Further to Randy's suggestions: use a long shutter time if you can- it helps! Also, I believe I have done the job looking at the waveform rather than a normal image, because you're looking at something you know.
Hi, can anyone tell me about experience with anti-gfp antibodies on glutaraldehyde fixed cells or tissue? I am looking for an antibody which is suitable for electron microscopical gold labelling. Someone out there who has already tried it with or without success? thanks reinhard . . . . . . . . . . . . . . . . . . . Dr. Reinhard Windoffer Fon: (00)49 (0)6131/39 3720 Universitaet Mainz Fax: (00)49 (0)6131/39 4615 Anatomisches Institut e-mail: windoff-at-mail.uni-mainz.de Becherweg 13 D-55099 Mainz Germany . . . . . . . . . . . . . . . . . . .
we have been informed that You work on the structural investigation of proteins by high resolution electron microscopy. According to recently found unpublished notes traceable to Ernst Ruska the resolution limitation of a transmission electron microscope due to spherical aberration can be overcome by placing the microscope into a highly inhomogenous magnetic field such as at the poles of the larger planets of our solar system. Additionally one can expect extremely stable cryo-conditions around 4 Kelvin in the outer regions of our solar system. The National Aeronautics and Space Agency of the United States of America has therefore launched a project to install a Philips CM200FEG inside an unmanned satellite to be sent to Jupiter where it will be held in orbit for the duration of the experiment. All manipulations such as alignment and specimen insertion will be carried out by a remotely controlled robotic arm.
We are currently seeking suitable specimens for these undertaking and would appretiate contributions from Your department in exchange for a co-authorship in all resulting publications.
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1922, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
Hello everyone, I am trying to make cross-section TEM samples of thin films (ZnO) on sapphire. So far I have tried to make the samples by the traditional method ie., glue two pieces face-to-face, polish to 15 micron, and then ion-mill. I have had some problems with this method. Some of the people I have contacted about this told me to try using the wedge polishing method. Has anyone out there used this technique for making cross-section samples of films on sapphire? I was informed by another person that this might not work as sapphire is very brittle when it is thin, and we might not get a very good wedge. I would greatly appreciate any hints that you can give.
Sincerely
Chandrasekhar (chandu) Gorla Rutgers University Dept. of Ceramic and Materials Science Piscataway, NJ 08855 TEl: (732)445-3663
____________________________________________________________________ Get free e-mail and a permanent address at http://www.amexmail.com
Unicryl is made by BBInternational and distributed in the USA by Vector Laboratories, Inc. (1-800-227-6666). They are a CA based company and are not open yet today. I did call there yesterday and asked for a method booklet on Unicryl and the individual who I talked to did not indicate that the resin was unavailable. In fact, I just recieved a promotional brochure on the resin yesterday which prompted me to look into it further.
For those that are not familiar with Unicryl, it is a resin that is supposed to be more all purpose. The company claims that it is good for light and electrom microscopy, immunolabeling, in situ hybridization and histochemistry. It is a one component resin,of low viscosity and can be polymerized with UV light at 4o C or at 55oC.
This all sounds good but I did want to benefit from others experience as to how it compares with other resins such as Epon generic, Spurr's, LR White, and the various lowicryls like HM20 (my favorite for low temp.ICC).
Debby Sherman, manager Microscopy Center in Agriculture Purdue University
--------------------------------------
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Dear Walter I feel there must be an easier way! However, I have despatched for your personal attention a live sample of the new mutant AIF strain of Ebola virus, which has a novel and enigmatic structure we are anxious to elucidate. I assume NASA has the appropriate containment resources to deal with the consignment on arrival. Although I don't expect to hear from you again immediately, I would appreciate it if you could keep me informed of progress with the experiment. Chris
------ Forwarded Message Follows -------
Hello Friends, Need your advice again! Can Spurr resin be etched like some of the Epon-type resins can for Immuno work? If so does anyone have a recipe? I have a difficult specimen, candida, that is great in Spurr, so-so in Epoxy, horrible in Lowicryl and this is getting frustrating. I don't know if Unicryl or LRwhite would work. The problem with the hydrophilic resins seems to be that the resin swells slightly as the sections are cut and the sharp distinction between the resin and the wall of the candida causes a split from the resin. The candida either fold over or fall out, leaving so few good ones that I can't proceed to immuno work and hope for any results. Thanks, Linda M. Fox lfox1-at-wpo.it.luc.edu
Hi Linda,
We used Unicryl resin for many years on several yeasts with good results. We do not remove the cellwall of the cells but "etch" it a little by an incubation with 0.1% meta periodate for 5 minutes prior to the dehydration. See the paper of Evert van Tuinen Journal of Histochem. & Cytochem. vol. 35/3, page 327-333 (1987).
You should also I have filed a formal complaint to the originator's postmaster of this message since it was signed by a bogus account from "rumble.csb.ki.se" and was sent in a way to attempt to both hide detection and also illegally represented an agency of the US Government.
It is obvious this was a joke, but it broke all the Listserver Rules, which everyone agrees to when they sign on.
Jokes I can put up with, but not changing addresses, attempting to hide and impersonating a government agency!
Does anyone know where one can find 100kV DC Cables? Multicore cables that will hold off the 100kV?
Your assistance is appreciated.
-Karl
-- "It wouldn't be research... if I knew what I was doing." +++++++++++++++++++++++++++++++++++++ ++ Karl R. Umstadter, Ph.D. ++ Los Alamos National Laboratory ++ P-23, PFX-I Research ++ MS H803, Los Alamos, NM 87545 ++ Office: 505.665.1509 ++ Fax: 505.665.4121 +++++++++++++++++++++++++++++++++++++
We have been using a LEO 435 VPi SEM with an Oxford Inst. ISIS 300 integrated system since December of 1996. Since day one, we have had a series of problems with this integrated system, apparently caused by the common operating systems (first Win 3.1, then WIN95). It appears that the two systems do not like being integrated under one computer, as the sharing of resources seems to cause lurking problems that randomly appear, and cause system failures. I would be interested if any other users of the ISIS 300 system have had any other similar problems. Oxford claims that we are alone with our troubles, but I find this very hard to believe.
The first incarnation of our system used WIN 3.1 as the operating environment. This seemed optimum for the LEO as we encountered no system problems here. The ISIS though, would constantly run out of system resources and crash, usually without warning and lock up the system. It turned out that the ISIS is a resource hog (perhaps due to being written in Visual Basic), and the upgrade to ver 3.2 did not help at all. We then upgraded the system to Win 95, and the resource problem with the ISIS appears cured, but we have uncorked a beast in the LEO. The LEO is being upgraded to a full win95 system soon, but I am seeing more and more bugs with the ISIS.
Am I alone in believing the ISIS to be a poorly conceived and executed system? We have had endless conflicts with the operating system, to the point of having to reinstall win95 several times after apparent simple crashes, which seem to render several ISIS modules unavailable after that. The user interface appears haphazard from module to module, with no user definable defaults to customize operation to our preferences. The Labbook system forces users to collect data according to the x-ray system parameters. We like to structure our data by customer and job parameters, and keep all x-ray and microscope data under one directory. This is impossible under the ISIS Labbook system.
I am greatly disappointed with the integration of these two products. The LEO has been a fine system, with the operating environment keyed to usability, and endlessly customizable by the user. The ISIS is user unfriendly, and rigid in structure. Unless the user follows the Labbook protocols exactly, the system is unavailable to them.
Has anyone heard of a replacement operating environment for the Oxford system? Our complaints to Oxford have been dismissed, and LEO is unable to effect changes to the abysmal Oxford operating system. I wonder if I am alone in my frustration with this unfriendly and quixotic x-ray system.
Mike Warfield Hughes Space & Communications M&P Laboratory
You didn't say if your were dimpling the sample before ion milling and you didn't say what your ion milling conditions were -these are important. 15 um is a lot of material to ion mill through and you can minimize or eliminate differential sputtering effects by using low angles. You should precision dimple the sample down to less than 5 um and you can probably get close to 1 um. Then ion mill. If you have an older ion mill, you are going to need a sector control stage. If you have a newer ion mill, use low angles but remember that you have to clear the rim of the dimple. This means that if you have a 100 um thick sample and dimple on one side, you can't get below about 4 degrees. Double-sided dimpling and thinner sample allows you to go lower.
You can do the Tripod technique but it takes an investment in time and some equipment to learn it. It is best to learn from someone who knows. South Bay Technology has a periodic short course taught by Ron Anderson. You should contact them to see what is involved.
Sapphire also works with the Small Angle Cleavage Technique. I've done ZnO on Si and glass coverslip substrates this way and it works well. You should be able to do it with sapphire easily enough. This technique has a lot of advantages, but one of the best is that it is very cheap. South Bay Technology has a kit for getting started on this technique called the MicroCleaving Kit. John McCaffrey and I have a detailed pictorial tutorial on how to do it in the "Specimen Preparation for TEM of Materials IV" book Vol 480 from MRS that should be able to get you started on it. I always try this technique first; if it works, it's great.
Good luck.
Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Hello everyone, I am trying to make cross-section TEM samples of thin films (ZnO) on sapphire. So far I have tried to make the samples by the traditional method ie., glue two pieces face-to-face, polish to 15 micron, and then ion-mill. I have had some problems with this method. Some of the people I have contacted about this told me to try using the wedge polishing method. Has anyone out there used this technique for making cross-section samples of films on sapphire? I was informed by another person that this might not work as sapphire is very brittle when it is thin, and we might not get a very good wedge. I would greatly appreciate any hints that you can give.
Sincerely
Chandrasekhar (chandu) Gorla Rutgers University Dept. of Ceramic and Materials Science Piscataway, NJ 08855 TEl: (732)445-3663
____________________________________________________________________ Get free e-mail and a permanent address at http://www.amexmail.com
We look after a number of Topcon / ISI systems and find that we have a repeat problems of the Power Transistors for the Bias, filament and EHT control blowing. These are the four transistors that sit in the small box on top of the EHT tank on the Sx SS and ABT range of E.M.'s We have tried a number of transistors and ways of mounting these, but they continue to give problems. Does anybody else have the same problem, but more importantly has any one solved this problem ?
Thanks Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290
The presence of microbes in the deep core is significant to considerations of life elsewhere. See the story for more information.
Scott
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
At 11:59 AM 4/1/98 +0100, you wrote: } Received: from rumble (rumble.csb.ki.se) by ondine (4.1/SMI-4.1) -=-snip-=- } Dear fellow researcher, } -=-snip-=-
Those wacky jokesters at the Department of Bioscience of the Novum Karolinska Institutet in Sweden. Always good for a few laughs. ;} )
} Can someone tell me a bit more about Unicryl? } I'm not a biologist, I work in a Geology department, would love a non- } viscous resin which would soak well into crumbly, porous rocks and } wouldn't polymerise until told to.
Ritchie -
I suggest that you try LR White, Hard grade. It works well for things like zeolite catalysts, for example. Check these refs for details:
Csencsits, R., Schooley, C.,& Gronsky, R. (1985) An improved method for thin sectioning of particulate catalysts. J.E.M. Technique 2:643-644.
Ulan, J.G., Schooley, C., & Gronsky, R. (1990) Microtomy of large particle zeolites for TEM. Mat. Res. Soc. Symp. Proc. 199:153-156
Ulan, J.G., Schooley, C., & Gronsky, R. (1990) Modified embedment procedure for microtomy of large particle zeolites. J.E.M. Technique 16:254-255
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
I would suggest that we could examine the Good Times virus or we may still have some Pilt Down Man bones in the basement.
Thanks for the opportunity to collaborate but I would prefer the air miles if NASA is in that scheme.
Malcolm Haswell University of Sunderland UK ----------
Dear fellow researcher,
we have been informed that You work on the structural investigation of proteins by high resolution electron microscopy. According to recently found unpublished notes traceable to Ernst Ruska the resolution limitation of a transmission electron microscope due to spherical aberration can be overcome by placing the microscope into a highly inhomogenous magnetic field such as at the poles of the larger planets of our solar system. Additionally one can expect extremely stable cryo-conditions around 4 Kelvin in the outer regions of our solar system. The National Aeronautics and Space Agency of the United States of America has therefore launched a project to install a Philips CM200FEG inside an unmanned satellite to be sent to Jupiter where it will be held in orbit for the duration of the experiment. All manipulations such as alignment and specimen insertion will be carried out by a remotely controlled robotic arm.
We are currently seeking suitable specimens for these undertaking and would appretiate contributions from Your department in exchange for a co-authorship in all resulting publications.
by pohl.acpub.duke.edu (8.8.5/Duke-4.5.3) with ESMTP id LAA23957; Wed, 1 Apr 1998 11:10:41 -0500 (EST) Received: (from saram-at-localhost) by soc11.acpub.duke.edu (8.8.5/Duke-4.4) id LAA00765; Wed, 1 Apr 1998 11:10:40 -0500 (EST)
Refs for virus pix:
Palmer EL, Martin ML. Electron Microscopy in Viral Diagnosis. CRC Press, Boca Raton. 1988. (Ebola in here)
Doane FW, Anderson N. Electron Microscopy in Diagnostic Virology. Cambridge Press, New York. 1987. (Marburg here)
Miller SE. Detection and identification of viruses by electron microscopy J Electron Microsc Tech (now J Microscopy Research and Technique) 1986;4:265-301.
Miller SE. Diagnosis of viral infections by electron microscopy. In Lennette EH, Lennette DA, Lennette ET (eds). Diagnostic PRocedures for Viral, Rickettsial, and Chlamydial Infections. American Public Health Assoc, Washington, DC. pp 37-78. (Ebola here)
Murphy FA;, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, Martelli GP, Mayo MA, Summers MD (eds). Virus taxonomy, 6th Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, New York. (Marburg and Ebola here0
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We use a simple ground glass screen with a magnifying lens built into a plastic housing which fits on the back of the camera in place of the Polaroid 545 back. It was cheap, and came from our local photographic supply house. It works fine.
Tony Garratt-Reed
At 02:45 PM 3/31/98 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
We use LRWhite alot. The stains rinse out very quickly. Use just 10 dips in 2 beakers of distilled water. then dry.
Bob Underwood Derm Imaging center
On Fri, 27 Mar 1998, Barbara Plowman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have been staining some LRWhite(medium) sections and I am getting very low } contrast with 30min. 3%UA and 10 min. Lead Citrate. I have tried reducing the } wash times and the KV to 60KV without any luck. Any ideas and/or advice? } } Barbara Plowman } University of the Pacific } School of Dentistry } 2155 Webster } San Francisco, CA 94598 } email:Bplowman-at-uop.edu }
In our diagnostic EM lab we currently use Paragon stain to stain our epoxy semi-thin sections, but it would be very nice if we had some kind of rapid H&E like stain that could give that kind of polychrome differentiation between the nucleus and the cytoplasm.
The Paragon stain is supposed to be a polychrome stain, but what we in effect see is more of a monochrome direct stain.
Does anyone know of a quick yet superior stain that might give us better results here?
What I would like to know is how Philips scored this cherry deal= to get their scope in orbit, and, in what way they reduced costs= in order to get below NASA's ceiling for non-competitive action?= Considering the application I suppose the vacuum pump system could= be deleted: Who needs a rotary pump when for backing vacuum you= can "vent to space"...
The question is would Balzers, Edwards and other members of the High= Vacuum Equipment Consortium initiate a Congressional Oversight Committee= Investigation into the matter? If so, our samples destined to be= launched with the probe could be lost for a decade or so in this= legislative morass!
Even with the reduction in maintenance-requiring systems did NASA= obtain a service contract through Philips or, in an effort to contain= costs, get one with another third party service provider? I pity= the poor service technician whose service region this unit falls,= or rather orbits, in. Then again, just think of the mileage reimbursement.= =20
The question lingering, however, will be if FedEx will be able to= deliver replacement parts still by P1 Overnight Express...
************************************************************ It's true- the inmates ARE running the asylum... ************************************************************ Laura Rhoads Electron Microscopy Facility Director Department of Biology Western Kentucky University 1 Big Red Way Bowling Green, KY 42101-3576
Folks, do any of you know of a supplier for ITO glass for making heated culture chambers Thanks Simon
----------------------------------------------------------------------- Simon C. Watkins Ph.D. Associate Professor Director, Center for Biologic Imaging University of Pittsburgh Pittburgh PA 15261 tel:412-648-3051 fax:412-648-2004 URL: http://sbic6.sbic.pitt.edu
Does any one have any info on a CITZAF type package that includes a thin film correction and is NOT CITZAF3 by John Armstrong?? We are interest in anything, Commercial or Freeware.
I am not currently part of the list, so please mail to me directly.
I am currently in the process of restoring a Zeiss Axiomat LM. We do not have the screw rings/adapters required for attaching a camera, and are still waffling over film vs. digital cameras. If I go with with film, do I absolutely need a low momentum shutter? Can anyone give me advice on where to look for the adapters? I know that it will be difficult to find parts for a twenty-something year old scope, but it seems to be in good working order. Thanks for any help
Charlie Ginsburg NCC Research Dept. Lombard IL cgins-at-yahoo.com
_________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
Hi, I just received a job announcement from the Mayo Clinic in Rochester, MN. by snail mail. They are looking for an Electron Microcopy Technologist to work at their Core Facility . Please contact Kaine Kerkhoff {kerkhoff.kaine-at-mayo.edu} if you are interested in this position. thought some of you might like to know, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------= =2E } =
} Fellow microscopists: } =
} We're trying to make a deposition of 2 micron gold particles on a sili= ca } substrate. } We cannot use an evaporator because we don't want a thin film: we want = to } deposit the particles directly on the substrate, keeping them isolated = and } without forming aggregates. } At first we dispersed the powder directly on the silica, but SEM analys= is } showed they formed aggregates. } Then we've tried making a suspension on etanol and then putting a dropl= et } of the solution directly on the substrate. But still it formed aggregat= es. } =
} Does anyone know of a simple method to make such a deposit (without for= ming } aggregates) ? } =
} Thanks in advance. } =
} Isabel Nogueira } Instituto Superior T=E9cnico } Departamento de Engenharia de Materiais } Av. Rovisco Pais } 1096 Codex } Tel.: 351 - 1 - 8418124/0 } Fax.: 351 - 1 - 8418120 } E-mail: isabeln-at-alfa.ist.utl.pt
There are reliable methods to make coloidal gold in suspension. The gold particles are very regular in size and show little aggregation. Also aggregates can be removed by centrifugation. Your problem would seem to be to deposit these particles onto the silica substrate without aggregation. I guess it should not be impossible by adjusting concentration and charge of the particles in suspension. Also, they probably adsorb to the film if you put the suspension briefly in contact with the substrate. If everything fails, may be they can be centrifuged into the silica. It is possible that the coloidal gold is more stable in solution than the gold you are using, but I do not know if it is possible to make it as large as you want. Usually the particles have up to 15nm. May be they can be grown after they are deposited in the silica.
Good luck
A.P. Alves de Matos Electron Microscopy Unit Curry Cabral Hospital Lisbon
I make a focusing screen for a homemade "roberts lens focuser" for a nikonos underwater camera, by grinding a 1.5" x 3" microscope slide using some grinding compound. Worked fine for that purpose, but you'd have to determine the plane of focus and make sure the glass you use will span the width. It may not have to cover the entire surface.
Alan Davis adavis-at-netpci.com
On Wed, Apr 01, 1998 at 12:22:11PM -0500, Tony Garratt-Reed wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Paula - } } We use a simple ground glass screen with a magnifying lens built into a } plastic housing which fits on the back of the camera in place of the } Polaroid 545 back. It was cheap, and came from our local photographic } supply house. It works fine. } } Tony Garratt-Reed } } At 02:45 PM 3/31/98 -0800, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Listers, } } } } I am in need of a type 545 Polaroid camera back that has a frosted } } glass where the film holder usually is. My SEM pix look OK on the screen } } but if I make a print from the neg. it looks slightly out of focus. } } I've been told that I need to focus the camera to match the screen } } & this fancy camera back with the glass helps you do that. I know they } } exist because I saw one on an ESEM. Does anyone know where I can buy one? } } I called Polaroid and they were confused. } } Can anyone help me with this problem? } } } } } } My future looks fuzzy & so do my prints, } } } } Paula :-) } } } } Paula Sicurello } } UC Berkeley } } Electron Microscope Lab } } psic-at-uclink4.berkeley.edu } } } } } } } } } Anthony J. Garratt-Reed } MIT Room 13-1027 } 77 Massachusetts Avenue } Cambridge, MA 02139-4307 } United States of America } } Ph: 617-253-4622 } Fax: 617-258-6478 } }
-- Alan E. Davis Marianas High School (Science Department) AAA196, Box 10001 adavis-at-netpci.com http://www.saipan.netpci.com/~adavis Saipan, MP 96950 15.16oN 145.7oE GMT+10 Northern Mariana Islands
By GFP, I presume you mean glial fibrillary acidic protein. If you don't, ignore the rest of this! If you do, then you're in luck. GFAP is one of the toughest antigens there is, surviving in OsO4 fixed tissue, embedded in resin and left in a cupboard for 15 years. I've done a lot of labelling for GFAP. The best antibody I've tried is Dako's polyclonal rabbit anti-cow. Others I've tried include Amersham monoclonal mouse anti-human and Sigma polyclonal rabbit; they weren't as effective. You can label glut fixed whole tissue - assuming, of course, that you either use vibratome sections or permeabilise the tissue. You can label any old resin sections - if you can see the filaments, then there should be enough GFAP to label. Pretreat with NaIO4 first, then immunolabel as usual. If you're interested I can send you the method(s) I use. I've labelled with 10nm gold conjugate and also 1nm with Ag enhancment. Both methods work well.
Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
It is important that the ground part of the glass is precisely in the focal plane. The ground glass should be facing the lens and sit on the film guides. A ground glass screen is easily made by grinding the face of a cut (perhaps larger than standard) microscope slide. Say 800 grid grinding compound (as used for grinding geol. section) does the job very well.
First the magnifier should be focused onto the ground screen.
Then press photo (no beam required) and use a line scan at low intensity as an "image" on the screen.
Adjust the distance between CRT and camera to bring the image into focus. Alternatively, the lens on the camera can be focused.
Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
____________________________________________ } We use a simple ground glass screen with a magnifying lens built into a } plastic housing which fits on the back of the camera in place of the } Polaroid 545 back. It was cheap, and came from our local photographic } supply house. It works fine. } } Tony Garratt-Reed } } At 02:45 PM 3/31/98 -0800, you wrote: ----------------------------------------------------------------------. } } } } Listers, } } } } I am in need of a type 545 Polaroid camera back that has a frosted } } glass where the film holder usually is. My SEM pix look OK on the screen } } but if I make a print from the neg. it looks slightly out of focus. } } I've been told that I need to focus the camera to match the screen } } & this fancy camera back with the glass helps you do that. I know they } } exist because I saw one on an ESEM. Does anyone know where I can buy one? } } I called Polaroid and they were confused. } } Can anyone help me with this problem? } } } } } } My future looks fuzzy & so do my prints, } } } } Paula :-) } } } } Paula Sicurello } } UC Berkeley } } Electron Microscope Lab } } psic-at-uclink4.berkeley.edu
INTERNATIONAL COURSES OF LIGHT MICROSCOPY, PHOTOMICROGRAPHY AND LASER SCANNING CONFOCAL MICROSCOPY GARGNANO (Lake of Garda) October 1998
The Course is a post-graduated theoretical/practical course, with propedeutical lectures and practical stages on microscopy, photomicrography and confocal microscopy. The course will take place in Gargnano (Lake of Garda) in October 1998.
All information and registration details (participation fee, date, special accomodation) at the following Web address.
http://imiucca.csi.unimi.it/endomi/micro.html
Thank you Paolo Castano
_____________________________________________________ Prof. Paolo Castano UNIVERSITY OF MILAN INSTITUTE OF HUMAN ANATOMY - CHAIR OF HUMAN ANATOMY FOR PHARMACY Via Mangiagalli, 31 - 20133 Milan (Italy)
Hello Friends, Need your advice again! Can Spurr resin be etched like some of the Epon-type resins can for Immuno work? If so does anyone have a recipe? I have a difficult specimen, candida, that is great in Spurr, so-so in Epoxy, horrible in Lowicryl and this is getting frustrating. I don't know if Unicryl or LRwhite would work. The problem with the hydrophilic resins seems to be that the resin swells slightly as the sections are cut and the sharp distinction between the resin and the wall of the candida causes a split from the resin. The candida either fold over or fall out, leaving so few good ones that I can't proceed to immuno work and hope for any results. Thanks, Linda M. Fox lfox1-at-wpo.it.luc.edu
Hi Linda,
We used Unicryl resin for many years on several yeasts with good results. We do not remove the cellwall of the cells but "open" it a little by an incubation with 0.1% sodium meta periodate (w/v in water) for 5 minutes prior to the dehydration. See the paper of Evert van Tuinen Journal of Histochem. & Cytochem. vol. 35/3, page 327-333 (1987).
Nestor J. Zaluzec wrote: } } Enough said ! } } You should also I have filed a formal complaint to the originator's postmaster } of this message since it was signed by a bogus account from } "rumble.csb.ki.se" and was sent in a way to attempt to both hide detection } and also illegally represented an agency of the US Government. } } It is obvious this was a joke, but it broke all the Listserver Rules, which } everyone } agrees to when they sign on. } } Jokes I can put up with, but not changing addresses, attempting to hide and } impersonating a government agency! } } Nestor
Dear Nestor and list members and NASA,
I'm sorry about any bad feelings I've caused and I'm turning myself in.
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
The problem you describe is something we saw with a S440i system about 3 years ago. As is usual, the SEM manufacturer and the ED manufacturer will both say the intergration works before you place the order and then both deny it is their system crashing after you have paid. ( please don't sue me for this !!!!??? ) Sorry to say it but you must simply de-integrate the two. Isis was designed around the HP type PC. So let it run on that. The Leo will run happily on the PC. We must remember that P.C. stands for Personal Computer. So documents and games work ......reasonably well !!???....... But if you load a P.C. with real work, it gets a bit of a speed wobble and crashes fairly often. The Leo, on it's own, should have 64Mbytes of memory fitted to make it work comfortably and ISIS needs the HP. We did this on the system here and found we have far less problems. With computers we always expect a few crashes.
Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290
-----Original Message-----
Hello to all,
We have been using a LEO 435 VPi SEM with an Oxford Inst. ISIS 300 integrated system since December of 1996. Since day one, we have had a series of problems with this integrated system, apparently caused by the common operating systems (first Win 3.1, then WIN95). It appears that the two systems do not like being integrated under one computer, as the sharing of resources seems to cause lurking problems that randomly appear, and cause system failures. I would be interested if any other users of the ISIS 300 system have had any other similar problems. Oxford claims that we are alone with our troubles, but I find this very hard to believe.
The first incarnation of our system used WIN 3.1 as the operating environment. This seemed optimum for the LEO as we encountered no system problems here. The ISIS though, would constantly run out of system resources and crash, usually without warning and lock up the system. It turned out that the ISIS is a resource hog (perhaps due to being written in Visual Basic), and the upgrade to ver 3.2 did not help at all. We then upgraded the system to Win 95, and the resource problem with the ISIS appears cured, but we have uncorked a beast in the LEO. The LEO is being upgraded to a full win95 system soon, but I am seeing more and more bugs with the ISIS.
Am I alone in believing the ISIS to be a poorly conceived and executed system? We have had endless conflicts with the operating system, to the point of having to reinstall win95 several times after apparent simple crashes, which seem to render several ISIS modules unavailable after that. The user interface appears haphazard from module to module, with no user definable defaults to customize operation to our preferences. The Labbook system forces users to collect data according to the x-ray system parameters. We like to structure our data by customer and job parameters, and keep all x-ray and microscope data under one directory. This is impossible under the ISIS Labbook system.
I am greatly disappointed with the integration of these two products. The LEO has been a fine system, with the operating environment keyed to usability, and endlessly customizable by the user. The ISIS is user unfriendly, and rigid in structure. Unless the user follows the Labbook protocols exactly, the system is unavailable to them.
Has anyone heard of a replacement operating environment for the Oxford system? Our complaints to Oxford have been dismissed, and LEO is unable to effect changes to the abysmal Oxford operating system. I wonder if I am alone in my frustration with this unfriendly and quixotic x-ray system.
Mike Warfield Hughes Space & Communications M&P Laboratory
At 02:45 PM 3/31/98 -0800, Paula Sicurello wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What you are looking for is a "focusing screen". It can be made following one of a number of materials, ranging from frosted glass to fresnel lenses to just a translucent piece of plastic --- anything which will allow you to catch the aerial image formed by the microscope. You can usually buy a piece of fresnel material at a science shop (most of the local malls have them now) or from a photographic store (I am not sure that they have one big enough to fit your Polaroid back. In any event, remove your film cassette and mount the screen in the plane where the film would be. If the adapter holding the camera system is adjustable, raise or lower the camera system until the image is in focus on the screen. (For more critical focus, try using either an 8x loop, available at any photographic store, or a simple 5x eyepiece from a light microscope). Once you have found this focal plane, lock the camera in position. You have now made your microscope and your camera system parfocal.
Being a light/confocal microscopist, I am unfamiliar with EM optics and how the image is formed at the film plane. In a light microscope, the depth of focus at the image plane is directly related to the magnification squared. As a result, using a low power objective gives the shallowest and most accurate focus in the image formed at the film plane and guarantees that all higher mags will automatically be in focus. }
Hope this helps.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
Garry, I, too, use the Paragon stain, but I do get a polychrome effect. I was wondering if you use a saturated Sodium Borate solution to enhance your staining. After drying the section on the hotplate, I put a 3 to 4 drops of Paragon stain and one drop of sodium borate on the slide (still on the hotplate). The combination of heat and sodium borate enhance the Paragon stain quite nicely. You might want to try this before adding H & E to your "to do" list.
Sharron G. Chism HT (ASCP) Electron Micropscopy Lab Harris Methodist Fort Worth Fort Worth, Texas ----------
In our diagnostic EM lab we currently use Paragon stain to stain our epoxy semi-thin sections, but it would be very nice if we had some kind of rapid H&E like stain that could give that kind of polychrome differentiation between the nucleus and the cytoplasm.
The Paragon stain is supposed to be a polychrome stain, but what we in effect see is more of a monochrome direct stain.
Does anyone know of a quick yet superior stain that might give us better results here?
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Dear Listers!
We have the problems with our JEM-4000EX: independent shut down after 10-30 minute of work. The possible cause is disturbance of logical schemes work of vacuum system. Does anybody else have the same problem, but more importantly has any one solved this problem ? Does anyone know how to make testing of vacuum system by CRT of microscope: how to load programme, where is check points and switchs e t.c.? I am not currently part of the list, so please mail to me directly.
Anton
gut-at-thermo.isp.nsc.ru
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We have a Durst Laborator 138 S enlarger. The lamp recently blew and we don't have a backup. We have tried to find a source for the special bulb but with no luck. Is Durst still in business? We have tried the Internet. The blown bulb is not the one that belongs in the enlarger. Thanks for any leads on Durst.
Researcher in the area of "Electron microscopy in plant research". Reference number 1716/98-4599.
At the Electron Microscopy Unit, Department of Plant Breeding Research, The Swedish University of Agricultural Sciences, Sval=F6v/Alnarp, Sweden. The EM Unit will be a common resource for the university's departments in Southern Sweden.
The candidate must have a doctor's degree, primarily not earlier than five years ago. Experience of work with plant material is a merit. Of importance are the scientific, pedagogic, adminstrative and other capacities of relevance for this occupation. Of importance also is the capacity to inform about scientific research in a popular way.=20
The appointment is first for two years, followed by another period of two years and the possibility for a further prolongation. The salary is individually determined (at least 20 000 Swedish crowns a month, before taxes). Women are engouraged to apply.
Together with the application, a short account on the applicant's scientific and pedagogic activities should be given. The application marked with the above-mentioned reference number, together with a certified C.V., merit and publication lists, and other documents (including scientific and pedagogic works) should be sent in two copies so that they reach the "Registrator, SLU, Box 7070, S-750 07 Uppsala, Sweden" at the latest on April 23, 1998.
For further information contact professor Waheeb Heneen, tel +46 418 667064 or fax +46 418 667081.
I work for VayTek, and would like to respond to Alex Black's question on 3D reconstruction using TEM.
Yes it's possible. You must aquire a series of images in sequence and equidistant from each other. They must be all the same size, resolution, and magnification.
I'm aware that this is a tall order. I've just completed doing a 3D reconstruction of some TEM images with great results. The process involved some very tricky TEM work, and some sofisticated image processing using three software packages.
I'm not at liberty to give you more details on the TEM work, but if you're interested in knowing more about the process, please e-mail me directly. I'll contact the lab doing this and ask them to contact you. I'll also fill you in on the software involved.
Best regards
Patrick Guerin Customer Technical Support Engineer VayTek, Inc. 305 West Lowe Avenue, Suite 109 PO Box 732 Fairfield Iowa 52556-0732 Tel : 1-515-472-2227 ext. 103 Fax : 1-515-472-8131 WWW.VAYTEK.COM E-mail : pguerin-at-vaytek.com
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Dear Sara, } } Does anyone have experience growing tissue culture cells on gold grids } with support films? We want to do IEM of filaments after digestion of } the cytoplasmic membrane, } The one time I tried this (and succeeded) I just spread the carbon- formvar-coated gold grids on the bottom of a culture dish, added PTK1 cells and let the cultures sit at 37 degrees. YMMV, if your cells do not grow over carbon-formvar; however, if you grow your cells on a treated culture dish to promote adhesion, treating the carbon-formvar film the same way should work (IMVVHO).
} Articles describing this technique mention specific kinds of reagents, } grids, etc. Are there better brands, methods? We'd appreciate any tips } to avoid a lot of trial and error. } } Source of gold grids? (supplier)
Probably makes no difference unless the "gold" in the grids contains different impurities, and if these affect cell growth.
} Type of support film? Formvar/collodion)
If the hydrophobicity, texture, etc. of the film matches that of the usual culture surface, you are likely to get the best results.
} Carbon coat? (heavy/light)
I'd reccommend it, unless you need uncoated formvar to match adhesion conditions.
} Method of sterilization? (UV/gas)
Probably irrelevant, unless the surface is affected adversely. Bear in mind that I am most definitely not an expert. Good luck. Yours, Bill Tivol
I know we should drop this subject but...........This is like dejavu. Several years back I was teaching the microscope structure/function section of an EM course and began thinking how to explain the theory of resolution, trying to make it interesting and not too complicated. My mind wandered (as it does quite often these days) and got to thinking about the effects of space and the resolution potential of an electron microscope, The Phillips engineer happened to be here and blew it past him. He wasn't sure what effects it would have but thought it interesting. He said the main limitation in current scopes was the quality of the lenses, but could we make better lenses in space then add that to the vacuum, cold, and magnetic/gravitational variable and get a significant increase in resolution? Just a thought, I realy don't want us to get into the physics of this.
Simon C. Watkins Ph.D. Associate Professor Director, Center for Biologic Imaging University of Pittsburgh Pittburgh PA 15261 tel:412-648-3051 fax:412-648-2004 URL: http://sbic6.sbic.pitt.edu
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As a class project, I am writing a proposal for a (fictitious) SEM lab for a university geosciences department. I am attempting to obtain the Alderson book on EM lab design, but would like additional references. Any citations would be appreciated.
Also, if any geoscientists have any suggestions for how they would like to see an SEM/EDS lab equipped, I would be happy to hear them.
with TCP; Thu, 02 Apr 98 17:04:05 EST Received: from [137.99.40.57] by oracle.pnb.uconn.edu with SMTP (Eudora Internet Mail Server 1.2); Thu, 2 Apr 1998 17:12:29 -0500 Message-Id: {v01510109b149acd24f9d-at-[137.99.40.57]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step. In some protocols a BSA buffer rinse is also used before the secondary antibody, and/or is added to the primary and secondary antibody solutions. In other cases it is omitted. My question, to immuno experts on the listserver, is whether there is a DISadvantage to adding the BSA after the initial blocking step. As I understand it BSA reduces non-specific binding, so it would seem prudent to add it whenever possible.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
In the mid-1970's NASA did send to universities a request for suggestions for experiments to put into space. At the time I was at Bristol and put it to my colleagues that would should submit a proposal to put a TEM into space. My argument was that all the instability problems would go away. No vibrations from vacuum pumps because there are no vacuum pumps - no vibration from anything else either. No instabilities in the lens coils because they would all be superconducting with no need for cooling equipment. No sample contamination because the vacuum is clean. It would make for perfect images. I was motivated by the thought that I might get to go into space to operate it (I was younger then). I could not persuade my colleagues to take me seriously. Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
You wrote: I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step.
If BSA is a problem i.e. failing to block non-specificity , there are several other blockers that can be used. Ovalbumin and fish gelatin are often "cleaner" blocking agents for gold labeling procedures. Also the purity of BSA can be a factor, I always buy a grade of BSA that contains greater than 95% BSA of MW 66,000. If I can be of further help, feel free to contact me off the listserver. Marge
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
We are looking at hi-res CCD cameras, preferably with video-rate output as well, for 300kV TEMS (1000 x 1000 pixel, 12 bit minimum), and would appreciate the benefit of other's experience regarding quality, ease of use. longevity, radiation problems, value-for -money, etc. TIA Sally Stowe ---------------------------------------------------------------------- Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: Box 475 ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 (0)2 6249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, Australia 2601 http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step. In some protocols a BSA buffer rinse is also used before the secondary antibody, and/or is added to the primary and secondary antibody solutions. In other cases it is omitted. My question, to immuno experts on the listserver, is whether there is a DISadvantage to adding the BSA after the initial blocking step. As I understand it BSA reduces non-specific binding, so it would seem prudent to add it whenever possible.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
I always use it in all solutions where antibodies and/or colloidal gold are used, and as an initial blocking solution. I don't know of any reason not to use it to reduce nonspecific staining, although cost might become an issue if you do your washes in between steps in very large volumes.
