The derived methodology is tested in several ways and finally applied for the quantitative analysis of iron in liver parenchymal cells of a porphyria cutanea tarda patient.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 183 - 193
EFFECT OF SPECIMEN THICKNESS AND CARBON CONTENT ON THE CARBON PEAK REGION IN EEL SPECTRA
R. DOOR, K. D. HABERLE & R. MARTIN Sektion Elektonenmikroskopie, Universitat Ulm, D-89069 Ulm, Germany
Summary
Amorphous carbon films of different thicknesses and combinations of carbon films with CaF2 films of different thicknesses were used to create a standard reference library of EEL spectra. The relationship between the size and shape of the carbon peak region and the low-loss part of the spectrum was determined. The steepness of the background of the C-K edge, the size of the carbon peak and the plasmon shoulder on top of the carbon peak were investigated as a function of the thickness of the films and of the relative amount of carbon after normalization of the zero- loss peak. The minimum at the C-K edge and the maximum of the carbon peak were considered as examples for a modelling of the carbon peak region in different specimens. Methodological aspects which affect the accuracy of the measurement (characteristic lines of the detector, calculation of specimen thickness, method of t/lamda measurement, mass loss, energy resolution) were examined. The results indicate that modelling of the whole carbon peak region, considering its dependence on mass thickness and carbon concentration, should be possible from our standard reference spectra without use of the low-loss region. The use of modelled standard reference spectra for background extrapolation in biological specimens with unknown calcium content is proposed.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 195 - 206.
APPLICATION OF RECORDING AND PROCESSING OF ENERGY-FILTERED IMAGE SEQUENCES FOR THE ELEMENTAL MAPPING OF BIOLOGICAL SPECIMENS: IMAGING-SPECTRUM
JEAN-LOUIS LAVERGNE, * + COLETTE FOA, # PIERRE BONGRAND, # DOMINIQUE SEUX ++ & JEAN-MICHEL MARTIN* * Ecole Centrale de Lyon, Laboratoire de Tribologie et Dynamique des Systemes, URA CNRS 855, Departement de Technologie des Surfaces, F-69131 Ecully Cedex, France + KODAK-PATHE, Laboratoire d'Analyses, Z.I. nord, F-71102 Chalon sur Saone, France # INSERM U 387, Laboratoire d'Immunologie, Hopital St Marguerite, F-13277 Marseille Cedex 9, France ++ Laboratoire de Developpement et de Pathologie des Tissus Dentaires, UPR CNRS 412, Faculte d'Odontologie, F-69372 Lyon Cedex 8, France
Summary
Computerized energy-filtered transmission electron microscope (EFTEM) permits the recording and the processing of energy- filtered images, allowing a part of an electron energy-loss spectrum for each picture element to be obtained. This method, called 'Imaging-Spectrum', uses a Zeiss CEM902 coupled to several image analysis systems. The actual configuration records sequences of 48 images, 256 x 256 pixels, in steps of the energy loss, þE . Processing these sequences results in part of a core- loss EELS-spectrum for each pixel. This approach produces elemental maps with a short processing time. We have implements three kinds of background calculation for the image subtraction. The influence of the irradiation dose and of the energy selecting slit width on the quality of the spectra is investigated. The method is applied to the analysis of some biological specimens (pericellular coat behaviour during adhesion between macrophages and red blood cells and location of calcite microcrystals in dental pulp cells). The Imaging-Spectrum method appears to be suitable for the analysis of large areas.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 207 - 223.
EVALUATION OF LANTHANIDE TRACER METHODS IN THE STUDY OF MAMMALIAN PULMONARY PARENCHYMA AND CARDIAC MUSCLE BY ELECTRON ENERGY-LOSS SPECTROSCOPY
H. FEHRENBACH, A. SCHMIEDL, F. BRASCH & J. RICHTER Abt. Elektronenmikroskopie, Zentrum Anatomie, Kreuzbergring 36, D-37075 Gottingen, Germany.
Summary
Lanthanum (La) has widely been used as a tracer to study the integrity of plasma membranes. With conventional transmission electron microscopy (cTEM), the absence of electron scattering deposits from the cytoplasm has generally been assumed to reflect an intact cell membrane. However, the application of electron spectroscopic imaging (ESI) and electron energy-loss spectroscopy (EELS) reveals that electron scattering deposits may be present which do not contain La. However, La could be detected in regions of pulmonary parenchyma and cardiac muscle that were devoid of electron scattering deposits. Therefore, to exclude misinterpretations based on CTEM the application of microanalytical techniques is strongly recommended for the study of the integrity of plasma membranes by means of La tracers. In addition, ESI and EELS are shown to distinguish between different tracers in simultaneous applications of La and terbium (Tb) which were used at the different faces of the pulmonary air-blood barrier. The analysis of the distribution of both tracers which form electron scattering deposits, indistinguishable by cTEM, may help us to understand the different functional significance of cellular alterations of both cellular borders of the barrier. As was shown for La, however, strictly controlled conditions are mandatory during the fixation procedure because an increase in the incubation time to more than 1 h in samples of pulmonary parenchyma may result in the occurrence of La deposits within the cytoplasm. In the absence of electron scattering deposits, the presence of La in glycogen granules and ribosome-containing areas of various types of alveolar septal cells even after 15 min incubation indicates that the absence of deposits does not necessarily correspond to the absence of the tracer.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 225 - 232.
