Here are two adresses of Raith GmbH, in the USA and in Germany : Raith INC. 6 Beech Road Islip NY 11751-4907 Phone (516)224-1764 Fax (516)224-2620 E-mail 73164.1330-at-compuserve.com
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We have tested these two FE-SEM and we want to have other opinions about the differences between these two apparatus. Did you see a big difference in resolution at low energy (1-2 keV). What about the practical use of the "In Lens" detector of the S4700 ? Does it really bring more information than the others ? I think there are some 6700F in Far East (and in the States ?), but no one in Europe. We are interested in all you can say about these two SEMs.
Sincerely Yours
J. Faerber IPCMS-GSI 23, rue de Loess 67037 Strasbourg CEDEX FRANCE
Jacques We have also evaluated these two microscopes, and I would be interested to compare notes with you, but not online. Chris
} } Dear collègues } } We have tested these two FE-SEM and we want to have other opinions about } the differences between these two apparatus. Did you see a big difference } in resolution at low energy (1-2 keV). What about the practical use of the } "In Lens" detector of the S4700 ? Does it really bring more information } than the others ? I think there are some 6700F in Far East (and in the } States ?), but no one in Europe. We are interested in all you can say } about these two SEMs. } } Sincerely Yours } } J. Faerber } IPCMS-GSI } 23, rue de Loess } 67037 Strasbourg CEDEX } FRANCE } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)0 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
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Thanks to everyone who responded to my cry for help.
To summarize my findings and your suggestions/comments; Kodak has discontinued the sale of transparent sleeves, CAT 150 3812, however, that item with the same catalogue number is now being sold by Tiffen (1-800-368-6257). Of course these sleeves may also be available from other outlets, eg. National Graphic Supply (1-800-223-7130). A number of microscopists are using and have suggested I try the polyview sheets which hold 6 TEM negatives and fit nicely into a 3-ring binder. These sheets, cat. #1310/3406 are sold by Negafile (1-888-881-6435) but again may also be available from other outlets, eg. Ted Pella (916-243-2200), in Canada (1-800-243-7765).
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Senior technician/manager for Electron Microscopy Laboratory Managerial position for state-of-the-art electron microscopy facility within the Center for Biologic Imaging, University of Pittsburgh Medical School. Duties include overseeing and conducting the processing and preparation of samples for Transmission and Scanning Electron Microscopy and specialized techniques including cryosectioning and immunoelectron microscopy. Incumbent will help with training and assisting students and faculty on equipment, oversee general lab procedures including budget, supply, purchase orders, photography and quality control. Prior experience in electron microscopy essential. Bachelor’s or Master’s degree or equivalent desired. The successful applicant will be responsible for directing the 3 other staff in the EM component of the center The Center is a nationally recognized imaging resource using and developing a complete array of imaging technologies using both light and electron optics. More details about the center can be found at our web site (http://sbic6.sbic.pitt.edu):. This position is within the EM component of the CEnter. We have a full array of prep equipment from freeze fracture to ultracryomicrotomes. Our microscope base includes two transmission and two SEM scopes including a new field emission gun SEM. If you would like further information please contact Dr. Donna Beer Stolz or myself. Please send applications electronically to dstolz+-at-pitt.edu
--------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor, Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 University of Pittsburgh Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-8330
Does anyone know what happened to MSA Certification? I stopped at their booth in August at MSA 2000 and was assured that things getting "back on track" but so far I've heard nothing from them.
J. Faerber wrote: ================================== To D. Sicard
Here are two adresses of Raith GmbH, in the USA and in Germany : Raith INC. 6 Beech Road Islip NY 11751-4907 Phone (516)224-1764 { { {NOT CORRECT Phone (631) 224-1764 { { {CORRECT Fax (516)224-2620 { { {NOT CORRECT FAX (631)224-2620 { { { {CORRECT E-mail 73164.1330-at-compuserve.com ====================================== There has been an area code change and the old numbers absolutely won't work !
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 ######################## ============================================
Does anyone already have a prepared procedure according to paragraphs 2.2 and 2.3 of MIL-STD-883E, Method 2018.3 they would share with me? What I am wondering is whether this procedure is detailed (this knob, that button, etc.) or more general in nature?
Based on the responses I did not receive from my earlier post about quantitative measurement of SEM resolution, it looks like paragraph 2 cannot be met. It states a resolution of 250A or less. Semantically, I'm sure it meant 250A or a lesser number. A resolution of 250A or less would tend to mean a poorer resolution than 250A and hence a larger number. Probably picky on my part.
Would appreciate any inputs regarding this MIL-STD.
I've recently started lowicryl (K4M) processing of mouse tissue. I am having good success with skeletal muscle, heart, kidney and lung but stomach, spleen, pancreas and adrenal had large holes and cut poorly. In addition, a yeast pellet that was resuspended in 2% agarose prior to processing was not well infiltrated.
Processing procedure: 3% paraformaldehyde/0.5% glut. fixation for 60 minutes with rotation (RT) Wash 3X for tem minutes each with PBS buffer Dehydrate 30% ETOH for 30 min. at 0C 50% ETOH for 60 min. at -20C 80% ETOH for 60 min. at -35C 100% ETOH (freshly opened bottle) for 60 min. at -35C 100% ETOH for 60 min. at -35C ETOH/K4M 1:1 for 60 min. at -35C ETOH/K4M 1:2 for 60 min. at -35C K4M for 60 min. at -35CC K4M overnight at -35C K4M 6-8 hrs at -35C
Embed with fresh resin Specimens are constantly rotated in freezer. All alcohols and resins are prechilled prior to use. Resin is mixeed by gently bubbling Argon gas to dissolve initiator. Polymerize at -35C overnight using UV light Raise temp to 0C and polymerize additional two days with UV Polymerize at room temp. for two days using UV
Any thoughts on what I'm doing wrong or things that I can change would be greatly appreciated.
Tom Januszewski Senior Electron Microscopist UT Southwestern Medical Center at Dallas Dallas, TX 75390-9039 214-648-7291 FAX: 214-648-6408 Email: tom.januszewski-at-utsouthwestern.edu
I have an old business card (how old I don't know) for Raith. The contact name is George Lanzarotta at (516) 293-0870. The address is (was?) 70C Carolyn Boulevard, Farmingdale, NY 11735.
Good luck, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
-----Original Message----- } From: Ladd Research [mailto:sales-at-laddresearch.com] Sent: Thursday, November 30, 2000 4:26 PM To: microscopy-at-sparc5.microscopy.com
Could anyone provide me with a phone number, e-mail or fax number for the company "RAITH USA" It is a Germany Company and I thought there was a distrbutor in the US
Thanks
D. Sicard --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Just got a new vacuum evaporator after many years of using our Balzers 400 for the same purpose. Course now we don't have e guns and must use a little tungsten basket and a bit of Pt wire. I did this many years ago but now I am having a problem I don't recall encountering back then. The tungsten basket breaks just as the wire melts. I suspect the energy on phase transition is cooling the tungsten and causing it to break but I could be way off there. Anyone else see this problem and know the solution?
TIA
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Giselle You probably should check out the Journal of Fuel or "Fuel". "Carbon" occasionally has articles linking the mineral content of the feedstock with the resultant char. The Northern Carbon Research Laboratories, U. of Newcastle-upon-tyne used to publish regularly in this field. Penn. State also has a very active Fuel Science Dept. Good Luck.
J. Roy Nelson, Ph.D. Material Testing Laboratory mtl-at-njcc.com
giselle melville wrote:
} } Good afternoon } } My name is Giselle. I am a graduate student working on my thesis. My } thesis topic is "Methods of preparation of petrographic thin sections for } Electron Microscopy". I have a collection of thin sections (chert) and I am } preparing the sections for analysis by a TEM 1200. The thin sections are } initially prepared by methods Ultramicrotomy and Ion Milling. TEM } parameters used will be electron diffraction and elemental composition to } obtain identification of the mineral and microstructures. I need more } information (references) on recent literature (1998-2000) on my subject or } similiar work done using these methods for analyis by TEM. } } I need your help!!! } } Thank you for your time.
I am helping a friend find a scope. I came across a Zetopan that may fit his budget. I use a Nachet DIC for viewing living organisms and find it very satisfactory. Could anyone help him with the versatility of a DIC Zetopan. I know my Nachett works as a bright field, DIC and sort of polarizing scope lacking a rotating stage and 1/4 and 1/2 wave plates. It is the scope I use all the time. Unless I am doing darkfield.
Having never been around a Zetopan I can't really answer his questions.
Thanks Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} } I my quest to find a better 'scope I have come across (with the help } } of one of the list members) a Reichert Zetopan DIC scope that *might* } } be available at in my price range. Previously I was just looking for } } a nice phase contrast job. I have worked with phase before but not } } DIC and would like to solicit other opinions as to whether this is } } the right scope for me. This is going to pretty much blow my budget } } so I want to be sure. } } } } I will be using this primarily for observing living organisms } } (protozoa, algae, etc). I have been looking at a lot of photos of } } protozoa taken using Nomarski DIC. While the photos are striking, I } } keep asking myself "but what does it really look like". The problem } } is the false 3D effect. I'm never sure what the part of the shape is } } real and what is generated. Doesn't this ever bother you? } } } } I have been told that it is possible to adjust out the 3-D effect but } } I'm not sure what kind of image you are left with. I would assume } } something in-between bright field and DIC. Anyone have any experience } } with this? } } } } I think if the Zetopan is unable to easily switch between a DIC image } } and a bright (or dark) field image then I may pass on it. I need the } } more traditional illumination as well to see what the real shape is. } } It really makes the Zetopan too special purpose for someone like } } myself. I think I would probably be happier with a less } } sophisticated scope that can do phase, bright, and dark field (and } } maybe oblique lighting as well). } } } } I would appreciate any thoughts you all may have. } } } } Bill } } -- } } Bill Tschumy } } Otherwise -- Austin, TX } } bill-at-otherwise.com
Rick A. Harris wrote: ================================================================ Just got a new vacuum evaporator after many years of using our Balzers 400 for the same purpose. Course now we don't have e guns and must use a little tungsten basket and a bit of Pt wire. I did this many years ago but now I am having a problem I don't recall encountering back then. The tungsten basket breaks just as the wire melts. I suspect the energy on phase transition is cooling the tungsten and causing it to break but I could be way off there. Anyone else see this problem and know the solution? ================================================================ There really is no perfect "solution". The precious group metals tend to alloy quite readily with tungsten, Au and Pd very quickly, and I believe Pt is quite similar in behavior.
You can reduce the magnitude of the "problem" by making sure that when you install the tungsten basket, the wire itself is under zero stress. This might mean a bit more time adjusting the holders and posts than you might be doing now. Also, ball up the Pt wire and press into the very bottom of the basket, this will enable you to get the most evaporated in the shortest possible time period, before the wire basket breaks. And you want to use more of a "flash" evaporation than a long slow continuous kind of evaporation.
Another approach, but one we would not recommend, is to use a thicker wire basket but many EM samples could be heat damaged from the additional radiant heat. 20 mil wire is our recommended basket wire diameter.
Hope this information will be helpful.
Disclaimer: SPI Supplies manufactures tungsten wire baskets for vacuum evaporation.
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 ######################## ============================================
Does anyone out there in microscopy land know of a computer database program ( preferably PC) that has the scientist in mind and not the business man. I have Excel, Access, and Project 2000 but they are orientated to calculate dollars and cent not milliliters and millimeters if you know what I mean.
Ron Austin Dept of Pathology LSU Medical Center Shreveport, LA rla-at- mindspring.com 318-675-4557
yes I am using databases from dbase II on cp/m systems
ricardo
----- Original Message ----- } From: "Vr. Richard Bejsak-Collorado-Mansfeld" {ricardo-at-ans.com.au} To: "Ronald Austin" {rla-at-mindspring.com} Sent: Sunday, December 03, 2000 11:16 PM
Greetings, Nomarski optics are widely considered appropriate for viewing living cells. While it is true that the "3-D" look might or might not be present in the cell (more about that later), one must remember that in phase contrast there is the famous "halo", representing an out of focus reverse-contrast image that certainly is not present in the cell.
Nomarski optics provide contrast by means of a gradient in optical path (= sample thickness times refractive index). That is anywhere in the sample where the local gradient in optical path is different than the background (which typically would have a zero gradient) the intenstity of the image is different than the background. When the gradient is positive the intensity is brighter than background, when the gradient is negative then the intensity is darker than background. The most obvious gradients of optical path for a living cell suspended in an aqueous medium is the change in thickness that happens at the cell's edges: at one edge the cell is getting thicker (= postive gradient, hence bright) and at the opposite edge the cell gets thinner (= negative gradient, hence dark). Consquently, the topographic appearance does relate to the sample in a real, predictable way.
Nomarski optics can be harder to adjust properly than phase contrast. In phase, you have the phase ring in the condenser and this has to be centered with the phase ring in the objective. That's all you have to do. For Nomarski, you have to have first crossed polarizers, you have to know where they are and be able to rotate at least one of them. THen, the system also has two Wallason prisms, one in the condenser and one above the objective. The Wallaston prism has an optical axis and this axis of each Wallaston must be parallal and at 45 degrees to the analyzer/polarizer alignment. Different implementations of Nomarski differ vastly in the accessibility of the elements and the ease of their aligment. In the good designs, it is no trouble at all. But of course, one should know where each one is, what it does and how it should be aligned.
Changing between nomarki and brightfied is no trouble, you just remove the analyzer or polarizer from the optical path.
One important fact about Nomarski arises from the optical axis of the Wallastons. This defines the direction used to assess the gradient in optical path. The optics cannot detect gradients that run perpendicular to the Wallaston's optical axis. This means that features revealed in the image depend on the orientation of the cell. A rotatable stage is therefore very desirable. What is more, depending on your purpose, and the types of cells your are looking at, the directional properties of Nomarski will be more or less of a problem.
I have no experience with a Zetopan. If at all possible have demonstration before you buy. Finally, in many scopes you can exchange conderser elements so that you can do either phase or Nomarski. Thus, you could expand your repertoire later, as needs arise. I would be useful to find out if the Zetopan lets you do that.
Hope this helps, Tobias Baskin
} } I am helping a friend find a scope. I came across a Zetopan that may fit } his budget. I use a Nachet DIC for viewing living organisms and find it } very satisfactory. Could anyone help him with the versatility of a DIC } Zetopan. I know my Nachett works as a bright field, DIC and sort of } polarizing scope lacking a rotating stage and 1/4 and 1/2 wave plates. It } is the scope I use all the time. Unless I am doing darkfield. } } Having never been around a Zetopan I can't really answer his questions. } } Thanks } Gordon } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger } } } } } I my quest to find a better 'scope I have come across (with the help } } } of one of the list members) a Reichert Zetopan DIC scope that *might* } } } be available at in my price range. Previously I was just looking for } } } a nice phase contrast job. I have worked with phase before but not } } } DIC and would like to solicit other opinions as to whether this is } } } the right scope for me. This is going to pretty much blow my budget } } } so I want to be sure. } } } } } } I will be using this primarily for observing living organisms } } } (protozoa, algae, etc). I have been looking at a lot of photos of } } } protozoa taken using Nomarski DIC. While the photos are striking, I } } } keep asking myself "but what does it really look like". The problem } } } is the false 3D effect. I'm never sure what the part of the shape is } } } real and what is generated. Doesn't this ever bother you? } } } } } } I have been told that it is possible to adjust out the 3-D effect but } } } I'm not sure what kind of image you are left with. I would assume } } } something in-between bright field and DIC. Anyone have any experience } } } with this? } } } } } } I think if the Zetopan is unable to easily switch between a DIC image } } } and a bright (or dark) field image then I may pass on it. I need the } } } more traditional illumination as well to see what the real shape is. } } } It really makes the Zetopan too special purpose for someone like } } } myself. I think I would probably be happier with a less } } } sophisticated scope that can do phase, bright, and dark field (and } } } maybe oblique lighting as well). } } } } } } I would appreciate any thoughts you all may have. } } } } } } Bill } } } -- } } } Bill Tschumy } } } Otherwise -- Austin, TX } } } bill-at-otherwise.com
I'm pleased to announce that a revised, up-to-date version of "Children's Microscopy: a Bibliography" will be mailed as a supplement to the December issue of Microscopy Today; watch for it. It has brief reviews and ordering information on books, videos, CD-ROMs and websites. If you can't use it yourself, PLEASE give it to a teacher, or a parent, or a Scout leader, or a librarian - anyone who can use it to introduce children to the microworld.
If you aren't a Microscopy Today subscriber {microtoday-at-mindspring.com} you can read the same text soon on Project MICRO's website (URL below). But it's a tightly packed 16 pages, so it will be easier to browse the printed version.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Does anyone out there have suggestions on histochemical stains (or other techniques) to reduce the autofluorescence of lignin? I'm using DAPI staining on wood (xylem) sections, which have highly lignified cell walls, and I would like to reduce background fluorescence. Thanks.
Rachel
****************************************** Rachel Spicer Biological Laboratories 3119 Organismic and Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138
It is relatively easy to program in Microsoft's Visual Basic 6 using databases. It is one of that program's strengths. You can tailor any properties that you want. It is easy to set it up to be compatible with Microsoft's Access, but you can have it set up for any ODBC database. You can actually set up a working database program by just using the Form Wizard that comes with VB6.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Ronald Austin [mailto:rla-at-mindspring.com] Sent: Saturday, December 02, 2000 10:28 PM To: Microscopy Society of America Subject: a data base program
--------------------------------------------------------------- --------- 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/MicroscopyListserv er/FAQ.html
Does anyone out there in microscopy land know of a computer database program ( preferably PC) that has the scientist in mind and not the business man. I have Excel, Access, and Project 2000 but they are orientated to calculate dollars and cent not milliliters and millimeters if you know what I mean.
Ron Austin Dept of Pathology LSU Medical Center Shreveport, LA rla-at- mindspring.com 318-675-4557
I've started wondering about what actually the point resolution of a TEM in phase-contrast imaging is.
} From Raleigh's criterion I would conclude that for a typical 120 kV TEM with a numerical aperture of about 20 mrad distances larger than about 2 Angstrom or 'density waves' with a wavelength longer than 2 Angstrom simply cannot be resolved.
In Scherzer defocus the Phase Contrast Transfer Function (PCTF) has its first zero at about the same resolution but there seems to be information at higher frequencies (even if it is inverted in contrast).
Which is the true resolution limit?
Yours sincerely,
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.csb.ki.se/em
Trying to cut serial sections of Spurr for 3D reconstruction. Anyone know of any tricks to make the sections stay together in long ribbons? Ellen Morgan
In the field of high-resolution transmission electron microscopy, the "resolution" is defined as the first zero in the phase contrast transfer function (PCTF) at Scherzer (or optimum) defocus as you point out. This definition is chosen because any higher-frequency information is phase inverted (as you also point out). However, it results in defining the resolution in terms of the spatial frequency at which transfer goes to zero, rather than in terms of the highest transferred spatial frequency.
I have suggested a definition in which the resolution is defined at the 70% pass limit within the Scherzer passband, giving d = 0.67 Cs^1/4 lambda^3/4 at an "extended" optimum defocus of sqrt(1.5Cs.lambda) instead of Scherzer's d = 0.707 Cs^1/4 lambda^3/4 at an optimum defocus of sqrt(Cs.lambda). Current practice seems to use the extended defocus with the crossover (zero-transfer) frequency to get d = 0.64 Cs^1/4 lambda^3/4 at the "extended" optimum defocus.
The information beyond the Scherzer cross-over (zero-transfer) frequency can be utilized to provide structural information. Because of the misphasings, this information is difficult to interpret directly. Originally, the way to get at this higher-frequency (smaller-spacing) information was to compare focal series of experimental images with images simulated from possible models. In this way, we "saw" the small tunnels in Nb12O29 (with a 2.5Angstrom spacing) with a JEOL 100B (Scherzer resolution of 3.5A) in a focal series obtained by Sumio Iijima (see "Resolution-limiting effects in electron microscope images", G.R. Anstis and M.A. O'Keefe, In 34th Ann. Proc. EMSA, Miami Beach (1976) 480-481). I used HRTEM image simulation programs such as my SHRLI (simulated high-resolution lattice image) code, or my later interactive TEMPaS (TEM processing and simulation) code (ported to the Mac as MacTempas) with great success, but only when the model was known or limited to several possible ones (for example "Inversion domains in GaN grown on sapphire", L.T. Romano, J.E. Northrup and M.A. O'Keefe, Appl. Phys. Lett. 69 (1996) 2394-2396). Currently, post-Scherzer information can be accessed by image reconstruction algorithms from focal series, or tilt series, or holography. I use the Philips/Brite-Euram software for focal-series reconstruction by Coene and Thust in my one-Angstrom microscope (OAM) project to extend the resolution of a modified Philips CM300FEG/UT from its native resolution of 1.7Angstrom to a super-resolution of 0.89Angstrom that can image the "dumbbells" in [110] diamond (see “The NCEM One-Ångstrom Microscope project reaches 0.89Å resolution”, M. A. O'Keefe in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000) 1192-1193).
For the equations that belong with a discussion of resolution, see "Resolution in high-resolution electron microscopy", M.A. O'Keefe, Ultramicroscopy 47 (1992) 282-297. For a (simple) explanation of the use of focal-series to obtain super-resolution see "Push TEM limits with super-resolution", Michael A. O'Keefe (1999), R&D Magazine October 1999, p79 or {http://www.rdmag.com/archives/basics/10microscopy.htm} . For details of the Philips/Brite-Euram software for focal-series reconstruction by Coene and Thust, see W.M.J. Coene, A. Thust, M. Op de Beeck and D. Van Dyck, Ultramicroscopy 64 (1996) 109-135 and A. Thust, W.M.J. Coene, M. Op de Beeck and D. Van Dyck, Ultramicroscopy 64 (1996) 211-230. In the above, I have dealt only with linear terms in the transfer of information from the specimen (more properly, the exit-surface wave from the specimen) into the HREM image. Non-linear terms can give rise to even-higher spatial frequencies in the image. However, the question of whether these can be termed "resolution" is doubtful (see "Resolution-damping functions in non-linear images", M.A. O'Keefe, in 37th Ann. Proc. EMSA, San Antonio, Texas (1979) 556-557).
Mike O'Keefe
Philip Koeck 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 } -----------------------------------------------------------------------. } } Hallo everybody, } } I've started wondering about what actually the point resolution of } a TEM in phase-contrast imaging is. } } } From Raleigh's criterion I would conclude that for a typical 120 kV } TEM with a numerical aperture of about 20 mrad distances larger than } about 2 Angstrom or 'density waves' with a wavelength longer than } 2 Angstrom simply cannot be resolved. } } In Scherzer defocus the Phase Contrast Transfer Function (PCTF) has its } first zero at about the same resolution but there seems to be } information } at higher frequencies (even if it is inverted in contrast). } } Which is the true resolution limit? } } Yours sincerely, } } 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.csb.ki.se/em
-------------------------------------------------------- Your mail has been rejected for the following reason(s): -------------------------------------------------------- OK, Nestor -- this time it's in plain text. -Mike
At 11:55 AM -0500 12/4/00, Ann Dvorak wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************************ Hi Ellen, I vaguely remember reading something about coating the sides of the block that will become the leading and trailing faces with something slightly tacky (like the adhesive dissolved off of 3M Magic Scotch tape). Once that has dried, the edges of the sections are supposed to stick together. I've never actually tried it, and I wonder how you do manage to separate the sections where you need to.
Does anyone out there remember this? Has anyone tried it?
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
The Microscopy Society of America is seeking an Archivist to manage the historical documents of our Society. This position is appointed by MSA Council for a period of three years.
We are looking for someone who is experienced in the storage and cataloging of (primarily) paper documents. Although this is a voluntary position (non-paying), the Society would provide funding for expendibles and day-to-day operations.