At 05:01 PM 4/2/98 -0400, MARIE CANTINO wrote:
} I have noticed that in EM immunogold-labelling protocols, BSA is often used } in buffer as an initial blocking step. In some protocols a BSA buffer } rinse is also used before the secondary antibody, and/or is added to the } primary and secondary antibody solutions. In other cases it is omitted. } My question, to immuno experts on the listserver, is whether there is a } DISadvantage to adding the BSA after the initial blocking step. As I } understand it BSA reduces non-specific binding, so it would seem prudent to } add it whenever possible. } } Marie } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology, U-131 } University of Connecticut } Storrs, CT 06269 } Ph: 860-486-3588 } Fax: 860-486-1936 } } } } } _____________________________________ Craig Lending SUNY Brockport Department of Biological Sciences Brockport, NY 14420
On Fri, 3 Apr 1998, SALLY STOWE wrote: } We are looking at hi-res CCD cameras, preferably with video-rate } output as well, for 300kV TEMS (1000 x 1000 pixel, 12 bit minimum), } and would appreciate the benefit of other's experience regarding } quality, ease of use. longevity, radiation problems, value-for } -money, etc. } Sally, I am also interested in this information, please forward anything not posted to the list. Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Dear Marie, I think the protocol you need to use depends on your primary antibody. If BSA fails to block aspecific binding, skimmed milk powder can be a better blocking agent. But whatever blocker I use, I include it in all the reaction steps. Armelle
Armelle Gollotte Biotechnology Department Scottish Agricultural College West Mains Road Edinburgh EH9 3JG Tel: (44) 131 667 1041 Fax: (44) 131 667 2601 a.gollotte-at-ed.sac.ac.uk
} If BSA is a problem i.e. failing to block non-specificity , there are } several other blockers that can be used. Ovalbumin and fish gelatin are } often "cleaner" blocking agents for gold labeling procedures. Also the } purity of BSA can be a factor, I always buy a grade of BSA that contains } greater than 95% BSA of MW 66,000.
In order to get a very 'clean' blocking I always use modified BSA-C (Aurion) It is more expensive than quality grade BSA and Cold Fish gelatin but I find it is much easier to use (you get it as a non-sticky solution) and it gives a very clean and efficient block. I have never had any problems using modified BSA-C.
Do any of you have benchmarks which you use when evaluating cameras, digitizing boards, or image analysis systems? Also, are you using any type of standardized slides for comparing systems?
Thanks
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
Do any of you have benchmarks which you use when evaluating cameras, digitizing boards, or image analysis systems? Also, are you using any type of standardized slides for comparing systems?
Thanks
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
At 09:18 AM 4/2/98 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
George,
Durst headquarters is at 10 County Line Road, Suite 29, Branchburg, NJ 08876. Phone is (800) 463-8778, and their fax is (908) 429-0777. You can find a web site for them at http://www.photoshopper.com/durst/index.html.
If this fails to help in your bulb search, try such distributors as Bulbman (sorry, don't have their address handy) or another of the specialists in lighting equipment and bulbs. They have catalogs available and the one's I've dealt with are very willing to track down obscure bulb types if you can provide them with descriptions, identifying numbers, applications, etc.
Good luck.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
First of all , I am no immuno expert. I use ovalbumin for initial blocking and wash grids on porcelain spot plat filled with buffer and use blocking step again before secondary antibody reaction. I see no merit of adding BSA or any other blocking agent during washing.
I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step. In some protocols a BSA buffer rinse is also used before the secondary antibody, and/or is added to the primary and secondary antibody solutions. In other cases it is omitted. My question, to immuno experts on the listserver, is whether there is a DISadvantage to adding the BSA after the initial blocking step. As I understand it BSA reduces non-specific binding, so it would seem prudent to add it whenever possible.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
I am preparing cross-sectional TEM samples of sapphire. I am trying to find a good way to measure the samples' thickness when I am under about 70 micron. Any ideas?
For ICC blocking I use gelatin (Sigma Gelatin, G-9382, from bovine skin, type III, approx. 225 bloom). This preference results from a sabbatical year (1985-86) I spent in the lab of Daniel Louvard, then at the Pasteur Institute in Paris. They had tried a good many blockers and decided on this gelatin (easy to obtain and use, few cross reactions, etc.). They blocked with 2% gelatin in PBS, and thereafter maintained 0.2% in washes and vehicle for primary/secondary antibodies. A 10% gelatin stock was stored in the refrigerator, and was warmed (for example under hot tap water) to liquify for use.
Kent
A. Kent Christensen Department of Anatomy and Cell Biology Medical Sciences II Building University of Michigan Medical School Ann Arbor, MI 48109-0616 akc-at-umich.edu Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
--------------------------------------
} } } } MARIE CANTINO {CANTINO-at-ORACLE.PNB.UCONN.EDU} 04/02/98 } 04:01pm } } } } } I have noticed that in EM immunogold-labelling protocols, BSA is often } used } in buffer as an initial blocking step. In some protocols a BSA buffer } rinse is also used before the secondary antibody, and/or is added to the } primary and secondary antibody solutions. In other cases it is omitted. } My question, to immuno experts on the listserver, is whether there is a } DISadvantage to adding the BSA after the initial blocking step. As I } understand it BSA reduces non-specific binding, so it would seem } prudent to } add it whenever possible. } } Marie } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology, U-131 } University of Connecticut } Storrs, CT 06269 } Ph: 860-486-3588 } Fax: 860-486-1936
What is Paragon stain, and where can I get it? It sounds like it's exactly what I need, and I'm sure it would be usefulto the rest of the list if it's as wonderful as it sounds. TIA. Lesley Weston.
On Thu, 2 Apr 1998, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Garry, } I, too, use the Paragon stain, but I do get a polychrome effect. } I was wondering if you use a saturated Sodium Borate solution to enhance } your staining. After drying the section on the hotplate, I put a 3 to 4 } drops of Paragon stain and one drop of sodium borate on the slide (still } on the hotplate). The combination of heat and sodium borate enhance the } Paragon stain quite nicely. You might want to try this before adding H } & E to your "to do" list. } } Sharron G. Chism HT (ASCP) } Electron Micropscopy Lab } Harris Methodist Fort Worth } Fort Worth, Texas } ---------- } From: Garry Burgess } To: 'Microscopy' } Subject: H&E Like Stain for Epon/Araldite } Date: Wednesday, April 01, 1998 2:25PM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } In our diagnostic EM lab we currently use Paragon stain to stain our } epoxy semi-thin sections, but it would be very nice if we had some kind } of rapid H&E like stain that could give that kind of polychrome } differentiation between the nucleus and the cytoplasm. } } The Paragon stain is supposed to be a polychrome stain, but what we in } effect see is more of a monochrome direct stain. } } Does anyone know of a quick yet superior stain that might give us better } results here? } } } Garry }
In a counterpoint to Mike Warfield's message, we have been very happy customers of Oxford (Link Systems in its previous incarnation) since 1985.
We have one integrated and one stand alone ISIS systems. The stand alone is a 2 y.o. ISIS 3.00 EDS replacement of an obsolete Noran on a 16 y.o. AMRAY 1600 SEM. The other unit is a brand new ISIS 3.2 integrated into an AMRAY 3200/C ECO SEM. (We also operate a 1985 vintage AN10 on a VG STEM from the era.)
I was intimately involved in evaluation and purchase of both ISIS systems. During the last five years, we have evaluated in exhausting detail every brand of EDS equipment and found the Oxford detectors to offer the best spectral resolution, and the software to be the most comprehensive and useful. The Oxford software worked flawlessly in all demos and in our lab. under Windows 3.1, 95 and NT 4.0.
We are an industrial lab serving several of our plants and many of the plants' customers. A high throughput, very high reliability, and good rapid service on our analytical equipment are paramount. In 1997, our old SEM was used to examine over 2,100 samples, each a unique specimen, and more than 90% required EDS analysis. During that time, the ISIS 3.00 performed very well and operated without a mishap.
As is the case in any new installation, we ran into initial problems, but Oxford has bent over backwards to help us out through both installations.
In the 1996 Noran upgrade, we purchased a used (demo) ISIS 3.00 analyzer sans the detector to inexpensively replace a third party kludge fix up of an obsolete Noran EDS system, but we were keeping the old detector as it seemed to work just fine. Following the installation, transient high noise levels were experienced in the EDS spectra. Over the next few weeks, Oxford engineers replaced every board on the system and, suspecting an incompatibility with the modified Noran detector, Oxford provided us gratis a brand new 30 mm2 thin window detector with 126 eV peak resolution. In the end, it turned out that an intermittent noise originated from a flaky pulse processor power supply, but Oxford let us keep the $20,000+ detector; well above and beyond the call of duty. Since the replacement of the power supply in February 1996, the system has operated flawlessly and it easily adapted to an upgrade from Windows 3.1 to 95.
In the ISIS 3.2 integrated installation on the AMRAY 3200/C ECO SEM, we ran into a problem of insufficient x-ray count rate. The detector resolution was just fine (118 eV), but we needed to acquire spectra for 200 seconds with a higher beam current for counts comparable to 50 s acquisition on the old SEM. It turns out that I goofed and specified a 10 mm2 detector for an operator used to working with 30 mm2. Also, the Oxford detector is aligned for a sweet spot at a 25 mm working distance (from the objective polepiece to the sample plane) and a tightly focused collimator prevented us from getting x-rays at working distance shorter than 20 mm or greater than 30. To compound the problem, an interference from a third party add-on, a Robinson back-scatter detector, required the nosepiece of the Oxford EDS detector to be withdrawn above the bottom of the objective polepiece when the Robinson was inserted. The combination of smaller detector area and a huge detector to sample distance killed the x-ray count rate. When we contacted Oxford with our dissatisfaction of the system performance, the response was initially slow and unfocused until we got past the newly expanded inexperienced service staff once it became clear that our problems went beyond the standard garden variety. An experienced service engineer flew in from outside his working area within 10 days of our initial call, and with an aid of a knowledgeable sales rep., helped to fully diagnose the problem. The collimator on the detector nose piece has been modified and we have tested it to our satisfaction at the sample to objective pole piece working distance range of 15 to 40 mm. Oxford has also promised to make in a short order a new EDS detector mounting flange to remove the interference between the EDS and the Robinson backscatter detectors.
Having been there, I empathize with Mike Warfield's frustrations with his equipment not working as well as it should. However, I think that not the full story has been exposed here and that the finger pointing at Oxford is less than fair.
Point #1:
Our older ISIS 3.00 runs just fine on a 66 MHz HP Vectra with 16 Mbytes RAM. It is a stand alone system and has had no computer related problems at all under Windows 3.1 nor 95. (We upgraded to 95 solely for better compatibility with our LAN.) The integrated ISIS 3.2 and AMRAY 3200/C run on a Micron 133 MHz P5 with 32 Mbytes RAM and 512 Kbytes L2 Cache under Windows NT4.0. No computer related problems have manifested themselves in this system either, though perhaps surprisingly the old system is somewhat more responsive. (I do wish we did succeeded in getting through to the SEM vendor that an integrated system requires a 300 MHz P5. Why pinch a few hundred dollars on a $300,000+ system?)
It occurs to a few users, and it seems to fewer SEM vendors, but when two machines (as in an integrated SEM & EDS system) are squeezed onto one computer, at least twice as much RAM and twice as high CPU frequency are required, and the perforce will still not be as good as of two stand alone systems. That is because, though your CPU may be screaming at 500 MHz, the RAM and the system buss still chug along at the same old 33 MHz while two suites of software are time-slice multitasked and interrupts from two sets of hardware are simultaneously serviced. Nevertheless, what little one may have to give up in responsiveness WHEN AN APPROPRIATELY SCALED COMPUTER is used for an integrated system is more than made up in convenience. (I just can't imagine simultaneously operating two keyboards, two pointing devices, and watching three monitors with any efficiency.)
Mike does not list the specifications of his PC, but I wonder whether it's a poky old 486 with 16 Mbytes of RAM which works just fine running either the SEM alone or the EDS alone but bogs down when asked to do both simultaneously. (If there is not enough RAM the system may be rally grinding to a halt while sloshing the programs back and forth between RAM and the "virtual RAM memory" on the hard drive.) A RAM boost or a PC upgrade might help here.
Note that this is an integrated EDS system onto the SEM by the SEM manufacturer, and the responsibility for sufficient computer resources for both systems lie with LEO and not with Oxford.
Point #2:
Windows 95 is a terrible operating system in general, and especially for real time multitasking. It does not provide sufficient system resources and it does not clean up after itself (just check the number of *.TMP files accumulated on your hard drive). If fixes suggested in Point #1 don't work, upgrade from 95 to NT4. If the software runs under 95 chances are good it will run under NT4. ISIS 3.2 works flawlessly on our integrated machine under NT4.
Again, the responsibility for choosing the appropriate operating system lies with LEO.
Point #3:
Some SEM manufacturers rely on the purchaser to choose the EDS system and will perform the integration as a courtesy to the purchaser. However, the satisfactory site installation and subsequent service contract on the EDS unit is the responsibility of the EDS manufacturer. This is the case with our integrated system.
Other SEM manufacturers purchase an EDS unit as a commodity and sell the SEM with an integrated EDS as one package. I imagine there is then just a single service contract and the SEM vendor is supposed to service both the SEM and the EDS. In principle this is a better deal because there should be less finger pointing, but in reality the SEM vendor's tech. support is not up to snuff on the EDS end and the user pays in frustration. I think the LEO integration falls in this second category.
Point #4:
I am surprised that Mike is expressing his dislike of the Oxford user interface at this point in the game. I think a more appropriate time frame would have been during the evaluation period, well before the whole system was purchased and paid for.
These opinions are mine alone and in no way reflect the views of my employer. I bear no interest in AMRAY, OXFORD, or LEO.
Valdemar Furdanowicz
valdemar-at-fast.net Homer Research Laboratories Bethlehem Steel Corporation
---------- } From: mdwarfield-at-mail.hac.com } To: Microscopy-at-sparc5.microscopy.com } Subject: LEO 435 VPi with ISIS 300 X-Ray Analysis System } Date: Wednesday, April 01, 1998 9:38 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello to all, } } We have been using a LEO 435 VPi SEM with an Oxford Inst. ISIS 300 } integrated system since December of 1996. Since day one, we have had a } series of problems with this integrated system, apparently caused by } the common operating systems (first Win 3.1, then WIN95). It appears } that the two systems do not like being integrated under one computer, } as the sharing of resources seems to cause lurking problems that } randomly appear, and cause system failures. I would be interested if } any other users of the ISIS 300 system have had any other similar } problems. Oxford claims that we are alone with our troubles, but I } find this very hard to believe. } } The first incarnation of our system used WIN 3.1 as the operating } environment. This seemed optimum for the LEO as we encountered no } system problems here. The ISIS though, would constantly run out of } system resources and crash, usually without warning and lock up the } system. It turned out that the ISIS is a resource hog (perhaps due to } being written in Visual Basic), and the upgrade to ver 3.2 did not } help at all. We then upgraded the system to Win 95, and the resource } problem with the ISIS appears cured, but we have uncorked a beast in } the LEO. The LEO is being upgraded to a full win95 system soon, but I } am seeing more and more bugs with the ISIS. } } Am I alone in believing the ISIS to be a poorly conceived and executed } system? We have had endless conflicts with the operating system, to } the point of having to reinstall win95 several times after apparent } simple crashes, which seem to render several ISIS modules unavailable } after that. The user interface appears haphazard from module to } module, with no user definable defaults to customize operation to our } preferences. The Labbook system forces users to collect data according } to the x-ray system parameters. We like to structure our data by } customer and job parameters, and keep all x-ray and microscope data } under one directory. This is impossible under the ISIS Labbook system. } } I am greatly disappointed with the integration of these two products. } The LEO has been a fine system, with the operating environment keyed } to usability, and endlessly customizable by the user. The ISIS is user } unfriendly, and rigid in structure. Unless the user follows the } Labbook protocols exactly, the system is unavailable to them. } } Has anyone heard of a replacement operating environment for the Oxford } system? Our complaints to Oxford have been dismissed, and LEO is } unable to effect changes to the abysmal Oxford operating system. I } wonder if I am alone in my frustration with this unfriendly and } quixotic x-ray system. } } Mike Warfield } Hughes Space & Communications } M&P Laboratory } }
I also have a Durst 138 S enlarger. I use both the Pullam light source (point)or 110V-200 Watt opale lamp (basically a giant light bulb). Durst is still in business. I called the supplier I use and they can order it from Durst for you. They are: Espinoza Photo Systems, 5213 A Clayton Road, Suisun, CA 94585 Phone (707) 428-0912. Ask for Patty. JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94022
At 05:08 PM 3/31/98 -0500, Heeschen, Bill (WA) wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Call the folks at Zygo. I am not sure if their complete software is available, but they may have a fragment available which would solve your problem nicely. Try either Bob Smythe at (860)704-5101 or Les Dock (860)347-8506 (general number).
Wyko Corp. also has extensive experience in this area. They were recently acquired by Veeco. The last number I have for them is (602)741-1297.
Best of luck.
Barbara Foster Consortium President Microscopy/Microscopy Education 125 Paridon Street - Suite 102 Springfield, MA 01118 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions in all areas of microscopy, sample preparation, and image analysis. Our goal is to help you optimize your microscopy.
Hi Simon, We've been buying ITO glass from Donnelly Optics Corporation of Tucson, Arizona at 1-888-250-4455 (or 520-806-3800, Sales dept.). Although our application is different we have been using 2 inch squares, the ITO having a conductivity/resistivity of about 100 ohms per inch. If you need only a few pieces of this glass I could arrange to get some to you, otherwise try contacting Donnelly.
Good Luck, Paul Gerroir ----------
Folks, do any of you know of a supplier for ITO glass for making heated culture chambers Thanks Simon
----------------------------------------------------------------------- Simon C. Watkins Ph.D. Associate Professor Director, Center for Biologic Imaging University of Pittsburgh Pittburgh PA 15261 tel:412-648-3051 fax:412-648-2004 URL: http://sbic6.sbic.pitt.edu
and add a few grains of Thymol to preserve it. LET IT AGE 2 WEEKS BEFORE USING!
} What is Paragon stain, and where can I get it? It sounds like it's exactly } what } I need, and I'm sure it would be usefulto the rest of the list if it's as } wonderful as it sounds. TIA. } Lesley Weston. } } } } On Thu, 2 Apr 1998, Chism, Sharron wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Garry, } } I, too, use the Paragon stain, but I do get a polychrome effect. } } I was wondering if you use a saturated Sodium Borate solution to enhance } } your staining. After drying the section on the hotplate, I put a 3 to 4 } } drops of Paragon stain and one drop of sodium borate on the slide (still } } on the hotplate). The combination of heat and sodium borate enhance the } } Paragon stain quite nicely. You might want to try this before adding H } } & E to your "to do" list. } } } } Sharron G. Chism HT (ASCP) } } Electron Micropscopy Lab } } Harris Methodist Fort Worth } } Fort Worth, Texas } } ---------- } } From: Garry Burgess } } To: 'Microscopy' } } Subject: H&E Like Stain for Epon/Araldite } } Date: Wednesday, April 01, 1998 2:25PM } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } In our diagnostic EM lab we currently use Paragon stain to stain our } } epoxy semi-thin sections, but it would be very nice if we had some kind } } of rapid H&E like stain that could give that kind of polychrome } } differentiation between the nucleus and the cytoplasm. } } } } The Paragon stain is supposed to be a polychrome stain, but what we in } } effect see is more of a monochrome direct stain. } } } } Does anyone know of a quick yet superior stain that might give us better } } results here? } } } } } Garry } } } }
I am faced with a problem of selectively staining teflon particles impregnated in nickel matrix. I would really appreciate any suggestions regarding the chemicals and/or the procedure to stain teflon particles.
We have made samples of sapphire using the wedge technique and it works = very well. Sapphire is a hard material, therefore, it is easy to control = how much material you are removing even though it takes longer to thin = it down. Since sapphire is transparent to light we glue a piece of = silicon to the sample and use the change of color of the Si piece as a = reference on how thin the sapphire sample is. =20
Microscopists: There is an actual H&E like stain. Use Celestin Blue B and Eosin after removing the plastic. A stain like this is almost indistinguishable from H&E and in fact microtomists may all be using Celestin blue B as the supplies of tropical logwood (source of dye for hematoxylin) dry up.
There is a lot that can be said about additives to incubation buffers, the composition of blocking media and washing procedures (and a lot more to come probably!). I don't want to clog everyone's mailbox: we offer two Newsletters (free) with in-depth information on the topic of specificity and non-specific results in immuno. Those who are interested are welcome to request Newsletters through our website http://www.aurion.nl. There is a number of FAQ's re. immunolabeling taken up as well.
Just in short: A distinction should be made between a blocking buffer and an incubation buffer. They serve different purposes: the blocking buffer serves to bind protein (BSA, Gelatine, Ovalbumine, Casein etc.) to sticky (hydrophobic) specimen areas. The incubation buffer and wash buffer should prevent background based on hydrophilic interactions (charges) by competition. Different additives may be in order. Many additives may work, although empirical finding sooner than logics seems to be at the basis of their application. An additive that seems to work fine with one set of primary antibody and secondary reagent seems to be less suited with different reagents. With this in mind we have developed Aurion BSA-c, which proves to be a great help with all kinds of antibodies and immunoreagents. } From a cost point of view it may be tempting to leave out additives during the washing steps, although during washing unreacted antibodies or labels may stick to the specimen when they're no longer prevented to.
We use poly-l-lysine coated slides for all our in situs. During the course of a month we will go through several dozen slides. Consequently we have to coat slides quite often. All attempts with commercially available slides have met with unsatisfactory results. We prefer the quality of slides we generate "in house". What I would like is some advice on storage of a large vol. of poly-l-lysine solution that can be accessed and used on a regular basis. We make up 4 L of .05 mg/ml poly-l-lysine solution in 10 mM Tris buffer in a large tray, store this is the freezer and thaw when need. This system allows us to coat several dozen slide simultaneously (we soak geletin coated slides in the PLL soln. for 10-30 min). The problem is that the solution appears to generate a ppt. during storage, a white "fluff".
Looking for all kinds of ideas
Thanks, Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
A friend gave my your e-mail. This is probably old news, however on the off chance this will help, here is an address. CH-Imports Ltd 3410 Deep Green Drive Greensboro NC 27401 Phone 910-282-9734 Fax 910- 288-3375 Their Catalog shows 3 items: Cedarwood Atlas Mardoc OG 71.50/KG Cedarwood Crude Kenya OG 41 /kg Cedarwood Forte USA OG 64.40 /kg Cedawood Virginia OG 33.50 /kg OG stands for Organically grown.? Larry Albright
Heaven is where: the police are British, the cooks are French, the mechanics are German, the lovers are Italian, and all is organized by the Swiss.
Hell is where the police are German, the cooks are British, the mechanics are French, the lovers are Swiss, and all is organized by the Italians. albrite-at-Plasma-Art.com 419 Sunset Avenue Venice CA 90291 310-399-0865 310-392-9222 FAX
"Fundamentals and Applications of Light Microscopy"
July 27-31, 1998 Waltham Massachusetts
A 5-day practical course on how to get maximum information from the light microscope, taught by experienced microscopy problem-solvers.
Hands-on experience, a firm foundation in the principles of LM, and knowledge of contrast enhancement techniques will be emphasized. A full range of reflected and transmitted light microscope and contrast equipment will be available for use by the students.
Although ideal for beginners, the course will also benefit experienced workers. Students are encouraged to bring their own samples.
For Further information contac: Mary McCann McCann Imaging 161 Claflin Street Belmont MA, 02178 Telephone:617-484-7865 Fax: 617-484-2490 E-mail address: mccanns-at-tiac.net URL: www.microscopyed.com
On Fri, 3 Apr 1998 scm5-at-ix.netcom.com (Sanjay Mehta) wrote:
} I am faced with a problem of selectively staining teflon particles } impregnated in nickel matrix. I would really appreciate any suggestions } regarding the chemicals and/or the procedure to stain teflon particles.
We have sometimes looked at PTFE under the electron microscope, with some of our preparations involving "Tetra-Etch", which is, I think, a solution of sodoum napthalene compound in an organic solvent (perhaps tetralin OR tetrahydrofuran, hence the name.) This turns the surface of the PTFE into a sort of carbon, which might take heavy metal stain (but we were using replication, so we did not stain).
If I knew what sort of method was required (SEM, TEM, etc.) I might be able to be more specific.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Dear Neil, would you, please, kindly provide us with the information, what sections = should be stained (paraffin, celloidin, if resin(s) which kind of??). = Either, I think it would be quite difficult to stain only ECM which = normally would be divided into pericellular matrix, territorial matrix = and interterritorial matrix, respectively ("chondrones"). If you're looking for "cartilaginous lacunae" in order to measure such = structures: believe me: they are artifacts of specimen preparation due = to wrong fixation/dehydration schedules......but, perhaps, your = colleague would be doing those measurements for a scientific evaluation = of artifact-induction by poor fix, dehydration, embedding criterions. If you like we could discuss that matter off the list.....
best wishes and good luck Wolfgang
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: Neil Hand[SMTP:mpznhand-at-unix.ccc.nottingham.ac.uk] Gesendet: Montag, 06. April 1998 06:49 An: HistoNet-at-Pathology.swmed.edu Betreff: Q: Cartilage staining, specific
This is a inquiry on behalf of a colleague (not working in Nottingham) I recently received.
"I am currently looking for an optimal staining or sectioning procedure = for samples of ear and articular cartilage, whereby the stain is confined to the extracellular matrix without staining (or very light staining) the cells and nuclei within the cartilaginous lacunae. The reason for this = is to be able to accurately assess the ratio of extracellular matrix to lacunar space by computerized morphometric analysis. Currently, unavoidable staining of cells is obscuring the measurements made by the = morphometric program, which uses gray scales and contrast to arrive at = values".
Further details I do not have. Any suggestions you may have on this = matter would be appreciated.
Neil Hand, Nottingham, England UK.
___________________________________________________________________ Neil Hand Department Histopathology, University Hospital, Nottingham NG7 2UH. work : Tel: (0115) 924 9924 extension 43725 ___________________________________________________________________
For everyone interested, here is the procedure we use to stain semi-thin sections with Paragon stain. Paragon Stain: Ethyl Alcohol ................ 1,028 mls Deionized water .......... 2,397 mls These combine to make up a 30% ETOH solution. (Set this aside for the moment.)
Toluidine Blue ............... 25 grams Basic Fuchsin ................ 9.25 grams Combine these in a 1 gallon (or 4 liter) container.
Add the 30% ETOH and mix well.
Allow the stain to stand for several days, then filter into an amber gallon (4 liter) bottle to protect the stain from light. (It will last a LONG time!)
Filter the stain again before use.
Staining Procedure: Transfer cut section onto a drop of deionized water previously place on a clean microscope slide.
Place the slide holding the section on a hot plate set at 200 degrees C. Allow the water to evaporate and the section to dry.
Drop 3 to 4 drops of filtered Paragon stain on the dry section. Add a drop of saturated solution of Sodium Borate. Continue to heat for 10 - 15 seconds or until the stain begins to show a metallic sheen.
Using a wash bottle of 50% ETOH, gently wash off all the excess stain.
Dry the slide with gentle heat using a blow dryer ... DO NOT DRY ON HOT PLATE!
At this point, maybe I should point out that I use Spurr's resin. I also keep a little Spurr's in the freezer to use as a coverslipping medium for my semi-thin sections. I hope this is helpful!
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Fort Worth Fort Worth, Texas
In a message dated 98-04-02 17:21:53 EST, CANTINO-at-ORACLE.PNB.UCONN.EDU writes:
{ { I have noticed that in EM immunogold-labelling protocols, BSA is often used in buffer as an initial blocking step. In some protocols a BSA buffer rinse is also used before the secondary antibody, and/or is added to the primary and secondary antibody solutions. In other cases it is omitted. My question, to immuno experts on the listserver, is whether there is a DISadvantage to adding the BSA after the initial blocking step. As I understand it BSA reduces non-specific binding, so it would seem prudent to add it whenever possible.
Marie } } Marie,
I agree with you, and this is the way I've always used BSA. Some Abs seemed to give more problems than others, or perhaps it was the phase of the moon, I was never sure.
I would put it in all solutions (including the buffers and the diluents for the Ab and the gold), only removing it from the final buffer and water washes at the very end after the immunolabeling has been completed. This seemed to work for me, and my reasoning was always, "if it ain't broke...."
Best regards, Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Cryostats / Microtomes / Tissue Processors / Embedding Centers / Slide Stainers / Glass Coverslippers / Microscopes (Representing Leica since 1967) / Fiber Optic Systems / Linear Measuring / Micromanipulation (Linear Encoded, Video) / Image Analysis, Archiving, Capture / Video / Video Printers (Cooled CCD, Digital, RGB, Super VHS, 3-chip) / Vibration Isolation Systems / Programmable Stages / Heating & Cooling Stages / Motion Control, Positioning and Measurement Systems
Dear Linda, } } What I would like is some } advice on storage of a large vol. of poly-l-lysine solution that can be } accessed and used on a regular basis. We make up 4 L of .05 mg/ml } poly-l-lysine solution in 10 mM Tris buffer in a large tray, store this is } the freezer and thaw when need. } The problem is that the solution appears to generate a } ppt. during storage, a white "fluff". } Have you looked at the "fluff" under the LM or EM? If it is poly- l-lysine, you may have to divide the PLL into aliquots before freezing & redissolve each aliquot before use. If it is something else--I can't im- aging from your description that it could be bacterial growth, but a brief examination would tell--you would have to identify it before deciding what to do. Good luck. Yours, Bill Tivol
} Is it possible for one week old paraformaldehyde (originally } in argon sealed ampules)-EM grade- to actually permeabilize membranes? } (by some glycol formation). All comments are welcome. } Are these membranes--such as plasma membranes, mitochondrial inner or outer, etc.--with which other microscopists have had preparative experience, or are they unusual in some respect? Since I might expect paraformaldehyde and/or some of its polymers to react with some membrane components, it is possible that the membranes could become permeable to some substances. What substance did they become permeable to? What was its molecular mass? Yours, Bill Tivol
Does anyone have a dysfunctional HummerVI sputter coater, which could be cannibalized or purchased as a surplus item. I need vacuum pump (built in, type G-50D made by ULVAC-Sinku Kiko or equivalent) for this coater. Thank you in advance for your reply.
Chris Terlecki
Applied Analytical Sciences 3303 Harbor Blvd. Ste. H-4 Costa Mesa, CA 92626
What are recommended conditions for staining Epon generic or Spurr resin sections with either the Celestin Blue/Eosin or Paragon stain? I would like to know desired concentrations, temperature, time, etc as starting points and any handy hints to improve success on a variety of tissues.
I have someone wanting to use a stain like this on rat liver, spleen and lung. He has looked at the material in parafin with H & E but wants to get better quality thick sections and then thin sections for TEM of selected areas.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University
--------------------------------------
Microscopists: There is an actual H&E like stain. Use Celestin Blue B and Eosin after removing the plastic. A stain like this is almost indistinguishable from H&E and in fact microtomists may all be using Celestin blue B as the supplies of tropical logwood (source of dye for hematoxylin) dry up.
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Please remove us from the list amrayinc-at-aol.com
I measure the thickness of my samples that I have mechanically thinned two ways. 1) a dial indicator mounted on a granite check block. Mine is digital inches/mm with mm to the third decimal. Look in a tool catalog. Zero on a mounting stub and slide the sample underneath. 2) a dimpler. Zero the dimpler on a mounting stub and then place the sample without wax on a measure the thickness. ----------
I am preparing cross-sectional TEM samples of sapphire. I am trying to find a good way to measure the samples' thickness when I am under about 70 micron. Any ideas?
If the HummerIV is anything like the III I once owned, a direct replacement is not required. I managed to "shoehorn in" a Welch pump. If I had it to do over again, I wouln't even put the pump in the box, but behind it. ...Still a fairly short path for a rough pump and the servicability would be decades better.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all,
Does anyone have a dysfunctional HummerVI sputter coater, which could be cannibalized or purchased as a surplus item. I need vacuum pump (built in, type G-50D made by ULVAC-Sinku Kiko or equivalent) for this coater. Thank you in advance for your reply.
Chris Terlecki
Applied Analytical Sciences 3303 Harbor Blvd. Ste. H-4 Costa Mesa, CA 92626
id sma016866; Mon Apr 6 17:04:15 1998 Received: from elba.wadsworth.org (elba [172.16.1.100]) by newton.wadsworth.org (8.8.6.Beta3/8.7.1) with ESMTP id QAA11665; Mon, 6 Apr 1998 16:58:17 -0400 (EDT)
Dear Isabel, } } We're trying to make a deposition of 2 micron gold particles on a silica } substrate. } We cannot use an evaporator because we don't want a thin film: we want to } deposit the particles directly on the substrate, keeping them isolated and } without forming aggregates. } At first we dispersed the powder directly on the silica, but SEM analysis } showed they formed aggregates. } Then we've tried making a suspension on etanol and then putting a droplet } of the solution directly on the substrate. But still it formed aggregates. } } Does anyone know of a simple method to make such a deposit (without forming } aggregates) ? } If the particles were all of like charge, they might not aggregate. I suggest first making the substrate hydrophobic, if possible, then use an atomizer to spray a suspension of the particles in water onto the substrate. You will want to charge the droplets as they are leaving the atomizer (un- less simply using the atomizer works). I'm sure there's a way to charge the droplets--that's what Milliken (sp?) did with oil droplets to measure the electron's charge. Another possibility would be to deposit a thin film of wax on the substrate (with a conducting material also mixed in the wax) and propel the particles so that they stick to the wax. Good luck. Yours, Bill Tivol
An important first question is: What sample preparation method are you using to prepare your sapphire samples? To measure the thickness of the sapphire it is often useful to mount sacrificial silicon pieces around yo= ur otherwise transparent sample. Transmitted light is then used to monitor the color of the silicon which can be correlated to a thickness value. Th= is is done quite frequently when tripod polishing samples and is a very effective means of measuring the thickness. Of course, you will not be able to measure the thickness until you reach about 10 microns where the silicon starts to become translucent. This technique can also be used to=
ensure that you are polishing evenly across your sample.
We do have a mouse pad that I could send to you that provides a thickness=
scale for silicon as seen through both daylight and a tungsten filament i= f that would be helpful. =
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Brad Tinkham } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
I am preparing cross-sectional TEM samples of sapphire. I am trying to find a good way to measure the samples' thickness when I am under about 7= 0 micron. Any ideas?
Hummer VI coaters were orignally designed to accept several brands of vacuum pumps including the smallest Leybold, Welch and the Varian pumps. They can be bought easily on the used equipment market or from pump reconditioning companies. If you can't find one to fit internally, remote mount the pump and use a plastic vacuum hose to connect it to the baseplate. It doesn't hurt the system to have the pump remote mounted and you may find a larger pump on surplus easier than small one.
Good luck, Steve Collins South Bay Technology East
What is the purpose of your poly-L-lysine??? If it is simply to get sections to stick, then you would do well to switch to Fisherbrand Superfrost/Plus Microscopy Slides, catalog #12-550-15, FisherScientific, Pittsburgh, PA 15219. They are specially made for this purpose.
We use them exclusively and never lose sections, frozen or epoxy. They are more expensive than plain slides, but considering your time and your storage problems, you might like the convenience.
I have no commercial interest in this product--just a satisfied customer.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Orcein stains cartilage on paraffin sections a pretty distinct deep purple. You might check if it's neccesary to retain the cells, as they might be removed with alkali treatment (KOH?). I don't know what alkali does to orcein staining, but it would be easy to check. Best of Luck, Charlie Ginsburg NCC
} "I am currently looking for an optimal staining or sectioning procedure for } samples of ear and articular cartilage, whereby the stain is confined to } the extracellular matrix without staining (or very light staining) the } cells and nuclei within the cartilaginous lacunae. The reason for this is } to be able to accurately assess the ratio of extracellular matrix to } lacunar space by computerized morphometric analysis. Currently, } unavoidable staining of cells is obscuring the measurements made by the morphometric program, which uses gray scales and contrast to arrive at values". } } Further details I do not have. Any suggestions you may have on this matter } would be appreciated. } } } Neil Hand, } Nottingham, } England UK. } } ___________________________________________________________________ } Neil Hand } Department Histopathology, University Hospital, Nottingham NG7 2UH. } work : Tel: (0115) 924 9924 extension 43725 } _________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
A postdoctoral position is open at Northwestern University to work on In-situ deposition of Cubic Boron Nitride films, combining Ultra-High Vacuum Electron Microscopy with in-situ growth. Some details about the available hardware can be found at: http://www.numis.nwu.edu http://www.numis.nwu.edu/SINBAD and see also Collazo-Davila et al, Appl Phys Letts 72, 314-316 (1998) A strong background in thin film growth is required, and additional background in TEM and UHV will be highly relevant. Interested applicants should send a short CV via email to ldm3-at-apollo.numis.nwu.edu including email addresses for references.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 email: ldm3-at-apollo.numis.nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Paula Sicurello wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Listers, } } I am in need of a type 545 Polaroid camera back that has a frosted } glass where the film holder usually is. My SEM pix look OK on the screen } but if I make a print from the neg. it looks slightly out of focus. } I've been told that I need to focus the camera to match the screen } & this fancy camera back with the glass helps you do that. I know they } exist because I saw one on an ESEM. Does anyone know where I can buy one? } I called Polaroid and they were confused. } Can anyone help me with this problem? } } My future looks fuzzy & so do my prints, } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu Paula,
You didn't say what kind of SEM. The Japanese ones all use Mimya cameras. Others may be Graphlex compatible. You might try Calumet Camera in the Chicago area. I don't have the address, but they specialize in 4x5 and 8x10 cameras and have the frosted glass/freznel lens combination and hoods, or shades to make viewing easier.
You want to be sure of the compatability because the frosted glass MUST sit at the plane of the film.
Ken Converse Quality Images 16 Creek Rd. Delta, PA 17314 third party SEM service
Has anyone out there done any xNGF staining en grid, and could you possibly give me some guidelines for fixation and embedding, i.e., can you osmicate and what resins might be advisable. Many thanks Grace
I am the senior anatomopathologist of Paediatric Institute of Research of the University of Trieste(Italy). Due to two international collaborative research projects we ought to send to UK and USA(Baltimora) our frozen section of intestinal biopsies. These sections will be analyzed for EmA(antibidies antiendomysium) in Celiac Disease with Immunofluorescenze indirect and for cytokines TNF-alfa,INF-gamma in IBD with ISH. I fix the sections with acetone-cholrophormio(1:1) and store at -20=B0C (they last 2-3 months). I would like to know if there i= s any other technique to fix sections in a way I could store them at 2-6=B0= C and preserve them before searching for antigen for a longer period(such as 5-6 months).Yours sincerly Kind regards dr. Angelo Citt=E0 Dept.Anatomia Patologica Istituto per l'Infanzia 34100 TRIESTE e mail: acitt-at-tin.it
SUMMER I 1998 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-Section B)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I 1998 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered four days per week (Monday through Thursday) between the hours of 8:00 am and NOON. Classes will begin on May 26 and end on June 25, 1998.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 per credit (for Nassau County residents or New York State residents with a certificate of residency).
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students.
If you have further questions, you should e-mail me directly at the address below.
For information about mail or telephone registration (Dial-a-Course) point your browser to http://www.sunynassau.edu/courses/sum98/sum98.htm. Telephone registration is only available until April 30, 1998.