DEMONSTRATION OF ALUMINIUM IN POLYPHOSPHATE OF LACCARIA AMETHYSTEA (BOLT. EX HOOKER) MURR. BY MEANS OF ELECTRON ENERGY- LOSS SPECTROSCOPY
I. KOTTKE * & F. MARTIN + * Universitat Tubingen, Botanishches Institut, Spezielle Botanik und Mykologie, Auf der Morgenstelle 1, D-72076 Tubingen, Germany + INRA, Laboratoire de Microbiologie Forestiere, Centre de Recherches Forestieres, 54280 Champenoux, France
Summary
Toxic aluminium species in acidified soil damage the rootlets of trees. The symbiotic association with ectomycorrhizal fungi may, however, protect the rootlets by sequestration of aluminium in polyphosphates. Electron energy-loss spectroscopy was used to identify the relative amount of aluminium in the polyphosphate of the mycelium of the ectomycorrhizal fungus Laccaria amethystea. A three-window method at the L-2,3 edge yielded results which were in agreement with the spectra obtained at the aluminium K edge. The aluminium net distribution images were recorded at 85 eV, the backgrounds at 70 eV and 60 eV, setting the energy selecting slit to 5 eV and the beam current to 15 microA. The method can be used to study the sequestration of aluminium in mycelia of ectomycorrhizal fungi under different stress situations.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 233 - 238.
ESI IN SITU ANALYSES OF EXTRACHROMOSOMAL RNP GRANULES
S. ABOLHASSANI-DADRAS, * G. H. VAZQUEZ-NIN, *# O. M. ECHEVERRIA, *# V. BOUTINARD ROUELLE-ROSSIER* & S. FAKAN* * Centre of Electron Microscopy, University of Lausanne, 27 Bugnon, CH-1005 Lausanne, Switzerland # Laboratory of Electron Microscopy, Department of Biology, Faculty of Sciences, National Autonomous University of Mexico, Mexico City, D.F. 04510, Mexico
Summary
Observation of unstained ultrathin sections of salivary gland cells of Chironomus thummi and C. tentans, by means of electron spectroscopic imaging (ESI), has revealed that phosphorus is distributed in two types of granular structures in the nucleoplasm of these cells. In addition to a specific type of premessenger ribonucleoprotein (RNP) particle, known as the Balbiani ring (Br) granule, ESI revealed a new type of phosphorus-rich small granular component. Examination of unstained sections by energy-filtering transmission electron microscopy (EFTEM) offers the opportunity of obtaining the signal from the specimen alone, thus avoiding the possible contributions of heavy metals present in any staining product.
I need a reviewer for a short manuscript on the use of electron microscopy in the examination of minerals and soils. If any subscribers are qualified and willing to help in this matter please contact me directly.
*************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
A recent posting listed the FTP site at STD.COM for obtaining the Internet Companion. It would be a great help if the posting individual could provide pointers to the directory where it is listed. The search engines always seem to be tied up when I try to log on, so this would be a help. Thanks.
Does anybody have any information regarding the thickness and roughness of nitrocellulose in amyl acetate (0.1%)? The procedure we used was to take 2 drops of the solution in a pasteur pipet and cast onto a very clean glass coverslip. This technique is used for a motility assay of actin filaments on myosin absorbed on the nitrocellulose layer. Any references or protocols would also be helpful.
Margaret (Peggy) Bisher
NEC Research Institute, Inc. 4 Independence Way Princeton, NJ 08540
Subject: Time:3:32 PM OFFICE MEMO LM Date:5/19/94 I am doing basic morphological studies on rat kidneys. I am looking at 1 micron epon sections that have been processed for standard electron microscopy. I have a few questions on what I am looking at, and I am looking for someone that I can talk to that might be able to answer some simple questions such as what kind of artifacts will you get when you perfuse, or is a perfused kidney even preferred over a biopsy, how much variation is there from rat to rat, etc., etc.. If anyone has experience working with rat kidney, I would really appreciate it if you could contact me. Thank-you, Jeanne
There is a new Macintosh software for electron diffraction and crystallographic calculations developed by Drs. J. M. Zuo and H.R. Zhu. Preliminary report can be found in last year MSA proceedings. This program plots crystal structure, stereoprojection electron diffraction pattern (SAD and CBED) and backscattered kikuchi pattern with a simulated microscope control panel. Intensity of kinematic approximation is used. The program also has calculator for complex numbers and vectors of real and reciprocal space, and functions for calculating electron and x-ray structure factors and Bragg angles etc. For more information contact
We have an Akashi/ISI EM-002A TEM (1988), one of only eight or nine made. I am keen to get in touch with any other owners of these instruments with regards to sorting out technical problems and obtaining spare parts. This would include owners of the EM-002B, which is similar in many ways. Yours faithfully,
Richard Easingwood EM Unit,School of Medicine University of Otago Dunedin New Zealand
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
I recently downloaded the Internet Companion successfully, as follows:
site: ftp.std.com dir: \ftp\OBS\The.Internet.Companion files: internet.companion (349 910 bytes) (Text version) internet.companion.Z (zipped version, Unix) COPYRIGHT Order.form Tracy.gif (I assume a .gif of the author, Tracey Laquey)
Note: if you are downloading to a DOS machine, you will have to save the files to a DOS-acceptable filename..(eg internet.txt) The ftp-site filename of {internet.companion} is OK in Unix, but not in DOS. [The same applies if you ever download uuencoded or zipped files. If the filename within the uuencoded file is more than 8 characters or contains unnacceptable characters, the file may not be uudecoded on a DOS machine. It is then necessary to change the filename within the compressed file (using any text editor) before uudecoding etc.]