If interested (or if you know of a qualified person), please contact me at:
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
A predecessor in our laboratory came back from the MSA conference a few years back with a sample from a vendor of something called Tacki Wax, which was supposed to serve this purpose. I think the idea was that the sections would be gently held together, but not so strongly that you couldn't work them apart as needed while collecting them. I've never tried it myself, as I don't need to do serial sectioning, but having a product name to go by might help your search.
Elaine Schumacher UOP 25 East Algonquin Rd. Des Plaines, IL 60017-5017 phone: 847-391-3403 fax: 847-391-3719 efschuma-at-uop.com
I am pleased to announce that the Live-cell Course continues to grow and prosper. Over 130 students from 23 countries have taken the course over the last 5 years.
Now I am organizing the next 11-day summer course on "3D Microscopy of Living Cells" for June of 2001. As in the past it will be at the University of British Columbia, in Vancouver, BC. The Fifth Workshop on 3D Image Processing will be held June 30 - July 2.
Please check out http://www.cs.ubc.ca/spider/ladic/course/bulletin.html if you would like to have a feeling for last years's course.
The information for 2001 is below.
Thanks for your help,
Jim Pawley
Sixth Annual INTERNATIONAL 11-Day Short Course on
3D Microscopy of Living Cells June 18 - 28, 2001
Fifth, Post-course Workshop on
3D Image Processing, June 30 - July 2, 2001
Organized by Prof. James Pawley, (University of Wisconsin-Madison) (SEE ADDRESS AT END OF MESSAGE)
in association with Dr. Elaine Humphrey UBC BioSciences Microscopy Facility: University of British Columbia Vancouver, BC, Canada
DATES
Applications must be received by March 1, 2001 Deposit due April 15, 2001 Registration 5:00 - 7:00 PM Sunday, June 17, 2001 First Lecture 7:30 PM Sunday, June 17, 2001 Live-cell Course ends, noon Thursday, June 28, 2001 3D Image Processing Course, June 30, - July 2, 2001
APPLICATIONS DUE BY MARCH 1, 2001
APPLICATIONS Applicants must complete a questionnaire to assess knowledge level, field of interest and proposed personal, live-cell, project. Enrollment will be limited to about 24 participants (exact number depends on number of 3D Systems available). Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with access to basic texts to read before the course begins. Application forms requesting information on field of interest and level of experience may be down-loaded from the WWW site at http://www.cs.ubc.ca/spider/ladic/course/reg1.htm, or obtained from:
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4
Additional information is available from: http://www.cs.ubc.ca/spider/ladic/course/bulletin.html
We expect to have at least 11, 3D microscope workstations for student use and there will be an international faculty of 17.
Application deadlines:
Application forms must be received for screening by March 1, 2000. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2000. In general, refunds of the deposit will not be possible. The remainder is due before Registration.
3D Course tuition (includes lunches and snacks): $2150 (US) Workshop Tuition (includes lunches and snacks): $850 (US)
Room / board about $40/day (US)
I hope that this includes all of the information that you need, but if not, please get back to me. -- **************************************** 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
Thanks for offering to put my 3D rendering question on the discussion group. My lab currently acquires and processes images using IPlab and a SPOT RT camera. We have MicroTome by VayTek for deconvolution, but this isn't much good without 3D rendering. Our primary application will be measuring the volume of nuclei and cells which have been fluorescently stained. Which 3D programs for a PC might work well for this?
Hello all, Any one with on advice on who in the Baltimore area can do chemlumanesence (spelling?) on live cells, realtime and time series? What equipment do we need, which company or labs are doing this.
I'll take a stab at this. Please correct me if I am wrong:
The point resolution refers to the resolution limit of a TEM for (as the name implies) two point like objects. A pointlike object is mathematically a delta-function and has a Fourier spectrum that is constant for all frequencies. So the question becomes: how many of the Fourier components can be transmitted without changing them, which in turn determines the minimum size of the object. The answer is then basically given by the first zero of the CTF. Until this point, the spatial frequencies are transmitted with the same phase and almost the same amplitude. For higher frequencies, the amplitude of the CTF oscillates, so that no reliable transmission can be achieved. The setting, where the first zero is furthest out in Fourier space is the Scherzer defocus. Thus the point resolution is determined by the first zero of the CTF at Scherzer defocus.
Line resolution refers to a periodic structure with very few Fourier components. It is therefore not necessary to transmit the entire Fourier spectrum, but only one specific component. Thus by changing the CTF through changing the focus, one can find a setting, where that specific frequency is transmitted and shows up in the image. The question therefore becomes: How well can I detect the structure. This is then in principle determined by the amplitude of the CTF. So the line resolution is then determined by the CTF envelope going below a certain threshold (I believe it is 20%, but I am not sure). That threshold is of course more or less arbitrary, but should give a reasonable estimate of the line resolution and allow comparison of the microscope resolutions.
Disclaimer: Please use this with caution. I have not been involved with EM theory for a few years. For a good account of high-resolution TEM, check out the book(s) by John Spence.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Philip Koeck [mailto:philip.koeck-at-csb.ki.se] Sent: Monday, December 04, 2000 8:45 AM To: 3dem list; MSA mailing list
Hallo everybody,
I've started wondering about what actually the point resolution of a TEM in phase-contrast imaging is.
} From Raleigh's criterion I would conclude that for a typical 120 kV TEM with a numerical aperture of about 20 mrad distances larger than about 2 Angstrom or 'density waves' with a wavelength longer than 2 Angstrom simply cannot be resolved.
In Scherzer defocus the Phase Contrast Transfer Function (PCTF) has its first zero at about the same resolution but there seems to be information at higher frequencies (even if it is inverted in contrast).
Which is the true resolution limit?
Yours sincerely,
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.csb.ki.se/em
For Light microscopy, we used a mixture of Weldwood contact cement from the local hardware store and toluene mixed 50/50. Place it on the leading and trailing edges of the block using a pin. We used a tool to pick up the sections I don't remember its name. It looked like a small metal trough.
For EM I used to make the block face area as small as possible focusing on a specific area of interest. Then I would cut the the leading and trailing edges parallel and then put the block on the ultra microtome and polish the edges with a glass knife set on zero advance. If the leading and trailing edges are smooth the sections stick together when coming off the block. I would make groups of ten sections. pick up with a tungsten wire loop (premade loops work better from EM venders of your choice) and place on a formvar coated slot grid.
Good Luck
Jon
{color} {param} 0100,0100,0100 {/param} On 4 Dec 00, at 15:23, Leona Cohen-Gould wrote:
} } Trying to cut serial sections of Spurr for 3D reconstruction. Anyone
} } know of any tricks to make the sections stay together in long
} } ribbons? Ellen Morgan
}
} ************************
} Hi Ellen,
} I vaguely remember reading something about coating the sides of the
} block that will become the leading and trailing faces with something
} slightly tacky (like the adhesive dissolved off of 3M Magic Scotch
} tape). Once that has dried, the edges of the sections are supposed to
} stick together. I've never actually tried it, and I wonder how you do
} manage to separate the sections where you need to.
}
} Does anyone out there remember this? Has anyone tried it?
}
} Lee
}
} Leona Cohen-Gould, M.S.
} Sr. Staff Associate
} Director, Electron Microscopy Core Facility
} Manager, Optical Microscopy Core Facility
} Joan & Sanford I. Weill Medical College
} of Cornell University
} voice (212)746-6146
} fax (212)746-8175
}
}
{nofill} Jon Ekman Associate Research Specialist Deptartment of Biological Sciences University of Wisconsin-Milwaukee phone W:414.229.6471 Web1 http://www.graffitimasters.com Web2 http://www.uwm.edu/~jekman
I would like to know whether there are any commercial labs that can provide up to 3000 um x 3000 um contact profilometry image maps such as the Veeco Dektak Profiler or the KLA-Tencor Profiler can provide.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Go to Rivlin and Raymond, 1987, Journal of Neuroscience Methods, 20:23-33, also see Raymond and Rivlin, 1987, Developmental Biology, 122:120-138. There you will find reference to the technique originally published by W. Fahrenbach, 1984, J. Electron. Microsc. Tech., 1:387-398. In the Rivlin paper you will find that a solution of Weldwood cement (rubber cement) was applied to the top and bottom edges of the trapezoid block face. This glue can be use undiluted or diluted in the appropriate solvent to obtain the desired workable concentration.
Linda Barthel, M.S. Research Associate II University of Michigan Department of Cell and Developmental Biology 4607 Medical Science Building II Ann Arbor, MI 48109-0616
There may be a position opening up in the Dallas/Ft. Worth Area for an experienced microscopist with polymers-TEM experience. If you might be interested please send me a resume. Jeff Day/wa5ekh-at-juno.com ________________________________________________________________ GET INTERNET ACCESS FROM JUNO! Juno offers FREE or PREMIUM Internet access for less! Join Juno today! For your FREE software, visit: http://dl.www.juno.com/get/tagj.
Does anyone have suggestions as to how to polish silicon wafers in cross section. I have a wafer (~0.5mm thick ) which has been randomly fractured with about a micron of metal on the surface and want to determine the metal thickness. We are trying various methods - first choice is by RBS but presently limited in max. energy so can not penetrate layer.
In my first couple of attempts at polishing (embedded wafer in thermoset) the silicon breaks quite easily producing a very rough surface. I assume I am being too aggressive and am trying slower more gentle procedure. Does anyone have a procedure for polishing cross sections of such brittle material?
The following comments are for the person who is trying to cut serial sections with Spurrs. I routinely cut serial sections with Spurrs without resorting to any kind of glue. Probably the most important thing is how the block is faced. I cut the trapezoid with a really long (.6 to .9 mm base) and the top of the trapezoid is almost the same length. What's really critical here is that those two cuts are parallel. The two sides of the trapezoid are very short (.1mm). I cut the sections with a Diatome knife and on a Reichert Ultracut E with a cutting speed of 0.8mm per second and a thickness setting of 80nm. I am doing this in a small room with a blasting air vent in the ceiling. I have cut up a cardboard box and stuck it into the ceiling tiles in such a way that the air is diverted away from the microtome. I can pick up 15 to 25 sections in a row on a 0.4 X 2mm copper slotted grid coated with just formvar or formvar/carbon. I have never been any good at making the support films my self, so I buy the slotted grids precoated from Ted Pella (catalog #s 01706, 01806, or 01816). Before picking up the ribbon, I dip a stick into chloroform and wave that over the sections, and they expand (or relax, depending on your point of view). I have a bunch of self locking tweezers and after picking up the sections, I leave the grid with the sections on it locked in the tweezers until they dry (if you put a wet slotted grid down on filter paper it might break the film). Good luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
} Trying to cut serial sections of Spurr for 3D reconstruction. Anyone know } of any tricks to make the sections stay together in long ribbons? Ellen } Morgan
Ellen -
I assume that you're talking about EM. not LM. Make sure that the top and bottom of your block face are REALLY smooth and clean - and as small as is consistent with the reconstruction. Viewed thru the microtome binocular, they should look glassy, not like sandpaper. Change your trimming tool (glass knife? Weck razor blade?) often.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Has anyone used a HP Laserjet IIIp with an EDAX PV9900, or know what drivers are needed, or where we can acquire them?
We are trying to use the IIIp because it has serial and parallel connections needed by the EDAX and by the Philips XL40 SEM with which we use it and we cant afford to buy a new one.
Reply in confidence to chris.smith-at-bbsrc.ac.uk if you don't want to clog up the list.
Many thanks, Chris, Plant Path., IACR-Rothamsted, UK.
Ed, I represent Allied High Tech Products. We have detailed procedures for preparing semiconductor device cross-sections. We also offer all the necessary products to accomplish it. I would be happy to speak to you and provide you with detailed procedures on how to prepare these samples. Since it can be quite detailed it would be best to do this offline. Please provide me with your phone number and I will call you.
Sincerely,
Ed
************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech Products 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (919) 846-9628 vm:(800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com
Equipment and Consumables for Metallurgical Sample Preparation *************************************************
-----Original Message----- } From: Ed Kurz [mailto:ekurz-at-mail.ims.uconn.edu] Sent: Tuesday, December 05, 2000 8:32 AM To: MICROSCOPY BB
Does anyone have suggestions as to how to polish silicon wafers in cross section. I have a wafer (~0.5mm thick ) which has been randomly fractured with about a micron of metal on the surface and want to determine the metal thickness. We are trying various methods - first choice is by RBS but presently limited in max. energy so can not penetrate layer.
In my first couple of attempts at polishing (embedded wafer in thermoset) the silicon breaks quite easily producing a very rough surface. I assume I am being too aggressive and am trying slower more gentle procedure. Does anyone have a procedure for polishing cross sections of such brittle material?
We have a two year old PGT Prism EDS detector/preamp/dewar assembly which we are thinking of selling. But I don't have much of an idea of the value of such a device.
Any thoughts?
Anyone interested?
Richard Shalvoy Arch Chemicals Cheshire, CT 203-271-4394
This is not a job application. Would like to seek your advice on how to analyse a polymer samples using TEM. We have tried on Poly-carbonate injection molded lense with hard coating. The purpose is to see the interface bonding of the hard coat on the lense. Unfortunately, when we microtome the samples, the sample curve up and we cannot manage to see anything. Would appreciate if you can share your experience on the sample preparation with us.
Thanks in advance of your help.
Regards, YH Koh (MQE Engineer) Motorola CGISS Penang, Bayan Lepas FIZ, Phase 3, 11900 Penang, Malaysia. Tel : 60-4-8504089 Fax: 60-4-6124903
-----Original Message----- } From: charles j day [mailto:wa5ekh-at-juno.com] Sent: Sunday, December 03, 2000 12:50 PM To: Microscopy-at-sparc5.microscopy.com
There may be a position opening up in the Dallas/Ft. Worth Area for an experienced microscopist with polymers-TEM experience. If you might be interested please send me a resume. Jeff Day/wa5ekh-at-juno.com ________________________________________________________________ GET INTERNET ACCESS FROM JUNO! Juno offers FREE or PREMIUM Internet access for less! Join Juno today! For your FREE software, visit: http://dl.www.juno.com/get/tagj.
"james patrick birrell" {jbirrell-at-students.uiuc.edu} wrote in message news: {Pine.GSO.4.10.10012041635310.22505-100000-at-ux12.cso.uiuc.edu} ... } I was wondering if anybody would know where I could obtain software to } facilitate the acquisition/analysis of images to be used to perform } wavefront reconstruction using images from a TEM. Specifically, I'm } looking for any software compatable with a JEOL 4000ex microscope with a } Gatan CCD camera. } Thanks, } James Birrell
Ammonium molybdate (AM) is suitable in the cases when uranyl acetate (UA) is bad (mostly if pH is a critical factor, UA pH = 5-5.5). In most cases AM gives less contrast than UA. AM gives you "pure" negative staining because do not chemically interfere with most biological samples. UA from another hand sometimes gives you mixed contrast, positive (DNA/RNA)&negative(proteins). For instance, ribosomes and many viruses shown mixed staining with UA. It's not bad because it may even enhance some structural details, but you have to remember about that. My own experience indicated that in most cases UA or other uranium salts (U-formiate or oxalate) after conditions adjusting (concentration, time, temperature, freshness, support film) has shown perfect results. If you have a problem with stain spreading, you may try different support films (carbon, formvar, parlodion). Very good results you may obtain using "double carbon" technique - this may solv even very worse cases. I also has have a good results with poly-lysine treatment of the grids prior sample adsorption. Personally, I don't like the glow-discharge. But it works in some way.
Sergey.
At 11:52 AM 12/5/00 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
EDAX has the HP laser printer driver for old DEC based systems. Call EDAX, or www.edax.com
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: chris smith (IACR-RES) {chris.smith-at-bbsrc.ac.uk} To: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com}
I found your post interesting. How do you make your edges parallel? Do you use a razor blade?
Dave
On Tue, 5 Dec 2000 09:47:29 -0500 Timothy Schneider {Timothy.Schneider-at-Mail.TJU.EDU} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } The following comments are for the person who is trying to cut serial } sections with Spurrs. I routinely cut serial sections with Spurrs without } resorting to any kind of glue. Probably the most important thing is how the } block is faced. I cut the trapezoid with a really long (.6 to .9 mm base) } and the top of the trapezoid is almost the same length. What's really } critical here is that those two cuts are parallel. } The two sides of the trapezoid are very short (.1mm). I cut the sections } with a Diatome knife and on a Reichert Ultracut E with a cutting speed of } 0.8mm per second and a thickness setting of 80nm. I am doing this in a } small room with a blasting air vent in the ceiling. I have cut up a } cardboard box and stuck it into the ceiling tiles in such a way that the air } is diverted away from the microtome. I can pick up 15 to 25 sections in a } row on a 0.4 X 2mm copper slotted grid coated with just formvar or } formvar/carbon. I have never been any good at making the support films my } self, so I buy the slotted grids precoated from Ted Pella (catalog #s 01706, } 01806, or 01816). Before picking up the ribbon, I dip a stick into } chloroform and wave that over the sections, and they expand (or relax, } depending on your point of view). I have a bunch of self locking tweezers } and after picking up the sections, I leave the grid with the sections on it } locked in the tweezers until they dry (if you put a wet slotted grid down on } filter paper it might break the film). Good luck, Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
{ { Would like to seek your advice on how to analyse a polymer sample using TEM. We have tried on Poly-carbonate injection molded lens with hard coating. The purpose is to see the interface bonding of the hard coat on the lens. Unfortunately, when we microtome the samples, the sample curves up and we cannot manage to see anything. Would appreciate if you can share your experience on the sample preparation with us. } }
We have looked at surface layers in polymer samples by cutting open a surface, then etching and looking, NOT at a cut section, but at the cross-sectional view of the remaining surface, either by SEM or by replication/TEM. Usually we etch our specimens, so before cutting open, a protective layer is applied over the outer surface to prevent this being etched downwards. Mainly we have looked at polyethylene or polypropylene specimens, using our permanganic etching technique, and different reagents would be needed for polycarbonate. However, it might be possible to replicate the cut surface directly and get useful information.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
The question was: how do I make the top and bottom edges parallel? I do it by eye. I use Gem single edge industrial blades from Ted Pella Cat# 121-1 (I'm sure these are available from many sources). One important point is that I spray them with 70% ETOH before using and wipe them off with a Kimwipe EXL. A word about the way I described the trapezoid face: it doesn't look like a trapezoid. What it looks like is a very thin shiny line (heck, I'm talking about putting 15 to 25 of these guys on a 2mm slot, you do the math). More good luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Thanks to all who responded to my question regarding conductive adhesives for salmon skin. I have ordered many of the products that were suggested and am looking forward to success (finally) :)
Thanks again and have a great holiday season!
Carla Aiwohi Western Fisheries Research Center Seattle, WA
We routinely use HEPES for our fixations with paraformaldehyde, glutaraldehyde and osmium. We see little or no difference from classical cacodylate buffers. I chose HEPES a long time ago and in retrospect, PIPES might have been a slightly better choice since HEPES has a pKa of 7.5 and PIPES has a pKa of 6.76. Since one generally uses aldehyde buffers at pH 7.0 to 7.4, either would seem appropriate but since I read that during fixation, H+ ions are generated and the solution tends to acidify, then having a buffer on the high side of the pKa would be better since you would have greater buffering power. Despite this "improved" logic, I have remained loyal to HEPES and never had a problem. Good luck.
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-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Would anyone happen to have the actual recipe for Gurr's buffer? I think it is some phosphate formulation - we've been hunting the recipe and have found some refs, but will have to interlibrary the journals/books and we thought we'd give the 'net a try, first.
I know we can buy the pre-made buffer, but we need such a tiny amount that we'd rather make it from scratch (plus we want to know what it is!).
I would be a bit concerned about your use of chloroform to flatten sections, especially in a small microtome room. We have used heat pens for the last decade or so, because apparently the chloroform we use these days is a lot more toxic than it used to be (as well as formaldehyde, glutaraldehyde, carbon tetrachloride etc).
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Timothy Schneider wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The following comments are for the person who is trying to cut serial } sections with Spurrs. I routinely cut serial sections with Spurrs without } resorting to any kind of glue. Probably the most important thing is how the } block is faced. I cut the trapezoid with a really long (.6 to .9 mm base) } and the top of the trapezoid is almost the same length. What's really } critical here is that those two cuts are parallel. } The two sides of the trapezoid are very short (.1mm). I cut the sections } with a Diatome knife and on a Reichert Ultracut E with a cutting speed of } 0.8mm per second and a thickness setting of 80nm. I am doing this in a } small room with a blasting air vent in the ceiling. I have cut up a } cardboard box and stuck it into the ceiling tiles in such a way that the air } is diverted away from the microtome. I can pick up 15 to 25 sections in a } row on a 0.4 X 2mm copper slotted grid coated with just formvar or } formvar/carbon. I have never been any good at making the support films my } self, so I buy the slotted grids precoated from Ted Pella (catalog #s 01706, } 01806, or 01816). Before picking up the ribbon, I dip a stick into } chloroform and wave that over the sections, and they expand (or relax, } depending on your point of view). I have a bunch of self locking tweezers } and after picking up the sections, I leave the grid with the sections on it } locked in the tweezers until they dry (if you put a wet slotted grid down on } filter paper it might break the film). Good luck, Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu
LM - Need help on staining semithin sections embedded in LR-white
I am doing anatomical studies on the seed coat and currently use toluidine blue with sodium hypochlorite prestaining and iodine/potassium iodite poststaining, described by Gutmann, M. : Improved staining procedures for photografic documentation of phenolic deposits in semithin sections of plant tissue. Journal of Microscopy 179:277-281 (1995) Can anyone suggest a better stain, specially one that is suitable for staining lignin ?
1. Apply a thin layer of M-bond on the surfaces of two pieces of your wafer and put these two surfaces together. 2. Use a clip to press these pieces of wafers as close as possible (not to break them). 3. Leave the sample to try (24 hours in air? or 1 hour at 120 C degree on a hot plate).
This is a good way to protect the thin "film" (your metal features) I think. The smaller the gap between these two pieces, the better. In this way you can get two cross section surfaces. Even after step 3, you may apply another thin layer of M-bond on the outside surfaces of the sample, so that you may get 4 surfaces for examination.
4 Embed the sample in a suitable epoxy as you did before, and grinding and polishing the cross sections carefully from coarse to fine.
When you observe the cross sections, it is easy to see what is the M-bond and what is the other thing you want to see.
Just a suggestion.
Good luck!
Zhenquan Liu
I think you may try the following way:
1. Apply a thin layer of M-bond on the surfaces of two pieces of your wafer and put these two surfaces together. 2. Use a clip to press these pieces of wafers as close as possible (not to break them). 3. Leave the sample to try (24 hours in air? or 1 hour at 120 C degree on a hot plate).
This is a good way to protect the thin "film" (your metal features) I think. The smaller the gap between these two pieces, the better. In this way you can get two cross section surfaces. Even after step 3, you may apply another thin layer of M-bond on the outside surfaces of the sample, so that you may get 4 surfaces for examination.
4 Embed the sample in a suitable epoxy as you did befoe, and grinding and polishing the cross sections carefully from coarse to fine.
When you observe the cross sections, it is easy to see what is the M-bond and what is the other thing you want to see.
Just a suggestion.
Good luck!
Zhenquan Liu
=================
Does anyone have suggestions as to how to polish silicon wafers in cross section. I have a wafer (~0.5mm thick ) which has been randomly fractured with about a micron of metal on the surface and want to determine the metal thickness. We are trying various methods - first choice is by RBS but presently limited in max. energy so can not penetrate layer.
In my first couple of attempts at polishing (embedded wafer in thermoset) the silicon breaks quite easily producing a very rough surface. I assume I am being too aggressive and am trying slower more gentle procedure. Does anyone have a procedure for polishing cross sections of such brittle material?