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Like Woody White, I replaced the original pump but with a larger model and placed this under the bench; much easier to service and less backstreaming of vapour. The first Hummer with a triode sputter head suffered from sooting-up of the sputter head. At the time I was able to demonstrate to the manufacturer that the sooting-up problem was caused by the mineral oil and could be avoided by using Fomblin rotation pump fluid. It's ancient history now but the problem seemed severe at the time and when I was replacing the pump for the third time I was consoled that "we have to replace over a hundred pumps." The original correspondent needs to replace the original pump, and well he might. However, some people may only need to have the right pump fluid and a decent size pump. The original pump's capacity was too low. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
_____________________________________________________________} } Does anyone have a dysfunctional HummerVI sputter coater, which could be } cannibalized or purchased as a surplus item. I need vacuum pump (built } in, type G-50D made by ULVAC-Sinku Kiko or equivalent) for this coater. } Thank you in advance for your reply. } } Chris Terlecki } } Applied Analytical Sciences } 3303 Harbor Blvd. Ste. H-4 } Costa Mesa, CA 92626 } } ph: 714-434-6894 } fax: 714-434-0294 } email: aas-at-pacbell.net }
Re: sputter coater pump _____________________________________________________________} } Does anyone have a dysfunctional HummerVI sputter coater, which could be } cannibalized or purchased as a surplus item. I need vacuum pump (built } in, type G-50D made by ULVAC-Sinku Kiko or equivalent) for this coater. } Thank you in advance for your reply. } } Chris Terlecki } } Excuse my ignorance re: details of pumping system for sputter coaters but some respondent have commented on backstreaming of pump oil. We use mechanical pumps on visible light microscope cathodoluminescence attachments using a cold cathode discharge and always use a simple foreline trap (copper gauze type - ca$300.00) to reduce the backstreaming. Yearly maintenance for trap is less than $50.00. Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1922, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
A customer in the Chicago, IL area has asked for help locating service for their HNU EDS system. They are having trouble getting service from the parent company, and would like to locate a third party service organization. Any information would be greatly appreciated.
Regards, Chris Wadelton Amray, Inc. (800) 591-8791
Advanced Surface Microscopy would like to buy used Digital Instruments NanoScope equipment of the following types: NanoScope II with AFM (contact mode) NanoScope III with contact and/or Tapping Mode AFM and other related equipment.
Please contact me offline, since I am not a regular subscriber to this listserver.
Thanks very much. Don Chernoff
Advanced Surface Microscopy, Inc. E-Mail: asm-at-indy.net 6009 KNYGHTON RD. Voice: 317-251-1364 INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA) web: http://www.a1.com/asm Fax: 317-254-8690 (note: "a1"= letter "a", numeral "1")
The AMRAYINC-at-aol.com address was removed from the listserver this morning because Amray is installing a new E-mail exchange system. The addresses will be changing, in the interum Amray is still available via E-mail at the above address as well as AMRAYSERV-at-aol.com (service) and AMRAYSALES-at-aol.com (sales).
Dear list, Similar question was discussed not so far, but I saw no reliable solution. I'm sorry if I simply missed it. The problem is to know diffraction orientation relative to the object feature. I can defocus diffraction spots and see images of the feature (edge for example) in each spot but I can defocus two sides and get opposite directions. Which one is true? Or there is another method? I do not need to calibrate rotation angle as it's always 0 (or 180deg?) at JEM2010. I want to know definitely if it 0 or 180. We need this to determine SiC and AlN polarity.
Hello to all, We have been using another department's double glass distilled water, and their still is down. Our building has a house "distilled" water tap. I called to find out the method by which this water is purified, and was told that the water is softened, passed through reverse osmosis, deionized, filtered, UV sterilized, and delivered to the building at about 15 megaohms of resistivity. This leads me to my question, what are the standards by which other labs (biological EM) gauge their water? Distilled vs deionized? Your opinions or a reference citing would be greatly appreciated.
-- Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
In the recent emails regarding frosted glass, noone has mentioned why these work well. I have been unable to find out why from several reference works, so does anyone know? Is it that the uneven surface exaggerates the out-of-focus appearance since there is then no even plane of focus on the glass?
Doug Darnowski
****************************************************************************** Douglas Darnowski Department of Crop Sciences 384 ERML 1201 West Gregory Drive University of Illinois Urbana IL 61801 work ph: (217) 244-6150 fx: (217) 333-4777 home ph: (217) 356-6606 fx: (217) 356-4454 email: darnowsk-at-staff.uiuc.edu
I think that you can use the shadow image in a convergent beam pattern. Underfocus the crossover and your shadow image in the BF disk lines up with the diffraction pattern.
You can check this with GaAs from a known wafer because the orientations are determined by etching the sample. The data sheet that comes with the wafer should let you know what the absolute directions are using the major and minor flats. You can prepare your sample very easily and maintain the crystallographic orientation of the sample with the small angle cleavage technique. John McCaffrey and I talk a little bit about this in our paper on SACT in the MRS TEM Sample Prep IV (Vol 480).
I may be mistaken, but I also thought that the 180 degree ambiguity was taken care of in the 2010.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Dear list, Similar question was discussed not so far, but I saw no reliable solution. I'm sorry if I simply missed it. The problem is to know diffraction orientation relative to the object feature. I can defocus diffraction spots and see images of the feature (edge for example) in each spot but I can defocus two sides and get opposite directions. Which one is true? Or there is another method? I do not need to calibrate rotation angle as it's always 0 (or 180deg?) at JEM2010. I want to know definitely if it 0 or 180. We need this to determine SiC and AlN polarity.
The frosting merely serves to scatter light from the film plane rather so that you can see the image. If it were not frosted, the light would pass thru - the glass would be invisible, and you would see no image.
If you could look at the image from the lens side you could focus the camera using an opaque material in place of the film. But since cameras are sealed up, they use the frosted glass so you can see an image from the back side.
At 11:30 AM 4/6/98 -0600, you wrote: } } In the recent emails regarding frosted glass, noone has mentioned why these } work well. I have been unable to find out why from several reference works, } so does anyone know? Is it that the uneven surface exaggerates the } out-of-focus appearance since there is then no even plane of focus on the } glass? } } Doug Darnowski
A student in my laboratory has a number of digital images of gels and transmission electron micrographs which she would like to include in her thesis. I know that prints from our inkjet printer have poor archival properties (they turn brown over a period of a year or so, depending on light, air exposure), but we do have access to a dye sublimation printers elsewhere. How long can we expect black and white dye sublimation prints to last in a thesis without discoloring or fading? Given that these printers haven't been around very long, does anyone actually know?
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
At 03:16 PM 4/7/98 -0400, MARIE CANTINO wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I posed this question to a Kodak rep at the last MSA in Cleveland. He stated the prints were archival; that is, they should last at least as long as properly fixed photographic material, at least 50 years.
Rick A. Harris, Director Microscopy and Image Analysis Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 752 3085 fax raharris-at-ucdavis.edu
} -----Original Message----- } From: MARIE CANTINO [mailto:CANTINO-at-ORACLE.PNB.UCONN.EDU] } Sent: Tuesday, April 07, 1998 12:17 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Archival properties of dye sublimation prints } } } -------------------------------------------------------------- } .... } } A student in my laboratory has a number of digital images of gels and } transmission electron micrographs which she would like to include in her } thesis. ... we do have access to a dye sublimation printers } elsewhere. How long can we expect black and white dye } sublimation prints } to last in a thesis without discoloring or fading? ...
My experience with dye-subs is that grayscale prints are usually a result of CMY printing, although two possibilities are (1) the CMY ribbon also had a "K" of black component, or (2) a monochrome "K" ribbon was used. If the prints were a result of "K" printing then they are not likely to change color, but they may fade while the paper stock remains white. If they are a result of CMY printing, they are liable to acquire a pink or green tint (... not bad though ...).
Lastly, and at least for Kodak dye-sub printer engines, there exists a ribbon/paper combination which lamenates and protects the image from UV ... or you can try to find UV protection, usually as a form of a spray, to apply to print.
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
It probably depends on what dye sub printer you have as to how well a print will stand up to the passage of time.
A Kodak printer we had put a plastic coat on top of the dye as the last step in the printing process.
An old Sony dye sub printer I had did not do this. The dyes would smear if any moisture at all got on them. Also after several months in the hallway under fluorescent light, the dyes would fade. If they were not exposed to light, they kept their color 4-5 years and I suspect they would keep much longer.
I do not know whether the plastic coating prevents the fading of the dye when exposed to light a number of months because I haven't tested any of these prints.
That is the extent of my knowledge on the subject. Hope it helps.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 73-384 CHS 951761 Los Angeles, CA 90095-1761
} ---------- } From: } CANTINO-at-ORACLE.PNB.UCONN.EDU[SMTP:CANTINO-at-ORACLE.PNB.UCONN.EDU] } Sent: Tuesday, April 07, 1998 12:16 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Archival properties of dye sublimation prints } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } A student in my laboratory has a number of digital images of gels and } transmission electron micrographs which she would like to include in } her } thesis. I know that prints from our inkjet printer have poor archival } properties (they turn brown over a period of a year or so, depending } on } light, air exposure), but we do have access to a dye sublimation } printers } elsewhere. How long can we expect black and white dye sublimation } prints } to last in a thesis without discoloring or fading? Given that these } printers haven't been around very long, does anyone actually know? } } Marie } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology, U-131 } University of Connecticut } Storrs, CT 06269 } Ph: 860-486-3588 } Fax: 860-486-1936 } } }
I would be happy to help. I was approached by HNU several years ago to sell their instruments, and after visiting their manufacturing facilities and meeting with their top brass, I decided that they were lacking in many respects and I didn't choose to go with them. I would be happy to help anyone, as best I can, who chose this manufacturer as their intrusion into the market was ill-planned.
They assumed that the US manufactured EDS detector combined with the German produced PC-based software would give them some market advantage, and they were right for a very short time. But the manufacturers who were more heavily invested in the art of EDS soon translated their DEC LSI-11 based software into PC based applications, and HNU was left out to dry as those manufacturers produced PC based applications that worked well and offered the market appeal of known market brands in the industry.
} Dear All, } } A customer in the Chicago, IL area has asked for help locating } service for their HNU EDS system. They are having trouble getting } service from the parent company, and would like to locate a third } party service organization. Any information would be greatly } appreciated. } } Regards, } Chris Wadelton } Amray, Inc. } (800) 591-8791 } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1992, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1992, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
Marie, Archival properties depend on your particular printer and the printing ribbon used.
We are on our 2nd generation dye-sublimation printer. Our first was purchased ~ 6 years ago when the technology was very new. Greyscale prints from this printer were made from an overlay of RGB color and tended to have a purplish cast to them. The colors did fade when exposed to UV light but were fine if kept in the dark the majority of the time. At that time there was no product with the transparent UV-protectant overlay available.
We recently upgraded with a Codonics 1660 dye-sub printer. Two of the main features I looked for before purchase was that there was a black and white ribbon available and a color ribbon that incorporated the UV protection overlay. This printer has both. The use of the black and white ribbon eliminates the chance of any misalignment of the color sheets to get true greyscale without hints of another color. Also available are RGB ribbons with (approx. $3/8x10" sheet) or without (approx. $2/sheet) the UV-protection laminate. We often use the less expensive color ribbon when output is not intended for long term use and then switch to the ribbon with overlay for final copies.
We have been assured that prints produced with the black and white or color + overlay ribbon are indeed archival but time will tell!!
Debby Sherman, Manager Microscopy Center in Agriculture Purdue University --------------------------------------
A student in my laboratory has a number of digital images of gels and transmission electron micrographs which she would like to include in her thesis. I know that prints from our inkjet printer have poor archival properties (they turn brown over a period of a year or so, depending on light, air exposure), but we do have access to a dye sublimation printers elsewhere. How long can we expect black and white dye sublimation prints to last in a thesis without discoloring or fading? Given that these printers haven't been around very long, does anyone actually know?
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
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Fellow Scanners,
I have a Sharp 2400 DPI Flatbed which I am thrilled with but Sharp has gotten out of the scanner market. So, whats the deal? Our ceramics department needs to purchase a flatbed for TEM negatives, slides, positives color B&W etc....Which one? The esteemed Dr. Mark Farmer from the University of Georgia has suggested a Umax; I know and respect Dr. Farmer and usually follow his recomendations so I would love to find the North East distributer for Umax or if anyone has another flatbed choice for our ceramics department I would appreciate a reply.
48 degrees, dreary, and it smells funny in several areas... just thought you'd like to know! John Grazul Rutgers University Electron Imaging Facility
High resolution (nanometer) imaging Protein folding force measures Protein binding force measures
This technique is still new enough that it requires a skilled microscopist, but is certainly breaking new ground in microscopy.
At 11:13 AM 3/27/98 -0700, Gillian Bond wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
you mention smudging with ink-jet inks. I have started using an Epson Stylus 600 and I notice that it doesn't smudge if you use the full glossy papers. I don't know about fading, though but it may be useful to use the glossy finish for more permanent results if your printer can use it and the cheaper papers for routine work.
Malcolm Haswell University of Sunderland UK ----------
Marie,
It probably depends on what dye sub printer you have as to how well a print will stand up to the passage of time.
A Kodak printer we had put a plastic coat on top of the dye as the last step in the printing process.
An old Sony dye sub printer I had did not do this. The dyes would smear if any moisture at all got on them. Also after several months in the hallway under fluorescent light, the dyes would fade. If they were not exposed to light, they kept their color 4-5 years and I suspect they would keep much longer.
I do not know whether the plastic coating prevents the fading of the dye when exposed to light a number of months because I haven't tested any of these prints.
That is the extent of my knowledge on the subject. Hope it helps.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 73-384 CHS 951761 Los Angeles, CA 90095-1761
} ---------- } From: } CANTINO-at-ORACLE.PNB.UCONN.EDU[SMTP:CANTINO-at-ORACLE.PNB.UCONN.EDU] } Sent: Tuesday, April 07, 1998 12:16 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Archival properties of dye sublimation prints } } A student in my laboratory has a number of digital images of gels and } transmission electron micrographs which she would like to include inb her } thesis. I know that prints from our inkjet printer have poor archival } properties (they turn brown over a period of a year or so, depending on } light, air exposure), but we do have access to a dye sublimation printers } elsewhere. How long can we expect black and white dye sublimation prints } to last in a thesis without discoloring or fading? Given that these } printers haven't been around very long, does anyone actually know? } } Marie } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology, U-131 } University of Connecticut } Storrs, CT 06269 } Ph: 860-486-3588 } Fax: 860-486-1936
by mail450.icon.co.za (8.8.8/8.8.8) with SMTP id RAA14537; Wed, 8 Apr 1998 17:03:02 +0200 (GMT) Received: by localhost with Microsoft MAPI; Wed, 8 Apr 1998 17:05:26 +0200 Message-ID: {01BD6310.86D0CF60.anaspec-at-icon.co.za} "'MSSA listserver'" {MSSA-at-UCTBC1.UCT.AC.ZA}
Hi all.
We were wondering if anybody out there may have a spare XP3 pulse processor we could by at a good price ? We have a specific need for high count rates off an Oxford detector. The XP3 has a fast process time of 2.5?sec which is ideal.
Thanks.
Luc Harmsen Anaspec, South Africa Technical support for E.M. operators, world wide. anaspec-at-icon.co.za TEL: ++ 27 (0) 11 476 3455 FAX: ++ 27 (0) 11 476 7290
Meeting Announcement San Francisco Microscopical Society
Thursday, April 9, 1998 Rockridge Branch, Oakland Public Library 7:00 P.M.
"Microscopic Pond Life" by Rick Ellis
Rick Ellis is a microscopist and photomicrographer whose talks we have all enjoyed in the past. This promises to be another fascinating evening as Rick takes us on an exploration of the rich and varied world of pond microorganisms. This is the type of microscopy that is readily available to everyone (especially in this year of El Ni=F1o). We will have a Saturday Workshop later this year to collect and examine pond critters, so this will be a good opportunity to see what you can expect to find, and how to prepare the samples for examination. Please join us!
Further Particulars:
http://www.microdataware.com/sfms
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
We recently purchased a Umax Powerlook III (a new model, about $2k U.S.). They call it a "42-bit" scanner through a bit enhancement technology, and it has a real resolution of 1200 x 2400 dpi. At any rate, it seems to work pretty good for both negatives and prints. The software is not overly intuitive and we received no book on how to use it but we have generally worked out the kinks and are fairly happy. It won't capture everything on your negatives since their grain size is smaller than the available resolution and their dynamic range is higher than what we can get (3.4, I think), but it works well for those times when you don't want to print or need to bring out a feature on the negative.
Their web address is : http://www.umax.com/ and it contains links to resellers in specific regions of the U.S. (and perhaps internationally; I didn't look).
Hope this helps.
Cheers,
John Vetrano
----------
Fellow Scanners,
I have a Sharp 2400 DPI Flatbed which I am thrilled with but Sharp has gotten out of the scanner market. So, whats the deal? Our ceramics department needs to purchase a flatbed for TEM negatives, slides, positives color B&W etc....Which one? The esteemed Dr. Mark Farmer from the University of Georgia has suggested a Umax; I know and respect Dr. Farmer and usually follow his recomendations so I would love to find the North East distributer for Umax or if anyone has another flatbed choice for our ceramics department I would appreciate a reply.
48 degrees, dreary, and it smells funny in several areas... just thought you'd like to know! John Grazul Rutgers University Electron Imaging Facility
I have used both Umax and Microtek brand scanners (SCSI I/O). No problems with either. They can be purchased from any number of computer suppliers like Computability, CDW, Microsystems Warehouse, B&H Camera, etc.
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Fellow Scanners,
I have a Sharp 2400 DPI Flatbed which I am thrilled with but Sharp has gotten out of the scanner market. So, whats the deal? Our ceramics department needs to purchase a flatbed for TEM negatives, slides, positives color B&W etc....Which one? The esteemed Dr. Mark Farmer from the University of Georgia has suggested a Umax; I know and respect Dr. Farmer and usually follow his recomendations so I would love to find the North East distributer for Umax or if anyone has another flatbed choice for our ceramics department I would appreciate a reply.
48 degrees, dreary, and it smells funny in several areas... just thought you'd like to know! John Grazul Rutgers University Electron Imaging Facility
Debby Sherman wrote: ========================================== Unicryl is made by BBInternational and distributed in the USA by Vector Laboratories, Inc. (1-800-227-6666). They are a CA based company and are not open yet today. I did call there yesterday and asked for a method booklet on Unicryl and the individual who I talked to did not indicate that the resin was unavailable. =========================================== The embedding resin, Unicryl(TM), is the product of the following:
British Biocell International Ltd. Golden Gate Ty Glas Avenue Cardiff CF4 5DX UK
SPI Supplies has imported Unicryl resin into the USA almost since its inception as a product and it is now also available from a number of the other suppliers of chemicals to microscopy laboratories, for example, Pella and EMS.
The debate on which embedding resin is or is not "better" will probably still be going on long after we are all gone from this word. At least some of our customers have reported excellent results with Unicryl but the one thing that does seem to stand out is that there is a perception among those who are sensitized (from a dermatitis standpoint) to this class of acrylic resins is that the Unicryl system is "milder" than the others. Now I will be the first to admit that this is not a very scientific sampling but if one should be listening to their customers, this is one thing at least some of them have been telling us.
I for one would welcome any kind of dialogue describing the experiences people have had with these different resin systems. If some are "better" or "worse" from a dermatological standpoint, it would be good to have the benefit of the collective experience of those working with these different media.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Hypothetically, if I had a fully loaded, 1 year old Kevex Sigma 4 system available (no detector), would anyone be interested in purchasing it and what would be a fair asking price. The system was purchased for $75,000 1 year ago.
Wayne Kaboord Materials Research Specialist Eaton Corporation Innovation Center 4201 North 27th Street Milwaukee, WI 53216 414-449-7783 WayneKaboord-at-eaton.com
id xma023997; Thu, 9 Apr 98 08:17:11 -0400 Received: from charlotte.cpt.afip.org (charlotte.cpt.afip.org [10.20.30.46]) by elon.cpt.afip.org (950413.SGI.8.6.12/950213.SGI.AUTOCF) via ESMTP id IAA16246; Thu, 9 Apr 1998 08:22:55 -0400 Received: from localhost (oliver-at-localhost) by charlotte.cpt.afip.org (950413.SGI.8.6.12/950213.SGI.AUTOCF) via SMTP id IAA05283; Thu, 9 Apr 1998 08:22:15 -0400
Please note that this is a personal observation and does not reflect the opinion of my office nor of any group associated with me.
I use a Umax Powerlook II as the workhorse machine in my lab, and have been generally happy. We will likely be moving to one of the newer machines soon. However, you should look at some reviews in the consumer press before making a final decision. Some folk in the lab next to me use an Agfa scanner, and they are happy with it.
The biggest criticism of the Umax scanners that I have seen in the press is that they sometimes do not give as great a depth or discrimination of tone as some competitors. For instance, if you place a sheet with 256 greyscale values in the scanner, it may only give you 200 values. I tried something like that with my scanner after I read the review and noticed the following:
If I took 256 values from black to white I got about 240 values out, which was pretty good. If, however, I took 256 values from dark grey to light grey, it did much more poorly -- about 150 values. In other words, there was no way to calibrate the illumination or sensitivity of the sensor to adjust for a dingy image.
In one sense, the scanner was "correct" in that I put in a dingy image and got dingy results. However, if one wants to take data acquired from a scanner and do image processing, one wants as much data as possible from that scanner. And that means that one would like to fill those 256 levels with whatever is available.
Ideally, one would want to be able to modify the illumination of the scanner to get a full 256 tone values out of an image, regardless of what the brightest and darkest regions are. The "histogram stretching" functions of the TWAIN driver software do *not* accomplish this; instead it simply does a stretch *after* data acquisition (at least as I have observed). Thus, you still get only, 150 grey values instead of 256, but they are stretched between 0 and 255.
This may not be a problem for you, but since I do image processing, I am very interested in getting as many tone levels as I can out of an image. I am *not* sure if competitors are all that much better, but you might want to look at some of the reviews which actually do quantitative measurements before you buy.
billo
On Wed, 8 Apr 1998, John Grazul wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Fellow Scanners, } } I have a Sharp 2400 DPI Flatbed which I am thrilled with but Sharp } has gotten out of the scanner market. So, whats the deal? Our } ceramics department needs to purchase a flatbed for TEM negatives, } slides, positives color B&W etc....Which one? The esteemed Dr. Mark } Farmer from the University of Georgia has suggested a Umax; I know } and respect Dr. Farmer and usually follow his recomendations so I } would love to find the North East distributer for Umax or if anyone } has another flatbed choice for our ceramics department I would } appreciate a reply. } } } 48 degrees, dreary, and it smells funny in several areas... } just thought you'd like to know! } John Grazul } Rutgers University } Electron Imaging Facility }
In response to Bill's comments, I have both an Agfa and two UMax scanners and I personally love the Umax's and would never buy another Agfa. Both give acceptable images and neither is consistently perfect. The difference is in the software. Trying to manipulate gain and black level on the Agfa is complicated and time consuming, whereas the Umax is fast and intuitive. My first UMax is still working 9 years later and still gives fine images. Dave
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Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
I am a faculty member at Brookdale Community College in Lincroft, NJ who operates a Zeiss 9C TEM which was donated to the College. Unfortunately, however, we do not have a knife maker or ultramicrotome for tissue preparation. Does anyone know of a used ultramicrotome and knife maker which can be purchased for a reasonable price? (or perhaps even donated to the College) Thanks in advance for your help.
Mike Palladino
Michael A. Palladino, Ph.D. Instructor of Biology Brookdale Community College Voice: (732) 224-2871 E-mail: mpalladino-at-brookdale.cc.nj.us
Currently I work as a corporate web designer and use Photoshop imaging software daily for the past 4 years. I am concerned as to how to combine my visual imaging skills with my interest in natural and physical science (so-so on medical). At 44, I am intimidated by the notion of schooling for many years. What is entailed and where can I ask about a microscopy program in New York City. I believe the visual aspect of scientific analysis is fascinating.
I am volunteering web pages for the invertebrate scientists at the American Museum of Natural History and have seen their electronic microscope equipment (I was asked to teach Photoshop so that the scientists could color the scans) and other imaging equipment and 3-d software. Thrilling!! I treasure my book on microscopic art (Felice Frankel). Also, I perform nature photography when away from the city.
Thank you in advance for any education or vocation advice you have time to give.
Renee Recker 16 W. 16th St. NYC, NY 10011 212-675-1665
some samples of web pages for Amer. Museum of Nat. History: http://www.renorex.com/invertebrateshome http://research.amnh.org/~mikkel
We have two Leitz metallographs and are having problems with the 450 watt xenon lamphouses on both.
One has operated fine for the last 15 years (same bulb!) Recently, I noticed an odor after about 15 minutes of continuous use, perhaps a burning phenolic. We replaced the transformer in the high voltage unit, but the odor persisted. Predictably, something finally blew and now the bulb will not ignite.
The other metallograph was fine and then one day the xenon bulb would not ignite. We replaced the bulb and it still fails to ignite (though the high voltage unit attempts ignition).
Are there any optical service engineers in the Chicago area knowledgable in xenon units. The bulbs are high pressure and hazardous to the uniformed. I suspect that there are either connection problems or blown components in the high voltage units.
If anyone can help, then please reply off-line or call me directly.
Thank you.
Alan Stone ASTON Metallurgical Services 773/528-9830
We have recently made some changes to our web site. Please visit us at www.rjlg.com. We would greatly appreciate any comments or opinions you may have pertaining to your visit.
Keith Rickabaugh Manager, Materials and Particle Characterization {krickabaugh-at-rjlg.com}
RJ Lee Group, Inc. 350 Hochberg Road Pittsburgh, PA 15146 ph: 724-325-1776 www.rjlg.com
Hey there imaging buffs 1) Is there a way to create the red/green 3D images using stereo pair TEM mages without buying an expensive 3D imaging program? 2) What thickness of sections would be needed for TEM work to get the effect of 3D depth? 3) What is the best angle? I would be using a Philips 410LS w/goniometer tilt stage. I'm going to hit the library but tips from the horses mouth are always much better. Thanks in advance.
Isabel Nogueira wrote: ================================================== We're trying to make a deposition of 2 micron gold particles on a silica substrate. We cannot use an evaporator because we don't want a thin film: we want to deposit the particles directly on the substrate, keeping them isolated and without forming aggregates. At first we dispersed the powder directly on the silica, but SEM analysis showed they formed aggregates. Then we've tried making a suspension on etanol and then putting a droplet of the solution directly on the substrate. But still it formed aggregates.
Does anyone know of a simple method to make such a deposit (without forming aggregates) ? =================================================== There are three ways I could suggest, not based on experience specifically with gold particles, but by extrapolation from the dispersal of other fine particles and powders:
a) Camphor/naphthalene dispersion method (I described this previously and it should be available in Nestor's archives), where by the powder to dispersed is added to a 60% camphor/40% naphthalene (the eutectic composition of the system) which becomes a liquid at a few degrees above room temperature. Putting a drop of the suspension on your substrate will result in its instantly freezing. Put substrate and all into a vacuum over night (mechanical pump is enough) and by morning, the camphor/naphthalene would have sublimed away leaving the gold particles nicely and finely dispersed. You would regulate the degree of coverage by controlling the loading of the eutectic solution and the amount deposited.
b) Dispersion in an organic solvent but add a small amount of Parlodion (R) or perhaps any of the other similar in composition materials (e.g. Collodion (R)). After dispersal on the substrate, you would remove the organics with a barrel-geometry plasma etcher. The gold particles should be left in situ and not agglomerate. The adhesion to the substrate with (a) might be better than with (b).
c) Variation on the above, but using water, which is then applied to the substrate while the substrate is below the water freezing point. The ice is then sublimed away over night in a freeze fracturing device, leaving the gold particles dispersed without agglomeration.
However, some powders just do not want to disperse, perhaps because they are to some degree aggregates and not simply agglomerates. Being able to use a method that can be tightly controlled, such as (a) or (b), is sometimes the only way to discriminate between different samples where their only real differences happen to be their degrees of aggregation (vs. agglomeration). Over the years we have seen on occasion, large (I call 2 um "large") gold particles as in thick film paste systems that have been doublets and triplets, etc. so it is not impossible that at least some of your problem could be that not all of the particles are in fact single individual particles.
Disclaimer: Structure Probe, Inc. performs these kinds of services for clients as a regular part of our business.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Could a local air conditioning/refrigeration/plumbing company help? How about an appliance repair shop? Brazing companies? Heat exchanger builders/rebuilders? Automobile radiator repair shop? Local technical university?
My $0.02
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
Conceptually it should be quite possible and straightforward.
We collect the left and right B/W images using a 4 to 6 degree tilt between the views for our SEM work. Much more than 6 degrees gets unnatural. It represents crossing the eyes too much.
Since I work with the SEM, I don't know what the sample requirements would be for TEM. I would think as long as you have some topographic relief you should be okay. Even layers of atoms might be enough if you are at high enough mag.
Once you have the images, you can easily change the hues for the one from gray to red and the other from gray to green or blue using programs like PhotoShop. Preparing the anaglyph simply requires merging the two colored images into one true-color image. Since one image would supply the red intensity and the other the green intensity, it should be straightforward - I can think of how I would write a program to do it. But maybe the Photoshop gurus can tell what buttons to push to make it happen.
At 06:02 PM 4/9/98 -0500, you wrote: } Hey there imaging buffs } 1) Is there a way to create the red/green 3D images using stereo pair } TEM mages without buying an expensive 3D imaging program? } 2) What thickness of sections would be needed for TEM work to get } the effect of 3D depth? } 3) What is the best angle? I would be using a Philips 410LS } w/goniometer tilt stage. I'm going to hit the library but tips from the horses } mouth are always much better. Thanks in advance. } } Rick Vaughn } RLVAUGHN-at-MAIL.UNMC.EDU ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-4563
I have had more success with SEM stereo pairs! The best results from TEM came from 0.1-0.5 um-thick sections. All tilted about 6 degrees. I have not done a lot of TEM because it was never very effective with normal ultrathin sections. It may be better with freeze fracture replicas and other specimens on formvar/carbon films such as direct prep. cells etc.
Take the stereo pair of images. Make good sized prints. Then get a friendly photgraphic person to photograph one print using a red filter onto colour slide film. Then double expose with the other print - in other words, photograph the second print onto the same film using a green filter. You then have effective stereo which you can show at meetings using red/green glasses and only one projector.
There is a convention which I forget regarding which way around you do this so that you hold the glasses properly. I'm replying to this from home, also my friendly photographer is skiing in Canada at the moment! Write back if you need the details (and if he survives Lake Louise or wherever he's gone this time!).
Keith Ryan Plymouth Marine Lab., UK
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Hey there imaging buffs 1) Is there a way to create the red/green 3D images using stereo pair TEM mages without buying an expensive 3D imaging program? 2) What thickness of sections would be needed for TEM work to get the effect of 3D depth? 3) What is the best angle? I would be using a Philips 410LS w/goniometer tilt stage. I'm going to hit the library but tips from the horses mouth are always much better. Thanks in advance.
I use Photoshop to do this in my bio TEM class, but you should be able to do it in NIH Image (can't beat the price). You could theoretically do on any section thickness, the effect depending on the magnification of the structure viewed. You make each image a different color, adjust the transparency and overlay them. You can view them on screen or output them to various hard copy media.
I usually have the students use their replicas (usually freeze-fractured yeast) since these have a 3-d topography, but I've also done this with chloroplasts in "thick" (about 250nm) section (don't stain them if you've used OsO4); they are just O.K. at 100kV. Depending on what you want to see, you can leach out a lot of background density by using KMnO4 and maybe work with thicker sections. More volts should also help as would an energy filter.
The best angle is dependent on the effect desired. To get an idea of the range of sizes and angles you can work with you might get a hold of Heuser, 1989 Journal of Electron Microscopy Technique 13: 244-263 and Steere, R.L. In Chapter 5 of Current Trends in Morphological Techniques Vol 2 CRC Press 1981. These deal mostly with conventional stereo pairs and replicas but the approach and geometry is similar .
cheers, John Heckman TEM Supervisor Center for Electron Optics Michigan State University} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hey there imaging buffs } 1) Is there a way to create the red/green 3D images using stereo pair } TEM mages without buying an expensive 3D imaging program? } 2) What thickness of sections would be needed for TEM work to get } the effect of 3D depth? } 3) What is the best angle? I would be using a Philips 410LS } w/goniometer tilt stage. I'm going to hit the library but tips from the horse s } mouth are always much better. Thanks in advance. } } Rick Vaughn } RLVAUGHN-at-MAIL.UNMC.EDU }
For the first question, here is a good starting point on the Web: http://www.tisco.com/3d-web/
There is a link called "how to make..." but it wouldn't come up. I don't know if that page is gone or just down temporarily. Basically you probably need to use Adobe Photoshop. We have used that quite successfully in making these images. There may be other ways, but if you want to do it digitally, you need a program that can separate the color channels.
My colleague with the good notes on the second two questions is out of town but if I don't see other answers forthcoming I'll have him contact you when he gets back. He images dislocation stereo pairs often.
Cheers, John Vetrano john.vetrano-at-pnl.gov
----------
Hey there imaging buffs 1) Is there a way to create the red/green 3D images using stereo pair TEM mages without buying an expensive 3D imaging program? 2) What thickness of sections would be needed for TEM work to get the effect of 3D depth? 3) What is the best angle? I would be using a Philips 410LS w/goniometer tilt stage. I'm going to hit the library but tips from the horses mouth are always much better. Thanks in advance.
I use an Agpha-Arcus ll and I'm very satisfied with the results it gives me, since we need grey scale for triblock copolymers. There is a newer vertion of the scanner which might have better features. Regards,
The plans for the CSMS/MIKMAS meeting are coming along. We will have everything from digital imaging workshops to a demo of Virtual TEM usage. There will be venders, and several of them will have live demo's of their equipment.
I hope to have the schedule out around next weekend.
For those of you who plan to attend, the directions to our facility are now on the web and can be found below:
I will also post the reults of the show and supper survey next week.
Please consider coming. We are going to set up a block of rooms at University prices at the Raddison Hotel just off of Neil Street in Champaign. Please give us until next Tuesday to complete the reservation.
Radisson Hotel #: 217-398-3400 The Hotel location is marked on the Web Page
If you have any Questions please call 217-244-1567 between 7:30am - 4pm.
**All those considering presenting, please contact me this week if you contacted me via mail , email or phone so far**
Lou Ann -- *************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/CSMS/csms.html
Personal Home Page: http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/LAM.html
I currently am trying to find a way to image composite fibers of 50 to = 500 micron in length and 10 micron in width with out them crossing and = touching. =20
The idea is to image, threshold, and then count and measure these = fibers. right now we have to go through each field and manully measue = them. This can be quite tedious and time consuming when there are = thousands involved. We can use the image analyser to do fields = automatically with some manual interaction.
Can anyone suggest a means of dispersing such fibers in the preperation = process that will not allow them to touch and overlap??? We can also = set the image analyser to randomly eliminate the ones that touch. This = is an alternative but would like to be able to measure them all, if = possible. Also this is done on just basic brightfield and does not = require any special illumination such as polarized light.=20
Any suggestions can be sent to this address and I will reply. Thank = You.
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Dear Fellow = Microscopist, {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I am in need of someone = assistance. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} I currently am trying to find a way = to image=20 composite fibers of 50 to 500 micron in length and 10 micron in width = with out=20 them crossing and touching. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} The idea is to image, threshold, and then count and = measure=20 these fibers. right now {/FONT} {FONT size=3D2} we have to go = through each=20 field and manully measue them. This can be quite tedious and time=20 consuming when there are thousands involved. We can use the image = analyser=20 to do fields automatically with some manual interaction. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Can anyone suggest a means of dispersing such fibers = in the=20 preperation process that will not allow them to touch and = overlap??? We=20 can also set the image analyser to randomly eliminate the ones that = touch. =20 This is an alternative but would like to be able to measure them all, if =
possible. Also this is done on just basic brightfield and does not = require=20 any special illumination such as polarized light. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Any suggestions can be sent to this address and I = will=20 reply. Thank You. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Cono = Passione {/FONT} {/DIV} {/BODY} {/HTML}
Funny you should ask this question. For the last two semesters I have been working on just this sort of thing with the SEM. I had fairly good success using Photoshop. The trick is making sure that your images are alligned vertically, and playing around with the color until it matches the color of the filters on the viewerlenses. As far as the tilt angle, that depends on the magnification and the degree of roughness of the surface being imaged. Generally speaking, a smoother sample will require more tilt (7-15 degrees) than a rough one (3-7 degrees). At higher magnification, too much tilt can mean too much parallax (displacement) and the result may be uncomfortable to view. Usually 4-10 degrees of tilt will produce sufficient parallax. If still unsure, you can do what I did and take a series of images, each at 3 degree differences in tilt. Just be sure that when you refocus the image after tilting that you do so without changing the magnification. On the SEM this means using the Z axis control instead of the objective lense control. Also be careful when you tilt that you maintain your field of view (tracing the outline of some major features with a grease pencil on the monitor screen works great).
I'm not sure of how you're planning on tilting your sample. Because you're using a TEM you may be best off tilting your beam instead of your sample. I've never tried this as our instrument wasn't able to do this, but it always seemed like a great way to go. I imagine there would be less refocusing problems. Less change in contrast too. Let me know how it works out if you try this method. There are two pretty good references on this technique you might want to check out. One is "The Perception and Measurement of Depth In the SEM", by A.Boyde in SEM 1979 Vol 2 page 67-78 (the part you want starts on pg 70). The other is "Introduction to Stereo Scanning Electron Microscopy" by Eric Chatfield in Vol 6 of Principles and Techniques of Scanning Electron Microscopy 1978. There are others I can give you if you want.
Another method you might try is horizontal displacement instead of tilting. The trick is to have enough displacement so as to produce sufficient parallax, but maintain enough similarity in the field of view that the brain can still fuse the two images. I didn't have much success with this technique. The above references describe this method as well.
Oh, heres a good one. While working on this little project of mine (which has now become somewhat of a recurring obsession) I discovered the National Stereoscopic Association (NSA). These people are very interested in any form of stereoimaging and would love to hear from you and help you in any way they can. They even have their own magazine Stereo World which is filled with you guessed it stereopairs. Its great if somewhat bizarre. And they have a cool website! Check it out at nsa-3d.org/nsa-membership.html and have your 3d glasses ready. You might try contacting Larry at larry-at-sapphire-star.com to see if he has any advice or suggestions.
Whew! I didn't realize I had so much to say! I hope this has been of some help. If you have any other questions or just want to discuss the frustrations of stereoimaging (and there are a few). Feel free to contact me at coopera-at-bigdog.engr.arizona.edu (the world's longest email address).
Good Luck and let me know how it turns out
Anne Marie Cooper
On Thu, 9 Apr 1998, Rick L Vaughn wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hey there imaging buffs } 1) Is there a way to create the red/green 3D images using stereo pair } TEM mages without buying an expensive 3D imaging program? } 2) What thickness of sections would be needed for TEM work to get } the effect of 3D depth? } 3) What is the best angle? I would be using a Philips 410LS } w/goniometer tilt stage. I'm going to hit the library but tips from the horses } mouth are always much better. Thanks in advance. } } Rick Vaughn } RLVAUGHN-at-MAIL.UNMC.EDU }
Cono Passione wrote: ================================================ I currently am trying to find a way to image composite fibers of 50 to 500 micron in length and 10 micron in width with out them crossing and touching.