Hello Ca imagers! We have been trying to image Ca transients in some cells (neutrophils and some mouse macrophages) following cross-linking of surface receptors but we have been running into some difficulties. Our hardware configuration is as following: We are using an intensified Hamamatsu CCD camera mounted on the side port of a Zeiss Axiovert. We are using the following filters: excitation 340nm 10nm FWHM or 350nm 10nm FWHM dichroic 375nm emission 405nm 45nm FWHM 490nm 45nm FWHM
The objective is a Zeiss 40X UltraFluar. We also use OD filters in the excitation path to minimize bleaching of the probe (usually 25 or 50% transmission). The dynamic range of the measurement, in terms of the ratio between the Ca-free and Ca-bound ratios, is only ~10, which may be insufficient to give good accuracy at Ca concentrations away from the Kd of the probe! This problem is compunded by the 8-bit accuracy of the imaging system (Image 1/FL, Universal Imaging Corp. using a Matrox video board). Does anybody have comments? Since most of the papers report values of [Ca]i and not the raw ratio values, I cannot judge whether we are doing something wrong! Is this a common observation? Comparable measurements with a spectrofluorometer on cells in suspension give us a dynamic range (as defined above) of ~40. Is anybody willing to share her/his experience? Any comments/suggestions will be greatly appreciated.
Thank you in advance!
************************************ *Massimo Sassaroli * *Dept. of Physiology and Biophysics* *Mount Sinai School of Medicine * *New York, NY 10029-6574 * *Tel: 212-241-9512 * *sassaroli-at-msvax.mssm.edu * ************************************
P.S. : Is there any other interest group dealing specifically with Ca measurement techniques? Thanks again
Hello, Last week the results of a poll on how people record their images was posted. After briefly scanning the replies, I deleted the file. Now I find myself wishing I hadn't. Could someone email the file to me? Is there an archival site where I could retrieve it from (seems to me I saw this listed here before). The reason I would like to recover this file is that we are contemplating interfacing a PC with our Hitachi S-4000 FESEM. Anyone who has any suggestions, please reply.
___________________________________________________________________ Randy Nessler rnessler-at-uiowa.edu The University of Iowa Central Electron Microscopy Research Facility
We have an Akashi/ISI EM-002A TEM (1988), one of only eight or nine made. I am keen to get in touch with any other owners of these instruments with regards to sorting out technical problems and obtaining spare parts. This would include owners of the EM-002B, which is similar in many ways. Yours faithfully,
Richard Easingwood EM Unit,School of Medicine University of Otago Dunedin New Zealand
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
We have an Akashi/ISI EM-002A TEM (1988), one of only eight or nine made. I am keen to get in touch with any other owners of these instruments with regards to sorting out technical problems and obtaining spare parts. This would include owners of the EM-002B, which is similar in many ways. Yours faithfully,
Richard Easingwood EM Unit,School of Medicine University of Otago Dunedin New Zealand
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
Message-Id: {1994May23.141113.859926163-at-ms.sjdccd.cc.ca.us} To: microscopy-at-anlemc.msd.anl.gov (microscopy list)
I am trying to subscribe to the NIH-Image Listserver and was told that the address was listserv-at-soils.umn.edu and the message should be subscribe NIH-IMAGE {your name}
However when I did that I got a return receipt from listproc-at-soils.umn.edu that indicated that there was no NIH-IMAGE to subscribe to. Does anyone know the correct address for subscription. Thanks Judy Murphy San Joaquin Delta College Dept. of Microscopy Stockton, CA 209/474-5284 murphy-at-ms.sjdccd.cc.ca.us
The message, "No matches to nameserver query," is generated whenever the ph nameserver fails to locate either a ph alias or name field that matches the supplied name. The usual causes are typographical errors or the use of nicknames. Recommended action is to use the ph program to determine the correct ph alias for the individuals addressed. If ph is not available, try sending to the most explicit form of the name, e.g., if mike-fox fails, try michael-j-fox.