Thanks, Ed
------------------------------------ Zhenquan Liu (Dr.) Arizona State University Room 146A, CSSS Tempe, AZ 85287 Tel (480) 965 4544 (o) (480) 775 7428 (h) Fax (480) 965 9004 (o) Email zhenquan.liu-at-asu.edu ------------------------------------
I have used PIPES as a substitute for cacodylate in most routine procedures simply because it has none of the precipitation problems of phosphate buffers. I understand that there may be more extraction with cacodylate in TEM so I would have thought that it may be a better buffer for SEM (less chance of shrinkage?). I would assume that HEPES was similar but have no experience with it.
These are my own feelings and I have seen no scientific evidence to support this - has anyone else?
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Bart De Pauw wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello listers, } } We use cacodylate buffer for our SEM samples. Is HEPES or PIPES buffer } an alternative for the toxic cacodylate ? Our samples are mainly animal } tissue. } } Bart De Pauw } Ghent University } Faculty of Veterinary Medicine } Department Morphology } Belgium
} To: Timothy.Schneider-at-Mail.TJU.EDU } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: MORE SERIAL SECTIONS } } If you are not limited by particular area in your sample, you may do the } trick, which warranty the parallelness of the edges. This trick did show } to me Prof. Borovyagim many years ago. The trick is: you do not form a } trapezoid, instead you make two cuts by razor blade, which owerlapping, so } you will have "/\"-shape without flat top. The top should be formed by } two overlapped cuts. There is no flat space on the top, which we usually } called "trapezoid". This two cuts will form a "line" which will be } paralle to the knife edge later. You have to form side fases of the } piramid as well. Finall, you will have something looks pretty like } triangle prizm. Not perfect prizm, of coarse. When you start cutting, } you have to carefully cut avay the top of the "prizm" to form } trapezoid. The final shape of the trapezoid will depends from orientation } of your cuts, but owerlapped cuts will form perfectly parallel sides. } } Another choice I friequently use: to use glass knife and trim on the } goniometer-head when you may tilt and rotate the sample. Make one face, } rotate 180 deg and make second face. } } At 09:24 AM 12/6/00 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I am currently in the process of setting up a histology/microbiology/EM facility. Does anyone have comments or suggestions on a good rotary tissue processor for light microscopy? My main concerns are cost, service availability, ease in operation(changing of fluids, etc.) and reliability. I may process 10 - 60 specimens a month, more as the lab is established. I appreciate any info.
I'm currently in the process of setting up a Histology/Microbiology/EM Lab. Having been away from histology for almost 30 years (been doing EM), I've gotten away from instrumentation associated with histology. Does anyone have a good recommendation for a rotary tissue processor? I've got info on Shandon and Tissue-Tek. Are there others out there? Thanks for any info.
} Date: Wed, 06 Dec 2000 15:16:31 -0800 } To: Timothy.Schneider-at-Mail.TJU.EDU } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: MORE SERIAL SECTIONS } } If you are not limited by particular area in your sample, you may do the } trick, which warranty the parallelness of the edges. This trick did show } to me Prof. Borovyagin many years ago. The trick is: you do not form a } trapezoid, instead you make two cuts by razor blade, which overlapping, so } you will have "/\"-shape without flat top. The top should be formed by } two overlapped cuts without "free" space between. There is no flat space } on the top, which we usually called "trapezoid". This two cuts will form } a "line" which will be parallel to the knife edge later. You have to } form side faces of the pyramid as well. Finally, you will have something } looks pretty like triangle prism. Not perfect prism, of coarse. When you } start cutting, you have to carefully cut away the top of the "prism" to } form trapezoid. The final shape of the trapezoid will depends from } orientation of your cuts, but overlapped cuts will form perfectly parallel } sides. This technique works if sample will permit. } } Another choice I frequently use: to use glass knife and trim on the } goniometer-head when you may tilt and rotate the sample. Make one face, } rotate 180 deg and make second face. If I have enough time, I prefer this } way, almost gives me the perfect pyramid. } } Sergey } } At 09:24 AM 12/6/00 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I'm currently in the process of setting up a Histology/Microbiology/EM facility. Being out of Histology for almost 30 years (been doing EM) I'm not sure of the best type of tissue processor to buy. I won't be doing that many specimens, so I'm leaning towards a rotary tissue processor. I have info on Shandon and Tissue-Tek. Are there others out there? Anyone having opinions on the 2 mentioned, I would appreciate any comments. I am looking for ease of use (changing solutions, fume safeguards), reliability on service and cost effectiveness. Thanks.
Peace be with you, Phil Rutledge (410)778-4136, 2120 prutledge-at-ars.usda.gov
I usually glue (with superglue or thin epoxy) 2-4 wafers together, metal coated surfaces facing each other. Then embed them or mount in microwise, cut with low speed diamond saw and polish, preferably with diamond. If you do not need nice pictures, just measurements, do not use fine polish, you will more or less ruin edges with it.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Ed Kurz [mailto:ekurz-at-mail.ims.uconn.edu] } Sent: Tuesday, December 05, 2000 8:32 AM } To: MICROSCOPY BB } Subject: Wafer Cross sections } } } -------------------------------------------------------------- } ---------- } 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 suggestions as to how to polish silicon } wafers in cross } section. I have a wafer (~0.5mm thick ) which has been } randomly fractured } with about a micron of metal on the surface and want to } determine the metal } thickness. We are trying various methods - first choice is by RBS but } presently limited in max. energy so can not penetrate layer. } } In my first couple of attempts at polishing (embedded wafer } in thermoset) } the silicon breaks quite easily producing a very rough } surface. I assume I } am being too aggressive and am trying slower more gentle } procedure. Does } anyone have a procedure for polishing cross sections of such } brittle material? } } Thanks, } Ed }
} Would anyone happen to have the actual recipe for Gurr's buffer? I think } it is some phosphate formulation ... } I know we can buy the pre-made buffer, but we need such a tiny amount that } we'd rather make it from scratch (plus we want to know what it is!).
Quite right too! (For making and wanting to know) I haven't heard of it so can't tell you the answer, BUT:
Does it matter what the exact formulation is? It is unlikely that either of the late Gurr Bros (Edward, and George T.) invented an original buffer. They were vendors of stains in Britain, about whom some older HistoNetters may well have entertaining anecdotes.
If a buffer does its job of stabilizing the pH at the correct value, and doesn't contain anything that would react with the other ingredients of the mixture, it shouldn't matter what you use. Phosphate buffers make a precipitate with many metal salts, including those of Ca, Co, Cu, Fe, Mg, and others that have insoluble phosphates. All the techniques books published since about 1955 have sets of tables for making buffers to cover a wide range of pH.
John A. Kiernan, Department of Anatomy & Cell Biology, The University of Western Ontario, LONDON, Canada N6A 5C1
Ted Pella Poly-L-Lysin 0.1% w/v aqueous solution, Cat # 18026 Float your grid with film on small drop of the solution for 1 min and than remove excess of the liquid by filter paper and put on the drop of water for 5-10 sec, blot it and immediately move it on the drop with your sample. Temperature - as necessary for your sample. It also helps to adsorb tuff samples. Surprisingly, it seems to me that it does not affect background.
Concerning "double-carbon" technique. It is more tricky. The idea is to sandwich your sample between two carbon films just before you finish the staining. I prefer the following technique. The approx. 1 ml plastic cap (have no idea what is that, you may use any suitable container) 0.7 cm diameter I filled with 0.5-1% aqueous UA in the way to make flat meniscus, so it will reflect the light. Than I will cut 3x3 mm piece of mica with carbon film and float carbon on the UA solution. Reflecting light will makes it visible under some particular angle (you have to practice a little bit). So, you have the plastic cap with UA and floated piece of carbon film on it and you can see carbon easily. Than you will adsorb sample on the regular EM grid with carbon. Than you have to move your grid into the cap with stain (avoid contact to the floated carbon, soak grid into the cap away from carbon). Looking through the carbon film, still floated on the UA, move the grid just under the film. So, you will see the grid through floated film. Than using one smooth movement you have to move grid vertically up picking up the carbon film. So, you will put grid just under the film (deep under the surface) and than move grid vertically up. The film will holds over the grid with drop of the stain solution. The last most important step - to remove excesses of staining solution: fix grid securely in the tweezer by rubber O-ring (or use "reverse-action" tweezer), small triangle shape piece of filter paper (I am using Whatmann 1M paper) insert between tweezer's "legs" (sorry, guys, I have no idea how it is called) as closer to the grid as possible. Filter paper chip should contact with excess of liquid holds by capillary force between the tweezer's "legs". Keep tweezer with grid and filter paper until grid will dry completely (at least 5 min). Enjoy the perfect spreading of the stain! There is a number of disadvantages in this method: -"double-carbon" will decrease the resolution; -sandwich will slightly flat the particles, expect to see 10-15% bigger size because of that.
Advantages: -sandwich will protect your sample from radiation damage; -such samples usually are more stable from all points of view; -because of thick carbon, you may use 400 mesh naked EM grids without "holey" support film (I still prefer to use the grids with my home-made "holey" film).
PTA vs AM. In my point of view, PTA does not have any serious advantages over the AM. Personally, I prefer AM. In some particular cases PTA may work better, I guess. If there is no chance to use UA, I would try U-formiate first and oxalate - next. U-formiate (UF) has pH around 6 or even higher (I don't remember the exact number). It gives you a very nice fine staining (I love it better than UA). The disadvantage of this stain - it is not stable. You have to prepare fresh solution every time. It is light-sensitive also (general room illumination is OK for 20-30 min, no direct light, please!). You could store it in the dark in the ice for couple of hours. U-oxalate (UO) is less stable than UF, you have to use it immediately. The beauty of UO - you may use it at pH 7. People's unsuccess with UF or UO mostly related to the storage problem, I believe. Most people don't understand how unstable those substances are. By the way, UA is light-sensitive also.
My staining conditions for all stains are: +4oC (if sample permits), adsorption time - 1-2 min; staining 0.5-1% aqueous solution - 1-2 min. I find, that it is better to adjust sample concentration rather than to play with adsorption time. Extended staining time will not dramatically improve the staining but may harm the sample's integrity (it depends, for IgG staining I am using 4 min 1% UA). For UA staining I also prefer to use my favorite "EM" buffer based on ammonium acetate with additives if necessary (50-100 mM ammonium acetate, pH 7.8 in simplest case). It is highly compatible with UA and gives good stain's distribution.
I am sorry if my description is not clear. Please, feel free to call me if any question. It's much simpler to do than read.
God luck Sergey
At 11:08 AM 12/6/00 -0500, you wrote: } Sergey, } Would you mind giving me the details of your treatment of grids with } poly-lysine and the double-carbon technique that you refer to in your } E-mail. I usually go to PTA if PH is a problem. How do you compare PTA } with AM? I am always looking for new methods to deal with some of the } negative stain. problems. } Thanks very much, } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: } dsherman-at-purdue.edu } 1057 Whistler Building } West Lafayette, IN 47907-1057 } } } On Tuesday, December 5, 2000 10:43 PM, Sergey Ryazantsev } {sryazant-at-ucla.edu} wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Ammonium molybdate (AM) is suitable in the cases when uranyl acetate (UA) } } is bad (mostly if pH is a critical factor, UA pH = 5-5.5). In most cases AM } } gives less contrast than UA. AM gives you "pure" negative staining because } } do not chemically interfere with most biological samples. UA from another } } hand sometimes gives you mixed contrast, positive } } (DNA/RNA)&negative(proteins). For instance, ribosomes and many viruses } } shown mixed staining with UA. It's not bad because it may even enhance } } some structural details, but you have to remember about that. My own } } experience indicated that in most cases UA or other uranium salts } } (U-formiate or oxalate) after conditions adjusting (concentration, time, } } temperature, freshness, support film) has shown perfect results. If you } } have a problem with stain spreading, you may try different support films } } (carbon, formvar, parlodion). Very good results you may obtain using } } "double carbon" technique - this may solv even very worse cases. I also } } has have a good results with poly-lysine treatment of the grids prior } } sample adsorption. Personally, I don't like the glow-discharge. But it } } works in some way. } } } } Sergey. } } } } At 11:52 AM 12/5/00 -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 } } } -----------------------------------------------------------------------. } } } } } } } } } Dear all , } } } Does any body use Ammonium Molybdate for Negative staining and also by } } } this chemical visualize structure better then Uranyl acetate. } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } } } }
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
We use the Shandon Citadel 1000. It's a good processor, but there's one disadvantage : there's evaporation of your fluids. There's no fume safeguard on the processor, so you have to put it in a fume hood.
Phillip Rutledge schreef:
} ------------------------------------------------------------------------ } 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'm currently in the process of setting up a Histology/Microbiology/EM facility. Being out of Histology for almost 30 years (been doing EM) I'm not sure of the best type of tissue processor to buy. I won't be doing that many specimens, so I'm leaning towards a rotary tissue processor. I have info on Shandon and Tissue-Tek. Are there others out there? Anyone having opinions on the 2 mentioned, I would appreciate any comments. I am looking for ease of use (changing solutions, fume safeguards), reliability on service and cost effectiveness. Thanks. } } Peace be with you, } Phil Rutledge (410)778-4136, 2120 } prutledge-at-ars.usda.gov
I did not publish this technique yet. So, you may refer to our ListServer. If you need more details - call me.
Sergey.
At 07:52 AM 12/7/00 +0000, you wrote: } Brilliant, Sergey. Have you published these methods anywhere? If } so, I would be interested to have the references. } Best wishes } Chris } } Date sent: Wed, 06 Dec 2000 22:18:27 -0800 } To: Debby Sherman {dsherman-at-purdue.edu} } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: Negative stain } Copies to: Microscopy-at-sparc5.microscopy.com } } } ------------------------------------------------------------------------ } } 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 Debby } } } } Ted Pella Poly-L-Lysin 0.1% w/v aqueous solution, Cat # 18026 } } Float your grid with film on small drop of the solution for 1 min and than } } remove excess of the liquid by filter paper and put on the drop of water } } for 5-10 sec, blot it and immediately move it on the drop with your } sample. } } Temperature - as necessary for your sample. It also helps to adsorb tuff } } samples. Surprisingly, it seems to me that it does not affect background. } } } } Concerning "double-carbon" technique. It is more tricky. The idea is to } } sandwich your sample between two carbon films just before you finish the } } staining. I prefer the following technique. The approx. 1 ml plastic cap } } (have no idea what is that, you may use any suitable container) 0.7 cm } } diameter I filled with 0.5-1% aqueous UA in the way to make flat meniscus, } } so it will reflect the light. Than I will cut 3x3 mm piece of mica with } } carbon film and float carbon on the UA solution. Reflecting light will } } makes it visible under some particular angle (you have to practice a } little } } bit). So, you have the plastic cap with UA and floated piece of carbon } film } } on it and you can see carbon easily. Than you will adsorb sample on the } } regular EM grid with carbon. Than you have to move your grid into the cap } } with stain (avoid contact to the floated carbon, soak grid into the cap } } away from carbon). Looking through the carbon film, still floated on the } } UA, move the grid just under the film. So, you will see the grid through } } floated film. Than using one smooth movement you have to move grid } } vertically up picking up the carbon film. So, you will put grid just } under } } the film (deep under the surface) and than move grid vertically up. The } } film will holds over the grid with drop of the stain solution. The last } } most important step - to remove excesses of staining solution: fix grid } } securely in the tweezer by rubber O-ring (or use "reverse-action" } tweezer), } } small triangle shape piece of filter paper (I am using Whatmann 1M paper) } } insert between tweezer's "legs" (sorry, guys, I have no idea how it is } } called) as closer to the grid as possible. Filter paper chip should } } contact with excess of liquid holds by capillary force between the } } tweezer's "legs". Keep tweezer with grid and filter paper until grid will } } dry completely (at least 5 min). Enjoy the perfect spreading of the } } stain! There is a number of disadvantages in this method: } } -"double-carbon" will decrease the resolution; } } -sandwich will slightly flat the particles, expect to see 10-15% bigger } } size because of that. } } } } Advantages: } } -sandwich will protect your sample from radiation damage; } } -such samples usually are more stable from all points of view; } } -because of thick carbon, you may use 400 mesh naked EM grids without } } "holey" support film (I still prefer to use the grids with my home-made } } "holey" film). } } } } PTA vs AM. In my point of view, PTA does not have any serious advantages } } over the AM. Personally, I prefer AM. In some particular cases PTA may } } work better, I guess. If there is no chance to use UA, I would try } } U-formiate first and oxalate - next. U-formiate (UF) has pH around 6 or } } even higher (I don't remember the exact number). It gives you a very nice } } fine staining (I love it better than UA). The disadvantage of this } stain - } } it is not stable. You have to prepare fresh solution every time. It is } } light-sensitive also (general room illumination is OK for 20-30 min, no } } direct light, please!). You could store it in the dark in the ice for } } couple of hours. U-oxalate (UO) is less stable than UF, you have to } use it } } immediately. The beauty of UO - you may use it at pH 7. People's } } unsuccess with UF or UO mostly related to the storage problem, I } } believe. Most people don't understand how unstable those substances } } are. By the way, UA is light-sensitive also. } } } } My staining conditions for all stains are: +4oC (if sample permits), } } adsorption time - 1-2 min; staining 0.5-1% aqueous solution - 1-2 min. I } } find, that it is better to adjust sample concentration rather than to play } } with adsorption time. Extended staining time will not dramatically improve } } the staining but may harm the sample's integrity (it depends, for IgG } } staining I am using 4 min 1% UA). For UA staining I also prefer to use my } } favorite "EM" buffer based on ammonium acetate with additives if necessary } } (50-100 mM ammonium acetate, pH 7.8 in simplest case). It is highly } } compatible with UA and gives good stain's distribution. } } } } I am sorry if my description is not clear. Please, feel free to call } me if } } any question. It's much simpler to do than read. } } } } God luck } } Sergey } } } } } } } } At 11:08 AM 12/6/00 -0500, you wrote: } } } Sergey, } } } Would you mind giving me the details of your treatment of grids with } } } poly-lysine and the double-carbon technique that you refer to in your } } } E-mail. I usually go to PTA if PH is a problem. How do you compare PTA } } } with AM? I am always looking for new methods to deal with some of the } } } negative stain. problems. } } } Thanks very much, } } } Debby } } } } } } Debby Sherman Phone: 765-494-6666 } } } Life Science Microscopy Facility FAX: 765-494-5896 } } } Purdue University E-mail: } } } dsherman-at-purdue.edu } } } 1057 Whistler Building } } } West Lafayette, IN 47907-1057 } } } } } } } } } On Tuesday, December 5, 2000 10:43 PM, Sergey Ryazantsev } } } {sryazant-at-ucla.edu} wrote: } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Ammonium molybdate (AM) is suitable in the cases when uranyl acetate } (UA) } } } } is bad (mostly if pH is a critical factor, UA pH = 5-5.5). In most } cases AM } } } } gives less contrast than UA. AM gives you "pure" negative staining } because } } } } do not chemically interfere with most biological samples. UA from } another } } } } hand sometimes gives you mixed contrast, positive } } } } (DNA/RNA)&negative(proteins). For instance, ribosomes and many viruses } } } } shown mixed staining with UA. It's not bad because it may even enhance } } } } some structural details, but you have to remember about that. My own } } } } experience indicated that in most cases UA or other uranium salts } } } } (U-formiate or oxalate) after conditions adjusting (concentration, } time, } } } } temperature, freshness, support film) has shown perfect results. If you } } } } have a problem with stain spreading, you may try different support films } } } } (carbon, formvar, parlodion). Very good results you may obtain using } } } } "double carbon" technique - this may solv even very worse cases. I also } } } } has have a good results with poly-lysine treatment of the grids prior } } } } sample adsorption. Personally, I don't like the glow-discharge. But it } } } } works in some way. } } } } } } } } Sergey. } } } } } } } } At 11:52 AM 12/5/00 -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 } } } } } -------------------------------------------------------------------- } ---. } } } } } } } } } } } } } } } Dear all , } } } } } Does any body use Ammonium Molybdate for Negative staining and also by } } } } } this chemical visualize structure better then Uranyl acetate. } } } } } } } } _____________________________________ } } } } } } } } Sergey Ryazantsev Ph. D. } } } } Electron Microscopy } } } } UCLA School of Medicine } } } } Department of Biological Chemistry } } } } Box 951737 } } } } Los Angeles, CA 90095-1737 } } } } } } } } Phone: (310) 825-1144 } } } } Pager: (310) 845-0248 } } } } FAX (departmental): (310) 206-5272 } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Dr Chris Jeffree } University of Edinburgh } Biological Sciences EM Facility } Daniel Rutherford Building } King's Buildings EDINBURGH EH9 3JH } Tel: +44 (0) 131 650 5345 } FAX: +44 (0) 131 650 6563 } } Inveresk Cottage, 26 Carberry Road, } Inveresk, Musselburgh, Midlothian EH21 8PR, UK } Tel: +44 (0) 131 665 6062 / Mobile 07710 585 401 } FAX: +44 (0) 131 653 6248 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
EMBO Practical Course on Electron Microscopy, Immunocytochemistry and Stereology for Cell Biology at EMBL, Heidelberg, Germany, Aug 30 - Sep 8, 2001
This is a course about the production of thin sections of biological material and their use in studying ultrastructure in the context of molecular cell biology. The course will introduce the techniques of cryosectioning, rapid freezing methods as an alternative to chemical fixation, freeze substitution and resin embedding. It will instruct participants in the sectioning of suitably prepared material and will then concentrate on the use of these sections for localizing specific molecules within cells. Ample time will be given to hands on practical work as well as formal and informal discussion of available techniques for the localization of subcellular molecules.
Further info: http://www.EMBL-Heidelberg.DE/courses/ElectronMicroscopy01/
**************************** Sylke Helbing Course and Conference Office European Molecular Biology Laboratory Meyerhofstr. 1 D-69117 Heidelberg
A general question whether anyone knows where I can obtain information on the metallisation or backside coating (through evaporation) of silicon wafers after backgrinding ? My limited previous metals coating experience has given me the impression that true metal bonding is the result of the formation of a intermetallic/segregated region - of both metallic constituents cooling to form what is basically a weld zone with no discernible porosity interface . A mechanical bond on the other hand would simply be the 'knitting' of rough surfaces . My interest includes the goal of successful metallisation and how it is assessed as normal mechanical preparation of a cross section can tear and smear the face causing difficulty in porosity / interface evaluation . Is FIB used or are there non destructive methods for example ?