The idea is to image, threshold, and then count and measure these fibers. right now we have to go through each field and manully measue them. This can be quite tedious and time consuming when there are thousands involved. We can use the image analyser to do fields automatically with some manual interaction.
Can anyone suggest a means of dispersing such fibers in the preperation process that will not allow them to touch and overlap??? We can also set the image analyser to randomly eliminate the ones that touch. This is an alternative but would like to be able to measure them all, if possible. Also this is done on just basic brightfield and does not require any special illumination such as polarized light. ================================================== This sounds like it could be an ideal application for SPI's "Tacky Dot Slides". If you are not familiar with this product line, you can get full details on our website given below.
You can chose between standard products with dots on either 500 or 1000 um "centers". If you went for the 500 um center to center dots, you could work with a dot size of only 15 um. That would ensure that only one fiber stuck per dot, but it would be theoretically possible for at least some small (but very small) population of fibers to be touching.
If you went for the 1000 um centers, the smallest dot size is 100 um. While you would be certain that no fibers on adjacent dots would ever be touching, there could be the possibility that there could be more than one fiber sticking on a given dot, giving another instance of fibers touching.
But despite these problems, it would seem that these "Tacky Dot Slide" products could either solve your problem completely or come very close to doing so. What you might need is a special order product with a smaller dot size on 1000 um centers.
Once mounted, the fibers could be imaged either by LM or SEM for the desired automated analysis.
Disclaimer: SPI Supplies manufactures Tacky Dot Slides (TM) as an exclusive licensee of E. I. DuPont de Nemours and Co., Inc. according to US Patent #5, 356,751.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
{smaller} I currently am trying to find a way to image composite fibers of 50 to 500 micron in length and 10 micron in width with out them crossing and touching.
{/smaller}
{smaller} The idea is to image, threshold, and then count and measure these fibers. right now we have to go through each field and manully measue them. This can be quite tedious and time consuming when there are thousands involved. We can use the image analyser to do fields automatically with some manual interaction.
{/smaller}
{smaller} Can anyone suggest a means of dispersing such fibers in the preperation process that will not allow them to touch and overlap??? We can also set the image analyser to randomly eliminate the ones that touch. This is an alternative but would like to be able to measure them all, if possible. Also this is done on just basic brightfield and does not require any special illumination such as polarized light.
{/smaller}
{smaller} Any suggestions can be sent to this address and I will reply. Thank You.
{/smaller}
{smaller} Cono Passione
{/smaller}
{/excerpt} { { { { { { { {
Dear Cono,
Other than an equally laborious process of aligning the fibers (you did not mention whether they were straight, curled, or crimped), one other approach is to find an image analysis system which has a special fiber module for analyzing fibers as they cross or touch. Companies to check on
I have students in my SEM course do an exercise in taking stereo pairs. It provides for a great show-and-tell at the end of the course. Since optimum angle depends on surface topography and magnification, I have different students do different angles and then c;compare end results. We usually use 7o as routine with 14o thrown in. The larger angle usually is overkill. It is very important to focus with Z-control and try to keep the image centered.
I have done a bit with TEM using a goniometer stage but not enough to give advise. It does work better if you have a rough surface specimen, either negative stain or sections in a type of resin that tears as you microtome (like lowicryls or LR White).
For reconstruction, I realign the images in photoshop, using the center of the image for alignment purposed. This is a critical step and must be done accurately for good final results. Using layers makes this routine. I put the original image (0 angle) in the first layer, cross-hairs on the second layer to mark my center point, and the second image on the third layer, etc. Reducing the transparency of the second image lets you easily see the cross-hairs and first image.
Using color channels, you can change the aligned images into your red and blue (or green) colors and adjust transparency to overlay them. However, it is easier to use an image analysis program such as NIH Image (free). I use ScanAlytic's program, IP Lab Spectrum which makes the conversion about a 20 second process.
The next problem is getting the final image into a form which can be shown to audiences. I have found that it is often difficult to get the right exposure using digital slide makers. The slides tend to come out too dark. You often also have color problems with some films. I have had the best luck just photographing the computer screen with a 35mm camera. I use Polaroid Presentation Film which gives you a "what you see is what you get" when photographing a computer screen. Kodak films are heavy on the blue in this instance and I haven't tried Fugi film (although it works great in some digital slide makers!). I use the option in the Adobe PhotoShop tool box of full screen mode which enlarges the image to maximum screen size and fills rest of monitor with a black border and then do an exposure series and get good slides every time.
Have fun with this...your audience will certainly appreciate it.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University --------------------------------------
Hey there imaging buffs 1) Is there a way to create the red/green 3D images using stereo pair TEM mages without buying an expensive 3D imaging program? 2) What thickness of sections would be needed for TEM work to get the effect of 3D depth? 3) What is the best angle? I would be using a Philips 410LS w/goniometer tilt stage. I'm going to hit the library but tips from the horses mouth are always much better. Thanks in advance.
Rick Vaughn RLVAUGHN-at-MAIL.UNMC.EDU
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I am searching for a hardware/software solution to control access and provide accounting information for electron microscope use. We operate a multi-user academic facility with five microscopes and other analytical equipment. Presently, instrument access is controlled by custom written software running on a very old PC. Once a user has logged in, the PC switches the "ready" signal of an individual microscope using a serial I/O interface card. Unless a user has logged in, the instrument is disabled. Our existing system is becoming less reliable and is not easily upgraded.
Are other members of the microscope community using a similar approach? Does anyone know of a commercial software package for this type of application? The ability to work in a network environment (web-based perhaps) would be a plus.
I'm conducting research for a short paper and presentation on Scanning Electron Microscopy due 21 Apr 97. Any sources or information you would recommend would be greatly appreciated.
} At 06:02 PM 4/9/98 -0500, you wrote: } Hey there imaging buffs } 1) Is there a way to create the red/green 3D images using stereo pair } TEM mages without buying an expensive 3D imaging program? } 2) What thickness of sections would be needed for TEM work to get } the effect of 3D depth? } 3) What is the best angle? I would be using a Philips 410LS } w/goniometer tilt stage. I'm going to hit the library but tips from the } horses mouth are always much better. Thanks in advance. } } } Rick Vaughn } } RLVAUGHN-at-MAIL.UNMC.EDU } ----------------------------------------------------
Rick,
Responding to questions 2 & 3 above:
When I got my on-site training after instalation of our Philips CM12, I was handed a huge compendium of TEM applications tailored for the CM12. There is a section on stereo-TEM in there which I will copy and send to you by snail mail, if you'll send me your address.
Within it, is a table and a graph giving suggested tilt angles for stereo TEM as a funtion of section [or sample] thickness and magnification, which was worked out long ago by Dr. Lee Peachey. The trend is, the higher the magnification, the smaller the tilt angle between stereo pairs should be, and the thicker the sample, the smaller the tilt angle should be.
In response to question #1, so far I haven't played around with the red/green method of stereo presentation. I make a pair of black and white slides, by digitizing my TEM negs and use PowerPoint to print them out to a Polaroid slide maker. Then I use inexpensive stereo slide viewers ($7 ea.,from Reel 3-D Enterprises, www.stereoscopy.com/reel3d. I have no commercial interest in Reel 3-D.) to view the pairs, which works quite well. I'm presently working out viewing bacteria in 3D that have had immunogold labeling applied to their surfaces, to better localize the location of the gold.
Good luck!
Gib
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
I would be most interested in your findings. Please summarize them to the list. I have thought of using Microsoft's scheduler program to handle signing up, but it would not do the accounting, nor does it appear readily accessible over the web. I bet there would be a few takers for such a product.
At 11:01 AM 4/13/98 -0400, you wrote: } } I am searching for a hardware/software solution to control access and } provide accounting information for electron microscope use.
1. I would like feed back from anyone that has purchased or worked with a Gatan slow scan camera that was purchased in the last year. How is life after the salesman? Does the system measure up to expectations? Satisfaction with tech support? Software/ upgrade problems? Comparison of PC platform to MAC. Things that really need to be improved. Any other comments are also welcome.
2. We are also interested in a side mounted video camera. Comments & experiences are welcome on this subject as well.
I am in need of the manual for an LKB Knifemaker model 7801B. At some point I have lost the manual and now I need to overhaul this well-used device. I recall it was just a few pages long. Could someone take the time to fax it or scan it for me? It can be faxed to:
530 754 7536
If it is easier to FTP it, please email me for the FTP address and password.
Thanks in advance.
Rick A. Harris, Director Microscopy and Image Analysis Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 752 3085 fax raharris-at-ucdavis.edu
Well Kiddies my sputter coater died. And after all the help you guys gave me to fix the grounding problem too. Sheesh! Anywho, the question now is:
Is the vacuum evaporator good enough (or better?) to do some sputtering? We can tilt & rotate while doing it ;-O. Will this be sufficient for people doing low mag. work (not going over 1000X)?
I have to send the poor ole beastie away to be repaired, I think the Pirani gauge broke. The vacuum sucks but the gauge does not read out, it just lies there all the way to the left (but this is Berkeley so I guess that's fitting).
Promising not to riot while awaiting replies,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Is anybody using stereo pairs projected on a screen as a teaching tool? If so, would you mind describing how the images are lined up?
I 've thought for years that stereo projection had to be one of the most valuable assets to any classroom. I remember that the basics included 2 Ektamatic projectors with crossed sheets of polaroid across their lenses to allow viewers to use crossed polaroid glasses. A special screen was required, but I can't recall the details. I think I remember that one image was mounted as usual, and the other adjusted to the correct stereo location using a special guide that was invented by Lee Peachey, I think, and sold by one of the EM suppliers.
That was many years ago, so if there's a better technique now, please let us hear about it. And/or correct me if I'm wrong.
I am faced for the first time with having to process histological paraffin sections for TEM, and have a few questions. (First I should say this is mammalian tissue, and we must use histology first to locate the region of interest, which would require too many blocks to search for by TEM).
I've seen posts/literature in the past regarding the process of xylene treatment and rehydration before starting TEM prep. at the OsO4 stage. The two questions I have are: 1. Since we can have new sections cut from the original block, how thick should they be, and should they be collected onto slides treated with some type of mold release agent? Is there a better substrate than glass slides for this process?
2. If using glass slides, is there a way to create a "well" or chamber around the section so that I don't have to use whole Coplin jars full of OsO4, Spurr's resin, etc. during processing? I would much rather apply the reagents to the area of the section only, similar to applying immuno-reagents. However, I doubt that the hydrophobic pens we use would work with solvents and Spurr resin. Any ideas?
Any other tips from experienced individuals would be appreciated!
Thanks as always, Karen
-- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
Dear Paul, } } would you mind describing how the images are lined up? } We've used them for presentations in-house and at MSA. We've just put someone in the audience with stereo glasses, projected the images, and adjusted until the "audience" saw optimal stereo.
} A special screen was required, but I can't recall the details.
It is one coated with aluminum rather than glass. Metals do not change the polarization upon reflection, but glass does. I don't know where they can be obtained; we guard ours jealously. Yours, Bill Tivol
Dear Gib, } } Within it, is a table and a graph giving suggested tilt angles for stereo } TEM as a funtion of section [or sample] thickness and magnification, which } was worked out long ago by Dr. Lee Peachey. The trend is, the higher the } magnification, the smaller the tilt angle between stereo pairs should be, } and the thicker the sample, the smaller the tilt angle should be. } One thing to note is that this table gives easily seen stereo, but not accurate apparent z-values. Depending on what you're going to do with the info, you may or may not want to use the table. Most computer programs are capable of working out the correct z-values if you tell them the tilt angles. Yours, Bill Tivol
Dear LISTERS-FRIENDS, Thank your very much to everyone for your kind replies on my problems with electron miocroscope JEM-4000EX.
There was problem in logical vacuum system. Namely the chip No. 4B of VAC SYSTEM PB in VALVE UNIT had been foulted. That was replaced on new one. Now everything is OK. Again thank very much for help. Looking forward to help YOU in future.
Sincerely yours Anton Gutakovskii Laboratory of Electron Microscopy Institute of Semiconductor Physics Novosibirsk, Russia
Any kind souls out there who could advise on layout and specs for utility needs and vibration/EM field interferences for a lab space that would include light microscopy, infrared microscopy, SEM, XRD, HPLC, sample prep, and offices?
I will summarize written e-mail replies for the group. Drawings or plans can be faxed to 413/458-2314.
This is a multi-part message in MIME format. --------------7A44FA4AB075D78DF1B1F64F Content-Type: text/plain; charset=koi8-r Content-Transfer-Encoding: 7bit
Dear LISTERS-FRIENDS, Thank your very much to everyone for your kind replies on my problems with electron miocroscope JEM-4000EX.
There was problem in logical vacuum system. Namely the chip No. 4B of VAC SYSTEM PB in VALVE UNIT had been foulted. That was replaced on new one. Now everything is OK. Again thank very much for help. Looking forward to help YOU in future.
Sincerely yours Anton Gutakovskii Laboratory of Electron Microscopy Institute of Semiconductor Physics Novosibirsk, Russia
--------------7A44FA4AB075D78DF1B1F64F Content-Type: text/x-vcard; charset=koi8-r; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Anton Gutakovskii Content-Disposition: attachment; filename="vcard.vcf"
In our expanding microscopy core facility, we would like to "Gait" (sp?) cells using an instrument called the Cell-dyne whcich used laser scattering and some morphological features to identify the populations on a fixed slide. Is any one very familiar with the Meridian Ultima system, and do you know if these similar features are incorporated into their software and/or if cells can be not only morphologically identified, but also categorized as to laser scatter, size, shape, re-located with x+y precision, etc, automatically? Any leads would be help ful in this regard. Thank you, Tom Baginski
I wish to receive notice about the book "Manual of microscopic Analysis of Feedstuffs. 3rd Ed. The Amer. Assoc. of Feed Microscopists, 1992, p. 73-93" or the articles in the same book, autors BATES L.e coll. Feed ingredient descriptions of animal origin.
} } } {kszaruba-at-MMM.COM} 04/13 5:53 pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear All,
I am faced for the first time with having to process histological paraffin sections for TEM, and have a few questions. (First I should say this is mammalian tissue, and we must use histology first to locate the region of interest, which would require too many blocks to search for by TEM).
I've seen posts/literature in the past regarding the process of xylene treatment and rehydration before starting TEM prep. at the OsO4 stage.
The two questions I have are: 1. Since we can have new sections cut from the original block, how thick should they be, and should they be collected onto slides treated with some type of mold release agent? Is there a better substrate than glass slides for this process?
2. If using glass slides, is there a way to create a "well" or chamber around the section so that I don't have to use whole Coplin jars full of OsO4, Spurr's resin, etc. during processing? I would much rather apply the reagents to the area of the section only, similar to applying immuno-reagents. However, I doubt that the hydrophobic pens we use would work with solvents and Spurr resin. Any ideas?
Any other tips from experienced individuals would be appreciated!
Thanks as always, Karen
-- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
Hi Karen,
I do not mess with paraffin sections. Once an area of intrest is determined, the corresponding area from the paraffin block is cut out, cut up into small pieces than processed for EM. I start with a 1% OsO4 in Toluene for a minimium of 4 hours; wash 3X with acetone than infiltrate in 1:1 Spurr's:Acetone overnight. Embed in the morning into 100% Spurr's, polymerize and section. If you do have to work with paraffin sections only, have them cut as thick as possible (} 5 um) and process on the slides. This is the fun part: when ready to polymerize, fill up a beem capusle with resin, invert it over the section (area of intrest), clamp, tape, hold the capsule to the slide, and place in oven. Next day place slide on a hot hot plate and gently rock the capsule until it pops off the slide. ( I am successful about 50% of the time) Note: before the slide goes into the oven, wipe off the bottom and any area away from the area of intrest of resin so that only the section is covered with resin.
Best of Luck, Ed Calomeni Dept of Pathology Medical College of Ohio Toledo, OH 43614 ecalomeni-at-mco.edu
Hello again, World, [sorry for bothering again I mistyped the name in the former message.]
Looking for the address and fax number of the Company called Coherent Radiation, Inc. I just spent half an hour browsing, not to avail. So sorry for bothering you but I have no other way than asking. Thanks,
--Yves MANIETTE Universitat de Barcelona http://www2.gol.com/users/scscope/maniette/ENTREE.HTM
Looking for the address and fax number of the Company called Coherent Technology, Inc. I just spent half an hour browsing, not to avail. So sorry for bothering you but I have no other way than asking. Thanks,
--Yves MANIETTE Universitat de Barcelona http://www2.gol.com/users/scscope/maniette/ENTREE.HTM
Other than glass slides?? Yes permanox slides are available. They do not embed in Spurr and can be more easily separated from the section.
Small volumes of fixative?? Yes, try a small slide mailer that is used for shipping slides, I have used these with all solutions, with the exception of propylene oxide, we infiltrate these in alcohol+ resin. Some small slide chambers are also made for autoradiography, but are pricey.
Using glass slides?? I have had pretty good success separating sections or cells from glass slides by inverting the glass slide over the beam capsules, so the excess resin will drain away. And immediately after removing from the oven, set the glass slide on a block of dry ice. Wait until bottom of block is frosty and snap block off slide.
Have you tried some of the other resins available for light and TEM?? I can be contacted off the listserver if you have any other questions, and will be glad to help.. Marge
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
Don't know what materials you are evaporating or if it is a low or high vac system, but it should suffice for less than 1000x. Coating grain size is typically larger but you won't see it. I have found that with my low vac evaporator, film thickness is much more difficult to control and I don't use if for high mag/resolution work if possible. If you are coating with carbon (vs Au or AuPd) the stopping power is less, but again, at low mag it won't matter. Carbon will, however, yield a more noisy signal than Au since it tends to adsorb more incident beam and liberate less BSEs and SEs.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Well Kiddies my sputter coater died. And after all the help you guys gave me to fix the grounding problem too. Sheesh! Anywho, the question now is:
Is the vacuum evaporator good enough (or better?) to do some sputtering? We can tilt & rotate while doing it ;-O. Will this be sufficient for people doing low mag. work (not going over 1000X)?
I have to send the poor ole beastie away to be repaired, I think the Pirani gauge broke. The vacuum sucks but the gauge does not read out, it just lies there all the way to the left (but this is Berkeley so I guess that's fitting).
Promising not to riot while awaiting replies,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
by primemail1.pcom.net (8.8.5/8.8.7) with SMTP id MAA16551 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 14 Apr 1998 12:45:07 -0400 Message-ID: {35339273.216A-at-pcom.net}
i'm having trouble distingushing between the two are there any special etches i can use ?
Fellow list members, BibMic - A bibliography of books relating to Materials Microscopy, previously published on paper in Materials Characterization 36(1996)105 is now available on the net at http://bibmic.metalmat.ufrj.br
It lists over 1000 books, and is searchable by author, title and keywords.
Hope you find it useful Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
} ---------- } From: Yves Maniette[SMTP:yves-at-giga.sct.ub.es] } Sent: Tuesday, April 14, 1998 8:10 AM } To: Microscopy List } Subject: Re: Coherent **Radiation**, INC.(Please forget about } previous msg) } =20 } = ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America=20 } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } = ---------------------------------------------------------------------- } -. } =20 } =20 } Hello again, World, =20 } [sorry for bothering again I mistyped the name in the former = message.] } =20 } Looking for the address and fax number of the Company called = Coherent } Radiation, Inc. I just spent half an hour browsing, not to avail. So } sorry for bothering you but I have no other way than asking. Thanks, } =20 } --Yves MANIETTE } Universitat de Barcelona } http://www2.gol.com/users/scscope/maniette/ENTREE.HTM } =20
Does anyone have an address, phone #, for a company called Genex Corp? Thanks for any help? Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children Dallas
I just know there is a association of microscopy in Internet. I work on the field for long time and I hope I can share the experience and knowledge with all of you. Please let me know if I need do something for joining , like fill a form.
Now I have a problem. I cut 50 micro meter paraffin section beautifully but I can not mount them well. There are a lot of wrinkles when the sections are dried. Maybe some experts can give me some idea. Thank you!
ou may want to be sure that your unit really isn't leak tight before sending anything away for repair. Can you put the gauge tube into an adapter (rubber stopper with a hole bored in it) and put this onto your mechanical pump and see if your gauge/pump combination works? If so then=
you have left out a sealing component while repairing the ground fault.
Keep smiling, it takes fewer electrons than frowning?
At last we have the printed course announcement for the RMC Materials Science Microtomy Course 1998.
This is the most comprehensive course available for those interested in doing polymers, metals, semiconductors, ceramics for TEM and SPM. =
The course dates are September 29-October 2, 1998 in Tucson,AZ. This is t= he fifth time the course has been held and is staffed by four experts in problem solving via ultramicrotmy and EM.
For complete details please see our web page; =
RMC-Scientific.com/microtomes/
Steve Miller Director of Sales, North America RMC 3450 S. Broadmont, Tucson, AZ 85713 Tel: 520-903-9366 Fax: 520-903-0132 Email: Steve.Miller-at-RMC-Scientific.com
} If you do have to work with paraffin sections only, have them cut as } thick as possible (} 5 um) and process on the slides. This is the fun } part: when ready to polymerize, fill up a beem capusle with resin, } invert it over the section (area of intrest), clamp, tape, hold the } capsule to the slide, and place in oven. Next day place slide on a } hot hot plate and gently rock the capsule until it pops off the slide. } ( I am successful about 50% of the time) Note: before the slide goes } into the oven, wipe off the bottom and any area away from the area of } intrest of resin so that only the section is covered with resin.
This is called the "pop-off" technique. We have posted a published paper delineating this technique in detail on our web site (address below). A second paper is available by snail mail.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
by hil-img-10.compuserve.com (8.8.6/8.8.6/2.10) id KAA15206 for microscopy-at-sparc5.microscopy.com; Wed, 15 Apr 1998 10:07:24 -0400 (EDT)
Giblab at GIBRALTAR STEEL wrote:
} I'm having trouble distingushing between the two are there any special } etches i can use ?
---------
After consulting with George Vander Voort (an authority on carbon steels and related etching techniques), I have discovered that there is not a good etchant to reveal retained austinite. An electrolytic method has be= en proposed in the past, but George told me he was never able to get good =
results from it. =
George's suggestion: =
Try Klemm's Reagent to tint etch the ferrite (if it's there). =
KLEMM's REAGENT: - Make up a stock solution of water saturated with Sodium Thiosulfate - Take 50ml of this stock solution and add 1gm Potassium Metabisulfite.
- Immerse sample (do NOT swab on the etchant). Watch the surface - of the steel, and remove from solution when the surface turns a - red/violet color. Rinse with water, ethanol and warm air dry.
Another option is to try Beraha's tint etchants listed on pg.643 of Georg= e Vander Voort's book, 'METALLOGRAPH Principles and Practice', published by McGraw Hill and available through ASM. This is not meant to be a book=
advertisement, but it is a handy reference if you are dealing with microstructural analysis on a regular basis.
Hope this helps. Scott D. Holt BUEHLER, LTD PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-6500 http://www.buehlerltd.com
I would like to receive information from anyone that has worked with a xenosput magnetron sputter coater. How is coating quality compared with conventional SEM coatings such as gold and chromium? Any other comments are also welcome. Thanks in advance.
Maria do Carmo Goncalves Institute of Chemistry University of Campinas Sao Paulo - Brasil e-mail: maria-at-iqm.unicamp.br
A position for a Senior Histology Technician is opening at Carolinas Medical Center in Charlotte, NC. Please address all inquiries to "Dr. Helen Gruber" {hgruber-at-carolinas.org} .
Thank you ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
I would like to receive information from anyone that has worked with a xenosput magnetron sputter coater. How is coating quality compared with conventional SEM coatings such as gold and chromium? Any other comments are also welcome. Thanks in advance.
Maria do Carmo Goncalves Institute of Chemistry University of Campinas Sao Paulo - Brazil FAX (55 19) 7883023 maria-at-iqm.unicamp.br
Without intent to re-open an old thread, I apologize to you all for comments made in haste and ignorance on this subject six months ago. I was visiting friends connected with Dartmouth last weekend, and was reminded of it by an article in the Dartmouth alumni magazine.
After reading the Aug. 97 Science article recommended by Robert Schoonhoven and some other references, including the glove tests showing astonishing permeability for normal lab gloves, I am now much more aware of how dangerous this stuff is compared to other mercury compounds. It's in the same league with VX nerve gas (which, in a ghoulish twist of bureaucratic humor, has an MSDS published by the Army).
Please forgive my erroneous pooh-poohing on this. It was a knee-jerk response to what looked at first like just another media scare story. I should know better by now than to react before having the facts.
With all this discussion of sterio pairs, I wondered if anyone was interested in a Wild M-5 sterio scope designed for viewing sterio pairs - scope has paired objectives that can be individually rotated to achieve convergence of sterio pairs for viewing as a 3-D image - I think it was designed for aerial photo and map viewing. I got it for a project I was doing a few years back and am done with it, so...... I am up for making someone a pretty good deal if you can use it.
Best to E-mail me at home spoefish-at-mindspring.com since the server here at work is often down for days (typical government stuff)
We use the 'pop-off' technique frequently in our lab (see Slap's post). If you must do it with sections, make sure they are 5 um thick or better. This will give you enough tissue to thin section.
We omit the OsO4 step, sense the ultrastructural morphology is very poor from formalin fixed paraffin embedded tissue.
(flame retardant: I understand some people have had good results using OSO4 with formalin fixed de-paraffinized tissue).
cya -gene
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
The Electron Microscopy Core Facility at the Mayo Clinic in Rochester, MN has an opening for an EM technologist to support both clinical and research projects. The laboratory offers expertise to collaborative projects which involve transmission and scanning electron microscopy. The laboratory is well equipped and has a history of excellent productivity and adequate funding. The successful candidate for this position will possess at least a bachelor's degree with experience in histology and/or electron microscopy. Additional experience in Immunology, Cell Biology, and Digital Imaging is desirable. Operating knowledge of transmission and scanning electron microscopes is preferred. The applicant must have excellent communicative skills and the ability to work well with a variety of personalities. The EM technologist interacts with all laboratory users in order to accomplish specific research and clinical goals with respect to electron microscopy procedures. Duties include: all aspects of specimen preparation for a variety of biomedical samples for TEM and SEM; operation of TEM and SEM; negative developing and printing; digital image capture, processing and archiving; and reporting. The technologist will also perform advanced research procedures including immunoelectron microscopy, x-ray microanalysis, and microwave processing. Mayo offers a competitive salary and benefits package. Candidates must be legally authorized to work in the United States. If interested please submit a cover letter and resume referencing job posting #98-818.col to:
Kaine A. Kerkhoff Mayo Medical Center Human Resources-OE 1 Rochester, MN 55905 Fax: 507-284-1445 Email: kerkhoff.kaine-at-mayo.edu
For more information about Mayo visit our homepage at: http://www.mayo.edu.
Jon Charlesworth, Coordinator Electron Microscopy Core Facility Mayo Clinic 1426 Guggenheim Building Rochester, MN 55905 ph: (507) 284-3148 fax: (507) 284-9349 email: charlesworth.jon-at-mayo.edu
Quick question.....has anyone ever experienced poor resin infiltration of tissue culture cells grown and processed in Permanox plates. I have performed this procedure a number of times with no problems and today, after looking at the sections on the TEM, the cells appear condensed and poorly infiltrated. I would be thankful for any shared experiences and solutions. Because this is such a specific situation, feel free to respond to me at my e-mail address. Thank You!
Sandy Perkins
Laboratory for Neurotoxicity Studies VA-MD Regional College of Vet. Med. VA Tech
We just got our Fuji Pictrography 3000 and the prints are flying out the door. I can't believe the quality. My question is: Does anybody have a great supplier (in regards to price) for the paper and donor material. I am paying $280 for the donor and $68 for a roll of glossy paper. Thanks. Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
At 11:30 15/04/98 -0300, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It is a vital part of our extensive high resolution scanning EM work where we hope to obtain featureless coating of insulating specimens.
The coating quality obtained with the xenosput sputtering chromium is as smooth as we can currently obtain with any technology. By comparison, gold sputtered in the conventional way is extremely rough. The only method which comes close is to sputter platinum at very low currents and pressures with a conventional magnetron sputter head.
Of course chromium CAN be sputtered in any vacuum system, but it will have a high content of chromium oxide due to residual gas. The Xenosput has the cleanest imaginable vacuum system in that a pre-coat sputter of titanium in the work chamber purges all gases except the xenon from the work chamber atmosphere. So you can apply a coat of pure metallic chromium which will not oxidise for several days.
The xenon is used as it has better characteristics for sputtering. Under xenon energetic neutral bombardment of the sputtered film is minimised and heat buid up in the specimen is reduced.
Hello All, I was wondering if anyone has had any problems with hardening of tissue when using lanthanum nitrate as an extracellular tracer? Also, is it advisable to use lanthanum in all processing solutions ie through osmium fixation, dehydration etc, or only in the initial incubation buffer? Any advice would be much appreciated.
Thanks in advance
--------------------------- Miss Peta Clode Zoology Department LaTrobe University Bundoora, Victoria Australia. 3083.
Has anyone come across an adhesive called " M Glue" ? It used to be = available 6-7 years ago and was superb for mounting small powders. I = think it was a water based adhesive which was "milky" at first and = cleared after 20 mins to leave a tacky surface which gave excellent = adherence and stability. We used to purchase it from the UK but I = think it was manufactured in S.Africa. We have been unable to find a = suitable replacement. Does anyone know of a similar product ?
Thanks,
Colin Reid
Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#c8e0d8} {DIV} {FONT color=3D#000000 size=3D2} Has anyone come across an adhesive = called "=20 M Glue" ? It used to be available 6-7 years ago and was = superb=20 for mounting small powders. I think it was a water based = adhesive=20 which was "milky" at first and cleared after 20 mins to leave = a tacky=20 surface which gave excellent adherence and stability. We = used to=20 purchase it from the UK but I think it was manufactured in = S.Africa. =20 We have been unable to find a suitable replacement. Does = anyone know=20 of a similar product ? {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Thanks, {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Colin Reid {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {BR} Electron Microscope = Unit, {BR} Trinity College=20 Dublin, {BR} Dublin 2, {BR} Rep. of Ireland. {BR} Tel: 353-1-6081820 {BR} Fax:=20 353-1-6770438 {BR} email: {A=20 href=3D"mailto:creid-at-tcd.ie"} creid-at-tcd.ie {/A} {/FONT} {/DIV} {DIV} {/DIV} {/BODY} {/HTML}
I have been asked to calculate the mfp (elastic and inelastic) of electrons in wool with increasing levels of a contaminant. At this stage I am going to consider the wool as a carbon then add in different levels of the contaminant. In the first cases this contaminant will be sulphur and bromine.
Is there any shareware out there (or even a macro or for something like Excel) which could perform these calculations and display the separate mfps. I only have access to PC's and Macs and can't get access to any compilers so the code would need to be executable!! I have seen the NTLAMBDA program in the archives and it is just what I want, but I have no way of compiling it!!
Any help would be greatly appreciated.
Colin Veitch
Instrumentation Scientist CSIRO Division of Wool Technology PO Box 21, BELMONT, Vic. 3216. Australia.
We are trying to find, to purchase or trade, working or not working, a LVC Sputter Coater once manufactured by Plasma Sciences, Inc. up until a few years ago. We believe that some of these units also could have carried the name of Energy Beam Sciences, Inc.
Please contact me off line if you have such a system that is excess to your current needs.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We have tried Temp-Fix but our powders are both temperature and solvent sensitive. At the moment we use "Sticky-Tabs" which do not give good stability. I am hoping that someone will know more about the original "M-Glue" to try and source a new supply. As with a lot of EM products, such as resins, they are used for other functions and the EM use is just an off-shoot. I know the company in the UK have tried to source the material and failed.
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
-----Original Message-----
Colin
NTLamdba will be running as a WWW applet in about a week. I'll shift around my "order of things" to convert and try to help you out. You will need a Java aware browser like Netscape The URL is:
http://tpm.amc.anl.gov/NJZTools
Just login next week and look for the EELS MFP Link to be activated.
} Hi All, } } I have been asked to calculate the mfp (elastic and inelastic) of } electrons in wool with increasing levels of a contaminant. } I have seen the } NTLAMBDA program in the archives and it is just what I want, but I have } no way of compiling it!! } } Colin Veitch
Is SEM considered radiation creating instrument that has to be registered? ******************************************************************* Pnina Ari-Gur, D.Sc. Materials Science and Engineering Western Michigan University Office: (616) 387-3372 Kalamazoo, MI 49008 FAX: (616) 387-6517 email: arigurp-at-wmich.edu or pnina.ari-gur-at-wmich.edu also: arigur-at-lab2.cc.wmich.edu *******************************************************************
In the state of Michigan all electron microscopes are considered radiation producing devices. They must be registered and readings taken once per year. There must be a registration tag on each instrument as well as the current inspection certificate. The Michigan Dept. of Consumer and Industry Services takes care of this. However, the registration and inspection is usually coordinated by a radiation safety officer at each university. I suggest you make a few calls at your university to get the complete story. The State can levy fines if you are not in compliance; however they are usually lenient, especially if you were not aware of the law. Stanley L. Flegler, Assistant Director Center for Electron Optics Michigan State University flegler-at-pilot.msu.edu
We are in the market for a dye-sub printer - primarily for SEM images, Electron Probe Maps, TEM images, transparencies, .... The Sony UP-D8800 was recommended but I would appreciate comments on other printers concerning the print quality, reliability, etc.
Thanks, steve rozeveld
} Steve Rozeveld } } Dow Chemical Co. } 1897 Bld. Door E-43 } Midland, MI 48667 } sjrozeveld-at-dow.com } % (517) 636-5167 } Fax: (517) 638-6443
Colin Reid wrote: =============================================== We have tried Temp-Fix but our powders are both temperature and solvent sensitive. At the moment we use "Sticky-Tabs" which do not give good stability. I am hoping that someone will know more about the original "M- Glue" to try and source a new supply. As with a lot of EM products, such as resins, they are used for other functions and the EM use is just an off- shoot. I know the company in the UK have tried to source the material and failed. ================================================ You might be thinking about "M-Bond 610". If that is indeed the one you are talking about, and it is used in several different ways around EM labs, you can find it on our website listed below. Its original use was indeed outside of EM, it was developed for the mounting of micro-strain gages on surfaces.
But for temperature and solvent sensitive powders, have you considered our Tacky Dot (TM) slides? This will enable you to mount your powders without exposure to solvents or temperature and the particles end up on orthogonal centers for easy viewing and analysis. These two are described on our website below. The advantage of the Tacky Dot slides over even M-Bond 610 is that there is no danger that your particles of interest will "sink into" the liquid adhesive while it is polymerizing. After all, even the M-Bond 610 has vapors and if your powders are indeed solvent sensitive, then they would or could be sensitive to the vapors from such solvents as well.
Disclaimer: SPI Supplies offers both types of products, as well as Temfix (TM), so we would have an obvious interest in promoting the use of these items.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The Midwest Microscopy and Microanalysis Society Materials Science Meet= ing will be held on May 22, 1998, hosted by Dr. Nigel Browning of the Depar= tment of Physics, University of Illinois at Chicago. In addition to a spea= ker slate of internationally renowned scientists, the meeting will showcase= the new electron microscopy lab at UIC with a tour of the facility followin= g the scientific sessions.
Invited presentations will cover the following topics: phase contrast imaging in TEM, scanning tunneling microscopy, electron energy loss spectroscopy, enery dispersive x-ray spectroscopy, and other surface analytical microscopic techniques.
The program will begin with an introduction at 8:45 a.m. and the first speaker at 9:00 a.m. There will be poster sessions and a buffet lunch.= The last scientific session will end at 4:00 p.m., with the facility tour following this session.
MMMS members will receive a mailing in the next few days that will incl= ude the meeting agenda and a map with travel information. Anyone else requ= iring information may contact me at (847) 935-0104 or by e-mail (preferred) = at jane.a.fagerland-at-abbott.com.
Steve asks: } ... } } } We are in the market for a dye-sub printer - primarily for SEM images, } Electron Probe Maps, TEM images, transparencies, .... } The Sony UP-D8800 was recommended but I would appreciate comments on } other printers concerning the print quality, reliability, etc. } } ...
You probably should have let us know with which computer you'll interface with ... much of what you're asking has as much to do with the software ... However, we use our dye-sub withing a situation of many different platforms all connected via TCPIP. We went with the Codonics NP1600 networking printer and haven't looked back. But if I were to wish for perfection, it would be that it printed with a CMYK ribbon rather than to create grayscale images with CMY. I don't know that any dye-sub will print grayscale without a tint if their ribbon doesn't have the K transfer. The only dye-sub I'm aware of with includes K with CMY is Tektronix ... I'd like to know of others. (... BTW ... All dye-subs will offer a K ribbon ... but switching between color and grayscale printing is not practical ...)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
There is a product called Microstik offered by Ted Pella (Cat. #16033) which sounds as if it may do what you need. I have only used it a couple times, but as I recall it did a good job of adhering fine particulates without drowning them.
No financial interest, etc.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
} But if I were to wish for perfection, it would be that it printed with } a CMYK ribbon rather than to create grayscale images with CMY. I don't } know that any dye-sub will print grayscale without a tint if their } ribbon doesn't have the K transfer. The only dye-sub I'm aware of with } includes K with CMY is Tektronix ... I'd like to know of others. (... } BTW ... All dye-subs will offer a K ribbon ... but switching between } color and grayscale printing is not practical ...)
One addendum to Michael Shaffer's comments:
We also are extremely well pleased with our Codonics 1660M dye sub printer.
We are using the Chroma Vista color dye sub media (CMY) and the b/w media for gray scale printing. They are supposed to be releasing a Direct Thermal media for b/w that does not use a transfer ribbon. Perhaps at that juncture, one could leave the color ribbon in place and use the Direct Vista paper for b/w printing since the medium is in the paper. Of course, this would require shutting off the color ribbon somehow. Nifty idea if they could pull it off. Time will tell.
JB
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Thanks to everyone who responded to my question! There were a lot of good suggestions, unfortunately it will be another couple weeks before I get to try them out.
Some of the suggestions were: 1. Use the paraffin tissue block itself instead of sections. [This is not an option on these samples, but will keep in mind for future.]
2. Make sure the sections are } 5 microns thick, as much as 20 microns.
3. Collect on: superfrost plus glass slides, Permanox slides, or ACLAR.
4. Or, process free-floating sections in vials (sections should be very thick and may have curling problems).
5. To reduce reagent volumes, add drops on top of section and cover with parafilm, or invert section over drops using toothpicks to prop up slide. Or, process in slide mailers, which are smaller than Coplin jars.