--------Unsent Message below:
Received: from anlemc.msd.anl.gov by nexus.uiowa.edu (1.38.193.4/931201) on Mon, 23 May 1994 23:41:13 -0500 id AA04787 with SMTP
Message-ID: {MAILQUEUE-101.940513102236.352-at-bmg.bhs.uab.edu} To: microscopy-at-anlemc.msd.anl.gov
Don, Signal Analytics Corporation makes several exellent digital imaging packages sold under the name IP Lab Spectrum for the Macintosh. Their forte is that the software is easy to install, configure, and use and the price is relatively (?) inexpensive, their base package starting around $1500. We have such a system up and running in our digital imaging facility at the University of Alabama at Birmingham and have had great success with it. Their address is:
Signal Analytics Corporation 440 Maple Avenue East Suite 201 Vienna, Virginia 22180
Phone 703-281-3277 Fax 703-281-2509 They are not on Internet yet but anticipate entering the network in the near future. I am currently working with the company to set up a user's group archive that will be accessable via Internet using Gopher. We hope to have this up an working in the next month or two. Any input from any interested parties would be greatly appreciated
Kevin McCarthy Kevin McCarthy Assistant Professor Department of Cell Biology University of Alabama at Birmingham Birmingham, Alabama 35294 Phone 205-934-9923/9924 Fax 205-934-7029
= = POSITION AVAILABLE = = Lab Manager for the Electron Microscopy Core Laboratory of the =Interdisciplinary Center for Biotechnology Research at the University =of Florida. = =DUTIES: Preparation and examination of biological specimens for = transmission and scanning electron microscopy in a facility that = serves the entire university community on a fee for service = basis. Collaborative research possible as part of the program. = Incumbent is expected to deal directly with the clients on = experimental design, execution of the project and preparation of = the data for publication, in consultation with the Scientific = Director of the laboratory. General lab management. = =QUALIFICATIONS: = experience in most aspects of biological electron microscopy. = Good communication skills, both oral and written. Ability to = deal on a one to one basis with investigators from a wide variety = of disciplines and backgrounds. = =STARTING DATE: Immediately = =APPLICATION DEADLINE: Open until position is filled. = =SALARY:$22,633 - 26,500. = =Inquiries by E-mail or full resumes to the address below = =********************************************************** =* Dr. Greg Erdos ** * =* Director, ICBR EMCL ** Phone 904-392-1295 * =* 218 Carr Hall ** FAX 904-392-8598 * =* University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * =* Gainesville, FL 32611 ** * =**********************************************************
To: MICROSCO:CENTRAL cc: DRStadden:R_D:Armstrong Subj: AO Spencer Stereoscope ------------------------------------------------------------------ I am interested in knowing if anyone has used a repair service for AO Spencer stereomicroscopes. We have a vintage 1959 model with a detatched prism, and find it nearly impossible to repair ourselves, using makeshift alignment procedures. There must be a better way. If you know of a good repair service, or a method for doing it that we may not have thought of, or have gone through a similar experience, I'd love to hear from you. If you've seen this message before, I apologize, but I got several undeliverable mail notices when I sent it out about a month ago and received no replies. Please reply to: DRStadde+aCENTRAL%ARMSTRONG-at-MCIMAIL.COM or phone: (717)-396-5109 THANKS!
Hi I received the J. of Microscopy, June issue summary, but I have no clue on how it was encoded so that I may be able to decode it and read it!! My mail is received by a VAX but it is automatically rerouted to a SGI workstation and, finally, to a Mac running Eudora v. 1.4. The header of the message does not include the encoding method. Thank you for your help!
Dear Nestor, After posting to sci.microscopy about the poll, I realized that it was mailed to me personally by the person who conducted the poll. I guess the reason that I received the results was that I had participated in the poll. As far as the below message, it is all I recieved. Was a file to be included? I have the address of another contributor to the poll, so I will see if they kept a copy of the file. ___________________________________________________________________ Randy Nessler rnessler-at-uiowa.edu The University of Iowa Central Electron Microscopy Research Facility
On Mon, 23 May 1994 MICROARCHIVE-at-anlemc.msd.anl.gov wrote:
} Randy } } I'm not sure if I sent you what you wanted, but from your description } it was the only thing that looked right in the archives.... } } Nestor Zaluzec } ANL EMCentee } r
I was told that there is a new Polaroid product called 'Polapan 400' which is a medium-speed (ISO 400), coaterless, black-and-white professional film. The Polaroid company claims that this film is a new choice for T52 and T53 applications.
Does anyone have used this new film before? Can I get some comments from you? I use a SEM and take a lot of T53 films on rocks and minerals. Thanks.
Long Liang Electron Microprobe and SEM Lab ARCO Exploration and Production Technology 2300 West Plano Parkway Plano, TX 75075 E-mail : LLIANG-at-is.Arco.COM
The polaroid polapan 400 film works fine as a replacement for type 52, we use it with the same settings as the type 52. I don't know how they compare as far as cost, we were given 8-10 sheets to demo. I'm sure they will provide you a sample if you ask.
Does anyone have any experience with or sources for the electropolishing of indium alloys, specifically InCd or InPb? I need to prepare TEM specimens of these materials, and keep all the steps taken during preparation below room temperature. Any suggestions will be greatly appreciated. ************************************************* F. Scott Miller smiller-at-umr.edu Electron Microscope Lab University of Missouri-Rolla voice: 314 341 4727 fax: 314 341 6934 *************************************************
A postdoctoral position is available in TEM characterization of high temperature superconducting (HTSC) materials in the Metals and Ceramics Division of Oak Ridge National Laboratory (ORNL). Job responsibilities would include characterization using advanced EM techniques (EDS, HREM, EELS, CBED, etc.) of a variety of oxide-based HTSC materials. Candidates *must* have a PhD in materials science or related field with a solid background *and* extensive experience in characterization of HTSC materials.