Thanks
Martyn Harris ESM LTD Cardiff Rd, Newport , Gwent South Wales , UK . Email : harrism-at-esm-semi.co.uk
I don't think the chloroform has gotten more toxic (unless I forgot about that fact, having used that technique for more years than I can remember....). Hopefully, a lot of us have gotten a tiny bit smarter about exposure to potentially harmful chemicals. Actually, over the years I have developed sensitivity to a number of chemicals in the EM Lab, and still tend to be a little careless about exposure to vapors - especially formalin, glut, osmium, xylene, acetone, EtOH. But I, too, have used the heat pens for well over 20 years (ever since the first cautery pens became available thru one of the EM suppliers), and urge individuals who do a lot of sectioning to switch to the heat pens.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
On Wed, 06 Dec 2000 17:20:07 +0000, Malcolm Haswell 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 } -----------------------------------------------------------------------. } } } Timothy } } I would be a bit concerned about your use of chloroform to flatten sections, } especially in a small microtome room. We have used heat pens for the last decade } or so, because apparently the chloroform we use these days is a lot more toxic } than it used to be (as well as formaldehyde, glutaraldehyde, carbon } tetrachloride etc). } } Malcolm Haswell } e.m. unit } School of Sciences } University of Sunderland } UK } } Timothy Schneider wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } The following comments are for the person who is trying to cut serial } } sections with Spurrs. I routinely cut serial sections with Spurrs without } } resorting to any kind of glue. Probably the most important thing is how the } } block is faced. I cut the trapezoid with a really long (.6 to .9 mm base) } } and the top of the trapezoid is almost the same length. What's really } } critical here is that those two cuts are parallel. } } The two sides of the trapezoid are very short (.1mm). I cut the sections } } with a Diatome knife and on a Reichert Ultracut E with a cutting speed of } } 0.8mm per second and a thickness setting of 80nm. I am doing this in a } } small room with a blasting air vent in the ceiling. I have cut up a } } cardboard box and stuck it into the ceiling tiles in such a way that the air } } is diverted away from the microtome. I can pick up 15 to 25 sections in a } } row on a 0.4 X 2mm copper slotted grid coated with just formvar or } } formvar/carbon. I have never been any good at making the support films my } } self, so I buy the slotted grids precoated from Ted Pella (catalog #s 01706, } } 01806, or 01816). Before picking up the ribbon, I dip a stick into } } chloroform and wave that over the sections, and they expand (or relax, } } depending on your point of view). I have a bunch of self locking tweezers } } and after picking up the sections, I leave the grid with the sections on it } } locked in the tweezers until they dry (if you put a wet slotted grid down on } } filter paper it might break the film). Good luck, Tim } } } } Timothy G. Schneider } } Director of Electron Microscopy } } Department of Pathology } } Room 229 Jefferson Hall } } Thomas Jefferson University } } 1020 Locust St. } } Philadelphia Pa. 19107 } } 215-503-4798 work } } 610-613-8170 cellular } } timothy.schneider-at-mail.tju.edu } }
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I have an old American Optical Spencer microscope (rescued from a trash bin). It is a binoc, three objective turret (with 4x, 10x, 20x) and built-in light source scope. I have loaned the scope to a young man who is being home schooled, and he has become really interested in microscopy, and aquatic biology. He is at the point that he needs at least a 40x objective, and all I have are the 3 that came with the scope. The only numbers I can find on the scope are 1063-1AA and 1235, and neither is specifically designated as the model or serial numbers. Can anyone direct me to a source for objectives for this type of microscope? These old scopes used a standard thread and tube length, so maybe there are non-manufacturer objectives that I can put on the scope. I haven't been able to look thru an Edmund Scientific catalog (there's one around here somewhere), but I would hope that either a collector or someone at Leica could provide some additional info.
Thanks in advance.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
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In principle, wave front reconstruction can be done by:
1) Off-axis holography: This methods need a special biprism for overlapping of two beams (one passed the specimen, the other one passed a hole). For the wavefront construction are two commercial programs available - one from Gatan and the other one from our company. The group around Prof. Hannes Lichte in Dresden is very active in this field - you should contact him, if you want to know details.
2) Focal variation: This method reconstructs the wave front from a focal series with very small defocus steps. For the reconstruction is at least one commercial program available - you should contact Andreas Thust in Juelich, Germany, if you want to know more about it. Also the group from Prof. Dirk van Dyck in Antwerp is very active in this field. The automatic acquisition of focal series is integrated in our programs and we are supporting also the JEOL-4000 remote control, but we are supporting only our cameras, no Gatan cameras. Sorry.
I apologize if I might have forgotten some references - I remember also a poster from Michael O'Keefe in Philadelphia from this year, which was related to this matter.
Regards, Ingo
++++++++++++++++++++++++++++++++++++++++++++++++ Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: www.tvips.com Email: ingo.daberkow-at-tvips.com
-------- Original Message --------
forwarded from the imaging news group
"james patrick birrell" {jbirrell-at-students.uiuc.edu} wrote in message news: {Pine.GSO.4.10.10012041635310.22505-100000-at-ux12.cso.uiuc.edu} ... } I was wondering if anybody would know where I could obtain software to } facilitate the acquisition/analysis of images to be used to perform } wavefront reconstruction using images from a TEM. Specifically, I'm } looking for any software compatable with a JEOL 4000ex microscope with a } Gatan CCD camera. } Thanks, } James Birrell
I have always understood that we use cacodylate buffer because it penetrates tissue faster and further than other buffers, taking the fixative with it. Plus, of course, the advantages that it does not form a precipitate with osmium, and that if you need to store tissues after fixing, then it discourages the growth of microorganisms. It is toxic, but surely not as much so as glutaraldehyde or OsO4, which we can't really avoid in EM. So, I would suggest we all go on using it, and take all the proper precautions for this and the other toxic agents that we need.
Lesley Weston.
On Wed, 6 Dec 2000, Malcolm Haswell 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 } -----------------------------------------------------------------------. } } } Bart } } I have used PIPES as a substitute for cacodylate in most routine procedures } simply because it has none of the precipitation problems of phosphate } buffers. I understand that there may be more extraction with cacodylate in } TEM so I would have thought that it may be a better buffer for SEM (less } chance of shrinkage?). I would assume that HEPES was similar but have no } experience with it. } } These are my own feelings and I have seen no scientific evidence to support } this - has anyone else? } } } Malcolm Haswell } e.m. unit } School of Sciences } University of Sunderland } UK } } Bart De Pauw wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hello listers, } } } } We use cacodylate buffer for our SEM samples. Is HEPES or PIPES buffer } } an alternative for the toxic cacodylate ? Our samples are mainly animal } } tissue. } } } } Bart De Pauw } } Ghent University } } Faculty of Veterinary Medicine } } Department Morphology } } Belgium } } }
I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 to the scopes in our facility. Both the previous director and I feel that we do not get our monies worth, so to speak, from this dewar. I had our local supplier look at it and they say it is fine. Once it gets to 1/2 tank,the remaining LN vaporizes and bleeds off within a day. The last time I had it filled, I calculated that I used about 50L of the 160L before it went dry. Once it got down to half, I dumped the remaining LN into a 34L dewar (and it is stll there). Does everyone have pretty much the same experience with these 160L dewars?
Thanks for your help, and Merry Christmas to all microscopists (who are the salt of the earth!)
Sincerely,
Todd
Dr. Todd A. Kostman Assistant Professor of Biology and Microbiology Director, Electron Microscopy Facility University of Wisconsin Oshkosh 800 Algoma Boulevard Oshkosh, WI 54901-8640 ph: (920) 424-3069 fax: (920) 424-1101 kostman-at-uwosh.edu
Yes, I have also found 160L dewars NOT to be cost effective with only a small user base. I switched to a 35L dewar many years ago. Even with the more frequent deliveries I'm saving money over the 160L.
************************************************ ### Free the Psoas ### **************
Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Geological Sciences ph.206-543-8393 Box 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
On Thu, 7 Dec 2000, Todd Kostman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear EMers, } } I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 } to the scopes in our facility. Both the previous director and I feel that } we do not get our monies worth, so to speak, from this dewar. I had our } local supplier look at it and they say it is fine. Once it gets to 1/2 } tank,the remaining LN vaporizes and bleeds off within a day. The last time } I had it filled, I calculated that I used about 50L of the 160L before it } went dry. Once it got down to half, I dumped the remaining LN into a 34L } dewar (and it is stll there). Does everyone have pretty much the same } experience with these 160L dewars? } } Thanks for your help, and Merry Christmas to all microscopists (who are the } salt of the earth!) } } } Sincerely, } } } Todd } } Dr. Todd A. Kostman } Assistant Professor of Biology and Microbiology } Director, Electron Microscopy Facility } University of Wisconsin Oshkosh } 800 Algoma Boulevard } Oshkosh, WI 54901-8640 } ph: (920) 424-3069 } fax: (920) 424-1101 } kostman-at-uwosh.edu } } } }
We have had a similar experience here, going to a 160L dewar after having to make a trip across campus with a 30L dewar everytime we needed LN2. The huge discount on the 160L container means we're paying about the same per liter actually used as we were before, plus we no longer have the aggravation of having to go and get it.
One discovery we made, however, is that by transferring LN2 from the 160L dewar to smaller, non-pressurized ones we are able to keep the nitrogen much, much longer. We have 10L, 50L and 30L containers in our lab, so now when a new 160L dewar is delivered, we fill up the smaller ones immediately, then use the 160L until it's gone. The smaller dewars will keep LN2 literally for weeks with minimal losses. Initially we were averaging 60 usable liters per 160 delivered, but now we are probably up way past 100 (I haven't actually figured it out.).
Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Todd Kostman [mailto:kostman-at-vaxa.cis.uwosh.edu] Sent: Thursday, December 07, 2000 12:29 PM To: Microscopy Listserver
Dear EMers,
I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 to the scopes in our facility. Both the previous director and I feel that we do not get our monies worth, so to speak, from this dewar. I had our local supplier look at it and they say it is fine. Once it gets to 1/2 tank,the remaining LN vaporizes and bleeds off within a day. The last time I had it filled, I calculated that I used about 50L of the 160L before it went dry. Once it got down to half, I dumped the remaining LN into a 34L dewar (and it is stll there). Does everyone have pretty much the same experience with these 160L dewars?
Thanks for your help, and Merry Christmas to all microscopists (who are the salt of the earth!)
Sincerely,
Todd
Dr. Todd A. Kostman Assistant Professor of Biology and Microbiology Director, Electron Microscopy Facility University of Wisconsin Oshkosh 800 Algoma Boulevard Oshkosh, WI 54901-8640 ph: (920) 424-3069 fax: (920) 424-1101 kostman-at-uwosh.edu
I don't agree. I seriously doubt that the diffusion rate of cacodylate and the "Good buffers" (i.e., the buffers like PIPES, HEPES, MES, that were designed by Dr. Good) were ever compared since they are from different eras. I am also not sure I understand how the diffusion rate of a buffer component "takes the fixative with it" at the molecular level. Arsenic buffers do inhibit bacterial growth but I have had bacteria grow in cacodylate buffers. Furthermore, if you have aldehydes or osmium in your buffer, you have all the anti-bacterial agent you need. Cacodylate buffers can give off toxic gases when acidified but more importantly, you are adding needlessly to the toxic waste stream. Aldehydes will eventually react or breakdown into non-toxic substances but the arsenic will remain a problem forever. Cacodylate was originally selected based on its pKa and non-reactivity with fixatives. Phosphate caused problems with calcium precipitation and Tris buffers had amino groups that react with the fixatives. The choices for buffers were very limited in the old days. Dr. Good made a significant contribution in his description of this family of buffers. Anatomists tend to be the most old fashioned scientists in the world.
} } } I have always understood that we use cacodylate buffer because it } penetrates tissue faster and further than other buffers, taking the } fixative with it. Plus, of course, the advantages that it does not form a } precipitate with osmium, and that if you need to store tissues after } fixing, then it discourages the growth of microorganisms. It is toxic, but } surely not as much so as glutaraldehyde or OsO4, which we can't really } avoid in EM. So, I would suggest we all go on using it, and take all the } proper precautions for this and the other toxic agents that we need. } } Lesley Weston. } } } } On Wed, 6 Dec 2000, Malcolm Haswell 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 } } -----------------------------------------------------------------------. } } } } } } Bart } } } } I have used PIPES as a substitute for cacodylate in most routine procedures } } simply because it has none of the precipitation problems of phosphate } } buffers. I understand that there may be more extraction with cacodylate in } } TEM so I would have thought that it may be a better buffer for SEM (less } } chance of shrinkage?). I would assume that HEPES was similar but have no } } experience with it. } } } } These are my own feelings and I have seen no scientific evidence to support } } this - has anyone else? } } } } } } Malcolm Haswell } } e.m. unit } } School of Sciences } } University of Sunderland } } UK } } } } Bart De Pauw wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Hello listers, } } } } } } We use cacodylate buffer for our SEM samples. Is HEPES or PIPES buffer } } } an alternative for the toxic cacodylate ? Our samples are mainly animal } } } tissue. } } } } } } Bart De Pauw } } } Ghent University } } } Faculty of Veterinary Medicine } } } Department Morphology } } } Belgium } } } } } }
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Hi Friends, Once a gain I need that voodoo that you do so well.
A colleague is asking how to embed, freeze and frozen-section (cross section) a rat sciatic perpheral nerve. Several attempts have failed to give adequate structural detail. Any tips and tricks are welcome.
We got similar results when we monitored nitrogen use 10 years ago, and again when we retested earlier this year. We tried two different 160 liter tanks, one of which was brand new, and never got more than about 80 liters out. I eventually gave up and went to four 50 liter tanks, which we receive at atmospheric pressure and tap with a withdrawal device that self pressurizes to about 9 psi. The device has a rubber (now teflon for better thermal properties) transfer hose, so there is little loss due to warming the hose. We usually get 40-45 liters out of these.
The losses, as I understand them, are from
1. passive losses from the tank itself (static evaporation rate) which are higher for the 160 liter tanks 2. losses due to cooling of the transfer tube (lower in the 50 liter tank with no phase separator) 3. depressurization loss, which is higher in the 160 liter tank pressurized to 20 psi than for the 50 L tank at 9 psi.
Obviously if you need high pressure or use high volumes, the 160 liter tank is better. Also, with the 50 liter tanks you have to plan ahead because it takes some time to pressurize the tanks, but otherwise the 50 liter option works for us.
Marie
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Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
we seem to be the exception. I expect to recover about 100L from a 160L dewar over a two to three week period. I'm sure we once got a near record 110L out but occasionally fall a little below the 100. We still therefore find the 160L storage dewar more convenient because the alternative would require more space, higher delivery cost and the need for multiple trolleys for several 25 litre dewars. We can normally get nitrogen almost to the bottom of the 160L dewar, providing that we don't leave less than about 10L in it (I'm guessing). The only down-side is that it is a pressure vessel and requires regular maintenance and testing to meet UK regulations, but this means that the spring loaded pressure relief valve is regularly checked (as well as the burst disk valve of course). It does however mean that if our 160L dewar ever fails its tests that we will probably just buy more 25L dewars instead of replacing it.
Good luck
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Todd Kostman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear EMers, } } I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 } to the scopes in our facility. Both the previous director and I feel that } we do not get our monies worth, so to speak, from this dewar. I had our } local supplier look at it and they say it is fine. Once it gets to 1/2 } tank,the remaining LN vaporizes and bleeds off within a day. The last time } I had it filled, I calculated that I used about 50L of the 160L before it } went dry. Once it got down to half, I dumped the remaining LN into a 34L } dewar (and it is stll there). Does everyone have pretty much the same } experience with these 160L dewars? } } Thanks for your help, and Merry Christmas to all microscopists (who are the } salt of the earth!) } } Sincerely, } } Todd } } Dr. Todd A. Kostman } Assistant Professor of Biology and Microbiology } Director, Electron Microscopy Facility } University of Wisconsin Oshkosh } 800 Algoma Boulevard } Oshkosh, WI 54901-8640 } ph: (920) 424-3069 } fax: (920) 424-1101 } kostman-at-uwosh.edu
Many thanks to all who replied to this query. I should have thought of the EDAX site, senile dementia is really kicking in now. Just as well I'm being thrown on the scrap heap at Christmas.
I do not know if I can help, but I have a an excellent 50x oil AO objective with a correction collar used on an AO MicroStar series compound microscope. This is the scope with a grey finish. The numerical aperture of this objective is 0.80 and with the correction collar exceeds the light gathering capacity and resolution of any dry 40x or 63x dry objective.
Do you have any items you can perhaps trade, i.e., Zeiss objective lenses?
Let me know what you think.
Ken ------------- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
Contact tel. number: 503-413-5391
On Thu, 7 Dec 2000, Roger Moretz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings: } } I have an old American Optical Spencer microscope (rescued from a trash } bin). It is a binoc, three objective turret (with 4x, 10x, 20x) and } built-in light source scope. I have loaned the scope to a young man who is } being home schooled, and he has become really interested in microscopy, and } aquatic biology. He is at the point that he needs at least a 40x objective, } and all I have are the 3 that came with the scope. The only numbers I can } find on the scope are 1063-1AA and 1235, and neither is specifically } designated as the model or serial numbers. Can anyone direct me to a source } for objectives for this type of microscope? These old scopes used a } standard thread and tube length, so maybe there are non-manufacturer } objectives that I can put on the scope. I haven't been able to look thru an } Edmund Scientific catalog (there's one around here somewhere), but I would } hope that either a collector or someone at Leica could provide some } additional info. } } Thanks in advance. } } Roger Moretz, Ph.D. } Dept of Toxicology } Boehringer Ingelheim Pharmaceuticals } } } } } } _______________________________________________________ } Tired of slow Internet? Get -at-Home Broadband Internet } http://www.home.com/xinbox/signup.html } } }
I receive my liquid nitrogen in 160 L tanks from our supplier and they generally last from 24 to 30 days with me filling up the EDX dewar twice a week (about 3-4 liters). I have had some tanks that only lasted two weeks and after I told our supplier about that they said the leak valve was stuck open and they replaced or adjusted the leak valve as needed. You should get your local supplier to inspect your tank and maybe replace or adjust the leak valve. I would not adjust it myself unless you really know what you are doing since an explosion is possible without proper pressure release. Terry Ellis Hallmark Cards Inc. e-mail: tellis2-at-hallmark.com
Imagine all day sectioning, it isn't hard to do. Putting sections on bare 200 mesh grids and heat fixing under a light to boot.
Imagine all the sections coming off while you do the stain. Oh oh You have to figure out what's wrong, while racking your brain. If you all can help, you'll help me from going insane.
Any suggestions will be greatly appreciated. Thanks is advance!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We have a post-doc that is interested in looking at tight junctions in cultured endothelial cells. He has found a vague reference for using silver stain to visualize endothelial cell junctions. Does anyone have any experience with this technique? Can you provide some technical details or point us to a reference? Thank you!!
ctfExplorer has been updated. New features/improvements: 1. Now it is possible to export graphs to text (tab-delimited). 2. List-tree with the microscopes is wider now. 3. Tab stops in dialogs arranged in a logical sequence. 4. Now it calculates the Lichte-defocus. (Useful for holography). 5. Additional slider-control is added: allows to change "magnification", i.e. convergence. Just to show people why it is not always a good idea to work at 1,000,000x and why some people do high resolution work at as low as 50,000x magnification.
Here is the link: http://clik.to/ctfexplorer (always use this link. it will direct you to the right location).
Enjoy,
Max Sidorov --- (until December 31 2000) TEM Applications Specialist FEI/Philips Electron Optics Eindhoven, The Netherlands e-mail: maxsidorov-at-bigfoot.com (mailto:maxsidorov-at-bigfoot.com)
----------Additional Info---------- Dear All: I wrote a piece of software which I believe would be of interest to the microscopy community. It's a highly interactive program which calculates/displays the contrast transfer function of TEMs. I know that there are similar programs floating around but ctfExplorer does not only 1d but also 2d calculations/display with 2-fold and 3-fold astigmatism imposed. There are other unique features to it. All parameters (defocus, voltage, Cs, etc) can be changed interactively.
DISCLAIMER: I do work for FEI/Philips Electron Optics. However, this software is not an official product of FEI and FEI is not responsible for its distribution/support. This software is freeware.
To the best of my knowledge, the software does not have any nasty bugs. It is tested under Windows 95/98 and Windows NT4.
Please give it a try. Please direct your suggestions and comments to maxsidorov-at-bigfoot.com I hope you'll find the software useful.
Here is the link: http://clik.to/ctfexplorer
Enjoy,
Max Sidorov --- TEM Applications Specialist FEI/Philips Electron Optics Eindhoven, The Netherlands e-mail: maxsidorov-at-bigfoot.com
Todd- I have not calculated how much LN2 I am able to deliver from my 160 L dewar, but it must be about 100 L since it will last me a month or more. Perhaps my dewar is of a better grade as it is also certified for liquid He use. I suspect that much of my LN2 loss is in cooling down the 10 L dewar I use to transport it to the EDX dewar. Are you using a high pressure, or low pressure dewar (mine has ~24psi max)? The high pressure dewars I have had have an internal coil for warming up some liquid to maintain the head pressure. If your dewar has this, there should be valves for closing off this pressure generation coil which will extend the storage time. By the way, what does your fill gauge read when the dewar is empty? These gauges are frequently very inaccurate. The fact that you could fit the remainder in a 34 L dewar when your gauge said 1/2 full, indicates to me that your gauge probably isn't accurate. I hope that something in there helps... Matt
Todd Kostman {kostman-at-vaxa.cis.uwosh.edu} on 12/07/2000 01:29:29 PM
To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} cc:
Dear EMers,
I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 to the scopes in our facility. Both the previous director and I feel that we do not get our monies worth, so to speak, from this dewar. I had our local supplier look at it and they say it is fine. Once it gets to 1/2 tank,the remaining LN vaporizes and bleeds off within a day. The last time I had it filled, I calculated that I used about 50L of the 160L before it went dry. Once it got down to half, I dumped the remaining LN into a 34L dewar (and it is stll there). Does everyone have pretty much the same experience with these 160L dewars?
Thanks for your help, and Merry Christmas to all microscopists (who are the salt of the earth!)
Sincerely,
Todd
Dr. Todd A. Kostman Assistant Professor of Biology and Microbiology Director, Electron Microscopy Facility University of Wisconsin Oshkosh 800 Algoma Boulevard Oshkosh, WI 54901-8640 ph: (920) 424-3069 fax: (920) 424-1101 kostman-at-uwosh.edu
Paula, I loved your poem! I too , had the same problem with sections falling off grids while staining, especially if using the thin bar grids. There's just not enough surface area for the sections to stick to. I solved the problem by putting the grids in the 60 degree oven overnight before staining. I haven't lost any sections since, and I cut both very large and small sections. The time in the oven doesn't seem to adversely affect the tissue.
Good luck, Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
Thanks to all who have already answered my plea. I think I need to make a few things clear. Me, in my effort to be witty forgot a few major details.
1. I'm cutting Spurr's embedded samples.
2. I'm cleaning the samples in a dilute acetic acid followed by an acetone chaser.
3. I've been heat fixing the grids/sections under a light for about 10 minutes.
4. I've been doing this for that past 10 months and things worked great until 3 weeks ago. I'm new to this area (Washington, DC) having been in California doing EM for years. Could this be a climate/weather related problem?
5. I will use formvar if I have to. Has anyone just dipped grids in formvar, instead of casting a film, to make the grids a little sticky? I inherited a bunch of filthy formvar with water & junk in it and it takes about a month to get supplies.
6. I haven't changed my supplier of grids or Spurr's components. So I think that things should be OK on the quality control end.
A lot of the suggestions I received are very good, but I have already tried most of them. Any bizarre rituals that I must do? I WILL NOT swing a dead cat over my head, but I might try other things. EM is basically voodoo, after all.
Thanks again,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
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Hi Paula, It is horrible to see naked grids come off the stain. We've probably all been there. Here are my auggestions:
1. Make sure that your grids are clean to start. You can dip them in acetone or ethanol (absolute) and then water and dry thoroughly.
2. Make sure that the sections have fully dried onto the grids. If you're not rushed, just let them dry overnight. If you are rushed (probably the usual situation), you can lay the grids out on a clean glass slide that is resting on a warm (NOT HOT) hot plate (around 35-40deg.C) for a few minutes.
3. And as wierd as this seems....make sure that the water you are using to make up your stains and to wash the grids is really clean (deionized or double distilled or microfiltered). I had a friend to used to come over to my building to fill a small carboy with water because my dept. had better water treatment than hers. She swore that her sections fell off when she used their water.
4. Maybe the simplest life saver....buy a "Coat-Quick G" grid coating pen. It deposits a small amount of the "secret formula" adhesive onto the grid bars, without blocking the spaces or causing problems with dirt/stain deposition. I frequently cut enormous sections (by EM standards) and this has been a blessing. I know that Electron Microscopy Sciences sells them. Other companies probably do too. Look in you favorite vendor's catalogue.
(Disclaimer: I have no financial interests in EMS)
Good luck,
Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Responding to the message of {sa30a2a7.040-at-gwise.gwumc.edu} from "Paula Sicurello" {patpxs-at-gwumc.edu} :
Paula,
Imagine........ a magic solution........
GRID CLEANING SOLUTION
Clean your uncoated grids each time you set up to do sectioning, because newly cleaned copper grids will oxidize within a day and sections will no longer stick to them very well during staining and rinsing.