Thanks again, Karen -- Karen Zaruba, kszaruba-at-mmm.com BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144
*The opinions above are my own, not necessarily my employer's*
I would greatly appreciate any information as to the possible health hazard of using halogen lighting as a light source in microscopy. Having scant resources at hand I built a light source using halogen bulbs. I was told that they are a source of UV radiation. My question is should this be of any concern?
Stanley L. Flegler responds: } } } ... } ... In terms of testing, they normally ask that it be tested at its } normal opering voltage. Stanley L. Flegler }
I figured the primary interest would be x-rays, but was looking to separate SEMs from TEMs because "normal operating voltages" are generally less than 30kV, rather than generally more than 100kV. Is there any rationale for claiming 30keV x-rays simply cannot escape the instrument???
Can anyone direct me to a source of those nifty lights/signs that go on the outside of a darkroom door with a switch inside to turn them on and off? I have looked several places with no luck. Even simple home made ideas would work for us.
Thanks
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Pnina Ari-Gur wrote: ================================================ Is SEM considered radiation creating instrument that has to be registered? ******************************************************************* This is a legal kind of question and the answer depends on the political jurisdiction in which you reside. In the USA, it is determined on a state- by-state basis, and I suspect in other countries, on a country by country basis.
In Pennsylvania, we are required to register each instrument with the state. We also are required to have someone radiation check each registered instrument. The requirements for NJ are different than they are for PA. So you have to contact the officials in your own state, usually located in some kind of a "radiation physics" kind of department, and find out about the legal requirements for your own specific situation.
But these requirements should not be lightly dismissed as some kind of bureaucratic boondoggle. There have been enough "horror stories" over the years and these are the kinds of stories that literally "drive" regulators and the promulgation of regulations:
a) On several occasions, a TEM "viewing glass" was broken, and when hearing the high price of a replacement from the manufacturer (which would have been lead glass), the user replaced the glass (without realizing what they were doing) with more ordinary silica glass, resulting in long term radiation exposure to the operator.
b) Some of the older instruments designed in the 1950's at a time before the hazards of radiation exposure were fully appreciated became recognized as "known emitters". The affected manufacturers apparently did various retrofits to those instruments once the hazards became known. I can myself remember operating one of these instruments but only while wearing a lead apron! But I would still be concerned about any pre-1960 instrument that has not been recently and regularly checked for radiation leakage, including the adjustment knobs on the lenses.
c) I have myself encountered at least on one occasion an SEM that had been modified with the placement of a "see through" port (to observe cathodoluminesence) which was not made of lead glass. In another instance, my admonitions caused an SEM user to not install a "port" plate with "Plexiglas".
Modern instruments seem to be made in a way that x-rays just can not escape during regular operation. And if the instrument is so far out of alignment that x-rays do escape, then it would also be out of alignment to the extent that it would not be possible to hold a high vacuum and therefore even get high voltage and a beam. So the end result is again, there is no chance of escape of x-rays.
Radiation badges seem to give some peace of mind with regard to the kinds of massive kinds of leaks described above. However, the leaks that might be occurring, such as when using adjustment knobs on the column, and which would result in exposure to the fingers and hands, would not be registering typically on a radiation badge. Hence one has to guard against the radiation badge becoming a false sense of security. And one should pay particular attention to radiation that might be emanating from the adjustment knobs on a column.
My firm offers no products or services in the radiation physics area. These are my own views and opinions as a result of 35 years in the world of EM. I am told by some that I over exaggerate the hazards of a modern SEM or TEM. But not everyone uses modern EMs and also, any laboratory is vulnerable to someone doing something dumb, like replacing a broken glass window with something other than the manufacturer's recommended replacement part (and without anyone even knowing about it). That is why I believe that any EM instrument, whether required by law or not, be checked periodically for radiation leakage, just to be sure.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The cheap (!) FARGO thermosub printers also print with CMY, CMY+Overlay (same ribbon) OR with a CMYK ribbon OR with a K ribbon (b/w). Resolution 300x600 max. We own one, it works well, but I don't have possibility to compare it with others, so can't state anything about that. We bought it because of tests in computer journals (don't remember which) and our limited money.
you wrote:
} But if I were to wish for perfection, it would be that it printed with } a CMYK ribbon rather than to create grayscale images with CMY. I don't } know that any dye-sub will print grayscale without a tint if their } ribbon doesn't have the K transfer. The only dye-sub I'm aware of with } includes K with CMY is Tektronix ... I'd like to know of others. (... } BTW ... All dye-subs will offer a K ribbon ... but switching between } color and grayscale printing is not practical ...)
Dr. Arthur Schuessler University of Heidelberg Zellenlehre Im Neuenheimer Feld 230 D-69120 Heidelberg Germany
We were inspected by the National Radiological Protection Board (our official Radtiation Advisers, in UK) who found nothing of concern but started us wondering. For about two years thereafter, we wore radiation monitoring badges initially which showed nothing. Then we posted badges at strategic place around the column (held on by sticky tape). We never found anything on them, although sensitive radiation counters could occasionally pick up counts from certain areas (such as around where the x-ray detectors enter the column!). The worst problem was the plaster (skim coating) on the wall of the room, the natural background in the building is relatively much higher!.
I would not think that you have any real problem. These bulbs are used generally in microscopy, slide projectors, car headlights and probably other uses too. All lamps will produce a spectrum of radiations but not excessively in the UV band unless designed to do so specifically, like high pressure mercury lamps for fluorescence microscopy. Most simple light microscopes don't incorporate UV filtering in their setup.
Common sense says don't spend long looking at it i.e. don't expose your eyes directly to the light!
I would be more concerned about the electrical safety. You need to make sure the unit checks out ok in that area. Don't risk electrocution!
In many states, it is. The funny thing is that the 'soft' radiation emitted by an SEM (should there be a breach in the shielding) is often not detectable by the survey equipment required. In most cases, you'll get higher readings from an EDS monitor.
As I recall (a fragile thing), Michigan is indeed a state that requires such testing. Your institution can likely provide the certification through their plant maintenance department.
TEM operators should be more attentive to these requirements. The higher operating voltages produce 'harder' radiation that can be more penetrating. In addition, there are more opportunities for emission through the column and chamber accesses.
Brings to mind a TEM manufacturer who, many years ago, supplied instruments with viewport glass that was not leaded per spec. Many old time TEM operators took to wearing their radiation badges after that.
} } Is SEM considered radiation creating instrument that has to be } registered? } ******************************************************************* } Pnina Ari-Gur, D.Sc. } Materials Science and Engineering } Western Michigan University Office: (616) 387-3372 } Kalamazoo, MI 49008 FAX: (616) 387-6517 } email: arigurp-at-wmich.edu or pnina.ari-gur-at-wmich.edu } also: arigur-at-lab2.cc.wmich.edu } ******************************************************************* } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Chuck made some interesting comments on radiation leaks from EM's. I am sure we have all come across very dangerous situations over the years. I can remember my first job in a hospital pathology lab where the Osmium was mixed on an open bench because the main operator had destroyed his sense of smell. Thankfully I moved on quickly !
There was one potential source of X-Rays that was not mentioned. With side arm insertion on TEM's if the arm is left out the beam may be switched on for alignment etc. ). This produces X-Rays at eye level. ( We use this to test our Geiger counter ! ) One to watch.
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
However, the leaks that might be occurring, such as when using adjustment knobs on the column, and which would result in exposure to the fingers and hands, would not be registering typically on a radiation badge. Hence one has to guard against the radiation badge becoming a false sense of security. And one should pay particular attention to radiation that might be emanating from the adjustment knobs on a column. {snip} --------------------------------------------------------------- If such a concern exists, TLD* finger rings are available.... *- ThermoLuminescent Dosimeter
I use them from time to time. Not to measure dose from my SEM. but to check the dose from some of the samples I must prepare!
Please allow me to inform you of a Minisymposium on SEMICONDUCTOR RADIATION DETECTORS which will be held during the 34th Conference on Microelectronics, Devices and Materials (MIDEM) in Rogaska Slatina, Slovenia, September 23-25, 1998.
In the framework of the minisymposia, several invited speakers will present different aspects of the chosen topic, thus offering the audience valuable complete information. The minisymposium invited speakers will be: Dr. Gerhard Lutz from Max-Plank Institute fur Physic, Munich, Germany; Dr. Joseph Kemmer, KETTEK Gmbh, Germany, Dr. Peter Weilheimer, CERN, Switzerland; Dr. Walter Bonvicini, Instituto Nazionale di Fisica Nucleare, Trieste,Italy
Complete information regarding submission of papers, topics of invited lectures, conference location etc. can be found on Web page http://pollux.fer.uni-lj.si/midem/conf98.htm
(Deadline for submission of papers is May 15th)
Hope to see you in Rogaska Slatina!
With best regards, Dejan Krizaj
*********************************** Dejan Krizaj, Ph.D. Laboratory for Electron Devices Faculty of Electrical Engineering University of Ljubljana Trzaska 25 1000 Ljubljana SLOVENIA
Email: dejank-at-fe.uni-lj.si ************************************ Disclaimer: This email was sent to addresses obtained through various sources. We apologize if you or your institution is not involved in activities covered by the Conference and Minisymposium. If, however, you know for persons that might be interested in this information, please forward them this message.**********************************
Please allow me to inform you of a Minisymposium on SEMICONDUCTOR RADIATION DETECTORS which will be held during the 34th Conference on Microelectronics, Devices and Materials (MIDEM) in Rogaska Slatina, Slovenia, September 23-25, 1998.
In the framework of the minisymposia, several invited speakers will present different aspects of the chosen topic, thus offering the audience valuable complete information. The minisymposium invited speakers will be: Dr. Gerhard Lutz from Max-Plank Institute fur Physic, Munich, Germany; Dr. Joseph Kemmer, KETTEK Gmbh, Germany, Dr. Peter Weilheimer, CERN, Switzerland; Dr. Walter Bonvicini, Instituto Nazionale di Fisica Nucleare, Trieste,Italy
Complete information regarding submission of papers, topics of invited lectures, conference location etc. can be found on Web page http://pollux.fer.uni-lj.si/midem/conf98.htm
(Deadline for submission of papers is May 15th)
Hope to see you in Rogaska Slatina!
With best regards, Dejan Krizaj
*********************************** Dejan Krizaj, Ph.D. Laboratory for Electron Devices Faculty of Electrical Engineering University of Ljubljana Trzaska 25 1000 Ljubljana SLOVENIA
Email: dejank-at-fe.uni-lj.si ************************************ Disclaimer: This email was sent to addresses obtained through various sources. We apologize if you or your institution is not involved in activities covered by the Conference and Minisymposium. If, however, you know for persons that might be interested in this information, please forward them this message.**********************************
Look into the Codonics 1600 and 1660. Quality is excellent, company support is great and cost per print is certainly comperable if not less than many other brands. It is built using a Kodak engine but has the most advanced software of any printer that I have heard of in this price catagory ($9000-13,000). This printer permits adjustment of gamma and contrast at the printer....a great feature when printing from drawing programs which do not permit these changes. We do a lot of labeling of images and gels in a drawing program and want to print directly with ability to adjust brightness and contrast of image if necessary.
Also it is very easy to send multiple files to a single sheet of paper with the printer doing the scaling to fit. You can instruct the printer to divid a sheet into 4-6 or more sections and then send that number of images to it. This is great for making quick hard copies of digital SEM images at low cost.
I have requested a number of feature updates since we purchased our printer 4 months ago and the company has been very receptive to addressing these issues. And if there is a mechanical problem with a printer that cannot be fixed over the phone, they are not opposed to drop shipping a new printer so your down time is minimal.
Debby Sherman, manager Microscopy Center in Agriculture Purdue University
--------------------------------------
We are in the market for a dye-sub printer - primarily for SEM images, Electron Probe Maps, TEM images, transparencies, .... The Sony UP-D8800 was recommended but I would appreciate comments on other printers concerning the print quality, reliability, etc.
Thanks, steve rozeveld
} Steve Rozeveld } } Dow Chemical Co. } 1897 Bld. Door E-43 } Midland, MI 48667 } sjrozeveld-at-dow.com } % (517) 636-5167 } Fax: (517) 638-6443
P.S. My apologies if I am covering an old topic.
}
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This came to me this morning via EduCom's edupage technology digest. Since CD-ROMs are most folks choice for archiving data and digital images, I thought this would be appropriate to pass on. Doug
DIGITAL ISN'T FOREVER "Digital information lasts forever, or five years -- whichever comes first," says a senior computer scientist at RAND Corp. The problem is that computer experts are finding out that under less-than-optimal conditions, digital tapes and disks, including CD-ROMs, can deteriorate in as little as five to 10 years. And the decay, although it happens gradually, isn't evident until it's too late, says the founder of Voyager Co., which makes commercial CD-ROM books and games. "CDs have a tendency to degrade much faster than anybody, at least in the companies that make them, is willing to predict." At the same time, as data is ported from an antiquated platform to a newer system, often there are bits that fail to make the transition. Sometimes it's just a matter of footnotes disappearing, but sometimes whole categories of data are lost. "It's like playing the child's game of Telephone. It doesn't take many translations from one media to another before you have lost significant aspects of the original data." (Business Week 20 Apr 98) ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
1. V1 doesn't work in auto mode -Control circuit is malfunction i.e. IC1(vac control),IC8, Tr5. V1 won't work in manual. -check V1 and solinoid valve, air pressure 2. if the control circuit and V1 working properly at 190 UA V1 should be open to get HVAC. 3. All valve will operate manually while they are running in Auto mode because we by pass the control circuit .
4. V5 doestn't operate in auto
-check airlock door swich
regards,
Paiboon Nuannin ---------------------------------------------------------------------------- Director Scientific Equipment Center Prince of Songkla University Hatyai 90112 Thailand
} } } ---------- Forwarded message ---------- } Date: Mon, 23 Mar 1998 10:03:47 -0400 } From: Brian McIntyre {mcintyre-at-optics.rochester.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: JEOL35 vacuum problem folowup } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } hi- } } thanks to all who sent info on about testing the vacuum system. } } seems i may have more problems: } } 1) the leak rate from HVac is {3uA/min } atm=250uA } Hvac=25uA on vacuum meter test points (OK) } } 2) V1 doesn't work in auto mode, and many times won't work in manual } } 3) DP heater warms up (light on) in about 20minutes } } 4) R4 vacuum control pot will not make V1 open when vacuum meter {190uA } while in auto mode } } 5) solenoid valve for the closing side of V1 will not release pressure } so that the opening side of the valve (when pressurized) will force the } valve to open.... most of the time (related to #2 above) } } 6) some of the valves (like v4) will actuate manually when the } controls are } in the auto mode (is this normal...or an indication that the logic is } all messed up?) } } 7) V5 (airlock) will not open in auto mode, but does in manual (as } does the } associated leak valve) } } i'm really confused about this since it seems to be a moving target with } many appearent "failures". is there one circumstance or condition that } would explain these observations???? } } thx in advance! } brian } } **************************************************************** } Brian McIntyre } Electron Microscopy Lab } Institute of Optics } University of Rochester } Rochester, NY 14627 } } 716-275-3058 } 716-244-4936(fax) } } } }
At 12:32 17/04/1998 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to know what is your EM. As a field engineer since 22 years, I can tell you NO SEM and NO TEM from any manufacturer has Xray leakage The problem is coming and at that point I agree with Mr Garber, when the people has modified or replacing original parts with local supply for example. Speaking about the microscope I know, Jeol, It's for example impossible to generate beam on a 2010 (200kv) with no sample holder, there is an opto coupler detector. Of course if you insert something that simulate the holder you can have the beam. But this is a wrong manipulation. Like to remove a Gamma source from it's lead castle. You can do it but you have to think about that because you have be warned! Same for electron microscopy. May be you can check the statistics about cancer in the EM environment, for myself since all that years, all my collegues and competitors are still living and I'm sure that cigarettes cause more painfull than EM radiation. This reply is not for starting a polemic but just to tell that ,we, manufacturers of microscopes are responsible people and it's also our health when we are working on theses machines. Cheers.
=================================== Jacky Larnould JEOL Europe SA Service Manager mailto:larnould-at-worlnet.fr mailto:larnould-at-mnet.fr WEB: {http://www.jeol.com} Phone:33 (0)4 67 72 28 26 Fax :33 (0)4 67 79 54 90
I am looking to purchase a video camera that can be mounted on a light microscope to display microscope sections via a video projector. The video projector is a Sharp model XGE1200U. The resolution of this projector is 800X600 for PC or 832X624 for Mac. I want to be able to project real time pictures from a light microscope using this same system. Last fall I tried this with an older video camera mounted on this microscope (640 x 480 resolution) but the images were too pixilated to be really useful. However, images projected from a Macintosh computer were fine. Does anyone know of a high line resolution camera (video rate) that they have used for a similar purpose that might help me out?
Kenneth A. Taylor, Ph.D. Office phone: 850-644-3357 Institute of Molecular Biophysics Lab phone: 850-644-4104 Florida State University EM room phone: 850-644-8769 Tallahassee, FL 32306-4380 Fax: 850-561-1406 E-mail: taylor-at-bio.fsu.edu Home pages: http://www.sb.fsu.edu/~taylor/ http://www.fsu.edu/~biology/faculty/Taylor/kat.html
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Leica, the first name in ultramicrotomy, Diatome US, and Electron Microscopy Sciences announce another in a series "ultramicrotomy of materials" workshops.
Bring Your Own Samples!!!
Focus on hands-on participation of the following:
Embedding of industrial samples Specimen trimming
Ultramicrotomy of hard materials SEM applications of microtomy
Ultramicrotomy of polymers Section collection & handling
Low temperature sectioning
Course Instructors:
Dr. Tom Malis Mr. Helmut Gnagi Materials Characterization Group Leader Hard Materials Specialist Canadian Federal Laboratory Diatome, Ltd. Switzerland
Mrs. Ani Issaian, MS Ms. Kathy Johnson California Inst. of Technology Materials Analyst-SEM Polymer Scientist Gates Rubber Co.-Denver
Where: University of Colorado, High Voltage EM Facility Boulder, CO
When: May 19-21, 1998
Tuition: $1,100.00 USD
For more information contact:
Paul DeGeorge 800-248-0665 X2279 Diatome US 215-646-1478
id sma001490; Fri Apr 17 14:08:01 1998 Received: from elba.wadsworth.org (elba [172.16.1.100]) by newton.wadsworth.org (8.8.6.Beta3/8.7.1) with ESMTP id OAA27471; Fri, 17 Apr 1998 14:01:50 -0400 (EDT)
Dear Jacky, } } I would like to know what is your EM. } As a field engineer since 22 years, I can tell you NO SEM and NO TEM from } any manufacturer has Xray leakage } The problem is coming and at that point I agree with Mr Garber, when the } people has modified or replacing original parts with local supply for example.
Here is where Charles Garber's points can be particularly well-taken. We had an old JEOL JEM200 (no longer in service) which had a radiation leak at the side-entry airlock. This leak was very directional, and with the operator seated, there was no exposure. However, if the operator were to stand up with the beam on, (s)he would, indeed, be exposed to x-rays. This occurred with the original equipment--there were no modifications. It must be added that this scope was manufactured before ANSI standards for radia- tion-producing equipment were in effect, and finding this sort of leak was part of the impetus for developing those standards.
} Speaking about the microscope I know, Jeol, It's for example impossible to } generate beam on a 2010 (200kv) with no } sample holder, there is an opto coupler detector. Of course if you insert } something that simulate the holder } you can have the beam. But this is a wrong manipulation. Like to remove a } Gamma source from it's lead castle.
This is a good example of an improvement of the safety of EM's. However, the old instrument would leak x-rays whether a specimen was in or not.
} You can do it but you have to think about that because you have be warned! } Same for electron microscopy.
Yes. There's only so much that the manufacturer can do. One must take responsibility for one's own safety. (Of course, someone must provide appropriate safety instructions, but it is up to the individual to follow them.)
} May be you can check the statistics about cancer in the EM environment, for } myself since all that years, all my } collegues and competitors are still living and I'm sure that cigarettes } cause more painfull than EM radiation.
Even if the EM produces some radiation, there is likely to be a larger effect from concrete walls (potassium-40) or living at altitude, which increases radiation exposure 24 hours a day, than from the equipment. And, yes, cigarettes are much more carcinogenic.
} This reply is not for starting a polemic but just to tell that ,we, } manufacturers of microscopes are responsible people and it's also our } health when we are working on theses machines.
I'd say that EM manufacturers are well above average in this respect. Yours, Bill Tivol
"ribbon doesn't have the K transfer. The only dye-sub I'm aware of with includes K with CMY is Tektronix ... I'd like to know of others. "
Silly, commment! The Kodak 8600 and 8650 series both can utilize the CMYK (and K only) ribbons. Afterall this is the engine upon which the N16xx's are built!
Next comment: ...and this is a really scary one. Since we couldn't afford the nice interface of the Codonics NP1600/1660 we just got delivery of a Kodak DS 8650 PS ($7k after current $1500 Kodak rebate). O.k. Last fall in desperation for something to print color we bought an Epson Photo-stylus for $325.00. After printing the exact same image (digital composed at true 800 dpi resoultion) to both the Kodak ($3.50 / page - 300dpi) and the Epson (glossy 'film' media $2.20/page - 720 dpi) a series of comparisons were made, by experience EM users and photographers here with the result being that (1) beyond 12" you can not see a difference other than the fact that the Espon print has a little sepia tint to it, (2) "Up-close / In-your-face" the ink dots are detectable in the Epson prints and there is a slight presence of the 'printer head lines' in the deepest solid black areas (i.e. print something solid black on a laser printer and look for horizontal lines), (3) final difference Kodak's extralife coating has an archive life expectancy of } 50yrs (well its lasted 5 days so far! We'll just have to wait and see.), (4) both printers take approximately the same amount of time to print (O.k., the Condonics is "the fastest on the market", and wasn't run in a side by side comparison).
Now on the cheaper papers (Media) for the Ink jets you get a very nice image quality (albeit not that'll match the dye-sub, but still very acceptible) you start dropping the price/page alot, i.e. $0.75/page, $0.24/page, $0.10/page (...then you hit laser printers, anyway). And for work prints, reviewer copies, etc. these are very good.
Lets see $14,000 printer = $325 Epsons x 43 units ....
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
In reference to your conversation on printers... You are right that the Epson printer has more bang for the bucks in my opinion. I have been selling the dye-subs for some years now. After doing umteen side by side demos you can hardly tell the difference between the two. i let the customer do the comparison... By far the Epson Stylus 800 and not the Epson photo Stylus is better. it does 1440 DPI.. I suggest the Photo glossy paper from Hammermill. Others such has the new paper from polaroid are sort of yellowish to start. That is kind of hard to notice unless you do a hands on comparison. The Epson Stylus 800 going now for about $350 with the paper running about a buck a print... Check it out, You'll be amazed....CP -----Original Message-----
"ribbon doesn't have the K transfer. The only dye-sub I'm aware of with includes K with CMY is Tektronix ... I'd like to know of others. "
Silly, commment! The Kodak 8600 and 8650 series both can utilize the CMYK (and K only) ribbons. Afterall this is the engine upon which the N16xx's are built!
Next comment: ...and this is a really scary one. Since we couldn't afford the nice interface of the Codonics NP1600/1660 we just got delivery of a Kodak DS 8650 PS ($7k after current $1500 Kodak rebate). O.k. Last fall in desperation for something to print color we bought an Epson Photo-stylus for $325.00. After printing the exact same image (digital composed at true 800 dpi resoultion) to both the Kodak ($3.50 / page - 300dpi) and the Epson (glossy 'film' media $2.20/page - 720 dpi) a series of comparisons were made, by experience EM users and photographers here with the result being that (1) beyond 12" you can not see a difference other than the fact that the Espon print has a little sepia tint to it, (2) "Up-close / In-your-face" the ink dots are detectable in the Epson prints and there is a slight presence of the 'printer head lines' in the deepest solid black areas (i.e. print something solid black on a laser printer and look for horizontal lines), (3) final difference Kodak's extralife coating has an archive life expectancy of } 50yrs (well its lasted 5 days so far! We'll just have to wait and see.), (4) both printers take approximately the same amount of time to print (O.k., the Condonics is "the fastest on the market", and wasn't run in a side by side comparison).
Now on the cheaper papers (Media) for the Ink jets you get a very nice image quality (albeit not that'll match the dye-sub, but still very acceptible) you start dropping the price/page alot, i.e. $0.75/page, $0.24/page, $0.10/page (...then you hit laser printers, anyway). And for work prints, reviewer copies, etc. these are very good.
Lets see $14,000 printer = $325 Epsons x 43 units ....
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Halogen bulbs in general use do produce a significant amount of UV radiation. The halogen lamps sold for home use now all have a simple UV filter to reduce the exposure. UV is readily filtered and as long as you are not looking at the light directly there should not be a problem. If you will be spending a lot of time looking at the light directly, that is not filtered by passing through some other material first, it might be a good idea to place a simple UV fiter in front of the bulb.
**** Keith Ryan wrote: I would not think that you have any real problem. These bulbs are used generally in microscopy, slide projectors, car headlights and probably other uses too. All lamps will produce a spectrum of radiations but not excessively in the UV band unless designed to do so specifically, like high pressure mercury lamps for fluorescence microscopy. Most simple light microscopes don't incorporate UV filtering in their setup.
Common sense says don't spend long looking at it i.e. don't expose your eyes directly to the light! **** Phil Dahlstrom PLDahl-at-aol.com
I have just completed putting the Microscopy & Microanalysis'98 Search Engine on-line at:
http://www.msa.microscopy.com
Just follow the links to the Microscopy & Microanalysis'98 Meeting Page. Using this engine you may search the program by Author, Keyword, Symposium and/or day of the week. The data base is not perfect, so if you find any errors please report them to me off-line.
The Program Committee has once again done an excellent job and it promises to be an great meeting as always. See you there....
My comment on the potential source of X-Rays from the side-arm of our TEM was not a recommendation for people to operate in this manner. For the record our TEM is 10 years old and the tip was given to us by an experienced TEM engineer from the manufacturer. We were testing for radiation and were not getting any counts ( except from the Manager's old watch ! ). He suggested this as a means to confirm that the test equipment was working. I have no idea whether any Jeol TEM's of a similar age would perform in the same way. Perhaps an owner of a JEM 1200 would like to test this ? I do agree that the manufacturers have been extremely safety conscious in the development of modern EM's. Older EM's unfortunately were not as safe. Also I am sorry to be the barer of bad news but some of your colleagues are not in the best of health, since a number of them that I have known personally have passed away over the last number of years. One of my own colleagues died from cancer in her 30's. Electron Microscopy, & associated preparative techniques, can be a hazardous environment. All we can do is be as safety conscious as possible and try and minimise the risks.
Colin Reid
Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
} Hello } } I would like to know what is your EM. } As a field engineer since 22 years, I can tell you NO SEM and NO TEM from } any manufacturer has Xray leakage } The problem is coming and at that point I agree with Mr Garber, when the } people has modified or replacing original parts with local supply for example. } Speaking about the microscope I know, Jeol, It's for example impossible to } generate beam on a 2010 (200kv) with no } sample holder, there is an opto coupler detector. Of course if you insert } something that simulate the holder } you can have the beam. But this is a wrong manipulation. Like to remove a } Gamma source from it's lead castle. } You can do it but you have to think about that because you have be warned! } Same for electron microscopy. } May be you can check the statistics about cancer in the EM environment, for } myself since all that years, all my } collegues and competitors are still living and I'm sure that cigarettes } cause more painfull than EM radiation. } This reply is not for starting a polemic but just to tell that ,we, } manufacturers of microscopes are responsible people and it's also our } health when we are working on theses machines. } Cheers. } } =================================== } Jacky Larnould } JEOL Europe SA Service Manager } mailto:larnould-at-worlnet.fr } mailto:larnould-at-mnet.fr } WEB: {http://www.jeol.com} } Phone:33 (0)4 67 72 28 26 } Fax :33 (0)4 67 79 54 90 }
We have been using an Epson 800 for a few months now and have produced images on Epson Glossy Film which have been accepted by customers in plac= e of conventional photos. Excellent quality but shame about the price (=A31.80/sheet) ! I have just tested some paper from a supplier ( "own = brand paper" ) here. The paper is 140g Glossy at a cost of 7p/sheet ( about 1= 0 cents ). The quality is excellent but it crinkles slightly. Still for the price !
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
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--Society for Ultrastructural Pathology--
Ultrapath IX will be held in beautiful Asheville, North Carolina, August 2-7, 1998. As in years past, a variety of topics will be covered and participation will be solicited from those attending. Those wishing to contribute to the scientific program are urged to contact Allan Tucker at the address below. Evening case presentations and poster board sessions will be held. A syllabus containing a summary of each presentation will be distributed to all participants. Among others, Dr. Feroze Ghadially of Canada and Dr. Doug Henderson of Australia are now confirmed speakers. The scientific program will emphasize the applications of immunocytochemistry, electron microscopy, in situ hybridization and related techniques in diagnostic pathology. The registration fee is $495.00 and includes the fee for attendance at all lectures and case presentation sessions, CME credit, syllabus, coffee breaks, five breakfasts, two luncheons, opening reception, excursion trip and two dinners.
The course will be held at the beautiful Grove Park Inn Resort in Asheville, NC (http://www.groveparkinn.com). The resort features an 18 hole championship golf course, as well as six outdoor and three indoor tennis courts, an indoor sports center, and indoor and outdoor swimming pools. The resort is located in the Blue Ridge Mountains of western North Carolina. Hiking, fishing, white water rafting, horse-back or mountain bike riding and other activities are available nearby. Daily jet service via the Asheville Regional Airport is available. A variety of recreational activities are planned for registrants and their families.
For further information, please contact either:
J. Allan Tucker, M.D. Department of Pathology University of South Alabama 2451 Fillingim Street Mobile Al 33617 Tel:334-471-7473 or 334-471-7799 Fax: 334-471-7884 or 334-660-5576 email: JTucker-at-usamail.usouthal.edu
or:
John Shelburne, M.D., PhD Department of Pathology Box 3712 Duke University and VA Medical Centers Durham NC 27710 Tel: 919-286-6979 or 919-286-6925 Fax: 919-286-6818 or 919-684-8689 email: shelburne.john-at-forum.va.gov
URL for Online Information: http://sup.ultrakohl.com/ultrapat.htm
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Opinions aside (and there are many!), the only true test of printer comparisons is for world-wide colleagues to take a digital image and find a vendor which can generate sample prints from the various print technologies available: inkjet, laserjet, dye-sublimation and hybrid. Also, relating the printer technology to what is applied in the darkroom will yield an understanding of what constitutes an acceptable image. Personally I have done the aforementioned with a series of digitally-captured TEM images. Basically, for our personal needs, TEM negatives have been printed using a high-contrast point light source. Diffuse-light sources generate softer edges (less contrast) which for us has been acceptable for black & white LM histology images but not for TEM. The technology of dye-sub printers (subliming the dye to a gaseous state which permeates the fibers of the print media) results in quality similar to using a diffuse-light source with TEM negatives--everything has a soft edge. Inkjets are high-contrast technologies and the high-resolution Epson (1440 dpi on photo-quality glossy paper) results in truly remarkable, near-photographic images (held at some distance away. Close examination reveals pixelation). The Epson also more closely matched in color balance a true TEM photographic print (print options set to BLACK ONLY--not COLOR). Finally, the best bang for the buck is outputted from the Fujix Pictrography, a hybrid type of technology that uses silver halide for image formation--so it acts like a Polaroid for image processing, but the results are darkroom photography! However, due to cost factors, we seem to be headed in the direction to print TEM image "thumbnails" on our dye-sub (Black Ribbon ONLY-not color) and generating manuscript prints on the Pictrography (both TEM and color LM). There--now I can get down off the Dye-Sub soapbox!
Best regards,
Walt Bobrowski Subcellular Pathology Parke-Davis Research Ann Arbor, MI 48105
The cheap FARGO PrimeraPro thermosub printer also prints with CMY, CMY+Overlay (same ribbon) OR with a CMYK ribbon OR with a K ribbon (b/w). Resolution 300x600 max. We bought one, it works well, but I don't have possibility to compare it with others, so can't state much about that. We bought it because of tests in computer journals (don't remember which) and our limited money.
wrote:
} But if I were to wish for perfection, it would be that it printed with } a CMYK ribbon rather than to create grayscale images with CMY. I don't } know that any dye-sub will print grayscale without a tint if their } ribbon doesn't have the K transfer. The only dye-sub I'm aware of with } includes K with CMY is Tektronix ... I'd like to know of others. (... } BTW ... All dye-subs will offer a K ribbon ... but switching between } color and grayscale printing is not practical ...)
Does anyone have experience with the Alps dry-ink printers? I'm interested in the Epson 800 or 1520 printers to take the burden off of our Tek Phaser IIsdx. They are listed at 720 dpi, yet with less bleed they might rival higher dpi inkjets, and the ink is water resistant.
TIA, -- Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax The box said "Requires Windows95 or better". So I bought a Macintosh.
Dear all, Please excuse the slightly non-microscopy topic of this posting.
I have an Epson Stylus Color 800 inkjet printer (with which I am very happy). The cost of Epson's own brand A4 OHP transparency slides is pretty high, though (} =A31 each). Are there aternative brands that work just as well but cost less or am I best of sticking to the manufacturers own brand?
Thanks for any advice, opinions etc. that you can give.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
It was not my original intention to post a summery but I have received more request for the summery than I have responses from users. The # of responses was small ( {10) but then I only asked for experiences on systems less than a year old.
Owners & users of video systems were generally happy with their systems. People with slow scan cameras that were in operational order were also generally content with the system performance. The catch here is "operational order" By far the major complaint in dealing with Gatan is that of timely delivery & getting online when promised. One respondent stated that delays were so great that they nearly lost their funding prior to delivery. Apparently field service leaves much to be desired. Salesman failing to return voice & e-mail was complaint. Salesman also seemed to be oblivious to the physical constraints of mounting their hardware in our confined spaces. I guess just like in a lot of other industries salesman are the problem, the 1st source of disinformation & false expectation.
A few comments on specific pieces: Model 789, a side mt. video camera "quality of the images is barely adequate" "not publication quality" " used primarily for demonstrations"
Model 673/III , video camera "expensive but extremely useful " "sensitive ...used for low-dose cryo" "low res & less than 8 bit gray scale, effectively"
Model 791, MultiScan camera " happy with hardware & software" " a very polished package"
MultiScan camera, model unknown. Gatan can't get software configured on G3 MAC, tech left & has yet to return.
Locally, Gatan has failed miserably in the tech support & software compatibility areas. I was a bit surprised at this report but knowing the source I considered it valid. My primary purpose for surveying the user group is to find out if this is a local or global problem. It seems global. Based primarily on this perception we have elected not to pursue a Gatan Multi-scan camera this year. Perhaps when Gatan is in better light we will reconsider, after all this type of system is a part of the modern hi-res TEM. In general people are happy with Gatan once installed & operational but the transition from salesman blowing air to operational seems riddled with delays. No one is impressed by the cost of the systems or upgrades. Again once operational there eq. performs as expected. For some applications Gatan is the way to go. Retrospectively I am sure most respondents will look closer at competitors next time around. For a lot of applications one can throw down 5-10K$ for a good digital negative scanner. Then there is a broad choice in software.
One thing I had hoped for but got no feed back on is Gatan's PC based software. I thought it had been released., maybe not. Can anyone address this subject?
Cell Vision Subscription Price List 1998 (Now included in Index Medicus and Medline)
Cell Vision publishes peer-reviewed scientific, medical and technical = articles in the fields of pathology, morphology, anatomy, histology, = microscopic sciences, etc., 6 times per year.
For individuals: $130 per year For institutions: $150 per year Shipping and handling: within USA add $6, international add $20.
Special price as a member of the International Society of Molecular = Morphology (Oklahoma City, OK, USA; information: Phone: (609) 802-0242, fax: (609) = 802-0245, e-mail: ismmcvjg-at-uscom.com)
Please make check payable to: Cell Vision, Institute of Molecular Morphology, 560 Fellowship Road, = Suite 407, Mount Laurel, New Jersey 08054, USA Phone: (609) 802-0242, fax: (609) 802-0245, e-mail: ismmcvjg-at-uscom.com
CELL VISION: NEW INTERNET WEB-SITE:
http://www.cbba.org/cbba/Default.htm Still under construction
Thank you for your interest. Yours respectfully
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
DISCLAIMER
Disclaimer: The views expressed in this e-mail message do not necessarily represent = the official views of the Inst. Anatom. Pathology LKA, from which this = message was conveyed. No commercial interest in products/product lines, company/-ies, = societies, if such names are mentioned or such are refered to. No = liability for incidentally incorrect information, no warranty: I'm = sharing only my daily experience.
I have some users who want to make high resolution scans of 4x5 TEM negatives. The 45 series scanners from Nikon, Polaroid, et al run US$8,000-9,000. However, I've been told that they are barely capable of 3.0 OD at 4"x5". The Imacon flatbed will deliver 3.9 at 4"x5", but it is about US$16,000. What are others using for scanning negatives?
TIA, -- Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax The box said "Requires Windows95 or better". So I bought a Macintosh.
We have an Alps printer and are very happy with it. We have it for the same reason you would like to have one-- to remove some of the pressure on our Tek Phaser IIsdx.
Patty Jansma Tel:520-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
On Mon, 20 Apr 1998, Glen MacDonald wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone have experience with the Alps dry-ink printers? I'm } interested in the Epson 800 or 1520 printers to take the burden off of } our Tek Phaser IIsdx. They are listed at 720 dpi, yet with less bleed } they might rival higher dpi inkjets, and the ink is water resistant. } } TIA, } -- Glen MacDonald } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, WA 98195-7923 } glenmac-at-u.washington.edu } (206) 616-4156 } (206) 616-1828 fax } The box said "Requires Windows95 or better". So I bought a Macintosh. }
I have an AMRAY 1830 SEM (with joystick for stage control) and EDAX 9900. Now I am thinking about getting a PC interface for the SEM and EDX using my very, very limited funding available. The features must include SEM image acquisition and storage(SEM control and image processing are not needed), stage control (in X,Y), unattended EDX acquisition and X-ray mapping(with Be window open/close control if possible). Anybody has such experience and would suggest a solution? Any response is appreciated.
Tong
-------------------------------------------------------------- Tong Wang email: tong-at-jlab.org MS 58, Jefferson Lab URL: http://www.ee.odu.edu/~xxu 12000 Jefferson Avenue fax: (757)269-7658 Newport News, VA23606 tel: (757)269-7334 --------------------------------------------------------------
I believe that TEM film has a density range of about 2.8-2.9. If this is true, any scanner with a range of over 3 should work.