*** DO NOT REPLY ELECTRONICALLY *** *** ELECTRONIC RESPONSES WILL NOT BE CONSIDERED *** *** RESPOND ONLY BY MAIL***
To apply, send a resume, publication list, and names of at least three references to:
G. Sims Oak Ridge National Laboratory P. O. Box 2008 Building 4500S, MS- 6116 Oak Ridge, TN 37831-6116
Oak Ridge National Laboratory is an Equal Opportunity/Affirmative Action Employer.
A postdoctoral position is available in TEM characterization of high temperature superconducting (HTSC) materials in the Metals and Ceramics Division of Oak Ridge National Laboratory (ORNL). Job responsibilities would include characterization using advanced EM techniques (EDS, HREM, EELS, CBED, etc.) of a variety of oxide-based HTSC materials. Candidates *must* have a PhD in materials science or related field with a solid background *and* extensive experience in characterization of HTSC materials.
*** DO NOT REPLY ELECTRONICALLY *** *** ELECTRONIC RESPONSES WILL NOT BE CONSIDERED *** *** RESPOND ONLY BY MAIL***
To apply, send a resume, publication list, and names of at least three references to:
G. Sims Oak Ridge National Laboratory P. O. Box 2008 Building 4500S, MS- 6116 Oak Ridge, TN 37831-6116
Oak Ridge National Laboratory is an Equal Opportunity/Affirmative Action Employer.
I use Polapan400 exclusively, the contrast is as good as Type 52, IMHO, and not needing coating is too good to pass up.
You can get a discount if you order the "bulk pack" - 10 boxes/case (no outer packing)
It does not, however, have a negative with it. If you need a good 4X5 neg, shoot sheet film!
James Long (no relation ;-))
On 24 May 1994, Liang, Long wrote:
} } I was told that there is a new Polaroid product called 'Polapan 400' } which is a medium-speed (ISO 400), coaterless, black-and-white } professional film. The Polaroid company claims that this film is a new } choice for T52 and T53 applications. } } Does anyone have used this new film before? Can I get some comments from } you? I use a SEM and take a lot of T53 films on rocks and minerals. } Thanks. } } } Long Liang } Electron Microprobe and SEM Lab } ARCO Exploration and Production Technology } 2300 West Plano Parkway } Plano, TX 75075 } E-mail : LLIANG-at-is.Arco.COM } } } }
We were just discussing that same problem yesterday with those same scopes. The solution in this department has been to put them in a closet and buy new ones. I found there is a closet full..... and I thought we ought to be able to fix them. I couldn't get the prism straight myself... so if anyone knows of an economical way to fix them, let us know. Thanks ... Nina Allen
Your local microscope repair facility should be able to fix those easily. If not, McBain Instruments in Chatsworth, Calif. certainly can. Their phone # is (818)998-2702.
Following the protocol of Bendayan I complexed Dnas, phospholipase and prteinase_K to colloidal gold but totally failed with RNase A using ph 9.3 and 7.9 and 5.6. I tried to use Sigma Cat. # R 5503 ribonuclease A from bovine pancreas described as being essentially protease and salt free. How ever when I added 0.2ml of a 5mg/ml pure water solution to 10 mls of colloidal gold it immediately turned blue/purple and begain to settle out justas if I had added a high concentration of salt.
Where did I go wrong???? ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
I am unable to achieve uniform illumination and focus on my prints, particularly with 35 mm (T-max 100) film. Does anyone know if this is a characteristic of the Beseler CB-7 enlarger or is it something to expect with any enlarger? My 50 mm lens (a Schneider Companon-S) gives a sharp grain (as seen with grain magnifier) in the center but considerably more light and focus degradation at the periphery. The pattern is not a simple circle. The 100 mm lens (Rodenstock Apo-Rodagon) gives better uniformity of illumination and focus but the image is not crisp. The lens apertures are 1-2 stops from wide-open. All exterior lens surfaces are clean. I am using the standard enlarger head with recently cleaned condensers and heat shield. The Kodak polycontrast filters are, unfortunately, below the focusing lens. If the Beseler is inadequate for biological EM lab work using 4 x 5, 3.25 x 4, and 35 mm films, does anyone have recommendations for a better enlarger?
Message-Id: {MAILQUEUE-101.940526141013.256-at-bunyip.ph.rmit.edu.au} To: microscopy-at-anlemc.msd.anl.gov
SEM : viewing polyethylene and other "plastics"
Is it possible to gold coat polyethylene in a standard uncooled gold sputter coater without heating the surface so as to modify it ? We are looking specifically at surface detail and do not wish to have it modified at all. We also wish to look at other probably higher melting point plastics as well. John Millar jjmill-at-rmit.edu.au
Rodney L Kuehn reported problems with evenness of ilumination and focussing on an enlarger. I think that the main problem is that the aperture used in the enlarger lens is simply too large (too small an f number). This results in a small depth of focus. If a larger f number is selected, such as f16 or f22, then the depth of field will increase significantly, and the focussing problem should disappear. The unevenness of the illumination is probably also a problem of aperture size and should also be cured by increasing the f number. I hope this is of use.