*ADD 40 ml CONCENTRATED HCL TO 400 ml DISTILLED WATER
*ADD 40 ml ACETONE TO THE ABOVE
*DILUTE TO 500 ml FINAL VOLUME
*SONICATE GRIDS IN THIS SOLUTION IN A 50 ml BEAKER FOR 1-2 MINUTES
*DISCARD USED SOLUTON DOWN DRAIN AND RINSE/SONICATE GRIDS FOR 30 SECONDS TWICE WITH 100% ACETONE
*DUMP GRIDS ONTO CLEAN FILTER PAPER TO DRY
* ITS TRADITIONAL TO COLLECT SECTIONS ON THE DULL SIDE OF THE GRIDS.
It works well for us here, and is used routinely every day. Good luck,
Gib
} } Happy holidays oh great wise ones of microscopy! } } (Apologies to John Lennon) } } } Imagine all day sectioning, } it isn't hard to do. } Putting sections on bare 200 mesh grids } and heat fixing under a light to boot. } } Imagine all the sections coming off } while you do the stain. } Oh oh } You have to figure out what's wrong, } while racking your brain. } If you all can help, } you'll help me from going insane. } } } Any suggestions will be greatly appreciated. Thanks is advance! } } Paula :-) } } Paula Sicurello } George Washington Univ. Medical Center } Dept. of Pathology, Ross Hall rm 505 } Electron Microscope Lab } 2300 Eye St. } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
I'm looking for methods of preparing compacted potassium chloride fertilizer for the SEM. We believe that grain size is indicative of compaction quality in our process, however we're not sure of the best way to prepare the samples. The compactor flake is approximately 2"x1"x0.25". Any advice would be much appreciated.
I have a freind that need then main prism slide containing the Wollaston prism for Reichert Zetopan DIC scope. This is the slide going in the upper body of the scope. He also needs the polorizor that fits over the light soruce but that is of much less importance beause it can be easily made. While the prism is constructed of unobtainium.
He is willing to pay a reasonable price for the part.
If any of you have a spare or left over you could make a very dissapointed man a lot happier.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
I have noticed that in some cases the kind of grid you use is quite important. I had similar problems with Gilder grids when using Spurr embedded plant tissue sections. Later I switched to Veco grids, the problem went away.
Good luck,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab and Fluorescence Imaging Facility Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
Sandy Hancock Try the following reference; Zand, Underwood, Nunnari, Majno and Joris, Endothelium and 'silver lines'. An electron microscopic study. 1982 Virchows Archiv of Path Anat. 395: 133-144 Good luck John
Thanks to everyone for the response. It will take me a little while to fully sort through everything, but I appreciate the help.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
On Fri, 8 Dec 2000 02:31:47 -0800 (PST), Ken Tiekotter wrote:
} Dear Roger, } } I do not know if I can help, but I have a an excellent 50x oil AO } objective with a correction collar used on an AO MicroStar series compound } microscope. This is the scope with a grey finish. The numerical aperture } of this objective is 0.80 and with the correction collar exceeds the light } gathering capacity and resolution of any dry 40x or 63x dry objective. } } Do you have any items you can perhaps trade, i.e., Zeiss objective lenses? } } Let me know what you think. } } Ken } ------------- } Ken Tiekotter, Adjunct Professor } The University of Portland } Department of Biology } 5000 N. Willamette Blvd. } Portland, OR 97203 } } Contact tel. number: 503-413-5391 } } On Thu, 7 Dec 2000, Roger Moretz wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Greetings: } } } } I have an old American Optical Spencer microscope (rescued from a trash } } bin). It is a binoc, three objective turret (with 4x, 10x, 20x) and } } built-in light source scope. I have loaned the scope to a young man who is } } being home schooled, and he has become really interested in microscopy, and } } aquatic biology. He is at the point that he needs at least a 40x objective, } } and all I have are the 3 that came with the scope. The only numbers I can } } find on the scope are 1063-1AA and 1235, and neither is specifically } } designated as the model or serial numbers. Can anyone direct me to a source } } for objectives for this type of microscope? These old scopes used a } } standard thread and tube length, so maybe there are non-manufacturer } } objectives that I can put on the scope. I haven't been able to look thru an } } Edmund Scientific catalog (there's one around here somewhere), but I would } } hope that either a collector or someone at Leica could provide some } } additional info. } } } } Thanks in advance. } } } } Roger Moretz, Ph.D. } } Dept of Toxicology } } Boehringer Ingelheim Pharmaceuticals } } } } } } } } } } } } _______________________________________________________ } } Tired of slow Internet? Get -at-Home Broadband Internet } } http://www.home.com/xinbox/signup.html } } } } } } }
_______________________________________________________ Tired of slow Internet? Get -at-Home Broadband Internet http://www.home.com/xinbox/signup.html
Paula Try this cleaning procedure for your copper grids: 1) Place your grids in a clean beaker, 100ml size will do. 2) Cover the gird with Glacial Acetic Acid, approximately 20cc (the idea here is to get the grim and grease off the grids not to eat the metal away) 3) Sonicate the grids in this acid for about 5 to 10 minutes. 4) Rinse in 100% acetone until the smell of the acetic acid is gone. 5) Do a final rinse in 100% ETOH (denatured is ok) and pipette the ETOH off and invert the beaker into a glass petri dish that has a clean whatman filter paper in it and place in your 60 degree over for an hour or more. The grids will fall on to the filter paper. Cover the bottom of the glass with a top. The grids will keep without father cleaning until they are all used up. I use a glass petri dish because there is less static charge with glass.
Ron Austin Dept. of Pathology LSU Medical Center Shreveport, LA rla-at-mindspring.com 318-675-4557
I'd just try a simple first shot approach of putting some of the particles on a sticky tab, and coating it with about 60A of Au/Pd. Then, take a look at it under the SEM. Based on this imaging, other approaches may or may not be necessary.
It may or may not make a difference relative to the type of SEM that you are using (FESEM vs. thermionic). I would think that these are rather large specimens. Either system should do a nice job.
gary g.
At 01:26 PM 12/8/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} From time to time, someone opens a grid storage box in our lab, and because of the static that has built up on the plastic, the grids fly out and are randomized. We have a Zerostat gun next to the TEM, but our laboratory/office space is fragmented worse than a SWAP file, and one can't simply reach for the ion gun every time one opens a grid box. Does anyone (a) have any simple solutions to this problem (b) know of a supplier of grid boxes made of a different plastic that doesn't charge up so horribly?
In the longer term, would there be scope for making grid boxes out of a plastic loaded with a filling to make it conductive enough for charges to slowly leak away, so avoiding the problem altogether?
And on a similar theme, does anyone know of storage boxes for SEM stubs into which the stubs can fit easily, i.e. not having to be rammed in?
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
Antistatic sprays are available from electronic components suppliers such as Farnell and RS Components. These are mainly intended to de-static the plastic covers of ammeters, etc. to stop them influencing the readings.
You could also try coating the interior surfaces of your box with a thin layer of carbon or gold next time you have the coater running.
Chris
Date sent: Sat, 9 Dec 2000 14:59:05 +0000 (GMT Standard Time) } From: "Robert H. Olley" {r.h.olley-at-reading.ac.uk} To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} Copies to: # {r.h.olley-at-reading.ac.uk}
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Robert Olley wrote: ================================================================ } From time to time, someone opens a grid storage box in our lab, and because of the static that has built up on the plastic, the grids fly out and are randomized. We have a Zerostat gun next to the TEM, but our laboratory/office space is fragmented worse than a SWAP file, and one can't simply reach for the ion gun every time one opens a grid box. Does anyone (a) have any simple solutions to this problem (b) know of a supplier of grid boxes made of a different plastic that doesn't charge up so horribly?
In the longer term, would there be scope for making grid boxes out of a plastic loaded with a filling to make it conductive enough for charges to slowly leak away, so avoiding the problem altogether?
And on a similar theme, does anyone know of storage boxes for SEM stubs into which the stubs can fit easily, i.e. not having to be rammed in? ================================================================= There are a number of grid boxes being molded today, some of which are being molded with plastics that have had added antistatic additives. The SPI Slide-A-Grid™ storage boxes (see website below) are molded with such antistatic additives. This same box is offered (but under other names) by some of the other leading firms offering these kinds of products to EM users . However, under certain conditions, apparently humidity related, even these boxes reportedly can develop a charge.
For SEM storage boxes, there are two approaches to the "base plate" that sits in the bottom of each box, the first being a "hard" plastic such as polypropylene and the other being with a "soft" plastic, specifically an elastomeric plastic. SPI Supplies has offered storage boxes with the soft plastic for some number of years, the advantage being that the mounts do fit in easily and do not have to be "rammed" in. I do not know of anyone else who is molding mounting plates out of the soft "elastomeric plastic" (but of course I could be wrong about that).
Note: One of the biggest sources of misfit between the storage boxes and the 3/8" (9.5 mm) round mounts is that boxes molded in the USA tend to be made for 3/8" diameter mounts and boxes molded elsewhere seem to be made for 10 mm diameter mounts. The problem arises when one tries to put a 10 mm round mount into a box designed for the 3/8"round mounts. If the "ramming" problem you described could be this kind of a problem, a mixing up of boxes/mounts might be the reason for the described difficulty. The flip- side of this problem is when 3/8" mounts are put into boxes molded in Europe to take 10 mm round mounts, and the result is, the mounts fall out when the box is tipped.
Disclaimer: SPI Supplies offers boxes for the storage of both TEM grids and SEM mounts, the details of which can be found on our website below.
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 ######################## ============================================
TECHNICAL OFFICER/SENIOR TECHNICAL OFFICER/CHIEF TECHNICAL OFFICER ELECTRON MICROSCOPE UNIT
If you have experience in electron microscopy and the use of computers, then we invite you to apply for this post in a fast-moving and ever-changing environment geared to servicing research needs.
The Electron Microscope Unit (EMU) primarily serves researchers in the faculties of Science, Engineering and the Built Environment and Health Sciences at UCT. The EMU aims to be the foremost provider of such services in the Western Cape. Currently, the Unit has two scanning and two transmission electron microscopes as well as a number of light microscopes and will take delivery of a cryo-TEM early next year.
Duties of the appointee will include instrument operation, sample preparation, user training and laboratory management.
The level of appointment will depend on the qualifications and the experience of the successful candidate. Therefore, the remuneration package, including benefits, is negotiable between R75 000 - R156 100 a year.
Please send a letter of application plus your CV (including the names, postal/email addresses, telephone/fax numbers of 2 referees) to Associate Professor Trevor Sewell, Director: Electron Microscope Unit, UCT, Rondebosch, 7701 by 8 January 2001; tel: (021) 650-2817; fax: 689-1528; email: sewell-at-uctvms.uct.ac.za
During my laboratory days I never worked in a room were static was that bad - and that included Canadian winters. Carpet and poor humidification can make a static problem worse. I never liked the boxes with a sliding lid for another reason: If they dropped when open from a modest height several grids may escape and that is really the same problem. Probably all of the EM suppliers carry grids boxes that hold 24 grids and have a rotating lid with two apertures. Only one grid at a time is can to escape. Presto: No more feral grids.
I'm sure that "smarter" stub boxes could be designed, but would people buy them at 2 or 3 times the price? We (and others) have a single SEM mount Storage/mailer and this is much better for holding pin type mounts. Wouldn't is be lovely if all EM manufacturers used the same (I vote for pin type) mounts. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Sunday, December 10, 2000 12:59 AM, Robert H. Olley [SMTP:r.h.olley-at-reading.ac.uk] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } From time to time, someone opens a grid storage box in our lab, and } because of the static that has built up on the plastic, the grids fly out } and are randomized. We have a Zerostat gun next to the TEM, but our } laboratory/office space is fragmented worse than a SWAP file, and one } can't simply reach for the ion gun every time one opens a grid box. Does } anyone (a) have any simple solutions to this problem (b) know of a } supplier of grid boxes made of a different plastic that doesn't charge up } so horribly? } } In the longer term, would there be scope for making grid boxes out of a } plastic loaded with a filling to make it conductive enough for charges to } slowly leak away, so avoiding the problem altogether? } } And on a similar theme, does anyone know of storage boxes for SEM stubs } into which the stubs can fit easily, i.e. not having to be rammed in? } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+ } } } }
On behalf of the Microscopy Society of America and the Microbeam Analysis Society, we invite you to attend Microscopy and Microanalysis 2001, August 5-9, in Long Beach, California. The program chair, Bob Price, and Co-Chairs Edgar Voelkl (MSA) and Inga Holl Musselman (MAS), have assembled another excellent technical program that continues the tradition of exciting and current presentations at the meetings. An outstanding list of invited speakers highlights the symposia in a number of key areas. Special symposia this year again cover a wide range, including "Microscopy and Microanalysis in the Real World," a special biological symposium honoring Dr. Inou*, and "Atom Location by Channeling Enhancement of X-Ray and EELS Signals" among others, plus a wide variety of applications in biology and materials research.
Both invited and contributed abstracts are critical to the success of these symposia, so we urge you to submit your work. Remember that the abstract deadline is February 15th, 2001.
A special Pre-meeting Congress, "Imaging Life: From Cells to Whole Animals," as well as several workshops and short courses on important, basic techniques will precede the meeting. Special symposia, tutorials and presentations sponsored by the Technologists' Forum, the MSA Education Committee, and various commercial exhibitors will be held during the course of the meeting. There are also Presidential Happenings, ceremonies for Award Winners, and the world's largest display of microscopes and related equipment.
Our Local Arrangements Committee, headed by Robert Koch, has arranged an excellent venue at the Long Beach Convention Center for both the scientific sessions and the exhibits. They have also arranged a memorable program of social events including a Sunday night reception on the Queen Mary, the annual golf tournament, a harbor cruise, and a large number of options for enjoying the city and the surrounding area (don't forget the theme parks!). With its rich history, wealth of educational institutions and active industry, Long Beach provides a wonderful context for this meeting, and we urge you to come to learn, teach, share and above all to enjoy. Whatever your connection to microscopy, we look forward to welcoming you to Long Beach in August at Microscopy and Microanalysis 2001.
Ron Anderson, President Microscopy Society of America
Richard Linton, President Microbeam Analysis Society
--------------------------
The program is as usual extensive and detailed information can be found on the WWW page.
I have fought static problems in wide variety of problems and three things help. Grounding with metal, raising the humidity to 50% and spraying the area or items with antistatic products designed for laundry. Downy was the one I used. Temporary relief can be had by spraying the area with water from Windex like spray bottle. This only last for a few minutes to an hour at most.
I am sure fabric antistatic spray would not be acceptable in many places in a lab. But floors and tables might benifit from them.
Spraying carpets with antistatic spray once a month work for us in 15 to 20% humidity.
Your mileage my vary. Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} From: "Jim at ProSciTech" {jim-at-proscitech.com} } During my laboratory days I never worked in a room were static was that bad - } and that included Canadian winters. Carpet and poor humidification can make a } static problem worse. } I never liked the boxes with a sliding lid for another reason: If they dropped } when open from a modest height several grids may escape and that is really the } same problem. } Probably all of the EM suppliers carry grids boxes that hold 24 grids and have } a rotating lid with two apertures. Only one grid at a time is can to escape. } Presto: No more feral grids. } } I'm sure that "smarter" stub boxes could be designed, but would people buy } them } at 2 or 3 times the price? } We (and others) have a single SEM mount Storage/mailer and this is much better } for holding pin type mounts. } Wouldn't is be lovely if all EM manufacturers used the same (I vote for pin } type) mounts. } Cheers } } } } } From time to time, someone opens a grid storage box in our lab, and } } because of the static that has built up on the plastic, the grids fly out } } and are randomized. We have a Zerostat gun next to the TEM, but our } } laboratory/office space is fragmented worse than a SWAP file, and one } } can't simply reach for the ion gun every time one opens a grid box. Does } } anyone (a) have any simple solutions to this problem (b) know of a } } supplier of grid boxes made of a different plastic that doesn't charge up } } so horribly? } } } } In the longer term, would there be scope for making grid boxes out of a } } plastic loaded with a filling to make it conductive enough for charges to } } slowly leak away, so avoiding the problem altogether? } } } } And on a similar theme, does anyone know of storage boxes for SEM stubs } } into which the stubs can fit easily, i.e. not having to be rammed in? } } } }
Kevex Quantum EDS system with Sesame drive is available for no charge for pick-up in Easton, PA. The detector is NOT included. System has Syquest drives and monitor is in need of repair.
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I am doing some bacterial counting work. My basic method involves taking some bacteria in a measured quantity of suspension and running it thru an anodisc (aluminum oxide) filter having .2 micron pores.
My problem is this: when mounting the filter on a slide, I find that the disc is almost dry before I can mount it. I have tried adding water and or oil below and above the filter. Water sometimes works but it is a very tricky matter because the water may redistribute the bacteria when applied. This affects my sampling since the bacteria may no longer be uniform.
The same problem occurs with oil. Furthermore, the oil, if applied to a filter with some water remaining, gives a blurry image, perhaps due to the oil-water mix.
I am also concerned with air bubbles. Is there a method by which one can prevent air bubbles under, over and in the pores of the filter?
I would *STRONGLY* recommend against using any anti-static spray on grid boxes themselves unless you are certain of the components of the spray. I remember one batch of grid boxes we had to throw away because the formulation of the plastic mix during manufactur was evidently wrong, and we got horrendous contamination of samples which had been stored in the boxes. We don't even touch the tweezers we are going to use for our samples with our bare hands! I would never spray some unknown material intended for some other, much less critical (in terms of cleanliness) application, anywhere near anything related to the samples themselves.
Tony G-R
} } Antistatic sprays are available from electronic components suppliers } such as Farnell and RS Components. These are mainly intended to } de-static the plastic covers of ammeters, etc. to stop them } influencing the readings. } } You could also try coating the interior surfaces of your box with a } thin layer of carbon or gold next time you have the coater running. } } Chris } }
} } } } } From time to time, someone opens a grid storage box in our lab, and } } because of the static that has built up on the plastic, the grids fly out } } and are randomized. We have a Zerostat gun next to the TEM, but our } } laboratory/office space is fragmented worse than a SWAP file, and one } } can't simply reach for the ion gun every time one opens a grid box. Does } } anyone (a) have any simple solutions to this problem (b) know of a } } supplier of grid boxes made of a different plastic that doesn't charge up } } so horribly? } } } } In the longer term, would there be scope for making grid boxes out of a } } plastic loaded with a filling to make it conductive enough for charges to } } slowly leak away, so avoiding the problem altogether? } } } } And on a similar theme, does anyone know of storage boxes for SEM stubs } } into which the stubs can fit easily, i.e. not having to be rammed in? } }
Apparently, Bravenet (who provides a quick link url for me) is down for updates (at least that's what they say). The quick link (http://clik.to/ctfexplorer) should start working again shortly. In a meantime you can use a direct link: http://www.ctfexplorer.f2s.com
Hope this helps, Max S
------------------- Dear All:
ctfExplorer has been updated. New features/improvements: 1. Now it is possible to export graphs to text (tab-delimited). 2. List-tree with the microscopes is wider now. 3. Tab stops in dialogs arranged in a logical sequence. 4. Now it calculates the Lichte-defocus. (Useful for holography). 5. Additional slider-control is added: allows to change "magnification", i.e. convergence. Just to show people why it is not always a good idea to work at 1,000,000x and why some people do high resolution work at as low as 50,000x magnification.
Here is the link: http://clik.to/ctfexplorer (always use this link. it will direct you to the right location).
Enjoy,
Max Sidorov --- (until December 31 2000) TEM Applications Specialist FEI/Philips Electron Optics Eindhoven, The Netherlands e-mail: maxsidorov-at-bigfoot.com (mailto:maxsidorov-at-bigfoot.com)
----------Additional Info---------- Dear All: I wrote a piece of software which I believe would be of interest to the microscopy community. It's a highly interactive program which calculates/displays the contrast transfer function of TEMs. I know that there are similar programs floating around but ctfExplorer does not only 1d but also 2d calculations/display with 2-fold and 3-fold astigmatism imposed. There are other unique features to it. All parameters (defocus, voltage, Cs, etc) can be changed interactively.
DISCLAIMER: I do work for FEI/Philips Electron Optics. However, this software is not an official product of FEI and FEI is not responsible for its distribution/support. This software is freeware.
To the best of my knowledge, the software does not have any nasty bugs. It is tested under Windows 95/98 and Windows NT4.
Please give it a try. Please direct your suggestions and comments to maxsidorov-at-bigfoot.com I hope you'll find the software useful.
Here is the link: http://clik.to/ctfexplorer
Enjoy,
Max Sidorov --- TEM Applications Specialist FEI/Philips Electron Optics Eindhoven, The Netherlands e-mail: maxsidorov-at-bigfoot.com
Microscopy Society of America Undergraduate Research Scholarship Program
With this year's call for applications the MSA Undergraduate Research Scholarship Program begins its 13th year providing funding for undergraduate research. To date over 65 projects covering a wide range of topics in the physical and biological sciences have received support through this program. Over the years nearly all the scholarship recipients have maintained a strong interest in imaging sciences and have gone on to graduate school, professional school, teaching, or industry positions.
The program, which is funded by MSA, is able to support approximately 30% to 40% of applicants. The maximal award from MSA is currently $3000 and helps to provide student stipends, supply costs, and limited travel expenses associated with the research. Additional support in the form of instrument use time, equipment purchases, etc. is generally provided by the studentís supervisor and/or through the sponsoring institution. Abstracts reporting the research results, are prepared by scholarship awardees, and published in "Microscopy and Microanalysis".
The program actively seeks external sources of matching funds in order to maintain the favorable levels of support both in terms of the number of projects supported and the level of support for each. We are extremely grateful for the matching support provided by MSA sustaining members and individuals. Their support has enabled the program to increase both the number of awards and the maximum amount of each award to $3000.
The MSA Undergraduate Research Scholarship Program is currently soliciting applications from students interested in conducting a research project which involves the use of any microscopy technique. Students must be sponsored by a member of MSA. The maximal award is $3000.
This is not limited to students in the USA or North America. Any undergraduate student anywhere is eligible to apply for the award, provided the person supervising their research is a member of MSA.
**The application deadline is Dec 31, 2000.**
Applications can be obtained from the MSA Business Office, businessoffice-at-msa.microscopy.com or call toll free at (800) 538-3672.
If you have any questions or require additional information regarding the program please contact either:
Dr. Ralph Albrecht, University of Wisconsin 1656 Linden Drive, Madison, WI 53706 (608) 263-3952 or 263-4162; (608) 262-7420 FAX; albrecht-at-ahabs.wisc.edu
Dr. Richard Ornberg, Monsanto Company Analytical Sciences Center, 800 N. Lindbergh Blvd., St. Louis, MO 63167 (314) 694-1184; (314) 694-6727 FAX; rlornb-at-ccmail.monsanto.com
FOR SALE: 1988 JEOL 100CXII with (+/-) 45degree tilt stage, dual sample holder, and Gatan camera system: a rugged, reliable transmission electron microscope in perfect working order, with water chiller: $30,000. Buyer is responsible for take-down, shipping, and set-up costs. A Kevex Delta I Class EDS analysis unit is also included, but currently not installed.
Feel free to come by for a test drive! 317 Egan Research Center Northeastern University Boston, MA 02115 For directions and to set up an appointment, call (617) 373-5909.
If interested in purchasing, please contact: Professor Terry K. Baker (617) 373-2123
RJ Lee Group's Bay Area Facility, a materials characterization and consulting laboratory, is seeking a motivated individual to actively manage projects related to the evaluation of materials and processes used in the electronics industry. Job applicants should have an MS or equivalent in physics, materials science or related discipline, at least two years of relevant work experience, and be amenable to working in a hands-on and fast-paced team environment. A working knowledge of the application and theory associated with sample preparation methods and corresponding analytical techniques such as optical microscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Auger spectroscopy and other surface analysis methods is critical. Strong oral and written communication skills are a must. Job duties would involve, but not be limited to: materials consulting, developing / evaluating analytical approaches to constructively investigate problems, and presenting laboratory results to a variety of technical as well as non-technical personnel. Salary is commensurate with experience. Interested candidates should forward qualifications along with salary history to: RJ Lee Group, Inc., Attn. Human Resources, 350 Hochberg Road, Monroeville, PA 15146.