Henk
} } I have some users who want to make high resolution scans of 4x5 TEM } negatives. The 45 series scanners from Nikon, Polaroid, et al run } US$8,000-9,000. However, I've been told that they are barely capable of } 3.0 OD at 4"x5". The Imacon flatbed will deliver 3.9 at 4"x5", but it } is about US$16,000. } What are others using for scanning negatives?
} TIA, } -- Glen MacDonald } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, WA 98195-7923 } glenmac-at-u.washington.edu } (206) 616-4156 } (206) 616-1828 fax } The box said "Requires Windows95 or better". So I bought a Macintosh. } } Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
We use flatbed scanners from either Perkin-Elmer or Zeiss. Both, however, are fairly expensive, say well over 50 k$ :-(. But, they deliver for that kind of money (of corse they'd have to ...). What kind of scans are the users interested in? And, on the OD, most scanners that I know of begin to run of headroom at 3 OD. J.
-- Jaap Brink, Ph.D. Biochemistry, One Baylor Plaza, Baylor College of Medicine, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Mon, 20 Apr 1998, Glen MacDonald wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have some users who want to make high resolution scans of 4x5 TEM } negatives. The 45 series scanners from Nikon, Polaroid, et al run } US$8,000-9,000. However, I've been told that they are barely capable of } 3.0 OD at 4"x5". The Imacon flatbed will deliver 3.9 at 4"x5", but it } is about US$16,000. } What are others using for scanning negatives? } } TIA, } -- Glen MacDonald } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, WA 98195-7923 } glenmac-at-u.washington.edu } (206) 616-4156 } (206) 616-1828 fax } The box said "Requires Windows95 or better". So I bought a Macintosh. }
About a month ago I posted a request for information regarding resellers = of used optical microscopes. Since then, I've gotten a number of = requests to summarize the information I received. So, without further = ado:
Websites:
LabX www.labx.com Labequip www.labequip.com Montana Microscope www.montanamicroscope.com Conneaut Lake Scientific www.wwweb-pro.com/cls Used Mechanical Equipment www.execpc.com/ume Lehman Scientific www.lehmanscientific.com Capovani Brothers www.capovani.com Scientific Equipment Exchange www.sci-equip-ex.com Bid Service www.bid-service.com
Plus, a web search of "used microscopes" using your favorite search = engine will turn up a few others.
Phone Contacts:
Martin Microscope Co (864) 242-3424 or (864) 859-2688 Bob Martin Mel Sobel 1-888-ALL-SCOPES or (516) 935-7794 Technical Instruments (415) 431-8231 Rick Staples=20 Bay Optical (415) 431-8711 Tom Henry ARC Instruments (606) 498-1345 Phil Hutcheson
There's a ton of junk equipment out there, but after searching long and = hard, we were able to come up with what we were looking for.
Many thanks again to all who responded to my original post for = information!!!
Tom
Thomas C. Isabell, Ph.D. Research Scientist E.A. Fischione Instruments, Inc. tc_isabell-at-fischione.com webpage: www.fischione.com
I have to buy a new vacuum gauge, (heads and controller), and my short list is either Leybold or Balzers (having eliminated Edwards because of their frequent model changes, which lead to "sorry, no service now available for that model" after not many years). Oh for the good old analogue-and-repairable-for-ever days! I have no experience with either brand, anyone got any good or bad news about either?
Confidentiality guaranteed for personal replies.
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
I haven't timed it for a file that size. An A4 page with two of my imag= es would be approx. 2MB and would print in about 2-3 mins at top resolution. Routinely I print the greyscale images on a 600dpi laser and only use the Epson for higher quality. The configuration of your computer will help = to a certain extent. I am lucky in that it is attached to my spare 200MHZ = PC so I don't have to sit and watch it.
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
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I have to buy a new vacuum gauge, (heads and controller), and my short list is either Leybold or Balzers (having eliminated Edwards because of their frequent model changes, which lead to "sorry, no service now available for that model" after not many years). Oh for the good old analogue-and-repairable-for-ever days! I have no experience with either brand, anyone got any good or bad news about either?
Confidentiality guaranteed for personal replies.
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
we decided on a different set-up: we purchased a MicroLumina camera with a high frequency lightbox. We wanted to scan not only TEM negatives but also X-ray films and a series of old 35 mm slides. Besides we are using the set-up for macro-photography as well. The camera is mounted on a photographic stand, which allows one to adapt the camera-film distance always to have the full size negative/object in view and therefore the full camera resolution on the object. The camera resolution is 3000x2600 pixels and sells for less the US$ 9k. Again, as the camera is adjusted to the object size, this resolution is available for a TEM negative as well as a 35 mm slide. The only drawback is each scan takes about 1-2 minutes as the camera is based on a line array.
Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
} ---------- } From: Glen MacDonald[SMTP:glenmac-at-u.washington.edu] } Sent: Monday, April 20, 1998 5:12 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Negative Scanners } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We use all three, Codonics, Epson Stylus, and Fargo Primera Pro printer. We are most happy with these printers in the order listed. Quality of print is very important to us, but time is, too. The two latter printers have taken upwards of 30 minutes to print many of our requested TEM and Confocal images. The Codonics never takes that long for tif, bmp, or postcript images. The Epson and Fargo both require that you have specialized software like Adobe,for example and drivers that will make their printer work. They print only a limited image type. The Codonics has all the conversion hardware/software built in and can be placed any where on the network ! A tremendous feature for us. Since the entire University can send prints to it, and they can use almost any format, bmp, tif, gif, eps, ...ETC. with almost no further need to adjust their image. Since we support 1000's of prints every year, and sometimes 1000 monthly, near grant writing time, we find the Codonics indispensable. We find the 300 pixel per inch resolution image is very similar to the 1200 dot per inch image you get off other printers, but there is no pixelation when looking closely at the image (if you started with a good image). The reproducibility is astounding. The first of 20 images looks just like the last one. And it takes about 20 minutes to do 20 images. None of the others would do that for us. For example, although the Epson comes close, it would takes us about 6 hours or more to do 20 images at high resolution, so we did not even try to do that. The other problem, is that there does not seem to be an easy way to send an image and get 20 prints from the Epson or Fargo printers, a software problem I guess. We can not have someone stand by and send a print every 30 minutes. Small things, but that costs us time. We now use the Epson for fast-low-res images (at 300 dpi) when needed since it has that capability and we can use plain paper. But the low cost per print is still about 35 cents. The Codonics, price per print has recently gone down for us, bulk buying, to about $2.00 print. We are hoping to see improvements on this possibility. After an extensive survey about two years ago and after multiple copies (over 800) we were overwhelming advised by research faculty (99%) to purchase the Codonics. We are very happy with it, and the service record is also outstanding. They have always found a way to get us support QUICKLY whenever we needed it. Supplies are readily available and are expected to be for a long time. KODAK is the major supplier for their machines and for Codonics with the Kodak engine. However, Codonics, does re-package the ribbon and paper and supports their product 100%. Feel free to contact me for a more detailed discussion if you wish. Technical Coordinator for Microscopy, Tom Baginski, 301 295 5691 or email:tombg-at-bictom.usuf1.usuhs.mil DOD/USUHS/BIC, Bethesda, MD
We have an ALPS MD-2300 which does dye sub and microdry printing. The dye sub technique produces images good enough for publication but is quite slow, requiring nearly 3/4 hr for an 8"x10" print. The standard microdry printing is fine for rapid display of CMYK images if one uses their special paper. The printer is finicky about other, less expensive paper, but can be gotten to work. I find their monochrome black non-dithered half toning a little annoying even at 600 dpi, but usable. Hope this helps.
Len } Does anyone have experience with the Alps dry-ink printers? I'm } interested in the Epson 800 or 1520 printers to take the burden off of } our Tek Phaser IIsdx. They are listed at 720 dpi, yet with less bleed } they might rival higher dpi inkjets, and the ink is water resistant. } } TIA, } -- Glen MacDonald } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, WA 98195-7923 } glenmac-at-u.washington.edu } (206) 616-4156 } (206) 616-1828 fax } The box said "Requires Windows95 or better". So I bought a Macintosh. } } Leonard Zablow Howard Hughes Medical Institute 722 West 168 St. New York, N.Y. 10032
Jon, A few years ago I ordered a Beseler negative carrier #8343, 4x5 anti-newton from Brandons, Inc. in Jacksonville, FL (800-874-5273). It has glass for supporting the film. I use it for my TEM negatives. I think they special ordered it for me. I would also suggest searching the web for Beseler and try to buy it directly from them.
best regards, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
We use all three, Codonics, Epson Stylus, and Fargo Primera Pro printer. We are most happy with these printers in the order listed. Quality of print is very important to us, but time is, too. The two latter printers have taken upwards of 30 minutes to print many of our requested TEM and Confocal images. The Codonics never takes that long for tif, bmp, or postcript images. The Epson and Fargo both require that you have specialized software like Adobe,for example and drivers that will make their printer work. They print only a limited image type. The Codonics has all the conversion hardware/software built in and can be placed any where on the network ! A tremendous feature for us. Since the entire University can send prints to it, and they can use almost any format, bmp, tif, gif, eps, ...ETC. with almost no further need to adjust their image. Since we support 1000's of prints every year, and sometimes 1000 monthly, near grant writing time, we find the Codonics indispensable. We find the 300 pixel per inch resolution image is very similar to the 1200 dot per inch image you get off other printers, but there is no pixelation when looking closely at the image (if you started with a good image). The reproducibility is astounding. The first of 20 images looks just like the last one. And it takes about 20 minutes to do 20 images. None of the others would do that for us. For example, although the Epson comes close, it would takes us about 6 hours or more to do 20 images at high resolution, so we did not even try to do that. The other problem, is that there does not seem to be an easy way to send an image and get 20 prints from the Epson or Fargo printers, a software problem I guess. We can not have someone stand by and send a print every 30 minutes. Small things, but that costs us time. We now use the Epson for fast-low-res images (at 300 dpi) when needed since it has that capability and we can use plain paper. But the low cost per print is still about 35 cents. The Codonics, price per print has recently gone down for us, bulk buying, to about $2.00 print. We are hoping to see improvements on this possibility. After an extensive survey about two years ago and after multiple copies (over 800) we were overwhelming advised by research faculty (99%) to purchase the Codonics. We are very happy with it, and the service record is also outstanding. They have always found a way to get us support QUICKLY whenever we needed it. Supplies are readily available and are expected to be for a long time. KODAK is the major supplier for their machines and for Codonics with the Kodak engine. However, Codonics, does re-package the ribbon and paper and supports their product 100%. Feel free to contact me for a more detailed discussion if you wish. Technical Coordinator for Microscopy, Tom Baginski, 301 295 5691 or email:tombg-at-bictom.usuf1.usuhs.mil DOD/USUHS/BIC, Bethesda, MD
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Listserve members, I will be traveling to P.R. of China in a few weeks and will, with my husband, be visiting Biology Departments at a university in Beijing and a university in Heifei where I will be meeting with microscopists. I would like to bring along a few inexpensive items which may be difficult to get in China for our hosts. One friend has already asked for double-sided scotch tape. Any suggestions for other small items useful in microscopy labs would be appreciated. Please reply directly to me as I don't know if this would be of general interest to the whole list.
Debby Sherman, manager Phone: 765-494-6666 Microscopy Center in Agriculture Fax: 765-494-5896 Purdue University E-mail: sherman-at-aux.btny.purdue.edu W. Lafayette, IN 47907 or: emcenter-at-btny.purdue.edu
by huey.cadvision.com (8.8.5/8.8.5/DCE/TRI) with ESMTP id HAA55576 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 21 Apr 1998 07:36:10 -0600 Message-Id: {199804211336.HAA55576-at-huey.cadvision.com}
We currently use an older Leaf 45 film scanner for film format from 35mm to 4 x 5. These scanners are often available from digital image equipment supply houses such as B & H in New York (212) 444-5008 0r Graphtronix (612) 461-5151. Although I have never had dealings with these companies myself, they seem to have good reputations in the digital imaging communities. A 4 x 5 scan on our leaf produces a file size of ~82Mb. The dpi is 1200 and the image is 6000 x 4740 pixels. These are for a landscape orientation. For portrait, the figures are 1200 dpi, 4760 x 4740 pixels and a file size of ~67Mb. The scanner accommodates a standard Beseler 4 x 5 negative carrier. If anyone in the research community is interested, we will be glad to scan a few negatives for experimental purposes. Just send the neg. along with either a Zip or Jazz disk and we will send back some scanned files. Since we do this for a commercial venture, please only academics or research inquiries. If you want a commercial price please contact us off of this list server at our e-mail.
Jon McGovern J. P. McGovern and Associates e-mail: jcgover-at-cadvision.com (403)291-3196 (403)291-1423 Fax
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Hi there
I have to buy a new vacuum gauge, (heads and controller), and my short list is either Leybold or Balzers (having eliminated Edwards because of their frequent model changes, which lead to "sorry, no service now available for that model" after not many years). Oh for the good old analogue-and-repairable-for-ever days! I have no experience with either brand, anyone got any good or bad news about either?
Confidentiality guaranteed for personal replies.
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
by mail.unixg.ubc.ca with smtp (Exim 1.71 #1) id 0yRgHA-0000SX-00; Tue, 21 Apr 1998 09:52:08 -0700 Message-Id: {2.2.32.19980421165018.008df1a0-at-pop.interchange.ubc.ca} X-Sender: mager-at-pop.interchange.ubc.ca X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Dear Jonathan, There is a special negative holder for Polaroid 4" X 5" negatives, made by Beseler. We use it for the TEM negatives, turned sideways. When the original broke, we purchased a new one from our local photography store. The other one we have looks homemade, before my time, which has a hinged holder consisting of two metal plates that fits the Beseler, with two glass plates held in cutouts. The negative is sandwiched between the glass plates. It holds up to 4" X 5". The disadvantages is that you must make sure your negative and 4 sides of glass are clean. You wrote: } Hi: } } We inherited a Beseler 45M enlarger, but no negative holder for 3 1/4" x 4" } TEM film. } } We use a crude, homemade holder now. A real one would be nice. Anyone have } an extra or know where I can get one? } } Thanks } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
I am looking for information to procure proceedings from the Pfefferkorn Conferences. The 1983 proceedings was published by Scanning Electron Microscopy, Inc. Does anyone have information on these proceedings?
I am interested in comparing volcanic ash from different sources. If you have ash samples that you would be willing to part with, please email me directly. Thank you!
Bob Willis ManTech Environmental Technology, Inc. willis.robert-at-epamail.epa.gov
you will find there representatives (addresses and phone/e-mailnumbers)
Another www-page I could offer to you is:
http://www.pfeiffer-vacuum.de=20
(German branch of PFEIFFER VACUUM TECHNOLOGIES, perhaps or certainly you = will find on their web-page connections to representatives in your area)
hope this helps, best of luck
Wolfgang MUSS
Dr. Wolfgang MUSS Salzburg General Hospital (LKA) Department of Anatomical Pathology,=20 EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: Ritchie Sims[SMTP:r.sims-at-auckland.ac.nz] Gesendet: Mittwoch, 22. April 1998 11:53 An: microscopy-at-sparc5.microscopy.com Betreff: Re: Vacuum Gauges TEM Hardware: vacuum SEM/TEM
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Hello
Does anyone from Edwards High Vacuum USA read this list? If so, please=20 contact me. Alternatively, does anyone have an email address for them?
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 =20 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 =20 Auckland New Zealand
I have a Leitz Laborlux D microscope at home and I'm trying to find a 'C' mount and a 2.5 or 3.3X photoeyepiece for it. If anyone has thes and is willing to part with them I'd like to buy them from you. Yes, I'm aware that I could probably get them from Leica but dollers (mine) are a concern.
TIA
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
This response is to an inquiry concerning a particular high resolution sputter coater and a reply by its first user, Mel Dickson. That posting is a few days old but requires a response and I sought and have included here comment from EMITECH (UK). There is nothing wrong with Mel's comments, except that without further discussion the topic is treated one-sidedly. Clearly, the Xenosput works well, but its suitable for very small samples only; its expensive to purchase and the Xenon gas which is heavier than argon and hence more efficient, is also rather expensive to buy. There is an amazing range of coating equipment on the market, all with their particular pros and cons.
"High resolution SEM has created a requirement for high resolution coating. Typically chromium is now used as the target material, primarily because chromium has a much finer grain size than gold. It also does not readily form coagulated islands, as is the case with gold and that is the limiting factor of conventional gold sputter coatings in high resolution SEM.
However, for Cr sputtering the vacuum system must be kept clean and the energy levels for sputtering increased to allow sputter cleaning of the target prior to sputtering. This has resulted in a new generation of sputter coaters like our Emitech K Series, which includes the K575 and the K675. The K575 can readily coat evenly an area of 76mm diameter, and the K675 which was designed specifically for the semi conductor market's 203mm wafers, can coat up to 253mm diameter. Such instruments are turbo pumped and fully automatic and employ a flushing and cleaning protocol to ensure a clean vacuum. They may also have a shutter assembly to protect the specimen during the sputter clean operation. This results in very good quality and repeatable deposition. As well as chromium targets other materials such as platinum should be available. Other desirable features of such instruments are a fully automatic control, with argon as process gas and nitrogen as purge gas. David Robinson, EMITECH Ltd"
Please note that ProSciTech has an interest in EMITECH equipment, since we distribute this equipment, however, we are only a regional distributor for this company (Australia, NZ, parts of SE Asia). Considering our profile in the home market, I don't see any commercial advantage from posting this item here. I just like to contribute to this topic which deserves fuller treatment. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** -----Original Message-----
The address is correct. Thanks for adding me to the list.
Hello: We are using MagOp Disks on Power Macs to archive images in our lab, and I am interested in knowing if anyone has found a Disk Repair Program which repairs or retrieves files from the occasional MagOp disk which starts to have problems.
I need information to educate the administration on how close to "cost neutrality" a microscopy facility can get. I currently supervise a facility with a part-time employee to assist in repairs. Much of the equipment is 20-25 years old and is constantly breaking down. I am the sole revenue generater with a few students who have been trained to use the microscope and have beamtime hours. Within a year or so of operation the facility went from generating $0.00 in revenue to ~$60K maybe 70K/year. This doesn't pay all the bills of an EM Facility with 2 TEMs and an SEM as well. My question is, is their any facility that is COMPLETELY cost neutral with revenue only for service work provided? Are the numbers I describe for a medium sized University reasonable? I would greatly appreciate any input. If their is a University that spends only what they bring in for doing samples in a multi-user environment please contact me I be happy to know how you did this.
Best Regards,
Kirk J. Czymmek, Ph.D. Biological Electron Microscopy Facility University of Delaware Newark, DE 19716 kirk-at-udel.edu
We would be interested in suggestions and collective experience with measuring the energy spread of a FEG-TEM by a method other than using a PEELS spectrometer, i.e. by some type of imaging/diffraction analysis technique. We have been using the standard method of measuring the contrast-transfer envelope and associated information limit by diffractogram analysis of amorphous Ge. However, after some work the accuracy of this method for resolving differences in energy spread even as large as 100% is not clear to us. Assessing the limits of the envelope function by seeing how well the microscope passes various fringe spacing in the 1.2 to 0.9 A range (i.e. out on the limits of the envelope function) seems reasonable, but what are the pitfalls? Other methods using beam tilt effects on diffractograms exist in the literature but remain to be checked out.
Input from you FEG-TEM artists out there would be appreciated.
Roy Christoffersen Materials Science Research and Engineering Center University of Houston 3201 Cullen Houston, TX 77204-5932 roy-at-bayou.uh.edu (713) 743-8273 FAX: (713) 743-2787
We have had good sucess from 1. Taking an EDTA tube of blood and spin it down 2. Take a long nosed pipet and collect the buffy coat area, roughly. 3. Take a Wintrobe tube ( tall skinny tube used in hematology) , place rough buffy coat in this tube) 4. Spin down the wintrobe tube. 5. Remove supernant, then collect Buffy coat with a thin glass pipet. 6. Layer gently in bottom of a vial of Karnovsky's and let set 20-30 min.
* At this point it should look like a fat miniture snake at the bottom of the vial.
7. Process gently through the rest of the EM procedure. __ should the buffy coat break up, place in ependorf tube and gently spin before each change in a microcentrifuge tube, low speed.
8. If the buffy coat is intact, by embedding time, cut with razor and seperate among embedding mold/capsules.
___ If the buffy coat is not intact, place in very small beam capsules, and put as concntrated of drops of samples as possible into the beam capsules.
__ Fill, close, and place in 1.5 ml ependorfs and spin in the microcentrifuge for 15-20 minutes at moderate speed. Polymerize as is in the capsule/ependorf assembly. At some point after fairly hard, razor off the eppendorf for better polymerization.
Hope this helps, just one way to try,
Lou Ann
Winnie Wrote: Hi There, Looking for a good technique for making a pellet of buffy coat from normal blood. Would appreciate any help. Thanks.
Winnie
***************************************-at-redfoot Lou Ann Miller Center for Microscopy & Imaging College of Veterinary Medicine Dept. of Veterinary Biosciences University of Illinois Rm 1108 Basic Sciences Bld 2001 S Lincoln Ave. Urbana, Illinois 61802
We are interested in buying a single- or double-tilt heating stage that = will fit a JEOL 2000 side-entry goniometer. I believe this goniometer is also = shared with the 1200, 200 and perhaps even their 100 series microscopes. I'm = not sure about the 4000. We would require an attainable temperature of 600=B0C. = At this time we are looking at buying a used stage, and probably won't have the = money to buy new. If you have a stage that fits this description and would = consider selling it, please contact me at the address below (email, snail mail, = phone or fax). =20
Thanks,
JSV *************************** John S. Vetrano Sr. Research Scientist Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
Does anyone know starting recipes for using cold water fish (teleost) gelatin as a blocking agent for immunocytochemistry? Does it work? Any pitfalls? We have a particularly sticky primary with lots of small punctate background appearing..controls with secondary alone are clean...presently using 0.5% BSA with 1% goat serum as block 1hr r.t....its not stopping it????
I am looking for software that can model the energy deposited by the electron beam in a thin film or bulk sample. Do any of the Monte Carlo codes do this?
I suspect that your punctate signal, when it is background indeed, may come from aggregated primary antibody molecules. In that case using more or different additives to the blocking buffer or incubation buffer will not be very helpful in improving the signal-to-noise ratio. Spinning down the antibody solution (stock, but preferably the diluted antibody) at high speed will help to get rid of aggregates. I think this is the easiest thing to try to begin with. Another explanation might be that there are indeed sticky sites in your specimen. But then one would expect different antibodies from the same animal source to give similar results. I don't know whether that is the case. If so, then fine tuning the blocking and incubation buffer composition is the way to a solution. You may find more info on our web site regarding background at http://www.aurion.nl, and in a previous series of messages to this Listserver sent in the first week of April.
Good luck,
Jan
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Managing Director Costerweg 5, 6702 AA Wageningen The Netherlands
We spin the anti-coagulated blood in a narrow test tube, then carefully remove as much as possible of the plasma without disturbing the buffy coat. Buffered 2% Glutaraldehyde is then very gently layered on top and the tube left to stand in the fridge for about a couple of hours. This gives a buffy coat which is embedded in solid plasma and can be removed from the tube with the help of a thin wooden stick or similar. The resultant disc can then be trimmed and the pieces processed to resin as you would for normal tissue. If you embed in flat moulds your specimen will have orientated layers of plasma, platelets, white cells and red cells.
It's a simple and effective technique, with only one centrigugation to give you a sample which is easily handled. The only downside is that some drugs (e.g. aspirin, I believe) may inhibit the action of glutaraldehyde on the plasma and prevent the conversion of the plasma to a solid.
James Ito Pathology Department Royal Hospital for Sick Children Yorkhill Glasgow Scotland. ---------- } From: Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: COLLECTING A BUFFY COAT FOR TEM } Date: 22 April 1998 21:50 } } Hi There, } Looking for a good technique for making a pellet of buffy coat from } normal blood. Would appreciate any help. Thanks. } } Winnie }
Its easy - if you have an administrator who does not just care about balancing the budget and finds the EM Manager easier to push about than some professor who presides over a poorly performing department. Your EM Unit costs the university real money and ought to be shut down if you supply nothing much in return. I trust that the credit side of your ledger shows that you have sufficient and happy users. In a university they would be mostly grad and some undergrad students and they require many services within an EM facility. Since the university collects money for these students, the unit should get notional or real credit. The "user pays" arguement is poor economics when the script dictates: Collect all the students funding and distribute this to administration, library and departments but give no credits to the electron microscope unit. Too often the EM user has already paid but nothing has gone to the EM Unit and the administrator is in fact looking for a second payment from the user. Kirk, our correspondent's problem is that few administrators are interested in good governance. Most just want to balance the budget along the path of least resistance. Been there, done that, I commiserate. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** } Dear Collegues, } I need information to educate the administration on how close to "cost } neutrality" a microscopy facility can get. I currently supervise a } facility with a part-time employee to assist in repairs. Much of the } equipment is 20-25 years old and is constantly breaking down. I am the } sole revenue generater with a few students who have been trained to use } the microscope and have beamtime hours. Within a year or so of operation } the facility went from generating $0.00 in revenue to ~$60K maybe } 70K/year. This doesn't pay all the bills of an EM Facility with 2 TEMs and } an SEM as well. My question is, is their any facility that is COMPLETELY } cost neutral with revenue only for service work provided? Are the numbers } I describe for a medium sized University reasonable? I would greatly } appreciate any input. If their is a University that spends only what they } bring in for doing samples in a multi-user environment please contact me I } be happy to know how you did this. } } Best Regards, } } Kirk J. Czymmek, Ph.D. } Biological Electron Microscopy Facility } University of Delaware } Newark, DE 19716 } kirk-at-udel.edu } } }
Does anybody have a spare puck (or 2) for a Complot Series 7000 Digitizer (Houston Instruments). After two buyouts of the original company the current manufacturer is not able to help out with this 10 year old (at least) tablet.
Alternately, we are in need of a replacement for the 24" x 36" digitising tablet we were using for measuring area and perimeter (via DOS software package) from projected histological images. We have tried to replace this system with a videomicroscope and image analysis package but cannot get a system to work as well, or as cheaply. Does anybody have a recommendation for tablets of this size, or perhaps 18x24, and some simple software (PC) to get the data we need? Our image analysis software (Optimas) will probably do it, but is a bit of overkill and I figure there must be something simple out there somewhere.
Thanks in advance --------------------------------------------- Rod Dilley Baker Medical Research Institute PO Box 348, Prahran 3181, Victoria, AUSTRALIA fax: +613 9521 1362 email: rod.dilley-at-baker.edu.au ---------------------------------------------
It works well, alone or in conjunction with serum, depending on the application and the nature of the secondaries. Start with a 2% (total protein) solution.
The only drawback is the thick nature of the stock gel. You will have to weight it.
good luck
Ramin Rahbari Tel. 734-622-3383
} -----Original Message----- } From: loudy.d-at-pg.com [SMTP:loudy.d-at-pg.com] } Sent: Thursday, April 23, 1998 12:25 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: cold water fish gelatin } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Does anyone know starting recipes for using cold water fish (teleost) } gelatin } as a blocking agent for immunocytochemistry? Does it work? Any } pitfalls? } We have a particularly sticky primary with lots of small punctate } background } appearing..controls with secondary alone are clean...presently using } 0.5% BSA } with 1% goat serum as block 1hr r.t....its not stopping it???? }
To all who have thus far replied and requested feedback regarding my query about microscopy facility revenue. I can't afford to FAX all the copies that I have received at this point but will mail them to all who have asked once they are compiled.
I will also provide a synopsis to submit to the listserver when I have had the chance to go through all the responses.
Incidently, all costs associated with the facility (salaries, repairs, supplies, incidentals) must be recovered via users fees according to the decree. Is salaries alone were covered, I could actually have enough money to repair or replace some of my more unreliable equipment.
Best Regards,
Kirk J. Czymmek Biological Electron Microscopy Facility University of Delaware Newark, DE 19716 kirk-at-udel.edu
} } Dear Collegues, } } I need information to educate the administration on how close to "cost } neutrality" a microscopy facility can get. I currently supervise a } facility with a part-time employee to assist in repairs. Much of the } equipment is 20-25 years old and is constantly breaking down. I am the } sole revenue generater with a few students who have been trained to use } the microscope and have beamtime hours. Within a year or so of operation } the facility went from generating $0.00 in revenue to ~$60K maybe } 70K/year. This doesn't pay all the bills of an EM Facility with 2 TEMs and } an SEM as well. My question is, is their any facility that is COMPLETELY } cost neutral with revenue only for service work provided? Are the numbers } I describe for a medium sized University reasonable? I would greatly } appreciate any input. If their is a University that spends only what they } bring in for doing samples in a multi-user environment please contact me I } be happy to know how you did this. } } Best Regards, } } Kirk J. Czymmek, Ph.D.
This is what I've been faced with for the last four years. I was able to work out a deal with the enginnering school to fund a portion of my expenses (diminishing to ~$50K/year) for classroom activities (lecture/lab for a variety of courses) with the remainder of the roughly $120K budget made up by direct charge-back to sponsored projects. I've been able to make it so far (barely), but my biggest problem is equipment replacement (which we all know is very expensive!). I think I'm currently losing some high end work to other installations with better equipment, but I hope to upgrade soon.
For reference I have these income components for the lab: Educational mission Sponsored project charge-back Unsponsored projects (gets taken from educational mission funds) Off campus "clients"
I'd like to secure some "harder" monies but those days are all but gone.
Let me know if you can come up with a better way...
Brian
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax) "Be well, do good work, and keep in touch"
I have a Philips 300 that I need to sell. She has been my teaching scope for ten years but non-the-less well cared for. It has the DP and backing tank up grade as well as a plethora of spare parts.
No reasonable offer will be refused... the esteemed Dr. Mark Farmer from the University of Georgia finished up his thesis work with this instrument, so just think of its historical value alone!
The instrument is located in the Electron Imaging Facility at Rutgers University. Please contact either through E-mail or phone {732-445-5308 est}
Wet, dreary, traffic, strange smells, thats life in the Garden State.
John Grazul Rutgers University Electron Imaging Facility
The 2-methyl butane is actally the cryogen (freezing agent)and the LN2 is what is used to bring it down the the proper temperature. The reason that agents like 2-methyl butane, propane, ethane and freons etc. are used is that they have a higher boiling point than LN2 (-196 oC). LN2's low boiling point creats a vapor barrier (Leidenfrost phenomena) to form around tissues that are immersed directly into it which prevents the rapid conduction of heat from the tissue to the cryogen. The best cryogens have high boiling points and and low freezing points.
hope that is the answer to you question
-- Begin original message -- } } Could someone tell me what exactly is the role of 2-methyl butane when used } in } snap freezing tissue in liquid nitrogen? I appreciate any insight ... } } Andrea Hooper } Dept of Pathology } NYU Medical Center
-- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
We are doing ICC of unfixed cell bits (synaptic densities) stuck to glass. In spite of blocking with bacitracin and BSA (after sticking the bits to glass freshly cleaned with conc nitric acid), there is a scattering of colloidal gold on the glass (but virtually no stuck gold with the seconday/gold conjugate alone). I am wondering if anyone has come across a blocking or glass cleaning regime that can be applied after sticking the cell bits to the glass that will inhibit the primary antibody from sticking to the glass and keep background under control. Many thanks....Tom Reese
A few hours after posting this last week we had a major power failure and our server was out many days. I have had some complaints of unavailability, and repost. Please excuse any inconvenience. ****************************************** Fellow list members, BibMic - A bibliography of books relating to Materials Microscopy, previously published on paper in Materials Characterization 36(1996)105 is now available on the net at http://bibmic.metalmat.ufrj.br It lists over 1000 books, and is searchable by author, title and keywords. Hope you find it useful Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
I have lost track of the vendor for my favorite 3 1/4 X 4 1/4 negatives sleeves. The package has the name RPS, catalogue # S-00650, Made in Japan, Plastine Film Preservers, Acid Free. I would appreciate learning who distributes these. I am not interested in any of the other brands or types. Thank you.
} } Could someone tell me what exactly is the role of 2-methyl butane when used in } snap freezing tissue in liquid nitrogen? I appreciate any insight ... } } Andrea -
I presume you're talking about plunge freezing. If you do this directly into liquid N2, the N boils, producing a layer of gaseous N around the tissue. That's a good insulator, so you get slow freezing. So you use an intermediate coolant that will stay liquid up to ~-80, for rapid heat transfer. When I was still in the lab (I'm retired) we used Freon 22, which is no longer permitted, or propane, which explodes if you're careless.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Dear Henk, } } I believe that TEM film has a density range of about 2.8-2.9. If this is } true, any scanner with a range of over 3 should work. } This is true for most EM images; however, ED patterns can have higher OD's. For quantitating ED intensities it is better to have as great a dynamic range as possible. Yours, Bill Tivol
Does anyone know of a book or a web site that is essentially an atlas of bacterial ultrastructure. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Assistant Director, The Biotechnology Program PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Responding to the message of {3.0.2.32.19980423103550.00701d5c-at-itsa.ucsf.edu} from Larry Ackerman {mishot-at-itsa.ucsf.edu} : } } } I have lost track of the vendor for my favorite 3 1/4 X 4 1/4 negatives } sleeves. The package has the name RPS, catalogue # S-00650, Made in Japan, } Plastine Film Preservers, Acid Free. I would appreciate learning who } distributes these. I am not interested in any of the other brands or types. } Thank you.
Larry, RPS (Reeves Photo Sales, Inc.) is at 9000 Sovereign Row, Dallas, Texas 75247, as of 3-4 years ago. I don't have their phone number. They also make really nice 35mm negative jackets.
Good Luck,
Gib
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
We are looking at some cells grown on Falcon cell culture inserts (essentially polyethylene terephthalate membrane filters). We fixed and osmicated, embeded in an epon-like epoxy resin. When we cut 0.5 um thick semi-thin sections, they wrinkle up something fierce due to the membrane. This makes it impossible to get a good LM photo. Anybody have any tricks? Falcon's tech bulletin was not very informative.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
We are planning on conducting some elemental analyses of 400-year-old glass beads. Would ZAF, Proza or Bence-Albee be the preferred correction method? We have a Noran Voyager with v2.7 software.
TIA
Bob Wise
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
This centrifugation/fixation works. We do this then cut the slender tube with a razor blade above and below the buffy coat (on a piece of Parafilm), making a short log with open ends. Then with a paper clip or applicator stick (depending on the diameter of the tube) we push out the packed buffy coat. If it tends to fall apart, we then encase it in 1% molten agar to keep it together. (You can encase it without washing out the glut, but don't fix the agar-encased pellet in glut of the other solutions won't infiltrate properly. If it sticks together, you can skip the agar. We then process the pellet or encased cells as a piece of tissue.
S Miller
On Thu, 23 Apr 1998, James Ito wrote:
} Date: Thu, 23 Apr 1998 09:29:17 +0100 } From: James Ito {james.ito-at-dial.pipex.com} } To: Microscopy-at-sparc5.microscopy.com, } Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } Subject: Re: COLLECTING A BUFFY COAT FOR TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We do this occasionally in my lab. } } We spin the anti-coagulated blood in a narrow test tube, then carefully } remove as much as possible of the plasma without disturbing the buffy } coat. Buffered 2% Glutaraldehyde is then very gently layered on top and } the tube left to stand in the fridge for about a couple of hours. This } gives a buffy coat which is embedded in solid plasma and can be removed } from the tube with the help of a thin wooden stick or similar. The } resultant disc can then be trimmed and the pieces processed to resin as } you would for normal tissue. If you embed in flat moulds your specimen } will have orientated layers of plasma, platelets, white cells and red } cells. } } It's a simple and effective technique, with only one centrigugation to } give you a sample which is easily handled. The only downside is that } some drugs (e.g. aspirin, I believe) may inhibit the action of } glutaraldehyde on the plasma and prevent the conversion of the plasma } to a solid. } } } James Ito } Pathology Department } Royal Hospital for Sick Children } Yorkhill } Glasgow } Scotland. } ---------- } } From: Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } } To: Microscopy-at-Sparc5.Microscopy.Com } } Subject: COLLECTING A BUFFY COAT FOR TEM } } Date: 22 April 1998 21:50 } } } } Hi There, } } Looking for a good technique for making a pellet of buffy coat from } } normal blood. Would appreciate any help. Thanks. } } } } Winnie } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
On Wed, 22 Apr 1998 hoopea01-at-endeavor.med.nyu.edu wrote:
} } Could someone tell me what exactly is the role of 2-methyl butane when } used in snap freezing tissue in liquid nitrogen? I appreciate any } insight ... }
Replies received so far are absolutely correct and on the nail. So I'm simply adding a couple of details for the general readers ...
(1) Being old fashioned, I still call 2-methyl butane ISOPENTANE, which might help if you're doing a literature search;
(2) If you have things that don't like being dunked in a hydrocarbon (rubbers and some plastics) it is possible to get quite powerful cooling by using a roughly equal mixture of acetone and methanol, which goes down to at least -110^C and has a higher boiling point than the isopentane. The b.p. of isopentane is twenty-something, which would make it hard to store in Arizona and similar places warmer than the UK.
Mixtures naturally have lower freezing points than the pure solvents, which is the principle of Dowtherm, an industrial heat transfer fluid which is mixture of Naphthalene and Diphenyl Ether.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Your blocking sems to be fine since gold conjugates do not stick. Have you tried to do the complete incubation on glass without cell bits on the surface? If you get the same background problem, you may have to purify or further dilute the primary. Are you using additives to the incubation buffer for the primary or only for the secondary? If there is no background on glass without cell bits, could it be that antigen leaks from the unfixed material?
Regards, Jan
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Managing Director Costerweg 5, 6702 AA Wageningen The Netherlands
---------- } From: wise-at-vaxa.cis.uwosh.edu } To: Microscopy-at-sparc5.microscopy.com } Subject: ZAF, Proza or Bence-Albee? } Date: April 23, 1998 7:42 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } To all, } } We are planning on conducting some elemental analyses of 400-year-old glass } beads. Would ZAF, Proza or Bence-Albee be the preferred correction method? } We have a Noran Voyager with v2.7 software. } } TIA } } Bob Wise } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html } } I understand Bence-Albee is the best way to go for minerals, and glass (old or not) is essentially a mineral, so there you go.
Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane. Its easy to store in a lab a small gas cylinder with a blunt needle on a bit of tubing as the outlet. With little gas flow rub the needle over the small metal cup that is cooled by liq N2. Soon you will have a couple of ml of liquid propane. Do this in a fumehood, which is a good idea when using solvents too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** } } On Wed, 22 Apr 1998 hoopea01-at-endeavor.med.nyu.edu wrote: } } } } } Could someone tell me what exactly is the role of 2-methyl butane when } } used in snap freezing tissue in liquid nitrogen? I appreciate any } } insight ... } } } } Replies received so far are absolutely correct and on the nail. So I'm } simply adding a couple of details for the general readers ... } } (1) Being old fashioned, I still call 2-methyl butane ISOPENTANE, which } might help if you're doing a literature search; } } (2) If you have things that don't like being dunked in a hydrocarbon } (rubbers and some plastics) it is possible to get quite powerful cooling } by using a roughly equal mixture of acetone and methanol, which goes down } to at least -110^C and has a higher boiling point than the isopentane. } The b.p. of isopentane is twenty-something, which would make it hard to } store in Arizona and similar places warmer than the UK. } } Mixtures naturally have lower freezing points than the pure solvents, } which is the principle of Dowtherm, an industrial heat transfer fluid } which is mixture of Naphthalene and Diphenyl Ether. } } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+ } } }
Tom I have had similar problems with membrane filters of all kinds. I assume its due tho the differences in compression and stretching of the two materials. I did'n get rid of the wrinkles entirely but by embedding in a flat mold with the sample elivated off the bottom so that there was resin nearly equal above and below, the sections laid flatter and straighter.
Rick Vaughyn
PS Sometimes changing the hardness of the resin also helped, usually harder.
Dear Bob, I would try both and see if they differ. Generally, if there are heavier elements around, the proza correction is better at correcting the matrix absorbtion correction for the light elements. I have no experience with the Bence-Albee. It is very useful, with glasses, to run a well-characterized standard glass first, and try the different methods. You wrote:
} } To all, } } We are planning on conducting some elemental analyses of 400-year-old glass } beads. Would ZAF, Proza or Bence-Albee be the preferred correction method? } We have a Noran Voyager with v2.7 software. } } TIA } } Bob Wise } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
Frank writes ... } } } } I understand Bence-Albee is the best way to go for } minerals, and glass } (old or not) is essentially a mineral, so there you go. }
Bence-Albee was a data reduction method for older computers ... e.g., a DEC PDP-5 with 4kb memory. It was a look-up table method of reducing elemental x-ray intensities to wt% oxides in the context of oxides. It was hardly computer intensive ... and many assumptions were made regarding simple hyperbolic modeling of the look-up beta factors. These assumptions hold true for select groups of elements, but many problems exists with the hyperbolic model. The other thing to realize is that the look-up table was created with ZAF ... that is, ZAF correction factors best fitted to the hyperbolic model. This would imply you'd do better with ZAF and no hyperbolic assumptions. Now ... lets talk about which "ZAF" ...
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - http://darkwing.uoregon.edu/~mshaf/ Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu or mshaf-at-oregon.uoregon.edu
I notice that propane is actually somewhat soluble in (liquid) water, and I wonder if it diffuses into the sample to cause artifacts, perhaps altering membranes or extracting lipid droplets, which are my main interest. I would assume that IF this happens, isopentane being less soluble in water would have less of this kind of effect. Has anyone noticed?
In what way is propane noticeably better? Just due to lower temp? ( I must have missed the discussion a few months ago)
Thanks Richard
} } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } Quite right, but now the discussion goes to: Which is the better cryoagent and that was a topic here a few months ago. Propane gas liquefied by cooling is a much, much better cryo-agent than is isopentane.
... } } (2) If you have things that don't like being dunked in a hydrocarbon } (rubbers and some plastics) it is possible to get quite powerful cooling } by using a roughly equal mixture of acetone and methanol, which goes down } to at least -110^C and has a higher boiling point than the isopentane. } The b.p. of isopentane is twenty-something, which would make it hard to } store in Arizona and similar places warmer than the UK. } } Mixtures naturally have lower freezing points than the pure solvents, } which is the principle of Dowtherm, an industrial heat transfer fluid } which is mixture of Naphthalene and Diphenyl Ether. } } } +------------------------------------------------------------------------+ } | Robert H.Olley
If you happen to have very little blood you can also use microhematocrit tubes. The tube is broken near the buffy coat and the cut tip immersed in glutaraldehyde until the buffy-coat hardens. It can then be removed and processed. For details of the method see: Moura Nunes. J.F., Soares, J.O. and Alves de Matos, A.P. - Micro-Buffy Coats of whole blood: a method for the electron microscopic study of mononuclear cells. Stain Technol. 54(5)257, 1979
A.P. Alves de Matos mtlopes-at-fc.ul.pt
---------- } From: Sara Miller {saram-at-acpub.duke.edu} } To: James Ito {james.ito-at-dial.pipex.com} } Cc: Microscopy-at-sparc5.microscopy.com; Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } Subject: Re: COLLECTING A BUFFY COAT FOR TEM } Date: sexta-feira, 24 de abril de 1998 2:03 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } This centrifugation/fixation works. We do this then cut the slender tube
} with a razor blade above and below the buffy coat (on a piece of } Parafilm), making a short log with open ends. Then with a paper clip or } applicator stick (depending on the diameter of the tube) we push out the } packed buffy coat. If it tends to fall apart, we then encase it in 1% } molten agar to keep it together. (You can encase it without washing out } the glut, but don't fix the agar-encased pellet in glut of the other } solutions won't infiltrate properly. If it sticks together, you can skip
} the agar. We then process the pellet or encased cells as a piece of tissue. } } } S Miller } } On Thu, 23 Apr 1998, James Ito wrote: } } } Date: Thu, 23 Apr 1998 09:29:17 +0100 } } From: James Ito {james.ito-at-dial.pipex.com} } } To: Microscopy-at-sparc5.microscopy.com, } } Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } } Subject: Re: COLLECTING A BUFFY COAT FOR TEM } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } We do this occasionally in my lab. } } } } We spin the anti-coagulated blood in a narrow test tube, then carefully } } remove as much as possible of the plasma without disturbing the buffy } } coat. Buffered 2% Glutaraldehyde is then very gently layered on top and } } the tube left to stand in the fridge for about a couple of hours. This } } gives a buffy coat which is embedded in solid plasma and can be removed } } from the tube with the help of a thin wooden stick or similar. The } } resultant disc can then be trimmed and the pieces processed to resin as } } you would for normal tissue. If you embed in flat moulds your specimen } } will have orientated layers of plasma, platelets, white cells and red } } cells. } } } } It's a simple and effective technique, with only one centrigugation to } } give you a sample which is easily handled. The only downside is that } } some drugs (e.g. aspirin, I believe) may inhibit the action of } } glutaraldehyde on the plasma and prevent the conversion of the plasma } } to a solid. } } } } } } James Ito } } Pathology Department } } Royal Hospital for Sick Children } } Yorkhill } } Glasgow } } Scotland. } } ---------- } } } From: Winnie Westbrook {ewwestbr-at-hsc.vcu.edu} } } } To: Microscopy-at-Sparc5.Microscopy.Com } } } Subject: COLLECTING A BUFFY COAT FOR TEM } } } Date: 22 April 1998 21:50 } } } } } } Hi There, } } } Looking for a good technique for making a pellet of buffy coat from } } } normal blood. Would appreciate any help. Thanks. } } } } } } Winnie } } } } } } } } } Sara E. Miller, Ph. D. } P. O. Box 3020 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-8735
I am trying to find an etchant for deep etching a Ni-W eutectic alloy (between 17.5 + 20.7 at% W).
Also, I would appreciate any suggestions for electrically thinning such a material.
thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
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Richard: Isopentane becomes very rubbery near liquid nitrogen temperature. The temptation is to warm it a little with a metal rod, reducing the temperature differential some more. Propane has greater freezing speeds making vitrification possible. These things where published 20 and more years ago when cryo fixation was developed. I would be surprised if propane penetrates the specimen to any extent because snap freezing is so rapid. Later the specimen is sublimed and the propane would be the first phase to go. Also propane has been used to widely that somebody would have noticed an effect on lipids - if that was greater than isopentane's. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** } I notice that propane is actually somewhat soluble in (liquid) water, and I } wonder if it diffuses into the sample to cause artifacts, perhaps altering } membranes or extracting lipid droplets, which are my main interest. I } would assume that IF this happens, isopentane being less soluble in } water would have less of this kind of effect. Has anyone noticed? } } In what way is propane noticeably better? Just due to lower temp? } ( I must have missed the discussion a few months ago) } } Thanks } Richard } } } } } "Jim Darley" {jim-at-proscitech.com.au} 04/24/98 04:43am } } } } Quite right, but now the discussion goes to: Which is the better cryoagent } and that was a topic here a few months ago. } Propane gas liquefied by cooling is a much, much better cryo-agent than } is isopentane. } } ... } } } } (2) If you have things that don't like being dunked in a hydrocarbon } } (rubbers and some plastics) it is possible to get quite powerful cooling } } by using a roughly equal mixture of acetone and methanol, which goes } down } } to at least -110^C and has a higher boiling point than the isopentane. } } The b.p. of isopentane is twenty-something, which would make it hard } to } } store in Arizona and similar places warmer than the UK. } } } } Mixtures naturally have lower freezing points than the pure solvents, } } which is the principle of Dowtherm, an industrial heat transfer fluid } } which is mixture of Naphthalene and Diphenyl Ether. } } } } } } +------------------------------------------------------------------------+ } } | Robert H.Olley }
"New Developments in Multi-photon Excitation Microscopy"
to be held, July 11 - 12, in Atlanta GA, prior to the MSA annual meeting
Organizer: Jim Pawley
Cost: Workshop $300 (all day, HANDS ON in PM) Symposium $50 (lectures only)
Multiphoton excitation microscopy is the latest wrinkle in 3D light microscopy. Compared to earlier methods such as widefield deconvolution and confocal microscopy, multiphoton excitation promises a number of advantages: reduced effects of specimen-induced light scatter, better dichroic efficiency and the confinement of the photodamage zone to only the plane of focus. These improvements add up to the ability to image features far below the surface of thick, transparent biological specimens and to do so with up to 1000x less phototoxicity and photodamage than is produced by "normal" confocal.
With 3D light microscopy being used increasingly to follow developments in living cells, embryoes and tissues and the recent introduction of several commercial multi-photon instruments, this field is displaying explosive growth. This Symposium will include not only talks by experts from all the leading labs in the field but will also include contributions of some who have just begun: people like you who have overcome the technical challenges of "femtosecond" lasers to get outstanding results!!
In addition, there will be talks by experts in fluorescent lifetime imaging, a techniques that makes even better use of the pulsed nature of the exciting light.
We expect to have at least 4 new, operating, multi-photon imaging workstations for use by those who sign up for the Workshop.
TO REGISTER, CONTACT
Annamarie Dowling, MSA Meeting Manager 7000 W. Southwest Highway Chicago Ridge, IL 60415 (708)361-6000 / FAX -6166 {MSA-at-tradeshownet.com}
AND SHE WILL SEND YOU AN APPLICATION FORM.
TENTATIVE PROGRAM
Saturday July 11, AM: The Foundations of Multiphoton
Dave Piston Vanderbilt University, "Two-Photon Excitation Imaging of In Vivo Glucose Metabolism"
Warren Zipfel Cornell University "TBA"
Dave Wokosin University of Wisconsin "Multi-mode Multi-photon Microscopy"
Vinod Subramaniam, Max Planck Institute, Goettingen "Multi-photon microspectroscopy and imaging of near-UV fluorophores by scanning near-field optical microscopy (SNOM)"
Sunney Xie Battelle Pacific Northwest Laboratory "Single-Molecule and Near-field Fluorescence Imaging with Two-Photon Excitation: New Damage Mechanisms"
Hans Gerritsen Utrecht University "Two-photon excitation fluorescence lifetime imaging."
Saturday PM: Manufacturer's presentations I
Saturday Evening: Reception.
Sunday AM: Doing it our way
Mark Cannell University of Aukland, NZ " Visualization and quantification of the transverse tubular system in Living cardiac cells by 2-photon microscopy."
Steve Potter, Cal Tech "Two photon time-lapse of dendritic spines"
Hadley Wilson Horch, Duke University "Setting up multi-photon microscopy: problems and solutions"
Montrose Johns Hopkins "Multi-photon imaging of pH and drug therapy in the gastrointestinal tract"
Rebecca Williams , Cornell University "TBA"
James Pawley University of Wisconsin "Photon efficiency and QE in 3D Microscopy"
FOR MORE INFO contact: James Pawley, 1117 Johnson Ave, Madison , WI. , 53706, USA, Tel: 608-263-3147, Fax: 608-265-5315 E-mail: jbpawley-at-facstaff.wisc.edu
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
Lou Ann -- *************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/CSMS/csms.html
Personal Home Page: http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/LAM.html
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I think that in general with issues of x-ray safety, the worst should always be assumed - so, x-rays may leak from an SEM.
In addition, I think there is also an argument to be made that SEMs could be more dangerous the TEMs. The higher the energy of an x-ray, the more penetrating it is and the longer the path over which it will deposit its energy. 30 kV x-rays could deposit more energy, and hence cause more damage, in a human body than 100 kV x-rays which having a shorter wavelength will interact less. I don't know what x-ray energy level is most dangerous to humans (perhaps somebody can enlighten) but I think it is wrong to assume that the higher the energy the more dangerous it is.
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
I am afraid that Larry's reply could not be "wronger". Some people will not know better and take the misleading information on board.
A TEM, when compared with a SEM potentially is the greater radiation risk. An SEM with a maximum of say 30kV is insignificant compared with a 200kV instrument. The wavelengths of electrons is changed with accelerating voltage but the K,L,M etc. X-rays generated by these electrons are in fact defined by the minimum electron energy required for their production. Regardless of the electron beam used to generate, the most powerful Cu X-ray is 8.978keV (the eV required to generate defines these X-rays). The minimum energy to generate the most powerful gold X-rays is 80.724keV. Those X-rays have far greater penetrating power and are rather more likely to leak from a badly modified instrument. The maximum X-ray generation occurs when the eV of the beam is about 2.8x the energy of a particular X-ray. So a 200kV instrument can almost reach maximum production of the most powerful K alpha Au X-rays. If only Al and C were used in an instrument (I know its not on) we would not generate such powerful X-rays. Unfortunately the light elements have very little X-ray stopping power. As a consequence in high kV instruments the manufacturers use in the very top aperture position an array of large, apertures with large, angled edges to stop the stray electron beam and the X-rays. Three or more apertures of different metals are used in that array to stop excess X-rays and to produce a "clean" beam for X-ray analyses. To do any harm X-rays must pass through the column. Happily that is easier prevented in the design of TEM and SEM than in X-ray diffractometers, which have far more movement of main components required. Any modern SEM or TEM, as provided by the manufacturers has so little leakage that microscopists should worry about the radiation received during a single flight at high altitude and not about sitting in front of these instruments. If you must modify an instrument, I suggest that an SEM is less likely to cause real grief than a TEM. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
-----Original Message-----
Postdoctoral/Senior technician opening:
An IMMEDIATE research position is available in the Department of Cell Biology, Baylor College of Medicine, Houston, TX, to join a team studying subcellular localization of steroid receptors and their cofactors, with a keen interest in nuclear/mitotic architecture. Moreover, particular interest will be centered upon covalent post-translational processing of wild-type and disease-associated receptors/cofactors in pathways leading to their turnover. In both cases, extensive effort will be focused upon live cell imaging using bioluminescence in multiple channels. High resolution studies at the ultrastructural level will focus on immunogold labeling and resinless-section microscopy. The successful candidate will have either/or extensive experience in cellular imagining, both by light and electron microscopic methodologies, strong skills in molecular cloning techniques and excellent communication skills (written and oral). This position can be appointed at the post-doctoral level, or as senior technician (BS/MS and } 7 years relevant experience). The annual salary range will be 25-30K, commensurate with experience.
These studies will make extensive use of the Department's Integrated Microscopy Core and the Computational and Visualization Biology Laboratory, containing state of the art hardware and software, including a new deconvolution-based wide-field optical workstation. High-speed data throughput is provided by a SGI Origin 2000 (equipped with 24 R10000 processors and } 1GB RAM), thus allowing nearly 'live' deconvolution imaging of four channels. Electron microscopy is supported by two TEMs, three ultramicrotomes and ancillary equipment. The Department offers a dynamic and intensive research environment with excellent opportunity for advancement.
Baylor College of Medicine is an Equal Opportunity/Affirmative Action/Equal Access Employer.
Interested applicants should forward (preferably electronically) a CV, letters of reference and research interests to:
Michael A. Mancini, Ph.D. Assistant Professor and Director Integrated Microscopy Core Department of Cell Biology Baylor College of Medicine Houston, TX 77030 mancini-at-bcm.tmc.edu 713 798 8592 713 790 0545 (fax)
Does anybody know if any or all the short courses at the M&M98 are still open? Or can anyone give me a phone contact? thank you Content-Type: image/gif Content-ID: {002401bd7146$92c6e680$6a8bd9ce-at-default}
I would think by now, most people on the Microscopy Listserver would know to go to the MSA WWW Site to find out information about M&M 98. But is seems that may not be the case. Here is the URL
http://www.msa.microscopy.com
To register or get information about the meeting contact the M&M98 Meeting Manager.
Contact information for both the Business Office as well as the Meeting Manager are also listed on the WWW site.
Microscopy & Microanalysis Meeting Manager
The Rebedeau Group 7000 W. Southwest Highway Chicago Ridge, IL 60415 Tel:(708) 361-6000; Fax (708) 361-6166 E-mail: MSAMeetingManager-at-MSA.Microscopy.Com
Mark Tobin wrote: ============================================ Has anybody heard of a material called Collodion being used for mounting materials for microscopy? I was asked by a colleague today and have not heard of it.
Any information, UK suppliers in particular, would be greatly appreciated == ========================================== Collodion™ is manufactured by nitrating with nitric acid and sulfating with sulfuric acid relatively ordinary cotton to make it soluble (as a result of the addition of soluble nitrate and sulfate groups). When dried, the solids become what is known as "gun cotton" because of its use as gun powder . We believe that Parlodion™ is very similar but it does have a different corporate and process origin and it unlikely to be exactly identical. At the very least, one could expect that the degree of nitrating and sulfating might vary between the several different manufacturers, thereby resulting in at least some subtle variation in the final properties. This could of course help explain why one researcher might have difficulty repeating someone else's work. However, most researchers report obtaining similar results, whether the application is for the casting of a film for TEM grids or for the making of replicas on metallurgical or ceramic types of surfaces.
We believe Collodion to be similar but not necessarily identical to the products called "Celloidin" and also, "LVN" , or "low viscosity nitrocellulose". And all of the mentioned products are supposed to be less explosive than the original "gun cotton".
These materials, e.g. all of the above mentioned products based on nitrocellulose nitrate, at one time, were also used widely as embedding resins, but have been replaced in most applications by more modern materials. However, for the embedding of really large samples, a technique called "double embedding" still requires a nitrocellulose type material. However, for specific kinds of samples, there might be a preference for one or the other of the nitrocellulose based materials.
Collodion, as well as the sister nitrocellulose based materials are flammable solids and require great care in their handling.
Disclaimer: SPI has offered both Parlodion and Collodion in both solid form as well as in solutions of 2% in amyl acetate for a long time. Both the solid material or solutions can be ordered either directly from SPI in the USA or from our distributor in the UK listed on our website. Our interest is to make sure that people understand that generically, these materials seem to be similar and while it is to the first approximation, if you have one "in hand", then you already have something that is very close to the other. Our other interest is to remind everyone that these are trade names of manufacturing companies and first use on a page should include the use of a "TM".
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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} I am afraid that Larry's reply could not be "wronger". Some people will not } know better and take the misleading information on board. } } A TEM, when compared with a SEM potentially is the greater radiation risk. } An SEM with a maximum of say 30kV is insignificant compared with a 200kV } instrument. The wavelengths of electrons is changed with accelerating } voltage but the K,L,M etc. X-rays generated by these electrons are in fact } defined by the minimum electron energy required for their production. } Regardless of the electron beam used to generate, the most powerful Cu X-ray } is 8.978keV (the eV required to generate defines these X-rays). The minimum
snips ..
} If you must modify an instrument, I suggest that an SEM is less likely to } cause real grief than a TEM. } Cheers } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } Great microscopy catalogue, 500 Links, MSDS, User Notes
Which actually misses the point I was trying to make:) I don't think that at any point I said that a TEM was safer than an SEM with regard to x-ray emissions. What I said, and the point that I was making, is that just because a SEM potentially generates less energetic x-rays than a TEM, it cannot be assumed that it is safer than a TEM (as Jim suggests).
As Jim points out, a SEM is just as capable as a TEM of generating plenty of x-rays. However, just because the HV is lower, it doesn't mean it is less dangerous. In addition, if people think that because of the lower HV, it is safer to modify SEMs, they will be heading for trouble - mainly because most people make an elementary mistake with regard to shielding for x-rays.
Many people think that to stop x-rays, you need a nice heavy metal, like lead. Which is correct as far as it goes but ignores the generation mechanism - electrons. Electrons, even at high kV have about as much penetrating power as a mosquito - high energy but no momentum. So, if you put a heavy metal in the way of an electron beam, it stops the electrons and generates lots of nice, highly penetrating heavy metal x-rays. The correct way to shield is a double layer of light elements and heavy elements - if electrons are going to hit anything, make sure it is aluminium or carbon. THEN, you have the lead to stop the Al x-rays - much safer and less lead needed.
The final point that I was raising is that not all x-rays are equally dangerous to people. I don't know the detail on this and enlightenment from someone who does would be useful. The energy of an x-ray (its wavelength) determines how strongly it interacts with matter. It is that interaction which causes damage. Something that passes straight through without interaction won't cause any damage - I guess we all have pleny of high energy neutrinos passing through us every day - with minimal effect. From this, very high energy x-rays, in themselves, would not be a direct danger. On the other hand low energy x-rays would be highly interacting and easily cause damage.
However, this observation need quantification - which I can't give. It would seem that there is an x-ray energy level which would cause most damage to a human. Below AND above that energy level would both be less dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will certainly be just as dangerous as TEMs. However, if the maximum danger energy level is 80 kV, then TEMs are a significantly greater potential danger.
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
{ Mark Tobin wrote: ============================================ { Has anybody heard of a material called Collodion being used for { mounting materials for microscopy? I was asked by a colleague { today and have not heard of it.
I have not heard to use Collodion as mounting material. However it can be used in replication techniques. To find out about this method you can read for instance:
D.H.Kay Techniques for E.M. (pages 60/61) G.Thomas Transmission E.M. on Metals (pages 134 & 179) A.M.Glauert in Practical Methods in E.M. Replica, Shadowing and Freeze-Etching Techniques (page 125)
Hans Brinkies Senior Lecturer SWINBURNE, University of Technology School of Engineering and Science Hawthorn, 3122, Melbourne - Australia Hbrinkies-at-swin.edu.au
Collodion is a 5% (I think) solution of celloidin in absolute alcohol and ether(diethyl). It is sold at this concentration for a number of simple applications. Fisher Scientific sells it, cat.#C408-500.
Scanning Probe Microscopy (SPM) Master Class Center for Interfacial Engineering University of Minnesota
DATE
Friday, May 22, 1998 and Saturday, May 23, 1998 Early registration deadline: April 24, 1998 Final registration deadline: May 8, 1998 Maximum Number of Attendees - 18
SPEAKERS
Professor Stuart Lindsay, Arizona State University Professor Andrew Hillier, University of Virginia Dr. Greg Haugstad, University of Minnesota (See enclosed interests/topics)
FORMAT
Friday morning (lunch provided) - speakers Friday afternoon - 4 hours on 6 experimental stations with expert assistance Saturday morning - 4 hours on 6 experimental stations with expert assistance
LOCATION
117/119 Smith Hall and 12 Shepherd Labs, University of Minnesota, Minneapolis, MN
PRICE
• Academic: $95 for early registration, $105 for late registration • Industrial: $295 for early registration, $325 for late registration • Price includes talks, instrument time with assistance, lunch
PAYMENT
With registration; print and mail attached registration card (TIFF).
CONTACT INFORMATION
Call Characterization Facility at 612-626-7594 to reserve spot. Attendees must bring samples and have prior SPM experience
HOTELS
Radisson Hotel 1-800-333-3333 615 Washington Avenue S.E. Minneapolis, MN 55414
Metrodome Days Inn 612-623-3999 2407 University Avenue S.E Minneapolis, MN 55414
Holiday Inn Metrodome 1-800-448-3663 1500 Washington Avenue So. Minneapolis, MN 55454
ABOUT THE SPEAKERS:
Stuart Lindsay is professor of physics at Arizona State University and Vice President of Research and Development at Molecular Imaging, a scanning probe microscope manufacturer. He received his Ph.D. in physics from the University of Manchester in 1976. In addition to numerous original-research articles, he has authored 16 review articles/book chapters and received 6 patents. His research interests are in biological physics: primarily biomolecular structure and electron transfer processes, as well as the development of new SPM instrumentation to enable these studies. Specific topics include the binding of regulatory proteins that switch certain genes on and others off. Prof. Lindsay will speak about in-fluid scanning force microscopy including MAC mode (intermittent contact under magnetic cantilever modulation), DNA imaging, and stiffness measurements within ordered fluid layers
Andrew Hillier is assistant professor of chemical engineering at the University of Virginia. He received his Ph.D. in chemical engineering from the University of Minnesota in 1995. His research interests focus on the chemical engineering aspects of materials and interfaces. This involves a range of topics including electrochemistry, materials chemistry, crystallization, catalysis, and the development and application of high resolution imaging techniques for in situ characterization of interfacial processes. Specific topics include the study of intermolecular forces and ordering phenomena in molecular systems, particularly during the crystallization of pharmaceutical products. Prof. Hillier will speak about resolution limits in scanning force microscopy, double-layer force measurements in electrolyte, and AC imaging with a thermally-driven cantilever.
Greg Haugstad is research associate in the CIE Characterization Facility at the University of Minnesota. He received his Ph.D. in physics from the University of Minnesota in 1991. His research interests are in polymer physics, with primary focus on energy-dissipative phenomena such as friction and viscoelasticity. Subtopics include the response of polymers to perturbative processes in SPM, i.e. changes in molecular-scale structure and/or properties induced by local shear/tensile forces. Another subtopic examines the mechanical/adhesive character of single or countable polymer molecules. A common thread in all of his work is the development of SPM methodologies to study nanoscale phenomena. Dr. Haugstad will speak about temperature-dependent scanning force microscopy of polymer friction/wear, and dynamic mode on polymer films with amplitude and phase imaging.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
URGENT: Application Deadline extended to May 8, 1998. ADJUNCT FACULTY NEEDED FALL Semester 1998 (Aug. 12 - Dec 18, 1998)
A San Joaquin Delta College Faculty member of the Microscopy Technology = Center is planning a Sabbatical Leave for the Fall '98 semester. San = Joaquin Delta Community College is currently searching for an EM = Instructor on an Adjunct or Temporary One Semester Contract. The = Contract will be at the discretion of the district. MINIMUM QUALIFICATIONS: Bachelor's Degree plus two years of directly related experience OR an = Associate Degree plus six years of directly related experience OR a valid = credential. DESIRABLE QUALIFICATIONS: Master's or PhD Degree in a Biological = Science; Experience in teaching Electron Microscopy COURSES TO BE TAUGHT: Introductory Techniques for Transmission Electron Microscopy (EM21) This is a lecture/lab course which includes beginning Transmission = Electron Microscopy dealing with the alignment and operation of the TEM, = vacuum techniques, photographic techniques, as well as the preparation of = particles and replicas for viewing in the TEM. Includes individual = training in the use of the TEM, preparation techniques, and written and = oral reports. (Lec - 2 hrs; Lab - 3 hrs/wk) Biological Ultrastructure (EM28) Course contents include specific information about the fine structure and = function of cells and tissue at the ultrastructure level. Videos, slides = and micrograph examination will be correlated with the lectures so that = students will learn to recognize the fine structure of cells and tissues = in relationship to their function. (Lec-2 hrs/wk) Current Microscopies: Optics, Theory and Application (EM30) Course contents include information related to the physical laws and = applications of the various types of current microscopies e.g. = TEM,SEM,FIB, AFM, and confocal microscopy, as well as other current = topics e.g. asbestos analysis, lab design, etc. (Lec - 2 hrs; Lab - 3 = hrs/wk) Advanced Techniques in Biological Electron Microscopy (EM37) Course contents include lecture and laboratory which covers advanced = techniques for biological specimen preparation in TEM including an = advanced research project.( Lec - 1 hr; Lab - 6 hr/wks) TERMS OF EMPLOYMENT: Adjunct / Non-tenured track position. APPLICATION: Contact Human Resources at 209/954-5056. DEADLINE: The Screening Committee will begin to review applications on = May 12, 1998
SJDC Microscopy Technology Program Information available at = http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html/
Human Resources San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207
id KAA21448; Mon, 27 Apr 1998 10:56:38 -0600 (MDT) Received: from nestor.NMSU.Edu by NMSU.Edu (8.8.8/NMSU-1.18) id KAA07444; Mon, 27 Apr 1998 10:57:46 -0600 (MDT) Received: from Microscope.nmsu.edu (microscope.NMSU.Edu [128.123.5.85]) by nestor.NMSU.Edu (8.8.6/8.7) with SMTP id KAA07872 for {microscopy-at-sparc5.microscopy.com} ; Mon, 27 Apr 1998 10:56:32 -0600 Message-Id: {3.0.2.32.19980427105655.0068f194-at-cnmailsvr.nmsu.edu} X-Sender: rtindell-at-cnmailsvr.nmsu.edu X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.2 (32)
Hi,
We are analyzing airborne particulates in an ongoing project and one question that has arisen concerns the possibility of separating the sample from the filter matrix. One particular sample was collected on a small fiberglass-type filter, making it nearly impossible to get into the fibers with the probe and get decent EDS done on particles. We have tried to take an overall spectrum of the filter, then subtract out a spectrum taken from an unused filter under identical beam/time/etc. conditions, but we're working blind as far as knowing if these results are giving us a real picture of what's there.
Is there a way of treating these filters, perhaps with solvents and sonication, that would give separate out enough particles to mount on a stub? We will try some things here, but someone with previous experience might be able to save us considerable time. The idea is to get an "overall" composition of airborne contaminants from the site, with additional analysis of individual particles, so hopefully what we separate from the filter would be representative of what was really in the filter. (Aye, there's the rub...)
Hopefully yours, Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Browsing in (Merriam) Webster's Collegiate and Hackh's Chemical dictionary reveals many interesting tidbits, though not necessarily useful ones. Of possible interest: collodion and pyroxylin are mostly the tri and tetranitrate of cellulose, gun cotton (main ingredient of smokeless powder) the hexanitrate. Terms first used ca. 1850-1880. No hint that collodion is a trademark, though Celluloid and Cellophane are or were. Use of ethyl alcohol + ethyl ether as solvents was possibly at one time superseded by the Cellosolve family (R-CH2CH2-OH), which combines the ether and hydroxyl functions in one molecule, but this group of solvents is now considered rather toxic.
Leonard R. Corwin Fort Dodge Animal Health Cyanamid Agricultural Research Center Quaker Bridge & Clarksville Roads PO Box 400 Princeton, NJ 08543-0400 609-716-2278 609-275-5239 fax corwinl-at-pt.cyanamid.com
{fontfamily} {param} New_Century_Schlbk {/param} RESEARCH ASSOCIATE POSITION AVAILABLE
A Research Associate position is available immediately to work on the 3-D structure of muscle and muscle/cytoskeleton proteins. One project aims to determine the structure of crossbridges in contracting insect flight muscle using fast freezing/freeze substitution procedures which trap force bearing myosin crossbridges with millisecond time resolution. Electron tomography is then used to produce 3-D images that preserve the variable structure of the crossbridges for subsequent analysis. Mechanical records on stiffness and tension are recorded up to the moment of freezing. Parallel time resolved X-ray diffraction data of the diffrent contractile states is also available with which to compare the EM data with native muscle structure. This project offers a unique opportunity to learn electron tomography, correspondence analysis of 3-D motifs and atomic modeling. The project involves primarily computing to obtain the 3-D images and computer modeling to interpret the structure. =20
The second project involves structure of cytoskeletal proteins on lipid monolayers, among these are alpha-actinins from many different species and tissues. The alpha-actinin crystals are large in extent and to date yeild structural information to at least 10 Angstroms resolution.=20 In addition, we have obtained crystals of a wide range of other proteins as well. Project is funded through January 2001. This project offers the opportunity to learn protein crystallization on lipid monolayers and methods for producing multiprotein complexes for 3-D imaging. =20
Applicants for both positions must have a PhD degree. Salary and relocation funds are negotiable based on relevent experience. =20
The successful applicant will become a member of the Structural Biology program at Florida State University that includes 3 protein X-ray crystallography groups, three macromolecular NMR groups, 2 EPR groups and 2 electron microscopy groups. Facilities for electron protein crystallography include a Philips CM300-FEG, a Philips CM120, 3 Gatan cryotransfer systems, a Gatan wide angle TV camera, a Perkin-Elmer PDS1010M densitometer, 2 surveying optical diffractometers. Inquiries and applications should be made to Dr. Kenneth Taylor, Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306-4380. Tel: (850) 644-3357. FAX (850) 561-1406. E-mail:=20 taylor-at-bio.fsu.edu. Applicants should provide a CV and the names and addresses of 3 former mentors or knowledgeable individuals who can provided reference letters. =20
Recent Pertinent Publications:
K. A. Taylor & D. W. Taylor. Projection image of smooth muscle {/fontfamily} {fontfamily} {param} Symbol {/param} a {/fontfamily} {fontfamily} {par= am} New_Century_Schlbk {/param} -actinin from 2-D crystals formed on positively charged lipid layers.=20 {italic} J. Mol. Biol. {/italic} {underline} 230 {/underline} , 196-205 (1993).
K. A. Taylor & S. Varga. Similarity of 3-D microcrystals of detergent solubilized (Na {smaller} + {/smaller} ,K {smaller} + {/smaller} )-ATPase from pig kidney and Ca {smaller} 2+ {/smaller} -ATPase from skeletal muscle sarcoplasmic reticulum. {italic} J. Biol. Chem.=20 {/italic} {underline} 269 {/underline} , 10107-10111 (1994). =20
H. Schmitz, C. Lucaveche, M. K. Reedy & K. A. Taylor. Oblique section 3-D reconstruction of relaxed insect flight muscle reveals the crossbridge lattice in helical registration. {italic} Biophys. J. {/italic} {underline} 67 {/underline} , 1620-1633 (1994). =20
H. Winkler & K. A. Taylor. 3D reconstruction by combining data from sections cut oblique to different unit cell axes.=20 {italic} Ultramicroscopy {/italic} {underline} 55 {/underline} , 357-371 (1994). =20
K. A. Taylor & D. W. Taylor. Formation of 2-D complexes of F-actin and crosslinking proteins on lipid monolayers: demonstration of unipolar {/fontfamily} {fontfamily} {param} Symbol {/param} a {/fontfamily} {fontfamily} {par= am} New_Century_Schlbk {/param} -actinin-F-actin crosslinking. {italic} Biophys. J. {/italic} {underline} 67 {/underline} , 1976-1983 (1994)
H. P. Erickson, D. W. Taylor, K. A. Taylor & D. Bramhill. Bacterial cell division protein FtsZ assembles into protofilament sheets and minirings; structural homologs of tubulin polymers. {italic} Proc. Nat. Acad. Sci. USA. {/italic} {underline} 93 {/underline} , 519-523 (1996)
Holger Schmitz, Mary C. Reedy, Michael K. Reedy, Richard T. Tregear, Hanspeter Winkler, Kenneth A. Taylor. Electron tomography of Insect =46light Muscle in Rigor and AMPPNP at 23=B0C. {italic} J. Mol. Biol. {/italic} 264, 279-301 (1996)
H. Schmitz, M. C. Reedy, M. K. Reedy, R. T. Tregear, H. Winkler, K. A. Taylor. Tomographic 3-D Reconstruction of Insect Flight Muscle Partially Relaxed by AMPPNP and Ethylene Glycol. {italic} J. Cell Biol. {/italic} {underline} 139 {/underline} , 695-707 (1997)
Kenneth A. Taylor, Jinghua Tang, Yifan Cheng & Hanspeter Winkler. The use of electron tomography for structural analysis of disordered protein arrays. {italic} J. Struct. Biol. {/italic} {underline} 120 {/underline} , 372-386 (1997)
} Mark Tobin wrote: } ============================================ } Has anybody heard of a material called Collodion being used for mounting } materials for microscopy? I was asked by a colleague today and have not } heard of it. } } Any information, UK suppliers in particular, would be greatly appreciated } ============================================
Collodion, as a mounting and particle manipulation medium for light and electron microscopy as well as elemental analysis, is widely discussed in the print Particle Atlas as well as the more recent Particle Atlas Electronic Edition. The sections are too extensive to quote here, but a couple of excerpts may be helpful:
} From Chapter 3, Particle Handling Techniques:
"Another mounting medium is flexible collodion. A dilute solution of particles in amyl acetate plus collodion is spread on a glass slide and allowed to dry, leaving the particles suspended in a clear film. Even though the refractive index of this medium is low, it disperses particles well. This medium is used primarily if there are very small particles ( {5 micrometers) to remove for further analysis. Collodion permits easy removal of single, very small particles by cutting out small squares of film after location microscopically."
Later, in the same chapter:
"A good example of how collodion can be used to separate particles from a matrix is a recent problem solved for a color TV manufacturer. Color TV tubes were being rejected because of tiny particles that produced green spots on the TV screen. A particle could not be picked up directly because it rested on a {1 micrometer aluminum film with a soft layer of phosphor beneath. The slightest touch of the needle would break the aluminum film, and the particle would be lost in the phosphor layer beneath. A 1 mm drop of collodion plus solvent was placed on the contaminated area and allowed to harden. The aluminum film, thus strengthened, was then lifted with a tungsten needle dipped in water. This wetted and held the loose phosphor beneath so that the collodion-aluminum film could be lifted, placed on a glass slide and softened with a drop of amyl acetate. An examination in transmitted light revealed a tiny hole in the aluminum film, with a 5 to 10 micrometer contaminant in the center. The particle was readily separated for microprobe and x-ray analysis with a No. 3 tungsten needle and just enough amyl acetate to allow the particle to flow slowly in the collodion."
For more information on the Particle Atlas Electronic Edition, contact me off-list at sshaffer-at-microdatawrae.com .