Dr Douglas J Arrell Mechanical Performance and Joining Institute for Advanced Materials 1755 ZG Petten Netherlands
Greg Erdos asked about precipitation problems during RNase / colloidal Gold complexing. Our experience suggests that concentrations of 0.5mg of most proteins per 100ml of gold (Frens method) is adequate. If the Sigma RNase is not completely salt free there may still be enough salt in there to aggregate the colloid. Try 0.01ml of the 5mg/ml protein solution per 10ml gold at pH5 - it may work - and still give you adequate label(l)ing.
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
We have used EMS for some time here without any great problems. The bu1 program does take some time to get used to, and building crystals using the space groups can be rather confusing. If the unit cell looks incomplete, try displaying a 2x2x2 block, as the structure may then become clearer.
Alternatively (the method I prefer) use the option which allows you to specify the lattice type (PFIC etc) and then the positions of the atoms in the motif. Thus by specifying F lattice and giving atoms at +- 1/8,1/8,1/8 the silicon crystal can be built very quickly. (I think, from memory, that the option needed is /b)
The bu1 program will provide a file containing all sorts of useful/useless information. In this is a count of the number of atoms in the unit cell. Whichever way you build the crystal, check this file to see that you have the right number.
George Burgess Materials Science & Metallurgy Cambridge CB2 3QZ
We have two versions of EMS, one running on a VAX and one on a PC. In both versions it works fine with silicon in x,y,z=1/8,1/8,1/8. If you look at the .prt file created by bu1, you can check if all atoms are there, and in the right place.
Anna Carlsson National Center for HREM Inorganic Chemistry 2 Chemical Center Lund University P.O. Box 124 S-221 00 Lund Sweden Phone: +46 46 108231 Fax: +46 46 104012 e-mail:Anna.Carlsson-at-oorg2.lth.se
} We were just discussing that same problem yesterday with those same } scopes. The solution in this department has been to put them in a closet } and buy new ones. I found there is a closet full..... and I thought we } ought to be able to fix them. I couldn't get the prism straight } myself... so if anyone knows of an economical way to fix them, let us know. } Thanks ... Nina Allen
If you want, you can contact Ron Roos at Leeds Instruments, 800-444-5333. He is supposed to be some kind of repair modification specialist. Leeds buys, sells, and trades all different kinds of older microscopes. Basic service is ~$60 an hour and parts. On site service is available. Other than liking the service this company provides, I have no connection with Leeds.
************************************************* _______________________ * * | | * * | _____ ______ | * Charles J. Butterick * |__| | | |__| * Electron Microscopy Center * | | * Department of Cell Biology and Anatomy * ______| |______ * Texas Tech University Health Sciences Center * | __ __ | * Lubbock, Texas 79430 * |__| | | |__| * USA * | | * Phone 806 743-2706 voice * | | * 806 743-2707 fax * | | * Email hecub-at-ttacs.ttu.edu * | | * * __| |__ * * |_______| *************************************************
The symptoms that Rodney Kuehn described for his enlarger sound to me exactly like a condenser out of alignment; they do not sound like a depth of focus problem. The Besslers that I am familar with have to be adjusted between 4 x 5 use and 35 mm use. If this is not done correctly, or if the bulb is not well centered, you get problems of focus and illumination that resemble those described by Kuehn. HTH, Tobias Baskin
If there is anyone out there still trying to nurse along a JEOL JEM100B TEM, we have just declared ours surplus. With the exception of a few parts that I have already committed, parts are free to a good home. If it's light, I'll ship it to you; if it's heavy, you need to pay the freight. Julian Smith III Biology Winthrop University Rock Hill, SC 29733 803-323-2111 (Vox) 803-323-2246 (Fax)
I was wondering if anyone has any information on film printers that can be hooked up to a computer (that is also connected to an SEM). Can anyone share their experiences (if any) with the use of these film printers with me and of course how much they cost. We had a refurbished Mirus film printer donated to us awhile back, but it didn't work right after awhile.
I hope that these are appropriate questions for this list. I sent almost the same questions to the NIH-Image list as well, but was not sure if that was the appropriate list either.
Thanks in advance for any information, Peling Melville
-------------------------------------------------------------- Peling Melville peling-at-amnh.org Interdepartmental Laboratories American Museum of Natural History
Lowicryl users, I have heard that HM20 resin is no longer available. Can anyone confirm this and/or offer any explanation as to why it is no longer sold.
Thank you.
Allan Mitchell
per Richard Easingwood, South Campus EM Unit University of Otago, Dunedin New Zealand.
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
I haven't kept up with this aspect of the field for the last few years, however, many moons ago when I was a younger lad, John Colby (of Magic IV fame) did some work on the shifts in light element K lines. Duncumb et al did work in the mid 60's, Holliday did work on carbide phases around the same time. These shifts are best measured by WDS (but as a uprobist you know this already). The chemical shifts are due to the fact that the emission lines are due to transitions of valence electrons to Kshell vacancies rather than deeper core levels (ie. L's or M's) to the K shell which we see in higher atomic number elements.
My suggestion is to look into some of the more recent books on Bulk X-ray analysis. Love and Scott, or the rewritten book by Reed (which should be on the market soon). You should also probably check out the last few years conference proceedings of the Microbeam Analysis Society.