RJ Lee Group is an Equal Opportunity Employer
Drew R. Van Orden, PE RJ Lee Group, Inc. 350 Hochberg Road Monroeville, PA 15146 (724) 387-1869 drew-at-rjlg.com
I am new to this group, so if this is question has been covered, please excuse. I usually do TEM on tissue from biopsies, etc., and now someone wants EM on cells grown in culture. I have done this for intact monolayers (very tedious), but these would be on trypsined-off and pelleted cells. I need to know if there is an easy way to handle them as an entire pellet, so I don't have to cfg. after each fixation/dehydration step, and so they will be compacted for sectioning. Thanks in advance for any advice. Marilyn Howton, Pathology Dept. West Virginia University
I would like to apologise for my humble trivial request to all highly qualified expert like you. I am running website www.coleoptera.org about beetles and I just open new section FAQ with info about collecting, preparation, techniques etc.
I just like to add section about mounting small insects into permanent slide. Is there anything so good as Canadian balsam?
Next question will be how to best prepare insects for SEM..
Any help deeply appreciated.
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
website: http://www.coleoptera.org listserver: coleoptera on www.egroup.com/group/coleoptera/info.html Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com).
Below is the result of your feedback form. It was submitted by (alineves-at-unisys.com.br) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, December 11, 2000 at 14:50:32 ---------------------------------------------------------------------------
Email: alineves-at-unisys.com.br Name: Aline Neves School: Federal University of Rio de Janeiro State: Rio de Janeiro
Question: How do I measure the retardation of a specimen in polarized light microscopy. I have the Berek conpensator to achive a quantitative measure, but exactly how do I do it? Where can I get more information about that?
Hi I insect morphology and use Nikon SMZ-2T binocular stereomicroscope to study the various insect body parts. I use micrometer inside the ocular lens to take measurements. I do not know how to calibrate these measurements. It will be nice if if someone can refer literature or material on internet for calibration and mathematical calculations.If my question is incomplete please let meknow so that i can provide more nformation. sandeep gupta
__________________________________________________ Do You Yahoo!? Yahoo! Shopping - Thousands of Stores. Millions of Products. http://shopping.yahoo.com/
I am working with human cell lines in suspension and process the cells in a pellet.
After the usual problems and much help from this list I worked out a procedure which works for me.
It is critical that the pellet is small - in my experience not more than 250 000 cells / pellet. This means you will not or hardly see the pelleted cells until they have been in OsO4. All the dehydration steps and infiltration steps are performed on a rotary shaker.
Here it is:
I transfer the cell suspension into conical BEEM capsules and spin them in home made adapters at 900 x g. (It is essential to use a swing bucket rotor, otherwise your cells smear along the sides of the container and float off during the dehydration steps.)
After carefully removing the supernatant with fine tipped mini pastetts I overlay the pellet with fixative, remove it again with pastetts and so on. As soon as the OSO4 stains the pellet one can see it and that makes the handling easier. I use glass pipetts with OsO4 and propylenoxide and the resin- propylen mixtures.
I usually infiltrate twice with epon-propylenoxide (1+2 and 1+1 respectively) for 45 minutes, then twice with pure resin. The tips of glass Pasteur pipetts have to be cut to widen the opening otherwise it is very difficult to remove the very viscous resin.
For sectioning I carefully flatten the very fine tip of the block with a razor blade. The cells are evenly distributed, there is no need to cut away surplus plastic. Just shape the block face to a trapezoid and it can go straight into the ultramicrotome.
Good luck - if you require more details feel free to ask.
Regards Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
Dear Marylyn Howton - one approach to fixing cells that have been stripped off is to spin them into a pellet(500rpm) and then take up the pellet in 2% agar - you can then take the pellet through deydration and embedding and all the cells remain in a pellet. You can fix the cells in the pellet but that is rather time consuming and it may be best to do the fixation steps first - spinning down after each and then make into a pellet. hope this helps Carole Elleman
your freind could try to contact Mr. Rasche in Germany: email: mikrovid-at-gmx.de homepage: www.mikrovid.com
best wishes, Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
Leica Microsystems Hernalser Hauptstr. 219 email: Joachim.Prutsch-at-leica-microsystems.com A-1170 Vienna Tel. +43 1 48899 - 235 Austria Fax +43 1 48899 - 350 ---------------------- Weitergeleitet von Joachim Prutsch/AUVIE/LMSCentral/Leica am 12.12.2000 14:11 ---------------------------
"Gordon Couger" {gcouger-at-couger.com} am 09.12.2000 00:12:26
An: microscopy-at-sparc5.microscopy.com
Kopie: (Blindkopie: Joachim Prutsch/AUVIE/LMSCentral/Leica)
Thema: Wanted Wollaston prisms for Reichert Zetopan DIC scope
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I have a freind that need then main prism slide containing the Wollaston prism for Reichert Zetopan DIC scope. This is the slide going in the upper body of the scope. He also needs the polorizor that fits over the light soruce but that is of much less importance beause it can be easily made. While the prism is constructed of unobtainium.
He is willing to pay a reasonable price for the part.
If any of you have a spare or left over you could make a very dissapointed man a lot happier.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Since you work in a hospital setting, get some unneeded serum from your clinical lab, pellet the sample cells, add 50-100 ul of serum to pellet, mix well pellet cells again and then overlay with 3% glut. Next day, remove plug of cells/serum, cut into em size tissue pieces and process as you normally would.
Best of Luck,
Ed
Edward P. Calomeni
Medical College of Ohio Department of Pathology 3000 Arlington Ave. Toledo, OH 43614
} } } "Marilyn Howton" {mhowton-at-hsc.wvu.edu} 12/11/00 03:38PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am new to this group, so if this is question has been covered, please excuse. I usually do TEM on tissue from biopsies, etc., and now someone wants EM on cells grown in culture. I have done this for intact monolayers (very tedious), but these would be on trypsined-off and pelleted cells. I need to know if there is an easy way to handle them as an entire pellet, so I don't have to cfg. after each fixation/dehydration step, and so they will be compacted for sectioning. Thanks in advance for any advice. Marilyn Howton, Pathology Dept. West Virginia University
Dear Nafisa Ghori: Ammonium Molybdate is especially useful to contrast membranes as in vesicles or liposomes. It also can be titrated to neutral pH whereas UA cannot.
Don Gantz Boston University School of Medicine Biophysics Dept.
Colleagues, a pathologist asked to use our Transmission Electron Microscope(TEM) and when I was going to help him load his copper grids, he said he would do it as it was a fecal preparation for viral studies. I found out that material on formar coated copper grids was not fixed. After he viewed (no photos taken) I pulled the copper grid holder and all items touched and ultrasonicated with absolute ethanol and then acetone and then placed items without touching in a 70oC oven for an hour. I feel that is enough, but would like to know if anyone else has been in this predicament. Thanks, Teresa
I am looking for a computer programm that simulates the contrast induced
by defects in crystals. The defects I am interested in are twins and steps in the twin planes. Which programms are available in which you can simulate many different types of defects and that allow to change the parameters easily?
Marilyn, I concentrate the cells by a gentle spin, and embed them in a drop of 2% noble agar. After hardening and trimming the blocks to a size of you usual biopsy you can process them without loosing any. A}
Alice Dohnalkova S&E Assoc., Dep. of Environmental Microbiology Battelle, Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
-----Original Message----- } From: Marilyn Howton [SMTP:mhowton-at-hsc.wvu.edu] Sent: Monday, December 11, 2000 12:39 PM To: Microscopy-at-sparc5.microscopy.com
I am new to this group, so if this is question has been covered, please excuse. I usually do TEM on tissue from biopsies, etc., and now someone wants EM on cells grown in culture. I have done this for intact monolayers (very tedious), but these would be on trypsined-off and pelleted cells. I need to know if there is an easy way to handle them as an entire pellet, so I don't have to cfg. after each fixation/dehydration step, and so they will be compacted for sectioning. Thanks in advance for any advice. Marilyn Howton, Pathology Dept. West Virginia University
"Re: EM on cell cultures" (Dec 12, 8:42am) References: {3A35E4DF.17130.27D219-at-localhost} X-Mailer: Z-Mail (4.0.1 13Jan97) To: Claudia Hayward-Costa {LS_S562-at-crystal.kingston.ac.uk}
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I wonder if you have heard of the Hohenberg technique described some 6 years ago in J.Microscopy; this technique makes life so much easier for people working with single cells in suspension, whether they are prokaryotes or eukaryotes.
H.Hohenberg could nicely demonstrate that the embedding and sectioning is very much facilitated and structures are much better preserved when: a) cells are taken up in cellulose capillaries, 200 micron in diameter; b) cells are high-pressure frozen (as the very first step for cryo-immobilization) c) cells are freeze-substituted but not dehydrated at RT. This technique has been used in the meanwhile by many other people with success, ... but still is not known to all working with single cells in suspensions.
Hohenberg et al (1994) J. Microscopy 175, 34-43
have fun! Reinhard Rachel
Dr. Reinhard Rachel Universitaet Regensburg Lehrstuhl fuer Mikrobiologie (Prof. Dr. K.O. Stetter) D - 93040 Regensburg Tel.: +49-941-943-4534 Fax.: +49-941-943-1824 e-mail: Reinhard.Rachel-at-biologie.uni-r.de
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id MAA02696 for dist-Microscopy; Tue, 12 Dec 2000 12:12:11 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id MAA02690 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 12 Dec 2000 12:11:41 -0600 (CST) Received: from amdext2.amd.com (amdext2.amd.com [163.181.251.1]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id MAA02681 for {MICROSCOPY-at-sparc5.microscopy.com} ; Tue, 12 Dec 2000 12:11:29 -0600 (CST) Received: from odyssey.amd.com (odyssey.amd.com [163.181.250.8]) by amdext2.amd.com (8.9.3/8.9.3/AMD) with SMTP id MAA13242 for {MICROSCOPY-at-sparc5.microscopy.com} ; Tue, 12 Dec 2000 12:10:20 -0600 (CST) Received: from 163.181.250.7 by odyssey.amd.com with SMTP (Tumbleweed MMS SMTP Relay (MMS v4.7)); Tue, 12 Dec 2000 12:10:19 -0600 X-Server-Uuid: d47a4d65-b66d-11d4-9a3a-00508be35655 Received: from 163.181.251.1 by iliad.amd.com with ESMTP (Tumbleweed MMS SMTP Relay (MMS v4.7)); Tue, 12 Dec 2000 05:11:07 -0600 X-Server-Uuid: d47a4d65-b66d-11d4-9a3a-00508be35655 Received: from ultra5.microscopy.com ([206.69.208.10]) by amdext2.amd.com (8.9.3/8.9.3/AMD) with ESMTP id FAA17843; Tue, 12 Dec 2000 05:11:07 -0600 (CST) Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id EAA01811 for dist-Microscopy; Tue, 12 Dec 2000 04:20:35 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id EAA01808 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 12 Dec 2000 04:20:05 -0600 (CST) Received: from mailhost1.rdg.ac.uk (sumh1.rdg.ac.uk [134.225.16.4]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id EAA01801 for {MICROSCOPY-at-msa.microscopy.com} ; Tue, 12 Dec 2000 04:19:53 -0600 (CST) Received: from sapc44 ([134.225.167.96]) by mailhost1.rdg.ac.uk with smtp (University of Reading Email Service) id {145mWD-0001n6-00} for MICROSCOPY-at-MSA.Microscopy.Com; Tue, 12 Dec 2000 10:18:45 +0000 Message-ID: {001b01c06425$23d800e0$60a7e186-at-rucsc}
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Dear Marylyn Howton - one approach to fixing cells that have been stripped off is to spin them into a pellet(500rpm) and then take up the pellet in 2% agar - you can then take the pellet through deydration and embedding and all the cells remain in a pellet. You can fix the cells in the pellet but that is rather time consuming and it may be best to do the fixation steps first - spinning down after each and then make into a pellet. hope this helps Carole Elleman
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Morning Marilyn,
Since you work in a hospital setting, get some unneeded serum from your clinical lab, pellet the sample cells, add 50-100 ul of serum to pellet, mix well pellet cells again and then overlay with 3% glut. Next day, remove plug of cells/serum, cut into em size tissue pieces and process as you normally would.
Best of Luck,
Ed
Edward P. Calomeni
Medical College of Ohio Department of Pathology 3000 Arlington Ave. Toledo, OH 43614
} } } "Marilyn Howton" {mhowton-at-hsc.wvu.edu} 12/11/00 03:38PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am new to this group, so if this is question has been covered, please excuse. I usually do TEM on tissue from biopsies, etc., and now someone wants EM on cells grown in culture. I have done this for intact monolayers (very tedious), but these would be on trypsined-off and pelleted cells. I need to know if there is an easy way to handle them as an entire pellet, so I don't have to cfg. after each fixation/dehydration step, and so they will be compacted for sectioning. Thanks in advance for any advice. Marilyn Howton, Pathology Dept. West Virginia University
You can use a tabletop airfuge with a very small rotor. Each tube holds about 0.2 ml liquid. You can spin your sample for about an hour at a very high speed and it will give you a very tight pellet. I used to spin the sample in fixative and once you are done spinning, you can process the pellet and do osmication, dehydration, infiltration and embedding.
Good luck,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab and Fluorescence Imaging Facility Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
I have been working with human red blood cells grown in cell culture that are infected with Plasmodium falciparum for the past 9 years. The way I handle them is to use normal fixatives (glut, OsO4, uranyl acetate) while they are in suspensions and spin them down in-between steps. Following fixation I spin them down in warm (45 degree C) 3% agarose (Sigma A5030 25gm, type 9 ultra low gelling temperature). When the agarose cools to room temperature the pellet becomes like Jell-O in the tip of the 15ml falcon tube. Then I simply cut off the tip and transfer the "glued together pellet" to a glass vial and continue with dehydration and infiltration steps as if I was handling a small hunk of tissue. Good luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Another person to try would be Dave Paschke, The Dawson Co., (617)484-7900. He had some parts for our Reichert MeF2 when I spoke to him a year ago...you never know.
Diane Ciaburri General Dynamics Pittsfield, MA 01235 (413)494-2847
"joachim.prutsch-at-leica-microsystems.com" 12/12/00 08:17 AM
To: Microscopy-at-sparc5.microscopy.com cc: Subject: RE: Wanted Wollaston prisms for Reichert Zetopan DIC scope
Dear Gordon,
your freind could try to contact Mr. Rasche in Germany: email: mikrovid-at-gmx.de homepage: www.mikrovid.com
best wishes, Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
Leica Microsystems Hernalser Hauptstr. 219 email: Joachim.Prutsch-at-leica-microsystems.com A-1170 Vienna Tel. +43 1 48899 - 235 Austria Fax +43 1 48899 - 350 ---------------------- Weitergeleitet von Joachim Prutsch/AUVIE/LMSCentral/Leica am 12.12.2000 14:11 ---------------------------
"Gordon Couger" {gcouger-at-couger.com} am 09.12.2000 00:12:26
An: microscopy-at-sparc5.microscopy.com
Kopie: (Blindkopie: Joachim Prutsch/AUVIE/LMSCentral/Leica)
Thema: Wanted Wollaston prisms for Reichert Zetopan DIC scope
I have a freind that need then main prism slide containing the Wollaston prism for Reichert Zetopan DIC scope. This is the slide going in the upper body of the scope. He also needs the polorizor that fits over the light soruce but that is of much less importance beause it can be easily made. While the prism is constructed of unobtainium.
He is willing to pay a reasonable price for the part.
If any of you have a spare or left over you could make a very dissapointed man a lot happier.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
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Reply to: RE: EM on cell cultures Dear Marilyn,
A good way to prepare cell monolayers for EM by embedding them as a pellet is as follows:
Fix the cells on the substrate by adding double strength fixative to the culture medium. This will contain serum which is good for the next steps. Leave the fixative on the cells for about 2-5 min and then carefully scrape off the monolayer using a small scraper specially made for this from either a small piece of teflon or a soft wooden stick previously soaked in buffer. The cells will come off the substrate in either a single monolayer (if there are tight junctions present, or as a powdery suspension. Quickly take the detached cells from the dish and centrifuge (I use full speed on the benchtop Eppendorf for about 2 seconds to get a pellet). If you do this without protein being present in the fixative (eg serum) the cells will not pellet but will stick to the sides of the tube (smearing). Once the cells have pelleted DO NOT resuspend them. Remove the fixative and medium and replace it with fresh fixative. Leave for the amount of time you want to fix for (eg 1hr). You will find that after the fixation is complete you will be able to remove the pellet from the bottom of the tube and treat it as if it was a piece of tissue. You should be able to cut it into smaller pieces for subsequent embedding. Follow your favorite protocol for this. I am sure that you will have pellets that do not fall apart. The secret is to fix as a pellet a.s.a.p. and then do not disturb until after fixation is complete. I think the fixative cross-links extracellular components of the cells together but this only happens if the cells are in close proximity as fixation occurs.
If you do prefer to process in the tube they were pelleted in, then a neat trick for speeding up all the steps is to centrifuge after each change of solution. You will see how quickly things go into the pellet if you centrifuge in the presence of osmium tetroxide. A 2 min centrifugation at top speed will cause even a large pellet to turn black all the way through. If you continue the processing in this tube then make sure the final infiltration in the resin is sufficient to go allthe way through the pellet. Be careful to remove all dehydrating solutions and propylene oxide from the tube. I tip them upside down for about half an hour before adding the final resin changes. You will soon realize if you did this successfully - a soft pellet will be the result of too much proylene oxide in the resin. I prefer to embed them after cutting into small pieces.
As they are a pellet of individual cells they will be much easier to embed in resin than a solid tissue piece. Any poor morphology you get will probably not be due to what you do them but what happens before you put them in fixative. Trypsin is not good for morphology, neither is transfection. Ccentrifugation or scraping prior to fixation will affect morphology too. It is a good rule to physically handle the cells as little as possible prior to fixation.
The biggest surprize is that they will have poor morphology if they have only been left to grow fror a day or two after passage. It is only after about three days that they really start to recover from this. Unfortunately, they will look really good in the light microscope so poor morphology will be your problem!
I hope this helps.
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Marilyn Howton 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 new to this group, so if this is question has been covered, please excuse. } I usually do TEM on tissue from biopsies, etc., and now someone wants EM on } cells grown in culture. I have done this for intact monolayers (very tedious), } but these would be on trypsined-off and pelleted cells. I need to know if there } is an easy way to handle them as an entire pellet, so I don't have to cfg. after each } fixation/dehydration step, and so they will be compacted for sectioning. } Thanks in advance for any advice. } Marilyn Howton, Pathology Dept. West Virginia University } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id A73530B01FE; Mon, 11 Dec 2000 21:27:17 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id OAA00529 } for dist-Microscopy; Mon, 11 Dec 2000 14:40:54 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id OAA00526 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 11 Dec 2000 } 14:40:24 -0600 (CST) } Received: from gateway.hsc.wvu.edu (gateway.hsc.wvu.edu [157.182.105.18]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id OAA00519 } for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 11 Dec 2000 14:40:13 -0600 (CST) } Received: from HSC-DOM4-Message_Server by gateway.hsc.wvu.edu } with Novell_GroupWise; Mon, 11 Dec 2000 15:39:02 -0500 } Message-Id: {sa34f516.000-at-gateway.hsc.wvu.edu} } X-Mailer: Novell GroupWise Internet Agent 5.5.3.1 } Date: Mon, 11 Dec 2000 15:38:37 -0500 } From: "Marilyn Howton" {mhowton-at-hsc.wvu.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Subject: EM on cell cultures } Mime-Version: 1.0 } Content-Type: text/plain; charset=US-ASCII } Content-Disposition: inline } Content-Transfer-Encoding: 8bit } X-MIME-Autoconverted: from quoted-printable to 8bit by } ultra5.microscopy.com id OAA00520 } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 273278220 } Status: U }
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Dear Listers,
Can anyone suggest a good reference or share a technique for preparing C-Pt rotary shadowed collagen molecules on mica for TEM?
Thanks.
---------------- Ann Hein Lehman EM Facility Manager Trinity College Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu
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Dear Marilyn,
I am working with human cell lines in suspension and process the cells in a pellet.
After the usual problems and much help from this list I worked out a procedure which works for me.
It is critical that the pellet is small - in my experience not more than 250 000 cells / pellet. This means you will not or hardly see the pelleted cells until they have been in OsO4. All the dehydration steps and infiltration steps are performed on a rotary shaker.
Here it is:
I transfer the cell suspension into conical BEEM capsules and spin them in home made adapters at 900 x g. (It is essential to use a swing bucket rotor, otherwise your cells smear along the sides of the container and float off during the dehydration steps.)
After carefully removing the supernatant with fine tipped mini pastetts I overlay the pellet with fixative, remove it again with pastetts and so on. As soon as the OSO4 stains the pellet one can see it and that makes the handling easier. I use glass pipetts with OsO4 and propylenoxide and the resin- propylen mixtures.
I usually infiltrate twice with epon-propylenoxide (1+2 and 1+1 respectively) for 45 minutes, then twice with pure resin. The tips of glass Pasteur pipetts have to be cut to widen the opening otherwise it is very difficult to remove the very viscous resin.
For sectioning I carefully flatten the very fine tip of the block with a razor blade. The cells are evenly distributed, there is no need to cut away surplus plastic. Just shape the block face to a trapezoid and it can go straight into the ultramicrotome.
Good luck - if you require more details feel free to ask.
Regards Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
Can anyone tell us where to buy a 12V 50W bulb part number HLWS5-A (by Narva) for Jena scope. Please respond to me directly at the address below.
Thanks
Owen P. Mills Electron Optics Facility Engineer Michigan Technological University Department of Materials Science and Engineering Rm 512 M&M Building Houghton, Mi 49931 Ph: 906-487-2002 FAX: 906-487-2934 Email: opmills-at-mtu.edu URL: http://www.mm.mtu.edu/~opmills/index.html
Here's a summary of the advice I received regarding my falling off section problem. It's long but read the last suggestion, it's my favorite.
Formvar coating the tradition way, or dipping the grid into formvar or 0.01% BSA. Dry by dragging grid across filter paper. Bake the sections on to the grid-use either a 60 or 35 degree C oven overnight. Heat fixing the sections using either a hot plate or slide warmer for 10 minutes. Use Coat-Quick G. Flame the grids with an alcohol lamp until the grid just changes color. When working with Spurr's resin, use Veco grids not Gilder. Make sure the water is utra-clean. Use double distilled, deionized, or microfiltered. Various cleaning techniques: 1. 100% acetone wash. 2. 0.1M Nitric acid dip, rinse with dd water, dry on filter paper. 3. 50% acetic acid, 50% acetic acid, 100% acetone-dip grid into each solution then dry well before use. 4. 40 ml concentrated HCL in 400 ml dd water, add 40 ml acetone, take volume to 500 ml with dd water. Sonicate grids in 50ml beaker for 1-2 minutes, discard solution and rinse 2 X with acetone (30 seconds each) dry on filter paper. Do this every day to prevent oxidation from building up on the grids. 5. 100ml clean beaker, cover grids with 20ml glacial acetic acid, sonicate for 5-10 minutes, rinse with 100% acetone until the smell of acetic acid is gone, rinse with 100% ethanol, pipette off the excess 100% ethanol, invert beaker onto filter paper in a glass petri dish. Place in a 60 degree C oven for about 60 minutes, the grids will fall to the bottom of the petri dish, cover the dish with the glass top, the grids will be good for a while. Glass is used to avoid the static that plastic can have. Put sections on the dull side.
Last but not least......
get a new job or get a technician.
I apologize for this being so lengthy, but I thought others might benefit from this information.
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Can anyone answer this question . Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Question: Is it possible to identify types of starch glue (rice, wheat, corn, potato, etc)under microscope? Is there any visuals available somewhere for comparisons?