Hello, I am in search of a good protocol for the negative staining and TEM of microtubules assembled in vitro in the presence of taxol. Specifically, I need to know the following: Concentration of MTs/taxol Fix used and concentration (if any) Time of fix Duration of sample application to grid Duration of uranyl acetate application to grid
Please cc response to my email address: rcm7-at-psu.edu
Thanks, Rich Moore Penn State University rcm7-at-psu.edu
#################################################### Rosemary Walsh Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:rw9-at-psu.edu ####################################################
Larry Stoter wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am afraid that Larry's reply could not be "wronger". Some people will not } } know better and take the misleading information on board. } } } } A TEM, when compared with a SEM potentially is the greater radiation risk. } } An SEM with a maximum of say 30kV is insignificant compared with a 200kV } } instrument. The wavelengths of electrons is changed with accelerating } } voltage but the K,L,M etc. X-rays generated by these electrons are in fact } } defined by the minimum electron energy required for their production. } } Regardless of the electron beam used to generate, the most powerful Cu X-ray } } is 8.978keV (the eV required to generate defines these X-rays). The minimum } } snips .. } } } If you must modify an instrument, I suggest that an SEM is less likely to } } cause real grief than a TEM. } } Cheers } } Jim Darley } } } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } Which actually misses the point I was trying to make:) I don't think that } at any point I said that a TEM was safer than an SEM with regard to x-ray } emissions. What I said, and the point that I was making, is that just } because a SEM potentially generates less energetic x-rays than a TEM, it } cannot be assumed that it is safer than a TEM (as Jim suggests). } } As Jim points out, a SEM is just as capable as a TEM of generating plenty } of x-rays. However, just because the HV is lower, it doesn't mean it is } less dangerous. In addition, if people think that because of the lower HV, } it is safer to modify SEMs, they will be heading for trouble - mainly } because most people make an elementary mistake with regard to shielding for } x-rays. } } Many people think that to stop x-rays, you need a nice heavy metal, like } lead. Which is correct as far as it goes but ignores the generation } mechanism - electrons. Electrons, even at high kV have about as much } penetrating power as a mosquito - high energy but no momentum. So, if you } put a heavy metal in the way of an electron beam, it stops the electrons } and generates lots of nice, highly penetrating heavy metal x-rays. The } correct way to shield is a double layer of light elements and heavy } elements - if electrons are going to hit anything, make sure it is } aluminium or carbon. THEN, you have the lead to stop the Al x-rays - much } safer and less lead needed. } } The final point that I was raising is that not all x-rays are equally } dangerous to people. I don't know the detail on this and enlightenment from } someone who does would be useful. The energy of an x-ray (its wavelength) } determines how strongly it interacts with matter. It is that interaction } which causes damage. Something that passes straight through without } interaction won't cause any damage - I guess we all have pleny of high } energy neutrinos passing through us every day - with minimal effect. From } this, very high energy x-rays, in themselves, would not be a direct danger. } On the other hand low energy x-rays would be highly interacting and easily } cause damage. } } However, this observation need quantification - which I can't give. It } would seem that there is an x-ray energy level which would cause most } damage to a human. Below AND above that energy level would both be less } dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will } certainly be just as dangerous as TEMs. However, if the maximum danger } energy level is 80 kV, then TEMs are a significantly greater potential } danger. } } -- } Larry Stoter } 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK } email: LPS-at-teknesis.demon.co.uk } Phone/Fax: +44 (0)1825 767967
Larry,
The main point that you have missed is that there is virtually nowhere for x-rays to escape from an SEM. The chief point of generation is the gun area and I have not seen any with less than about 1/2" of steel (often 301 stainless) that must be penetrated. 30keV won't do it.
By the time you get down to the chamber, you are generating very few x-rays.
You know the x-ray film that the dentist exposes through your cheek, Jaw and teeth in 1/25 to 1/10 second? He's using a 70keV 15-25mA source. I'm not sure what the target metal is, but it may be copper. When I use that film to align a wavelength spectrometer, I am exposing it at a distance of about a foot from the beam with an absorbed current of 100nA on copper at 30keV. (This is a very high current for an SEM. A typical current for EDS work is 200-400pA and high res imaging is 1-10pA.) exposure is on the order of 5-10 MINUTES!
Most SEMs don't have anything but steel around the chamber. Some have glass ports for specimen airlocks, but the HV is generally interlocked. I had a customer in Canada who made a Plexiglass port and often ran 12-18 hour EDS scans. He put a film badge inside the metal cover for this port and left it there for a month. He got no report of dosage back after turning in the badge.
Some day I plan to survey an SEM with a light element detector, then survey the color monitor that goes with the EDS. I'm betting that there is nothing detectable from the SEM and considerable emission from the monitor. Any takers?
TEMs ARE inherently dangerous because they CAN generate very hard x-rays that are difficult to contain, and you are looking directly at the impact point of a lot of very energetic electrons. The column is also far more complex, making it harder to shield.
You're correct about harder x-rays spreading their energy over a greater volume of tissue, but you miss the point that the soft x-rays can't leave the system. Your color TV at home is probably a lot more dangerous than any SEM I've ever seen.
Ken Converse Quality Images Delta, PA third party SEM service
} The main point that you have missed is that there is virtually nowhere } for x-rays to escape from an SEM.
want a bet:)
} The chief point of generation is the gun area
so what do all those electron streaming down the column do when they hit apertures, valves, etc. Create fluffy bunnies?
} and I have not seen any with less than about 1/2" of steel } (often 301 stainless) that must be penetrated. 30keV won't do it. } } By the time you get down to the chamber, you are generating very few } x-rays.
I'd agree that the greatest intensity of x-rays is likely to be generated around the gun - SEM or TEM. However, x-rays can be generated elsewhere. And remember this question related to modifications - if somebody starts making modifications anywhere they risk letting x-rays out.
} You know the x-ray film that the dentist exposes through your cheek, Jaw } and teeth in 1/25 to 1/10 second? He's using a 70keV 15-25mA source. } I'm not sure what the target metal is, but it may be copper. When I use
I know my dentist runs for cover everytime he x-rays my teeth. I can't help wondering as I sit there with the tube against the side of my head and the dentist 20 feet away behind a lead-lined wall if there is an international conspiracy by dentists to take over the world:)
} that film to align a wavelength spectrometer, I am exposing it at a } distance of about a foot from the beam with an absorbed current of 100nA } on copper at 30keV. (This is a very high current for an SEM. A typical } current for EDS work is 200-400pA and high res imaging is 1-10pA.) } exposure is on the order of 5-10 MINUTES!
But you keep assuming that everything is working correctly, in an unmodified SEM. I bet the guys at Three Mile Island just KNEW there was nothing to worry about.
} Most SEMs don't have anything but steel around the chamber. Some have } glass ports for specimen airlocks, but the HV is generally } interlocked. I had a customer in Canada who made a Plexiglass port and } often ran 12-18 hour EDS scans. He put a film badge inside the metal } cover for this port and left it there for a month. He got no report of } dosage back after turning in the badge.
Which I for one find very worryinging.
} Some day I plan to survey an SEM with a light element detector, then } survey the color monitor that goes with the EDS. I'm betting that there } is nothing detectable from the SEM and considerable emission from the } monitor. Any takers?
Try leaving your film badge on top of the TV at home over the week end. A standard SEM will certainly allow fewer x-rays to escape than any TV or monitor. However, we aren't talking about standard SEMs - we are talking about ones that users modify.
Which reminds me - if you ever get the chance, try a radiation check in a long haul airliner. I had a colleague who one took a flight from Germany over to California, to do some radiation checks on an instrument before it was shipped to Germany, He hand-carried his monitoring instruments in the cabin. In mid-Atlantic, the inside of the plane broke the German radiation exposure limits - not by just a little bit but by a factor of more than double.
} TEMs ARE inherently dangerous because they CAN generate very hard x-rays } that are difficult to contain, and you are looking directly at the } impact point of a lot of very energetic electrons. The column is also } far more complex, making it harder to shield.
Please, point out precisely where I said that TEMs weren't dangerous. I'm making a point that SEMs as well have the potential to be dangerous and that dismissing this danger because the kV is lower and won't harm you, might just be a triffle foolish. The english language is actually capable of greater subtleties than simple right/wrong arguments.
} You're correct about harder x-rays spreading their energy over a greater } volume of tissue, but you miss the point that the soft x-rays can't } leave the system.
In principle, always provided that the original manufacturers configuration has not been modified - which is where the discussion started.
} Your color TV at home is probably a lot more } dangerous than any SEM I've ever seen. } } Ken Converse } Quality Images } Delta, PA } third party SEM service
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
I wonder if somebody out there can help. We are looking for a power regulator board for a Cambridge S200 SEM, part number 852594. Leo can make one up for us, but it will take up to six months to deliver as it is an old model. Please contact us if you can supply.
Luc Harmsen Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Looking for source/supplier to get Freon 12. This is required for HT Tank } of JEOL TEM 2000FX. Any help will be greatly appreciated. } } Zia ur Rahman } Materials Characterization Facility, } University of Central Florida, } Orlando, Florida, } USA } } zur-at-mmae.engr.ucf.edu } Hi,
The original freon gas used in the gun and HT tank of JEOL microscopes is no longer used as it is a CFC and environmentally damaging. JEOL reccomend that it be replaced with SF6 (sulphur hexaflouride) gas and certainly contacted UK users with details of a modification to do this. The SF6 has to run at a slightly higher pressure: 0.18 to 0.20 kg/cm2 in the tank and 2.8 to 3.0 kg/cm2 in the gun for our 2010 on SF6 compared to 0.05 to 0.15 kg/cm2 in the tank and 1.75 to 1.95 kg/cm2 in the gun for freon. This just involves a change of the gas fittings on the tank to enable a SF6 cylinder to be used for the gas fill, however, the gun requires a change in gas fittings, a new (higher range) pressure gauge and replacing the overpressure valve (or fitting one if not fitted). You also need to be careful when checking for gas leaks as the pressure is higher then previously used, make sure that all pipes and fitting that you reuse are good enough for the higher pressure.
Either contact JEOL or sort it out yourselves, we have converted 2 JEOL 4000s and a 2010 successfully and will convert our 200CX when it next needs to be serviced in either the tank or gun. We have found that the characteristics of the HT have changed a little. If we have a flashover there is no longer a carbon track to be cleaned but the HT set prefers to be allowed to recover for a while before applying the maximum voltage again (usually 10 mins or maybe 1 hour for large flashover at 400KV).
Make sure that you read the safety data for SF6 gas, before you convert your machine, and take the appropriate local action. A local (to you) company makes a very good leak detector for freon or SF6. Leak-seeker model L-790a from CPS Products Inc. Hialeah, FL 33013 (Phone (305) 687-4121). (No interests in JEOL or CPS)
Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
We are analyzing airborne particulates in an ongoing project and one question that has arisen concerns the possibility of separating the sample from the filter matrix. One particular sample was collected on a small fiberglass-type filter, making it nearly impossible to get into the fibers with the probe and get decent EDS done on particles. ... Is there a way of treating these filters, perhaps with solvents and sonication, that would give separate out enough particles to mount on a stub? ---------------------------------------------------------------------------- ------------------------------- You mentioned that this is an ongoing project so I assume you will be taking more samples. I would highly recommend that further sampling for airborne particulate be done with polycarbonate membrane filters. Gelman and Nuclepore are two brand names that come to mind. The environmental services labs which monitored for asbestos used these extensively. The polycarbonate membrane is solid film with sub-micron, radiation induced holes etched in them which makes them ideal for generating carbon film replicas for TEM analysis. The particles of interest remained embedded in the carbon film while the filter was dissolved away. In contrast, larger particles can be analyzed directlyon the filter in the SEM or easily picked off of the filter surface.
Regarding your fiberglass filter, I'm sorry I can't help you too much. Whatever you know about the air particulate should guide you. Is it mineral or organic?
Dear Larry, Ken, et al., } } } The chief point of generation is the gun area } The chief points of generation in our high-voltage TEM are below the accelerator between the intersystem valve and the 1st condenser aperture.
} so what do all those electron streaming down the column do when they hit } apertures, valves, etc. Create fluffy bunnies? } Mostly, they create brehmsstrahlung (continuous energy spectrum) radiation.
} } and I have not seen any with less than about 1/2" of steel } } (often 301 stainless) that must be penetrated. 30keV won't do it. } }
The half-thickness for absorption is on the order of 100 mg/cm^2, which is about 0.1 mm of steel. A negligable fraction of 30 keV x-rays will get through 1 cm.
} And remember this question related to modifications - if somebody starts } making modifications anywhere they risk letting x-rays out. } The modification must involve a sufficiently short path so that a significant portion of the radiation can pass through. A relatively thin port would do the job, however.
} I know my dentist runs for cover everytime he x-rays my teeth. I can't help } wondering as I sit there with the tube against the side of my head and the } dentist 20 feet away behind a lead-lined wall if there is an international } conspiracy by dentists to take over the world:) } The major reason (s)he runs for cover is that (s)he exposes many x-ray films per day; whereas you are subjected to this procedure about once per year. In your case, the benefits outweigh the exposure risks, but the dentist receives no benefit from x-ray exposure, so trys to keep that exposure "as low as reasonably achievable". } } But you keep assuming that everything is working correctly, in an } unmodified SEM. I bet the guys at Three Mile Island just KNEW there was } nothing to worry about. } They were more-or-less correct. The guys at Chernobyl probably felt the same way, but were definitely incorrect. The estimated biolog- ical effect of Three Mile Island is about one excess cancer in the total exposed population. TMI was a disaster only to the utility company. Chernobyl was the real disaster with many people killed from acute radia- tion sickness and millions more exposed to significant radiation doses, which will cause latent effects including cancer and genetic abnormalities.
} } Most SEMs don't have anything but steel around the chamber. Some have } } glass ports for specimen airlocks, but the HV is generally } } interlocked. I had a customer in Canada who made a Plexiglass port and } } often ran 12-18 hour EDS scans. He put a film badge inside the metal } } cover for this port and left it there for a month. He got no report of } } dosage back after turning in the badge. } } Which I for one find very worryinging. } We have put thermoluminescent dosimeters, sensitive to millirad doses, around our microscope and have seen only background radiation. There is more radiation from the concrete in the building than from the microscope. Since we are always concerned about the effects of the modifications we have made over the years, we have taken care not to provide leakage pathways and have monitored this closely. However, lack of this sort of attention can, indeed, lead to situations where there are radiation leaks. Yours, Bill Tivol
Dear Larry, et al., } } } A TEM, when compared with a SEM potentially is the greater radiation risk. } } An SEM with a maximum of say 30kV is insignificant compared with a 200kV } } instrument. The wavelengths of electrons is changed with accelerating } } voltage but the K,L,M etc. X-rays generated by these electrons are in fact } } defined by the minimum electron energy required for their production. } } Regardless of the electron beam used to generate, the most powerful Cu X-ray } } is 8.978keV (the eV required to generate defines these X-rays). The minimum } Most of the x-rays generated are brehmsstrahlung x-rays, which have a continuous energy spectrum. A 200 keV electron can produce an x-ray with the same energy (it is stopped by a large mass--like a heavy atom--which conserves the momentum, and the energy is carried by the photon). The argument about characteristic x-ray generation, while correct, is not the whole story. Another complication is that characteristic x-rays are emit- ted isotropically, while brehmsstrahlung x-rays are mostly forward-directed. Since, however, electrons can be elastically scattered in various directions before producing brehmsstrahlung, the distrubution of such radiation produ- ced by a microscope depends on what the electrons are likely to hit as they go through the column.
} The correct way to shield is a double layer of light elements and heavy } elements - if electrons are going to hit anything, make sure it is } aluminium or carbon. THEN, you have the lead to stop the Al x-rays - much } safer and less lead needed. } Beryllium is also very good in this regard. One needs the lead to stop not only the Al x-rays, but the potentially more energetic brehms- strahlung x-rays.
} The final point that I was raising is that not all x-rays are equally } dangerous to people. I don't know the detail on this and enlightenment from } someone who does would be useful. The energy of an x-ray (its wavelength) } determines how strongly it interacts with matter. It is that interaction } which causes damage. Something that passes straight through without } interaction won't cause any damage - I guess we all have pleny of high } energy neutrinos passing through us every day - with minimal effect. From } this, very high energy x-rays, in themselves, would not be a direct danger. } On the other hand low energy x-rays would be highly interacting and easily } cause damage. } The half-thickness for the absorption of tens-of-keV x-rays in tissue is on the order of a centimeter. Any x-radiation which is pro- duced by a microscope will, therefore, penetrate far enough to lose most of its energy in the living part of a person, and would be almost comple- tely absorbed before passing through that person. A 1 MeV x-ray has a half-thickness of about 10 cm in tissue, so most of its energy will also be absorbed before passing through. These values come from a figure in Friedlander et al., Nuclear and Radiochemistry.
} However, this observation need quantification - which I can't give. It } would seem that there is an x-ray energy level which would cause most } damage to a human. Below AND above that energy level would both be less } dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will } certainly be just as dangerous as TEMs. However, if the maximum danger } energy level is 80 kV, then TEMs are a significantly greater potential } danger. } Unfortunately, the danger vs energy is a pretty flat curve with a broad maximum. Any scope (or other equipment) from which x-rays escape has the potential to be dangerous. All the more so since most of us work around the equipment for a large part of the day. Yours, Bill Tivol
could anybody send me an electronic version of the article "de Ruijter (1995) Imaging properties and applications of slow-scan charge-coupled device cameras suitable for electron microscopy, Micron, 26, 247-276" asap or indicate the web address where it can be found. Thanks a lot. With best regards,
Dmitry Cherny
Current address: MPI for Biophysical Chemistry, dept. of Molecular Biology am Fassberg 11, D-37077 Gottingen, Germany tel: +49(0) 551 201 1765; fax: +49(0) 551 201 1467 e.mail: dtcherny-at-mpc186.mpibpc.gwdg.de dtcherny-at-img.ras.ru
Wanted to put in my 2 cents worth before this thread closes. Obviously both sides have their points, and have made the larger, more important point, which is if you don't know how sharp the fangs and claws are, don't play with the cat. This goes for many pieces of equipment around the EM lab, not just limited to the electron microscopes but also including sputtering devices, ion mills, evaporators(electron beam especially), and critical point driers, to name only a few. All are capable of inflicting injuries as we have seen in the past. Repairs and modifications should be done by, or at least in consultation with experts such as Jim, Kenneth, Larry or the manufacturer, who recognize the dangers and can give advice to avoid them. A couple of additional things, I wanted to mention; 1. Any time that the high voltage exceeds 10 kV, regular glass is no longer capable of shielding against emitted x rays. Lead glass or metal is necessary. 2. It needs to be mentioned, although we all realize it, the ability to adequately detect radiation decreases as the energy decreases. A Geiger counter that is ok at 100kV does not usually truly indicate the magnitude of the radiation at 10 kV. Special equipment is needed at this level. I also understand film badges and dosimeters also fall off in their ability to detect low energy x-rays and special devices are needed. 3. Antidotally, I have never detected a radiation leak in a SEM but easily can find some area in most TEMs that will give some counts. (Just to add some perspective, you can usually detect some counts above background, around color computer monitors which usually operate around 30 kV and at about an order of magnitude higher current then SEM columns.) In the TEM's the radiation mostly emanates as beams. Usually the manufacturer covers these areas with lead shielding. This brings up the 4th item, make sure that all parts are returned to the microscope and in the proper position while doing repairs or cleaning. The innocent looking washer or plate may be shielding the manufactured included to prevent just these same radiation beams or leaks. Turning on a TEM without the lap shield in place or looking into the oil fill hole on top of an SEM gun while the HV is operating can subject you to unnecessary radiation exposure. We have several shields on an older microscope that can shift out of place easily during cleaning and need to be checked frequently.
Dear Randy, I don't know of any way to dissolve fiberglass, short of HF, so you should use a dissolvable filter in the first place. Nucleopore filters are my preference, since they are flat, as the particles sit up on top instead of down in the mesh and their EDS background is just polycarbonate (C and O). They can be dissolved, if necessary, in a Jaffe washer in chloroform for 48 hours. You wrote: } Hi, } } We are analyzing airborne particulates in an ongoing project and one } question that has arisen concerns the possibility of separating the sample } from the filter matrix. One particular sample was collected on a small } fiberglass-type filter, making it nearly impossible to get into the fibers } with the probe and get decent EDS done on particles. We have tried to take } an overall spectrum of the filter, then subtract out a spectrum taken from } an unused filter under identical beam/time/etc. conditions, but we're } working blind as far as knowing if these results are giving us a real } picture of what's there. } } Is there a way of treating these filters, perhaps with solvents and } sonication, that would give separate out enough particles to mount on a } stub? We will try some things here, but someone with previous experience } might be able to save us considerable time. The idea is to get an } "overall" composition of airborne contaminants from the site, with } additional analysis of individual particles, so hopefully what we separate } from the filter would be representative of what was really in the filter. } (Aye, there's the rub...) } } Hopefully yours, } Randy } } } Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } } rtindell-at-nmsu (work) } nrtindall-at-zianet.com (home) Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
Thanks for reminding us of the issue of brehmsstrahlung.
snips ...
} } However, this observation need quantification - which I can't give. It } } would seem that there is an x-ray energy level which would cause most } } damage to a human. Below AND above that energy level would both be less } } dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will } } certainly be just as dangerous as TEMs. However, if the maximum danger } } energy level is 80 kV, then TEMs are a significantly greater potential } } danger. } } } Unfortunately, the danger vs energy is a pretty flat curve with } a broad maximum. Any scope (or other equipment) from which x-rays escape } has the potential to be dangerous. All the more so since most of us work } around the equipment for a large part of the day. } Yours, } Bill Tivol
Which does add considerable weight to the point I was making - just because the kV of an SEM is lower than that of a TEM, it should not be assumed that it is less dangerous.
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
} Larry, } } The main point that you have missed is that there is virtually } nowhere for x-rays to escape from an SEM. The chief point of } generation is the gun area and I have not seen any with less than } about 1/2" of steel (often 301 stainless) that must be penetrated. } 30keV won't do it. } } By the time you get down to the chamber, you are generating very few } x-rays. } } You know the x-ray film that the dentist exposes through your cheek, } Jaw and teeth in 1/25 to 1/10 second? He's using a 70keV 15-25mA } source. I'm not sure what the target metal is, but it may be copper. } When I use that film to align a wavelength spectrometer, I am } exposing it at a distance of about a foot from the beam with an } absorbed current of 100nA on copper at 30keV. (This is a very high } current for an SEM. A typical current for EDS work is 200-400pA and } high res imaging is 1-10pA.) exposure is on the order of 5-10 } MINUTES!
Ken,
You and I come from the common point of ETEC SEMs. I try to use a wavelength alignment using 400nA, although that can be hard to achieve. At that point in the system, you are dealing only with a small subtended angle of the x-ray radiation emitted from the sample, after passing through a magnetic apeture that removes any electrons.
Regardless, consider the case where an ETEC has lost the small aluminum plug that covers the rectangular hole machined at the top of the objective lens. I would not leave that uncovered as it offers a path for a wide angle of x-rays emitted by the absorption of electrons in the electron-optics.
Regarding exposure to any ionizing radiation, such as x-rays, care must be taken to minimize long-term exposure to them. As an analogy, the current law suits brought by airline flight attendants regarding secondary tobacco should be seriously compromised by their constant exposure to the higher background radiation levels at the flight levels they encountered.
Ionizing radiation is an insidious problem, particularily at the energies present in an EM. It becomes not a problem of either-or, but a statistical problem of how-much, how-long. Normally unmeasured levels of radiation can over the long term produce potential problems.
Your later comments about the EDS monitor are correct. A typical survey of the equipment will show a substantial emission from the EDS monitor, and little or no emission from the SEM. However, you can not infer that this result means that there is no potential problem from the SEM.
Computer monitor manufacturers have recently realized the emissions from their equipment, and have taken steps to reduce them. SEM manufacturers have always been aware of the problems, and should have taken mechanical design steps to prevent a problem. But when those systems are being modified by the user, it is the user who must be aware of the problems inherent in those modifications.
} Most SEMs don't have anything but steel around the chamber. Some } have glass ports for specimen airlocks, but the HV is generally } interlocked. I had a customer in Canada who made a Plexiglass port } and often ran 12-18 hour EDS scans. He put a film badge inside the } metal cover for this port and left it there for a month. He got no } report of dosage back after turning in the badge. } } Some day I plan to survey an SEM with a light element detector, then } survey the color monitor that goes with the EDS. I'm betting that } there is nothing detectable from the SEM and considerable emission } from the monitor. Any takers? } } TEMs ARE inherently dangerous because they CAN generate very hard } x-rays that are difficult to contain, and you are looking directly } at the impact point of a lot of very energetic electrons. The } column is also far more complex, making it harder to shield. } } You're correct about harder x-rays spreading their energy over a } greater volume of tissue, but you miss the point that the soft } x-rays can't leave the system. Your color TV at home is probably a } lot more dangerous than any SEM I've ever seen. } } Ken Converse } Quality Images } Delta, PA } third party SEM service } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Hi - Perhaps I am missing something vital in my understanding of X-ray generation in EMs. Why should the bremsstrahlung be lower in SEM? I thought that bremsstrahlung is the result of electron interaction with the nucleus rather then electrons. A high continuum (versus defined X-rays) is typical of a spectrum generated by low atomic number elements. There is not much difference in the of elements used in SEM versus TEM.
Larry's argument that a SEM is potentially as dangerous as a TEM is at odds with my understanding of the subject and no valid argument has been advanced to change my view. I note that normally the various radiations are well contained in both instrument types, but consider what is happening within a 200kV TEM and a 30kV SEM.
The maximum energy of Pt (78; apertures) X-rays is 73.4keV and that of lead (82; shielding) is 88keV. It requires about 2.8x of the X-rays defining energy to maximise the production of those X-rays. In the 30kV SEM maximum emission of Zn (30; brass parts) with maximum of 10.4keV is possible; some emission of Mo (42; apertures) with maximum energy X-rays of 20.0keV. Clearly, a 200kV beam is sufficient to generate at lot of much more powerful - and I understand that these are also the most dangerous - X-rays.
If my understanding is wrong, still nothing would change to make the SEM as or more dangerous than the TEM. The TEM's powerful X-rays and primary and secondary electrons in turn exite other decreasingly powerful X-rays in a cascading reaction. Consequently the TEM not only generates powerful radiation within but also greater numbers of X-rays of numerous energy levels down to the ultra-soft.
As far as quantity of radiation is concerned: It is true that TEM's are commonly operated on 15 microamp emission and SEM at 80, but . . . . Without knowing actual numbers I am certain that a TEM operated at 200kV and 15uA produces not only rather more powerful X-rays but also greater total numbers of X-rays of all sorts of intensities when compared with a SEM operated at 30kV and 80 mA emission.
Many years ago I had to assume the additional role of an analyst. I am mostly self-tought in the relevant physics; I am happy to learn and not worried that I might be proven wrong. I think it is important that electron microscopists have some understanding of this topic. My conclusion is: If you are going to modify an SEM think and consult. If you are going to modify a TEM, think and consult twice. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au ***** } Thanks for reminding us of the issue of brehmsstrahlung. } } snips ... } } } } However, this observation need quantification - which I can't give. It } } } would seem that there is an x-ray energy level which would cause most } } } damage to a human. Below AND above that energy level would both be less } } } dangerous. Now, IF that maximum danger energy is around 10 kV, SEMs will } } } certainly be just as dangerous as TEMs. However, if the maximum danger } } } energy level is 80 kV, then TEMs are a significantly greater potential } } } danger. } } } } } Unfortunately, the danger vs energy is a pretty flat curve with } } a broad maximum. Any scope (or other equipment) from which x-rays escape } } has the potential to be dangerous. All the more so since most of us work } } around the equipment for a large part of the day. } } Yours, } } Bill Tivol } } Which does add considerable weight to the point I was making - just because } the kV of an SEM is lower than that of a TEM, it should not be assumed that } it is less dangerous. } } Regards, } } -- } Larry Stoter } 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK } email: LPS-at-teknesis.demon.co.uk } Phone/Fax: +44 (0)1825 767967 } } }
We are attempting to prepare TEM cross-section samples of LaAlO3 single crystal subtrate with a few thousand angstroms of YBCO film deposited on the surface. So far we haven't been successful using ion beam thinning techniques.
Any help would be greatly appreciated.
Thanks in advance
Fred Pearson email: eoptics-at-mcmaster.ca
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
We've been very successful preparing YBCO on CeO2 on sapphire using the small-angle cleavage technique (see M.W. Denhoff and J.P. McCaffrey, "Epitaxial Y1Ba2Cu3O7 Thin Films on CeO2 Buffer Layers on Sapphire Substrates", J. Appl. Phys. 70 (7), p. 3986 (1991)). There is an excellent recent update of the technique in: S.D. Walck, J.P. McCaffrey, "The small-angle cleavage technique: an update", Mat. Res.
Soc. Symp. Proc. Vol.480, pp.149-171 (1997).
If you can't get a copy of the MRS proceeding, drop me a note offline and I'll send you some material plus our newest video on the technique, featuring Igor and Dr. Cleavinstein!
Cheers John
John P. McCaffrey Institute for Microstructural Sciences National Research Council of Canada M-50 Montreal Rd. Ottawa, Ontario K1A 0R6 Canada
Tel: 613-993-7823 Fax: 613-990-0202 email: john.mccaffrey-at-nrc.ca ---------------------------------------------------------------------------- -- REPLY FROM: McCaffrey, John (IMS) Microsoft Mail v3.0 (MAPI 1.0 Transport) IPM.Microsoft Mail.Note
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Fellow Microscopists:
We are attempting to prepare TEM cross-section samples of LaAlO3 single crystal subtrate with a few thousand angstroms of YBCO film deposited on the surface. So far we haven't been successful using ion beam thinning techniques.
Any help would be greatly appreciated.
Thanks in advance
Fred Pearson email: eoptics-at-mcmaster.ca
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
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I have ordered a read write cd rom to back up my digital images. The unit I ordered is backordered for another month and I am reading some reports that it is not a good unit. I have a chance to change my order for another brand which is a La Cie 4X CD recorder, from Teac, I believe. Does anyone have any experience with this make our could anyone give me some advice or comments on what they are using. Thank you very much. Gary Zajic gzajic-at-amgen.com
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RE} TEM:YBCO Sample Prep 4/29/98
I made a similar cross-section sample with YBCO film on LaAlO3. The trick is ion-milling only from the back side of the film (substrate side). To do this, you need a pair of shields mounted on both top and bottom plates of the ion mill holder, leaving an opening about 90 degree. When mounting sample, put substrate side towards opening and film side away from opening. The ion milling time is much longer than normal, but it gives great results.
Chengyu Song National Center for Electron Microscopy Lawrence Berkeley National Laboratory 510-486-6751
by orac.early.com (8.8.7/8.8.7) with SMTP id QAA17562 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 29 Apr 1998 16:59:33 -0400 (EDT) Message-ID: {004a01bd73b1$1e052060$1cd6aacc-at-rafael}
I work in a materials testing lab (minerals, steels, refractories, polymers, papers and pigment coatings) using SEM, TEM and EDS methods. We are forging into the ISO Guide 25 culture and need to be involved in proficiency tests and/or round robins. We currently report elemental composition and particle size data. I've never been involved in this type of testing before and will appreciate info and comments about the process.
Nan Laudenslager Specialty Minerals, Inc. nhl-at-early.com Phone 610-250-3094 Fax 610-250-3206
This note is in response to a recent posting in this list server quoting sections of an April 20th Business Week article addressing the lifetime of data on digital media. The author of the posting indicated that CD storage was a popular choice for the readers of the list server, so I wanted to clarify some statements in the BW article.
First off, the quoted BW comments about the lifetime of CD media address CD-ROM, not recordable CD-R. The differences between the two media are significant (see http://www.kodak.com/daiHome/techInfo/permanence1.shtml for lots of information on the permanence, care, and handling of CD-R media).
Secondly, although some CD-ROM media have shown limited life in accelerated keeping studies, studies published by 3M substantiate an estimated life of their CD-ROM media of over 50 years (William P. Murray, NMLBITS, Newsletter of the National Media Laboratory, 2(2), 4, October 1992).
Thirdly, Kodak has invested considerable effort and expense to develop a CD-R medium with excellent data life which extensive accelerated testing allows us to estimate as exceeding 100 years when properly stored (Doug Stinson et al., NMLBITS, Newsletter of the National Media Laboratory, 9(1), 1, January/February 1995).
The bottom line is that data longevity is a media quality parameter. Like anything else, some CDs (and some CD-Rs) are made to last longer than others. If data longevity is important to you, then choose a supplier that meets your needs.
Job Vacancy in the Department of Materials Science and Engineering at Lehigh University
The Microscopy Center at Lehigh University is looking for an innovative individual to develop and maintain one of the top facilities in the nation for electron microscopy. Specific responsibilities include developing microscopy techniques related to computation and developing remote operation of microscopes for teaching and research. In addition, individual will maintain instrument and computer equipment and make networking and image transfer a standard for all our instruments.
To qualify, you should have a BS in computer science, electronics or electrical engineering and at least three years related experience. Experience with electron microscopy or other scientific instruments required. Good communication and interpersonal skills, a background in laboratory and classroom environment and PC/MAC software preferred. Overtime may be required.
For consideration, please send resume before May 15, 1998 to: Dr. Alwyn Eades, Materials Science and Engineering Department, Lehigh University, 5 East Packer Avenue, Bethlehem, PA 18015. Lehigh University is committed to recruiting, retaining, and tenuring women and minorities.
Sharon L. Coe Conference Coordinator Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem, PA 18015 Phone: 610/758-5133 Fax: 610/758-4244 e-mail: slc6-at-lehigh.edu http://www.lehigh.edu/~inmatsci/Microscourses.html
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have ordered a read write cd rom to back up my digital } images. The unit I ordered is backordered for another } month and I am reading some reports that it is not a good } unit. I have a chance to change my order for another } brand which is a La Cie 4X CD recorder, from Teac, I believe. } Does anyone have any experience with this make our could } anyone give me some advice or comments on what they are } using. Thank you very much. } Gary Zajic } gzajic-at-amgen.com } }
The stability and longevity of images stored on CD-R media is a constant worry both for myself and I am sure a good number of microscopists world-wide. We have been archiving images using CD-R for five years now and have approx. 25,000 images stored at present. In the archive I have had no failures yet, but a small number of discs which I have supplied to customers have failed within a year. The CD's in question were stored in normal conditions and the only difference was that they were carried around. There was no visible damage on any of the failures. The situation at present is that we are considering installing a CD duplicator to make back-up copies of the archive. It worries me when I see lifetimes of 50-100 years mentioned in relation to CD-R media. It places a great deal of faith in dye technology which in my experience should be treated with caution. Unwittingly people reading about lifetimes like this may store important data which may be irretrievably lost.
Colin Reid Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message-----
I am an honors student, whose project is on the growth mechanism of filiform corrosion in Aluminium. I'm finishing up my literature search, and as such am covering all my bases. Is there anyone, who is doing or has done work in this field who can recommend texts papers, internet sites, videos etc that they found useful and informative.
We are currently investigating the possibility of replacing our Cambridge S-250 SEM with a new instrument. All of my experience the past 22 years as a hands-on user has been with conventional SEM's equipped with tungsten filaments but not having cryo-stages. EDS has been an extensively used option. I welcome any comments from users or vendors comparing tungsten, LaB 6, and field emission electron beam sources. Our samples are 60-70% textiles, the remainder biologicals (fungi, botanical, etc.).
Thank you in advance.
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
Kovex Corporation announces the introduction of a new Confocal technology designed specifically for the industrial materials market. Please see our Web site at Kovexcorp.com or contact us directly at 612-486-9830/ kovex-at-spacestar.net for more details on KovexVision.
Bruce E. Batten, Ph.D. Kovex Corporation 3711 Lexington Ave. Shoreview, MN 55126 phone- 612-486-9830 fax- 612-486-9785
{HTML} Kovex Corporation announces the introduction of a new Confocal technology designed specifically for the industrial materials market. Please see our Web site at Kovexcorp.com or contact us {B} directly {/B} at 612-486-9830/ kovex-at-spacestar.net for more details on KovexVision.
} I have ordered a read write cd rom to back up my digital } images. The unit I ordered is backordered for another } month and I am reading some reports that it is not a good } unit. I have a chance to change my order for another } brand which is a La Cie 4X CD recorder, from Teac, I believe. } Does anyone have any experience with this make our could } anyone give me some advice or comments on what they are } using. Thank you very much. } Gary Zajic } gzajic-at-amgen.com }
We recently purchased a 4x12x CD-R from a different OEM (ClubMac) which also uses the Teac mechanism, and have had no problems with it. The system only a few months old, so I can't speak to long-term reliability or maintenance, but it was easy to set up and use and has yet to unsuccessfully burn a CD. We use Astarte Toast software from a PowerMac 8100/100 with an AV hard drive dedicated to CD mastering.
-Paul
Paul Voyles Loomis Laboratory of Physics University of Illinois at Urbana-Champaign 1110 W. Green St. Urbana, IL 61801 pvoyles-at-physics.uiuc.edu
I use a Yamaha CDR 4260, 2x4x6x. They make some of the most reliable drives on the market. "Smart and Friendly" uses the Yamaha drive for their product line.
} We are using a "Smart and Friendly" 2X6 CD recorder CD-R 2006 Plus and } find it ideal for backing up and saving digital images. } } --------------------------- } Miss Peta Clode } Zoology Department } LaTrobe University } Bundoora, Victoria } Australia. 3083. } } Ph (03) 9479 2177 / 2279 } Fax (03) 9479 1551 } --------------------------- } } On Wed, 29 Apr 1998, Gary H. Zajic wrote: } } } } } I have ordered a read write cd rom to back up my digital } } images. The unit I ordered is backordered for another } } month and I am reading some reports that it is not a good } } unit. I have a chance to change my order for another } } brand which is a La Cie 4X CD recorder, from Teac, I believe. } } Does anyone have any experience with this make our could } } anyone give me some advice or comments on what they are } } using. Thank you very much. } } Gary Zajic } } gzajic-at-amgen.com } } } }
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
CD-RW media are not readable on regular CD drives. Only CD-Rs can be read on regular CD drives.
Michael Bode Soft Imaging System Corp.
} ---------- } From: Peta Clode[SMTP:pclode-at-zoo.latrobe.edu.au] } Sent: Wednesday, April 29, 1998 9:16 PM } To: Gary H. Zajic } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: CD Burner } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } We are using a "Smart and Friendly" 2X6 CD recorder CD-R 2006 Plus and } find it ideal for backing up and saving digital images. } } --------------------------- } Miss Peta Clode } Zoology Department } LaTrobe University } Bundoora, Victoria } Australia. 3083. } } Ph (03) 9479 2177 / 2279 } Fax (03) 9479 1551 } --------------------------- } } On Wed, 29 Apr 1998, Gary H. Zajic wrote: } } } } ---------------------------------------------------------------------- } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } -. } } } } I have ordered a read write cd rom to back up my digital } } images. The unit I ordered is backordered for another } } month and I am reading some reports that it is not a good } } unit. I have a chance to change my order for another } } brand which is a La Cie 4X CD recorder, from Teac, I believe. } } Does anyone have any experience with this make our could } } anyone give me some advice or comments on what they are } } using. Thank you very much. } } Gary Zajic } } gzajic-at-amgen.com } } } } } }
Hello world, I would like any recommendations for the purchase of a new critical point dryer. We do biological materials (botanical, animal, microbiological, etc.), but not by the Gallon!! So, something nice and small and user friendly is preferable! Thanks!! Tracey Tracey M. Pepper Bessey Microscopy Facility Iowa State University Ames, IA 50011
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