Message-Id: {MAILQUEUE-101.940526162512.448-at-ahabs.wisc.edu} To: microscopy-at-anlemc.msd.anl.gov
Proteins below their isoelectric points are positively charged. Addition of the positively charged protein to the gold collapses the negative charge layer which keeps the gold sol in suspension. Raise the pH of the gold and the problem should disappear. Unfortunately, it is sometimes impossible to resuspend the gold in a buffer of much lower pH than that at which it will conjugate to the protein.
Scott Simmons University of Wisconsin
HELP!!\
Following the protocol of Bendayan I complexed Dnas, phospholipase and prteinase_K to colloidal gold but totally failed with RNase A using ph 9.3 and 7.9 and 5.6. I tried to use Sigma Cat. # R 5503 ribonuclease A from bovine pancreas described as being essentially protease and salt free. How ever when I added 0.2ml of a 5mg/ml pure water solution to 10 mls of colloidal gold it immediately turned blue/purple and begain to settle out justas if I had added a high concentration of salt.
Where did I go wrong???? ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
You said you had problems with focusing and uneven illumination with your enlarger. What type of enlarger and light source are you using? The aperture has very little to do with an image being out of focus. If you use a focusing aid to focus on the emulsion grains, make sure the focusing aid is in focus for your eyes. Check the focusing knob and make sure it stays put when you focus. Sometimes the focusing knob will have a tendancy to slip after a period of time if it is an older enlarger. You may be able to tighten the knob or have to have it replaced. Also, make sure the height adjustment knob is locked in place. Make sure the easel is nice and flat on the enlarger base. As far as uneven illumination, if you use a point light source make sure it is properly aligned in the enlarger head. Make sure your condenser lenses are clean. Make sure the lens you are using is the proper lens for the film. 35mm.......50mmlens 21/4.......90mmlens 4x5.......150mmlens Make sure the condenser lens matchs up with the film and lens you are using. Make sure the lens is in properly on the lens board and the board is in properly. Sometimes I have that problem with my enlarger. Someone else will change the lens and not get it in place properly. I use a Durst 45 enlarger and putting the lens in wrong is easy to do. F-stops has very little to do with uneven illumination or focus. F-stops are primarily for better depth of focus. If the image is out of focus nothing will help to bring into focus unless you use something like f:64 for a long enlarging time dependant on the density of the neg. If you a use point light source, you don't have to worry about f-stops anyway. Hope this helps! If you have any problems, you can reach me at: Phone: (410) 455-3582 Fax: (410) 455-3875 Email: prutle1-at-gl.umbc.edu
To: Rodney L Kuehn {kuehn002-at-maroon.tc.umn.edu}
Dear Rodney:
A common problem in our daily operation is unleveled upper condenser. This is particularly troublesome in the old Omega enlargers lacking clips for holding the upper (collecting) condenser in place. If the enlarger is moved, etc. the condenser will occasionally slide just enough to cause the problem you describe. This is less troublesome with enlargers that have motorized adjustments. I correct the problem by measuring the distance between the upper rim of the enlarger condenser housing and the glass edge of the condenser with a metric ruler. It takes some times to move it around until is all even. Then you must ensure that it will not move when returning it to the enlarger housing.
This happens here all the time. Agree, it is a pain.
***** ************ ************** ************ *Cesar D. Fermin, Ph.D |Fax (504) 587-7389 *Tulane Medical School |Answ. Mach.(504) 584-2618 *Pathology/SL79 |Secretary (504) 584-2436 *New Orleans, La 70112 | Lab (504) 584 2521 ***** ***************** ***********************
= =i am from the Sedat and Agard labs where more most of these observations have =been made, and the notion that one needs to wait until the 7th cleavage =division is entirely bogus. i believe it is everyone's experience that histone =incorporation into incorporation can be observed after a single round of =DNA replication. = =you may have some technical problems depending how deep your early clearvage =stage nuclei are in the embryo. i suspect that a conventional, wide-field =microscope would probably work ok (especially with a decent digital camera), =nut you could always use a confocal if necessary. certainly, being across the =Bay from Agard and Sedat's lab, there is no excuse for you not to call them, =arrange for a visit for some technical advice, and maybe even try some data =collection on their system with your embryos. = =i have no experience with the new molecular probe dyes. this would be another =good question for sedat (or perhaps a few other groups at ucsf like andrew =murray's or tim mithchison's). let me know what you find out... = ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
VCR Group uses the Ion Tech ion guns in its ion mills. You can contact them at VCR Group, Inc., 250 East Grand Avenue, Suite 31, South San Francisco, CA 94080, (415) 875-1000, FAX (415) 875-7111.
Russ Cook Electron Microscopy Center for Materials Research Argonne National Laboratory
There might be a chemical solution meeting your requirement, but I think it is hard to find. For my personal opinion, the best way to thin heterostructural cross-section TEM specimen is firstly mechanical grinding and polishing down to less than 10 um, then uses ion milling for final thinning.