We are trying to do some cryo TEM of organic solvents based samples, and the grids we have used so far were not very satisfactory, some solvents obviously influenced the formvar (and maybe the carbon ?) of the grid making it beam sensitive. We are planning on purchasing another type of grid (pure carbon lacey film, silicon dioxide or maybe silicon nitride) and I would like to know if anybody has tried anything for that kind of application. Any suggestion or comment most welcome !
Thanks to all how ansered my questions about these two FE-SEM. Its a pitty that I heard about MSA Listserver only two weeks ago. It would have been interesting to become these informations a bit sooner. It was very fruitfull to have some return of experience and offline discussions with some. Amazing was that amost only Hitachi owners had something to tell. And they are all pleased about their instrument. True that the Jeol is new, and much different from the former.
We have seen that our conclusions are similar that a lot of others, and so in a few words, what we can say about these two SEMs:
The Jeol 6700F seems to have at low primery energy (1-2 keV) and short working distance (1-3 mm) the best resolution and the easiest way to become nice images. Charging effect are weak, and magnetic samples have no visible propensity to "fly" in the chamber (magnetic nano powder, for exemple) ! The limitation now, is that this good resolution is lost very fast with increasing the working distance (6-8 mm). And the BSE working distance is 6-8 mm, analytical working distance 15 mm. Shure that in BSE or in X ray microanalysis, higher energy will bee used. But...and tilt ? And the vacuum chamber is limited in avaible ports.
On the other hand, the Hitachi may have the capacity to do so good, but it needs more experience of the manipulator and more time. One is shure : the "In Lens" detector of the Hitachi works well (is probably the best) at short and long working distances, gives a realy surface information, and with the last improvement, gives the possibility of composition contrast at low energy (schould be less sensitif to charge effects too, but we didn't be convinced).
So, mesured with a ladle (do you say so in english ? "Mesurer à la louche", in french !) The Jeol seems to be best for ultra high resolution at low energy and short working distance. The Hitachi seems to be best if you want to have the best images possible at analytical distance (12 mm), backscatered detector on place and the two "secondary" dectectors together. And is also capable to reach ultra high resolution. We didn't make detailed tests at high energy (} 15 keV). All instruments seems to be comparable at high energy, and then you don't observe the real surface.
But, and now I'll conclued, the two apparatus are VERY good, and it is VERY difficult to make a choice. If you have monney, buy one of eatch ! We have choiced the Jeol one, but whith some regree for Hitachi's "In Lens" detector, versatility and reliability.
I've said nothing about Leo's 1500 serie and Philips XL30-S-FEG, witch are also interresting. We have test them, but it would be to long to speak about these now. But cold FEG seems to give better high resolutions images,and is to be prefered if you don't need the long term stability of Shottky FE, for intensive XRay microanalysis fot exemple. Gemini's optical concept is very interresting, but more "classical" design give as good results.
I'm still intersted on other opinions or test results, and also avaible for questions, true MSA list server or on my direct mail.
By and thanks to all
J. Faerber IPCMS-GSI 23, rue de Loess 67037 Strasbourg CEDEX
We are trying to do some cryo TEM of organic solvents based samples, and the grids we have used so far were not very satisfactory, some solvents obviously influenced the formvar (and maybe the carbon ?) of the grid making it beam sensitive. We are planning on purchasing another type of grid (pure carbon lacey film, silicon dioxide or maybe silicon nitride) and I would like to know if anybody has tried anything for that kind of application. Any suggestion or comment most welcome !
subject: J. C. Russ, R. T. Dehoff, Practical Stereology, 2nd Edition, Plenum Press, New York, 2000 (!?!), isbn 0-306-46476-4
Some folks on this list have an early draft of my forthcoming (more on that below!) book on stereology, which was distributed on the CD with the Image Processing Tool Kit and also handed out in notes form at the various image analysis workshops that I’ve taught for the past many years. Bob Dehoff (who teaches those workshops with me) and I completed the final version of that book in December, 1999, and sent it in to the publisher. It has taken them a very long time to get around to doing anything with the book, and they are still apparently having a lot of internal technical problems (not worth going into here - but *.doc files and *.tif files on a CD seem to be a little too high tech for these people, and they clearly don’t have any idea what a *.pdf file is good for). At the M&M meeting in Philadelphia, quite a few people asked me about the book and at that time Plenum was promising that it would be available in September. Well, of course, it wasn’t, and still isn’t and frankly I’m getting tired of apologizing for it. None of the delay has anything to do with the authors, it is strictly production difficulties and a high level of technical incompetence at the publisher’s end. Anyway, to make a long story less long, I’ve put a copy of the entire book on the web in the form of a pdf file. In order to keep the download times reasonable (it is still 7.1 MB) the graphics are jpeg compressed, but I think still useful and legible. Anyone who wants to download a copy is welcome to it. When the book eventually (!) comes out the on-line version will evaporate. You may want to purchase a copy to get the index and somewhat better graphics, but I will leave that to your conscience. For the present, you can find a link to it at {http://members.aol.com/ImagProcTK/updates} (look toward the bottom of the page under "Data Analysis") or on my web page {http://members.aol.com/DrJohnRuss} (go to the "links" section). In the on-line file, the contents are linked to the book pages and of course you can use Acrobat Reader to search for any word in the text.
} I am looking for a tissue histology stain for human tongue } epithelium for use with tetramethylrhodamine-labeled probes. The stain } must not } fluoresce under 546nm wavelength light and also must not quench } tetramethylrhodamine fluorescence. Haematoxylin, eosin, and methylene } blue each } fail to meet these requirements. Does anyone have any suggestions? } } Dennis M. Walling, M.D. } Assistant Professor } Division of Infectious Diseases } Department of Internal Medicine } The University of Texas Medical Branch } 301 University Boulevard } Galveston, TX 77555-0835 } } Telephone: (409) 747-2361 } Fax: (409) 772-6527 } E-mail: dwalling-at-utmb.edu }
The Materials Characterization Group in Dow’s Corporate R&D Analytical Science laboratory has a full-time professional level opening for a particle analyst/light microscopist at Dow’s facility in Midland, MI. The primary responsibilities include working with partners to support research and production projects across a broad range of products. Good written and oral communication skills and the ability to work both independently and in a team environment are extremely important.
Key responsibilities will include: * Strong problem solving skills * Operation of particle size characterization instrumentation * Operation of light microscopes and supporting image analysis software packages * Method development * Documentation of work * Compliance with safety and quality programs
Qualifications: * A candidate with a BS or MS degree in material science, chemistry, or physics with experience in light microscopy and particle size characterization. * Experience with common commercially available particle size measurement instrumentation - single-particle, ensemble and fractionation based. * Experience in the general area of polarized light microscopy, digital image acquisition, and quantitative image analysis.
Please e-mail or send your resume and cover letter, with reference to this ad to: Email: {R&D-at-Dow.com} or The Dow Chemical Company, Employee Development Center, Workforce Planning, Midland, MI 48674. Email respondents must list Job 00622LUSA/JMK-JB and their last name as the first and second items on the subject line. Only those selected for an interview will be contacted.
We are an equal opportunity employer and offer a competitive compensation and benefits package including 401k, stock purchase, tuition reimbursement and performance incentives. The Dow Chemical Company is the fifth largest chemical company in the world with annual sales of US$20 billion. Dow manufactures and supplies chemicals, plastics, and agricultural products for customers in 164 countries and employs approximately 43,000 people worldwide. For more news and information about Dow, visit our web site at www.dow.com.
Qualifications (education, certification, language, etc.) and experience required:
A candidate with a MS or Ph.D. degree in polymer science, material science or chemistry is preferred with prior experience in electron microscopy. Good written and oral communication skills and the ability to work both independently and in a team environment are extremely important.
Job Overview:
The Surface/Microscopy/X-ray (SMX) Group in Dow's Corporate R&D Analytical Science Laboratory has a professional level full time opening for a polymer microscopist. The primary responsibilities include working with partners to support research projects involving new and existing products in Dow's polymer businesses.
Key responsibilities will include:
1. Extensive problem solving. 2. Microscopy preparation technique experience including 3. Ultramicrotomy and cryo-ultramicrotomy. 4. Operation of light microscopes, scanning and transmission electron microscopes. 5. Interpretation of images. 6. Documentation of work. 7. Compliance with safety and quality programs. 8. Active member of project and SMX work teams.
We are an equal opportunity employer and offer a competitive compensation and benefits package including 401k, stock purchase, tuition reimbursement and performance incentives. Please e-mail or send your resume and cover letter, with reference to this ad to: Email: {R&D-at-Dow.com} or The Dow Chemical Company, Employee Development Center, Workforce Planning, Midland, MI 48674. Email respondents must list Job 00623LUSA/JMK-JB and their last name as the first and second items on the subject line.
Only those selected for an interview will be contacted.
Can anyone tell me where I can find information on remote SEM? Is it possible to install remote SEM capabilities on an older existing instrument (JEOL 35C)? Does this technology work for LOM also?
Sam O. Mancuso Special Metals Corporation Research & Development New Hartford, NY 13413 smancuso-at-specialmetals.com
In my opinion, light microscopy is probably most useful in differentiating starch grains, although SEM is sometimes used.
By using a light microscope with two polarizing filters, it is possible to view a "cross" or "x-shaped" pattern within the starch grains. (This has sometimes been referred to as a "Maltese Cross".) The appearance of this cross can be an indicator of starch type. Also, by rotating one of the polarizers you can observe the cross as it comes into view, rotates a bit, then disappears again. Measuring the rotation arc of the polarizer within which the cross is visible gives you another criterion.
Other ways of telling starches apart are, of course, their size and shape. Sizes should be determined by averaging a number of grains, since they can be quite variable---in fact, the amount of variation in size is yet another criterion. Shapes range from round to various types of ovals and irregular ovoids. SEM excels at this, allowing very precise measurements.
If you really want to get involved, it's possible to set up heated water baths and sample a slurry of starch grains as the temperature rises, then count the melted versus unmelted grains in the light microscope. Melted starch grains are amoeba-like blobs, instead of nicely defined shapes. Different starches have different melting characteristics.
There is a two-volume set of work by Edward Tyson Reichert (1913), named "The Differentiation and Specificity of Starches in Relation to Genera, Species, Etc." It is loaded with methods and photos.
Here is a web site you may want to look at: http://www.siu.edu/~ebl/amylose.htm. I used to do starch research in the lab that puts this page out, back in "the old days".
Hope this gets you started. Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
You will need to carefully define what you mean by "Remote SEM". Do you mean true TelePresence or just TeleObservation.
You should begin by looking at the Microscopy and Microanalysis Proceedings for 1995, 1996, 1998, and 2000 there are a number of full session as well as articles in there by the various groups in the world working in this area.
Viewing images on-line is do-able in virtually any imaging system, with sufficient hardware upgrades, but remote "TeleOperation" can be a much more involved proposition particuliarly for an older instruments.
You might consider getting a copy of the MSA Education Committee Video Tapes made during the Tutorials presented at the Microscopy and Microanalysis 2000 meeting.
I did a presentation therein called Microscopy and Microanalysis over the Internet, which discussed a pretty full range of options. I don't know if 2000 meeting videos are ready yet, but you can contact Greg Erdos for that information at the above WWW page.
There are a number of vendors which can interface older instruments to computer systems for image acquisition which the minimum starting point. This can then, in principle, be adapted to TeleObservation. You should talk to the following companies: 4Pi, Quartz, Gatan , EmiSpec (to name a few there are more so please check the exhibitors list from the M&M 2000 meeting) as well as literally any company that manufacturers EDS equipment (a long list....). Most of the Electron Optical manufacturers also offer or will soon offer this as options on their newer instruments but in all likelihood for your older instrument you will have to go to the accessory market arena.
There is also going to be a session at the Microscopy & Microanalysis Technologist's Forum at the 2001 Meeting which will discuss the issue of TelePresence, you may wish to attend that meeting in Long Beach August 5-9, 2001. http://www.microscopy.com/MSAMeetings/MM01/MMHomePage.html
Nestor Your Friendly Neighborhood SysOp
Disclaimer: I have no commerical interests in the companies mentioned herein, but you might say I have a mild vested interest in "TelePresence" and it's application to Research and Education ;-)
As always my TPM Lab is open to the world.... http://tpm.amc.anl.gov. your welcome to visit anytime...
J. W. Robinson Mechanical, Automotive, and Materials Engineering University of Windsor Windsor, Ontario N9B 3P4
Phone : 519-253-4232, ext 2598
Could anyone send me any information on the L. M. Simard company, maker of the tri-mag sputtering source? I need to order replacement parts and the last phone number I have for them is no longer in service. Any help will be appreciated.
Ooops - guess I need to type more carefully. The correct URL for downloading the Practical Stereology textbook is {http://members.aol.com/ImagProcTK/update.html} . The link there goes to a grad student's computer (someone who had enough storage space that I could use for a while), and from time to time that machine may go off line (when he is doing some "real" work), so if you don't get through at first be patient and try again in a few hours or the next day.
We just installed a Tecnai 12. I am not sure what is a reasonable charge for users to use the TEM. We are at university and a self-supported EM suite. Could you please let me know your opinion or the user fee (hourly rate) charged at your work place? especially at Canadian universities.
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There is a freeware/shareware application WinVNC which basically allows remote control over a network of any other PC and the applications running on that PC. While not a perfect solution, it works well running PC-driven SEMs, such as the JEOL 5500/5600/5900. We have experimented with it in the JEOL UK Application Lab and, considering it is 'free', are happy with it.
Addressing your specific problem, you would first have to convert your JSM-35 to a PC driven SEM. I don't know about the US but in the UK, there are several 3rd party companies offering such conversions. If you don't get any information via the list, contact JEOL Inc.
Once such a conversion has been done, I see no reason why you shouldn't control it over a LAN with WinVNC.
The same applies to any equipment driven by a PC - connect to a LAN and use WinVNC. In fact, I'm told it will work with any platform connected to the LAN - Unix, Mac, etc.
I have a collaborator who wants to do IHC on colon tissue. She just bought some Technovit 8100 but doesn't know how to use it. I use Technovit 7100 for regular histology on plant tissue but I have no protocol experience with 8100, IHC or animal tissue. Does anyone have any suggestions or recommended protocols for doing IHC with Technovit 8100? This is for light microscopy.
w'ere short about 3 plate holders for our Jeol 100cx TEM camera film (3&1/2 by 4"). At $25 apiece new, we were wondering if anyone had some spare? Or if anyone knows which websites would have these sorts of things second-hand?
cheers Liz McKenzie
******************************************************* Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
Post Doctoral Position open in a multiple disciplinary laboratory at NIH We use molecular biology, biochemistry and image analysis (confocal and electron microscopy) techniques for studies on lipid transport in cells.
I am searching for someone who has training in electron microscopy to investigate cholesterol mobilization in cells.
The position is ASAP,
The position is for a person not more than 4 years post PHD.
Those interested should send their CV's as inserts to the E-mail.
Thank You
-------------------------------------------------- Dr. Joan Blanchette-Mackie Chief; Section of Lipid Cell Biology NIDDK / NIH Building 8, Room 427 8, Center Drive Bethesda MD 20892 - 0850
Hello, I am a venture capitalist located in Milwaukee WI and am in need of some basic information on Atom Probe Microscopy because I am evaluating a potential investment in a company that manufactures such a device. What is it and how is it different than TEM and SEM. What are the applications it is used for? Are there companies that sell them? What do they cost? Any references you can direct me to will be appreciated. Thank you.
Daniel Broderick Managing Director Mason Wells Biomedical 770 N. Water St. Milwaukee WI 53202 414 765 7830 fx 414 765 7850
I have EELS spectra from a material contain a mixture of Ce3+ and Ce4+ and wish to determine the relative amounts of each oxidation state. To do this I have collected spectra from a Ce3+ standard and a Ce4+ standard and propose to least squares fit these to the spectrum from my mixed state sample.
I presume this type of thing has been done before and there are probably any number of ways to implement the fitting using Excel or a KaleidaGraph or some purpose written software. I thought I'd ask if anyone would like to make their solution available to me or even just offer suggestions.
The ideal solution would be if the fitting could be done within EL/P or even DigitalMicrograph.
I look forward to any replies. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
The answer to this very much depends on what you are charging for. To find an answer, you need to identify (a) the net costs you wish to recover: e.g. capital costs (to be recovered over what period?, at what rate of interest?), running costs, salaries of direct (em technician) and indirect (clerical, admin.) support staff including overheads, any charges for space occupancy and services etc., less any subsidies (b) make a realistic estimate of the maximum number of hours per year that a charge can be recovered from. The answer is a/b. Simple, isn't it? Well, no. In practise you will probably find that b is dependent on a/b since all categories of users are sensitive to price, but you will have to model this differently for different categories of users.
Our charges per hour for self-help access to a Philips CM120 Biotwin are as follows: Departmental £15, Other departments £21, other publicly-funded research organisations £35, industrial £70. Non-attenders incur full charge for the period booked. These numbers were not exactly arrived at using rocket science. We charge extra for all materials consumed (film, processing) and for direct user-support from a technician. We do not include capital costs recovery in our charges.
I would strongly advise imposing a special concessionary charge on users who insist on tipping their specimens and retaining circlips into the guts of the Compustage! An electronically-operated trap door beneath the operator's seat is also an attractive option.
Chris
} From: "Ping Li" {pli-at-is.dal.ca} To: {Microscopy-at-sparc5.microscopy.com}
I've noted no response, so I better offer something. The technique was more commonly used to visualize stretched DNA fibers and you may find more references using the Kleinschmitt technique about 30 years ago.
I have used fixed shadowcasting and negative staining on collagen too and both methods "work". The advantage of rotary shadowcasting seems to be that it makes it easier to follow the continuity of fibers (up-down, round and about). Rotary shadowcasting is usually executed at a shallow angle of 6-12 degrees and so it builds up more metal at the near vertical surfaces of the fibers.
Sticky-tape the very edge of the prepared specimen grid to a flat surface (half of a microscope slide). A bit of tape may be required on two sides to assure that the grid lies flat. Arrange the grid(s) a the rotation center of a rotator within the vacuum chamber and locate the rotator to be between 60 and 100mm from the evaporation source and a little lower than the source to achieve an angle of say 10 degrees. The angle could be checked with a 10 degree card segment.
The source could be C-Pt pellets or 0.1mm dia. Pt wire (try 40mm) wrapped around a 1.5mm carbon cylinder about 5mm long that has been fashioned with a suitable C rod sharpener. The "loaded" C rod is spring tensioned against a second C rod that retains a flat end, but one that is reduced in size to about 3mm dia. using a conventional (but clean, e.g. no pencils) pencil sharpener.
Evacuate the system. Pt/C evaporation gives very fine granularity; about the best short of special equipment, but a good vacuum ( { 5x10-5 torr) is essential. Prior to evaporation start the rotator and adjust the speed to 30 to 60 rev/ minute. Increase the ampere control to achieve red heat, then back off to give the vacuum a moment to recover. Then increase ampere to a white heat and observe using a dark shield until the fashioned carbon tip has evaporated. Hope this helps. This technique is much easier demonstrated than written about. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, December 13, 2000 5:57 AM, Lehman, Ann [SMTP:Ann.Lehman-at-trincoll.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } Can anyone suggest a good reference or share a technique for preparing C-Pt } rotary shadowed collagen molecules on mica for TEM? } } Thanks. } } ---------------- } Ann Hein Lehman } EM Facility Manager } Trinity College } Hartford, CT 06106 } v. 860-297-4289 } f. 860-297-2538 } e. ann.lehman-at-trincoll.edu
If you are a material scientist and have experience in microscopy and the use of computers, then we invite you to apply for this position at the Bridgestone/Firestone Research, Inc. in Akron, Ohio.
Our Microscopy Analysis Laboratories provide research and service in microscopy for Bridgestone/Firestone Research, and, in general, Bridgestone/Firestone (BFS) associates, as well as extend service to non-BFS users. Currently, our laboratories have one 'state-of-the art' TEM/STEM/AIA, one SEM/EDX, as well as one AFM/STM and a number of LOM-s and ancillary equipments.
Duties of the appointee will include sample preparation, instrument operation, analysis, and reporting. The level of appointment will depend on the qualifications, and the experience of the successful candidate.
Please send a letter of application plus your CV (including the names, postal-e-mail addresses, telephone/fax numbers of 3 referees) to: Dr. Pat Sadhukhan Bridgestone/Firestone Research, Inc. Phone: 330-379-7518 1200 Firestone Parkway Fax: 330-379-7530 Akron, OH 44317-0001
Has anyone come across radiation damage to SrTiO3, either knockon or radiolytic?
------------------------------------------------------- Laurence Marks Director, Center for Transportation Nanotechnology Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
Dear Ping, When we installed our TEM, we calculated the hourly cost of the service contract, based on a 40 hour work week, and used that for a basis for our charge to other users at the University. It is currently 50 dollars per hour. The charge for commercial users is based on what commercial consultant firms charge, in our area about 200 dollars an hour. There are, in my experience, very few commercial users of TEM. At 04:45 PM 12/13/00 -0400, you wrote:
} We just installed a Tecnai 12. I am not sure what is a reasonable charge for } users to use the TEM. We are at university and a self-supported EM suite. } Could you please let me know your opinion or the user fee (hourly rate) } charged at your work place? especially at Canadian universities. } } Thank you. } Ping } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
As a late follow-up to a somewhat recent string on liquid nitrogen safety, I thought I would relate something that happened just this morning. While transferring LN2 from one dewar to another using a funnel, the funnel literally exploded just as the container was filled to the top. The explosion was pretty forceful, sounding like a gunshot and scattering sharp plastic shards all over the room and leaving behind a startled, but otherwise unhurt technician (yup, it was me).
It was a standard-issue, large size plastic lab funnel, nothing special. We had used the same funnel for over a year with no problems whatsoever. I don't have a clue as to why it blew up or how there was enough pressure generated to cause the explosion, but the remainder of the funnel looks a lot like the hat Jughead used to wear in the Archie comics. It is now on our bulletin board with a tastefully done joke notice about my recent brush with eternity, courtesy of the other technician, Cheryl, who shall remain anonymous.
Anyway, WATCH IT if you use the same low-tech LN2 transfer protocol we do (or did).
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
I am looking for comments from individuals who are currently using a PC based program or spreadsheet to treat data from Al or Au ring SAED calibration measurements. I would also like to be able to index minerals such as asbestiform silicates. I am aware of older Fortran based diffraction programs but I need a PC based system that someone is currently using.
On Wed, 13 Dec 2000 DrJohnRuss-at-aol.com-at-sparc5.microscopy.com wrote:
} Ooops - guess I need to type more carefully. The correct URL for downloading } the Practical Stereology textbook is } {http://members.aol.com/ImagProcTK/update.html} . The link there goes to a } grad student's computer (someone who had enough storage space that I could } use for a while), and from time to time that machine may go off line (when he } is doing some "real" work), so if you don't get through at first be patient } and try again in a few hours or the next day.
So far, today, none of the links get one to the download site.
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The fee depends on the purpose of charging.
If you just want to pay the cost of maintenance for the users inside the university, then I think that you just make a calculation based on the costs on liquid N2, N2 gas, films, darkroom expenses, fees for service contract, and the other consumption (everything related to the machine, except the salary of the professional staff). Also the fee could be quite different from university to university. Recently I have been two universities, the same machine, JEOL 2000FX, one charges user USD$35 per hour, the other charges user only USD$20. As I know, when you charge user at high price although it is reasinable, then you will get less users to use, finally you get less "income" to cover the cost. If it happens, you may have to lower the price to encourage users to use. This happened to one university I used to work.
Yes, people always charge those commercial companies at much higher price, 3 - 5 times?