We are considering purchasing either a Hitachi S4200 or an Amray 1845 FE-SEM. The SEM will be used in a research environment with multiple users and a wide variety of samples ranging from metallic alloys including hard magnets to ceramic powders. We will either purchase a Link ISIS or Noran Voyager EDS system. We would like comments from users on their likes versus dislikes of these systems, in particular the day to day performance in terms of SEI and BSE resolution, analytical integrity (i.e., beam stability, spatial resolution (beam scatter), and XRF behavior of the finial imaging system) and reliability of the vacuum system. Also, performance integrity (down time), willingness of service representative to trouble shoot and timeliness and performance of service personnel. Any other suggestions on what options or requests you would have done differently on the machine you have purchased would also be helpfull.
I am using an LKB cryoultramicrotome to demonstrate antigens in skeletal muscle and would appreciate any comments and suggestions from biologists using this technique to improve techniques.
Dr Terry A. Robertson Telephone: 61-9-3462935 Department of Pathology Fax: 61-9-3462891 University of Western Australia Nedlands WA 6009 Australia Email: troberts-at-eosin.path.uwa.edu.au
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A couple of years ago, the company PLANO made for us fluorescent screens on glass disks to capture TEM pictures on a video camera. The glass (dia 12 cm, 1cm thick) was set as a window at the bottom of the observation chamber. The coating uniformity was good enough to the level of in situ experiment recording. It was produced without (or with very little binding agent), thus it was very sensitive to fingerprints and scratches, but relatively resistant to burning. We lightly coated the powder layer with aluminum in a standard vacuum coater. For some they used their own powder and our P22G one for the others.
Ask for Mr. Noli, PLANO W. Plannet GmbH, Marburger Strasse 90, D-35043 Marburg 7 Phone ..+64.21.41077 =46ax ..+64.21.51173
Best regards Philippe Buffat
} Hello microscopists, } I need your help on the following point: } I URGENTLY would like to make a fluorescent screen (50 micrometers thick of } Y2O2S-Eu powder on circular glasses -diameter 36 mm and 53 mm). I have the } fluo powder and the glasses. I don't have the know how. } Do you know the address-phone-Fax-email of any commercial company } (France-Europe-Rest of the world) who could make it? } Thank you in advance } Don't bother anyone with this. Please reply directly to me. } Yours, } Pierre } -------------- } Pierre Trebbia, LASSI/GRSM, Universite de Reims, F51062 Reims cedex, France } Email : pierre.trebbia-at-univ-reims.fr } Phone : (33) 26 05 33 62 } FAX : (33) 26 05 32 50
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Institut Interd=E9partemental de Microscopie Electronique Address: EPFL-I2M, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 (Central European Time) Fax: +41(21)693 44 01 ______________________ Eudora 2.0.2 __________________________________
User involved in cryosectioning muscle samples for TEM. Would appreciate any suggestions from users actively involved in this techniqe. Having trouble demostrating anti-dystrophin antibody (nova castra dys1) with immunogold methods after fixing tissue in 2% paraformaldehyde for one hour as suggested by suppliers. Any helpful hints would be appreciated.
Terry Robertson
Dr Terry A. Robertson Telephone: 61-9-3462935 Department of Pathology Fax: 61-9-3462891 University of Western Australia Nedlands WA 6009 Australia Email: troberts-at-eosin.path.uwa.edu.au
Dear EM microscopists, We will be studying gecko oviducts with TEM and SEM. If anyone has any information regarding fixation of this lizard tissue we would like to hear from them.We are particularly interested in appropriate the buffer/fixative osmolarity to use.
Mark Gould
per R Easingwood
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
Although this may sound trivial, I refer to an experiment we had couple of years back. A researcher was studying bacteria and wanted to have immuno-gold labelling on it. She processed the samples according to the latest recipes but there was no labelling on the samples. After a month or so she showd the samples to me asking why. We checked the specimen processing step by step and finally I asked if she had mixed the immuno-gold bottle thoroughly. When the answer was no, we mixed it well and the problem was solved.
I am curious wether anyone else has been unsuccesfully trying to find gold.
With wishes for a good summer,
Jouko Maeki
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Jouko K. Maeki Navigare necesse est... Laboratory Manager, Ph.D. Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND University of Turku Tel.: + 358 21 633 7318 INTERNET: jouko.maki-at-utu.fi FAX.: + 358 21 633 7380
More on CPD. Currently I am buying a package which includes CPD 030 and magnetron sputter coater CSD 050 from BALZERS Australia , originally Balzers Lichtenstein, at a very reasonable price of Aus $19,700. CPD has got incorporated a refrigerator unit and apparently performs very well. Worth considering , perhaps? Cheers, Wis Jablonski EM Unit
M. J. Kramer Having used both types (manufacturer wise) I would recommend getting the Hitachi. I currently use Hitachi, Zeiss and JEOL scopes in my lab. Service from all three is outstanding. I used Amray many moons ago and even the service rep had problems getting replacememt parts that worked. If they corrected their problems with parts and service, I wouldn't know. The three I use have been around for a long time and Amray is still relatively new ( in my opinion) to the field. I think they have only been around for about 20 years. Hitachis' FE scope is a beautiful scope to work with. I tried it once at a demo and it seemed easy to use and the photos were very nice from low kv to higher kvs. The scope, as a whole felt "right".
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