Dear Ping, When we installed our TEM, we calculated the hourly cost of the service contract, based on a 40 hour work week, and used that for a basis for our charge to other users at the University. It is currently 50 dollars per hour. The charge for commercial users is based on what commercial consultant firms charge, in our area about 200 dollars an hour. There are, in my experience, very few commercial users of TEM. At 04:45 PM 12/13/00 -0400, you wrote:
} We just installed a Tecnai 12. I am not sure what is a reasonable charge for } users to use the TEM. We are at university and a self-supported EM suite. } Could you please let me know your opinion or the user fee (hourly rate) } charged at your work place? especially at Canadian universities. } } Thank you. } Ping } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
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As a late follow-up to a somewhat recent string on liquid nitrogen safety, I thought I would relate something that happened just this morning. While transferring LN2 from one dewar to another using a funnel, the funnel literally exploded just as the container was filled to the top. The explosion was pretty forceful, sounding like a gunshot and scattering sharp plastic shards all over the room and leaving behind a startled, but otherwise unhurt technician (yup, it was me).
It was a standard-issue, large size plastic lab funnel, nothing special. We had used the same funnel for over a year with no problems whatsoever. I don't have a clue as to why it blew up or how there was enough pressure generated to cause the explosion, but the remainder of the funnel looks a lot like the hat Jughead used to wear in the Archie comics. It is now on our bulletin board with a tastefully done joke notice about my recent brush with eternity, courtesy of the other technician, Cheryl, who shall remain anonymous.
Anyway, WATCH IT if you use the same low-tech LN2 transfer protocol we do (or did).
Dear Randy, It is possible that internal stress was present in the funnel, and upon cooling (this time) it built up past the ability of the funnel to adjust. This happens in an altogether different temperature range to quarz high-pressure Hg lamps. Teflon does not become too stiff at 77 K, so this shouldn't happen if you use a teflon funnel. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
FWIW, I no longer use a funnel (would be S.Steel if I did). A number of years ago I experienced the same scenario. My HDPE funnel exploded with great force sending shrapnel ricocheting off the walls...
A bit of spilled nitrogen is not particularly hazardous (don't pour in your shoe :) compared to the high speed funnel shards.
Woody White McDermott Technology
----------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi Listers, } } As a late follow-up to a somewhat recent string on liquid } nitrogen safety, I } thought I would relate something that happened just this } morning. While } transferring LN2 from one dewar to another using a funnel, the funnel } literally exploded just as the container was filled to the top. The } explosion was pretty forceful, sounding like a gunshot and } scattering sharp } plastic shards all over the room and leaving behind a startled, but } otherwise unhurt technician (yup, it was me). } } It was a standard-issue, large size plastic lab funnel, } nothing special. We } had used the same funnel for over a year with no problems } whatsoever. I } don't have a clue as to why it blew up or how there was } enough pressure } generated to cause the explosion, but the remainder of the } funnel looks a } lot like the hat Jughead used to wear in the Archie comics. } It is now on } our bulletin board with a tastefully done joke notice about } my recent brush } with eternity, courtesy of the other technician, Cheryl, who } shall remain } anonymous. } } Anyway, WATCH IT if you use the same low-tech LN2 transfer } protocol we do } (or did). } } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } }
Just one point on your comments relating to charge in the Hitachi 4700. In any SEM with an in lens detector the signal will be made up of SE. Should you use the lower detector in the Hitachi you will find an increase in the contribution of BSE and therefore a much lower charge rate.
Training many people in the use of the 4500 and 4700 I find they tend to become upper detector addicts, when the lower detector often has much to offer, particularly with samples that tend towards charging.
Hope this helps?
Steve Chapman
Senior Consultant Protrain Tel & Fax - 44 (0)1280 814774 E-mail- protrain-at-emcourses.com Web Site - www.emcourses.com Protrain for SEM, TEM & EDX Training World Wide with our own full time Professional staff
At 4:45 PM -0400 12/13/00, Ping Li wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
*********************************
Ping: I run the EM Core facility for our medical college. I had to assure my dean that my fees were in accord with other, similar facilities in the NY-metro area. I called around to many of my colleagues and got their fee schedules. My fees are about average...higher for some things, lower for others. Briefly, our per hour fees for the EM are $50 if the person has been trained and works alone, $75 if they need my assistance. Film is billed at $1 each, which just barely covers the cost...I'll probably have to raise that price this year.
If you'd like to see my whole price list (I have a fee for every service available in the facility), you can go to to the website: http://www.med.cornell.edu/research/cores/index.html Go to the bottom of the page, click on the up-down arrows and scroll to the Electron Microscopy Core.
I am responsible for covering all of the operating costs of the lab, with the exception of my salary (thank heaven!). I am the sole personnel in the facility for now, and handle about 100 clients/year. I have received approval for a part-time tech, and will then offer histology (paraffin embedding/sectioning, cryostat sectioning) as a service rather than just having the equipment here for people to use on their own.
Good luck. Setting up a fee schedule is no easy matter....be prepared for loads of complaints about the cost, and people asking if everyone pays the same rate. Everyone here pays the same fees, from the Sr. Assoc. Dean of Research to the newest Assist. Prof. That way no one gets their nose out of joint!
good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I think the point about hazards associated with LN2 that do not directly involve the cold liquid itself is worthy of some note.
Over the 15 years we've been maintaining a LN2 cooled beryllium crystal, there have been 3 incidents where luckily no injuries were sustained. Two involved breaking the 4-L dewer -- once when a student smacked the top edge of the dewer with the LN2 hose nozzle and once when a technician tripped and fell with a full dewer. The first merely resulted in a very loud bang with lots of fog, etc. as described in another post (no aluminum foil, though) and very thankfully, the technician who fell was unhurt by either the fall or the shattered glass. Again, lots of fog and running rivulets of LN2 down the hallway.
The third incident was just a close call and one the was totally unanticipated. Our dewer has a large cork in the top which is replaced after the dewer is filled. After filling, the technician reached down to pick up the nearly full dewer and, in the process, caused the LN2 to slosh and a bit evaporated causing the cork to 'fire' out of the mouth of the dewer (large 'pop gun' effect) and graze her face. She was not seriously hurt beyond a small scrape on her temple...but, had she been hit in the eye at such close range.... Anyway, since that graze incident, we who are responsible for filling the EDAX dewer, wear glasses or goggles and are very careful where the dewer is 'aimed' when we pick it up and carry it.
We have had similar occurrences here. Both old, and new funnels have exploded. It is my guess that it is internal stress in the plastic which causes the event (Or, should I say the rapid release of the stress from being cooled to below minus 200 degrees centigrade?)
What we have done, is to use only stainless steel funnels for LN2.
I'd say the use of a plastic funnel with LN2 is a no no anyway. It probably just shattered from being at that extreme temperature for a certain length of time, which is why the "explosion" coincided with the other dewar being topped off.
-----Original Message----- } From: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Thursday, December 14, 2000 10:20 AM To: 'microscopy-at-sparc5.microscopy.com'
Hi Listers,
As a late follow-up to a somewhat recent string on liquid nitrogen safety, I thought I would relate something that happened just this morning. While transferring LN2 from one dewar to another using a funnel, the funnel literally exploded just as the container was filled to the top. The explosion was pretty forceful, sounding like a gunshot and scattering sharp plastic shards all over the room and leaving behind a startled, but otherwise unhurt technician (yup, it was me).
It was a standard-issue, large size plastic lab funnel, nothing special. We had used the same funnel for over a year with no problems whatsoever. I don't have a clue as to why it blew up or how there was enough pressure generated to cause the explosion, but the remainder of the funnel looks a lot like the hat Jughead used to wear in the Archie comics. It is now on our bulletin board with a tastefully done joke notice about my recent brush with eternity, courtesy of the other technician, Cheryl, who shall remain anonymous.
Anyway, WATCH IT if you use the same low-tech LN2 transfer protocol we do (or did).
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From root Thu Dec 14 18:01:51 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id RAA05412 for dist-Microscopy; Thu, 14 Dec 2000 17:15:12 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id RAA05407 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 14 Dec 2000 17:14:42 -0600 (CST) Received: from tmpsmtp102.honeywell.com ([199.64.7.102]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id RAA05400 for {microscopy-at-sparc5.microscopy.com} ; Thu, 14 Dec 2000 17:14:30 -0600 (CST) Received: from 131.127.249.22 by tmpsmtp102.honeywell.com (InterScan E-Mail VirusWall NT); Thu, 14 Dec 2000 16:02:58 -0700 Received: by TMPCN197 with Internet Mail Service (5.5.2650.21) id {YBYJW3NY} ; Thu, 14 Dec 2000 16:13:27 -0700 Message-ID: {3348B4357E7FD4118DAE00508B074950607E0D-at-tmpex177.wins.allied.com}
I'd say the use of a plastic funnel with LN2 is a no no anyway. It probably just shattered from being at that extreme temperature for a certain length of time, which is why the "explosion" coincided with the other dewar being topped off.
-----Original Message----- } From: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Thursday, December 14, 2000 10:20 AM To: 'microscopy-at-sparc5.microscopy.com'
Hi Listers,
As a late follow-up to a somewhat recent string on liquid nitrogen safety, I thought I would relate something that happened just this morning. While transferring LN2 from one dewar to another using a funnel, the funnel literally exploded just as the container was filled to the top. The explosion was pretty forceful, sounding like a gunshot and scattering sharp plastic shards all over the room and leaving behind a startled, but otherwise unhurt technician (yup, it was me).
It was a standard-issue, large size plastic lab funnel, nothing special. We had used the same funnel for over a year with no problems whatsoever. I don't have a clue as to why it blew up or how there was enough pressure generated to cause the explosion, but the remainder of the funnel looks a lot like the hat Jughead used to wear in the Archie comics. It is now on our bulletin board with a tastefully done joke notice about my recent brush with eternity, courtesy of the other technician, Cheryl, who shall remain anonymous.
Anyway, WATCH IT if you use the same low-tech LN2 transfer protocol we do (or did).
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
I am currently using Crystal Studio (check google to find the source) to generate diffraction patterns for the amphiboles and others. Although it has some strange characteristics the patterns are reasonable. Be advised that the patterns are based on x-ray diffraction intensities - the actual SADs have different intensity distributions as you might expect.
Email me if you need help indexing the asbestos silicates.
Tom McKee wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Good Morning MSA group } } I am looking for comments from individuals who are currently using a PC } based program or spreadsheet to treat data from Al or Au ring SAED } calibration measurements. I would also like to be able to index minerals } such as asbestiform silicates. I am aware of older Fortran based } diffraction programs but I need a PC based system that someone is currently } using. } } Thanks Tom McKee
-- Gordon Nord Small Business Network Design and Construction Macintosh and Windows - Solutions and Conflicts
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I would like just to point that it is not good idea "preheat" carbon rods in the presence of sample. All contaminations from rods will contaminate the sample first and than you will shadow that. Shadowing will develop not only structural details but contamination ether. What I did, I preheat rods without samples, than open the chamber, install the samples and pump down the system again. Even when I done carbon film preparation, I prefer to use the same technique. It takes more time. In general, shadowing quality extremely depends from sample quality and sample preparation procedure. Most air-dried protein samples demonstrate very poor structural integrity preservation. Buffer composition is also important (you will shadow not only the "sample" but salts from buffer as well).
Sergey
} Date: Thu, 14 Dec 2000 20:58:32 +1000 } From: Jim at ProSciTech {jim-at-proscitech.com} } Subject: RE: Rotary shadowing } To: "'Lehman, Ann'" {Ann.Lehman-at-trincoll.edu} , } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } Reply-to: "jim-at-proscitech.com" {jim-at-proscitech.com} } Organization: ProSciTech } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
We have been doing some rotary shadowing to produce replicas on biological samples for examination under the TEM. For shadowing we used V-shaped tungsten wire and pure carbon for support,
They are a few problems we encountered. Every run is a trial and error process. This is especially so with the setting of current to vaporize the metal and the carbon. Every run is a different value although the voltage is constant. At each run we try to follow the prefer parameter but somehow the replica would look different from the previous one. Although, it is better to observe the colour of the glow on the metal and carbon but that is very hurting to the eye. I wonder if anybody has a definite value for the current used to vaporize 0,1 mm pt wire/tungsten and pure carbon rod in the vacuum chamber of 10 -5 torr.
Josephine NUS
-----Original Message----- } From: Jim at ProSciTech [mailto:jim-at-proscitech.com] Sent: Thursday, December 14, 2000 6:59 PM To: 'Lehman, Ann'; Microscopy-at-sparc5.microscopy.com
Josephine This message is only partially related to your topic, but, anyway. This is my comment.
I would say, it's very unlikely to have good reproducibility in your set-up. The easiest and probably cheapest solution is to use thickness monitor. It's relatively cheap (prices vary from $1500 to $5000), precisely measure the thickness starting from a few angstroms, easy to work. You may control the shutter by the thickness monitor in order to close the shutter at some point, say 2 nm.
More "solid" solution is to use electron guns for shadowing. I am using electron guns for decade and they shown outstanding ability to evaporate many substances: W, Pt/C, C in my case. Electron gun evaporated carbon films have outstanding stability (in compare with thermal evaporated) under the beam. I would mention that I forgot the time when I have a problem with drift. Last time it was, probably 15 years ago when I was looking for good technique for carbon film preparation. Additional reason to switch to the Electron Gun - sample thermal damage. Electron guns produces less infra-reds than regular set-up.
Another issue, I would like to comment is a level of vacuum in the evaporators. The level 5x10-5 torr in my point of view is not acceptable for ANY applications related to biology. The abequate range started from 2x10-6 torr I believe. I would point that practically all commercially available evaporators are able to pump system down to the good 10-6 even without LN2 trap. 10-5 is a result of bad evaporator's maintenance. Replace diffusion oil on SANTAVAC 5 (and forget the problems with DP), check the O--rings for cracks, replace old O-rings (do you know how old your system?), keep system clean, run system at least for 24 h a month. I just could not believe that films produced at 10-5 torr would not contaminated by hydrocarbons from oil, from that a problem, many colleagues described: bad hydrophilic properties of the films.
Sergey
} Date: Fri, 15 Dec 2000 12:20:19 +0800 } From: "Howe L. C. Josephine" {michowej-at-nus.edu.sg} } Subject: FW: Rotary shadowing } To: "'microscopy-at-sparc5.microscopy.com'" {microscopy-at-sparc5.microscopy.com} } Cc: "'jim-at-proscitech.com'" {jim-at-proscitech.com} } X-Mailer: Internet Mail Service (5.5.2650.21) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Pt coating is not as fine as simultaneous Pt - C. Support film (carbon backing) is only needed if you need to dissolve a thick biological specimen; for thin fibres (say unstained biological material under 50nm) a replica is rather cumbersome. I expect that you also have problems with Pt forming an amalgam with W and so varying amounts are evaporated. That is another advantage of using Pt - C. Another problem is that for C evaporation you do best at around 30 Volts and for metal, you achieve much better control with about 10 volts. The volt settings on some evaporators can be changed but the variable that you are using is changing amperes not volts. I expect that the control of one of your electrodes is very touchy. So take a pick, you have several problems and you may be able to eliminate one or two. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, December 15, 2000 2:20 PM, Howe L. C. Josephine [SMTP:michowej-at-nus.edu.sg] wrote: } We have been doing some rotary shadowing to produce replicas on biological } samples for examination under the TEM. For shadowing we used V-shaped } tungsten wire and pure carbon for support, } } They are a few problems we encountered. Every run is a trial and error } process. This is especially so with the setting of current to vaporize the } metal and the carbon. Every run is a different value although the voltage is } constant. At each run we try to follow the prefer parameter but somehow the } replica would look different from the previous one. Although, it is better } to observe the colour of the glow on the metal and carbon but that is very } hurting to the eye. I wonder if anybody has a definite value for the current } used to vaporize 0,1 mm pt wire/tungsten and pure carbon rod in the vacuum } chamber of 10 -5 torr. } } } Josephine } NUS } } -----Original Message----- } From: Jim at ProSciTech [mailto:jim-at-proscitech.com] } Sent: Thursday, December 14, 2000 6:59 PM } To: 'Lehman, Ann'; Microscopy-at-sparc5.microscopy.com } Subject: RE: Rotary shadowing } } ------------------------------------------------------------------------ } The Microscopy ListServer-Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe-Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I've noted no response, so I better offer something. The technique was more } commonly used to visualize stretched DNA fibers and you may find more } references using the Kleinschmitt technique about 30 years ago. } I have used fixed shadowcasting and negative staining on collagen too and } both methods "work". The advantage of rotary shadowcasting seems to be that } it makes it easier to follow the continuity of fibers (up-down, round and } about). Rotary shadowcasting is usually executed at a shallow angle of 6-12 } degrees and so it builds up more metal at the near vertical surfaces of the } fibers. } Sticky-tape the very edge of the prepared specimen grid to a flat surface } (half of a microscope slide). A bit of tape may be required on two sides to } assure that the grid lies flat. Arrange the grid(s) a the rotation center of } a rotator within the vacuum chamber and locate the rotator to be between 60 } and 100mm from the evaporation source and a little lower than the source to } achieve an angle of say 10 degrees. The angle could be checked with a 10 } degree card segment. } The source could be C-Pt pellets or 0.1mm dia. Pt wire (try 40mm) wrapped } around a 1.5mm carbon cylinder about 5mm long that has been fashioned with a } suitable C rod sharpener. The "loaded" C rod is spring tensioned against a } second C rod that retains a flat end, but one that is reduced in size to } about 3mm dia. using a conventional (but clean, e.g. no pencils) pencil } sharpener. } Evacuate the system. Pt/C evaporation gives very fine granularity; about the } best short of special equipment, but a good vacuum ( { 5x10-5 torr) is } essential. Prior to evaporation start the rotator and adjust the speed to 30 } to 60 rev/ minute. Increase the ampere control to achieve red heat, then } back off to give the vacuum a moment to recover. Then increase ampere to a } white heat and observe using a dark shield until the fashioned carbon tip } has evaporated. Hope this helps. This technique is much easier demonstrated } than written about. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, December 13, 2000 5:57 AM, Lehman, Ann } [SMTP:Ann.Lehman-at-trincoll.edu] wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer-Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe-Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Listers, } } } } Can anyone suggest a good reference or share a technique for preparing } C-Pt } } rotary shadowed collagen molecules on mica for TEM? } } } } Thanks. } } } } ---------------- } } Ann Hein Lehman } } EM Facility Manager } } Trinity College } } Hartford, CT 06106 } } v. 860-297-4289 } } f. 860-297-2538 } } e. ann.lehman-at-trincoll.edu
OK Folks, I am open to suggestions. The computer with the file on it is still down, and I'm getting tired of all the nagging emails. Who can help me? The file is 7.5 MB. Who has enough space and reasonable access and is willing to host the file? The only conditions are that when the printed version of the book eventually appears, the electronic on-line version will need to be removed. In the meantime, anyone who wants to download the pdf file is welcome. If you are interested in providing host space, please contact me (off the list, please). Also note that there has to be some way for me to get the file to you - many mail systems won't accept such a large attachment.
John Russ John_Russ-at-NCSU.edu or DrJohnRuss-at-AOL.com
Email: William.SHort-at-cpe.amedd.army.mil Name: William Short
Question: One of my tasks is to implement the Verein Deutscher Ingenieur 3492 Method for analyzing asbestos fibers and other indoor pollutants using the Scanning Electron Microscope and an Energy Dispersive X-ray Analyzer. At this point I am nearly completed with the implementation. We have a nice old Hitachi S-450 model SEM and we use the KEVEX EDX to get our spectral data off the unknown samples. I am putting together the paperwork and reports to tell our customers what it is that we analyzed from their sample submissions. I give a Concentration result, the instrument Sensitivity determined by the filter area and air volumes, the raw data, fiber types ( i.e. asbestos, inorganic, calcium sulfate, or other), and a 95% Confidence Interval using the Poisson distribution. Also included are the fiber dimensions, voltages, a picture of the fiber morphology, scale in nanometers, and a copy of a spectral spot return from the EDXA. My problem is that I don't know if I'm producing the correct results for the Confidence Interval calculation using the Poisson function. Aren't there special considerations for the results obtained from getting 0 fibers, 1 fiber, 2, fibers, and 3 fibers ? Can you help me with this area or will I just have to "punt" ? The VDI 3492 Method is being used in Europe for this analysis to comply with the Federal Governing Standards for Host nations.
Any information that you could give me would be greatly
On the subject of metal shadowing, might I recommend the excellent treatise by Willison and Rowe (my ex-head of department!) "Practical Methods in Electron Microscopy - Volume 8" - One of the excellent Glauert edited series.
I am currently in the process of ordering Kodak electron microscope film 4489 (3.25 x 4 in.), however, it seems that I can no longer order the film from Kodak Canada Inc. Kodak is still producing the film but only in the U.S. I guess. This may not seem problematic but the price of the film from the Canadian distributers is at least two times as much (almost three times in one case) as the price paid the last time it was ordered.
I believe that AGFA produces a similar type of film but I do not know the product name or who to contact. If anyone has information on distributers, of either Kodak or the AGFA film, with reasonable prices I would appreciate hearing from you. Alternatively, if anyone has another favorite TEM film that they use and would like to share that information I would be happy to hear from you.
Please send me the information offline at the following address: smithh-at-em.agr.ca
Thank you in advance! Heidi Smith Microscopy Technician Agriculture and Agri-Food Canada Kentville, NS, Canada
I've been running the Core EM Facility at Harvard Medical School in Boston for about 5 years now. When I started the lab, I had absolutely no clue what to charge, so I checked out what other Facilities were charging. After a little research (but still feeling pretty clueless) I started charging 30$/ beam hour for Scope time, which I think was was below average in 1995. There weren't that many EM labs in the area so I took an average of whoever I got hold of across the country..
My idea was to charge a little less to get people interested. Which I think worked well (I had good financial support for a few years, until things took off.. I don't know if your lab there is new or already established..) Now that people are hooked, we have an average use of 50-60 hour/month on 2 scopes. I'm charging 31.50$/hour for Med. School users and 45$/hour for outside, for assisted Scope use I charge 63$/hour for Med. School and 94$/hour for outsiders. With our current use, which has been pretty stable for the last year, the income from scope use covers the service contracts and supplies..
I think my rates are lower than average, but I have a huge user group. I have two half time technicians and the Facility income pretty much covers the operating costs now (..not including my salary..!)
I have a full fee-for serivce pricelist at http://www.hms.harvard.edu/core/em.html
Good luck with the EM lab!
Maria
} At 4:45 PM -0400 12/13/00, Ping Li wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________
Maria Ericsson Harvard Medical School EM Facility 220 Longwood Avenue Boston, MA 02115 (617) 432 1698 maria_ericsson-at-hms.harvard.edu http://www.hms.harvard.edu/core/em.html
Florida Society for Microscopy-Florida Chapter of the American Vacuum Society & Surface Analysis 2001
Annual Symposium
Technical Meeting March 12-14 - Short Courses March 12-15 University of Central Florida; Student Union Building; Orlando, Florida
Meeting Highlights * Technical Sessions: Microscopy in the Physical and Biological Sciences, Surface Science, Thin Films, Focused Ion Beam Techniques, Biomaterial Interfaces, Environmental Science, Metrology, and Electronic Materials * AVS & Vendor Sponsored Short Courses * Job Fair * 3rd Annual FIB Users Workshop, and Vacuum Educators Workshop * Student Poster Competition (over $5000 in prizes can be awarded) -plus the MSA Traveling Poster Display * Equipment Exhibit with Surface, Analysis, Vacuum, and MicroscopySciences Vendors * No registration fee for Symposium or Equipment Exhibit
Confirmed Invited Speakers to Date * Donald Baer Pacific Northwest National Lab * Alice Dohnalkova Pacific Northwest National Lab * Klaus Edinger University of Maryland * Robert Hull University of Virginia * Bill Lamberti Exxon * Eero Ristolainen Tampere University of Technology * Bruno Schueler Physical Electronics * Catherine Taylor Wright State University * Edgar Voelkl Oak Ridge National Lab
Abstract Submission The abstract deadline is January 5, 2001 for hard copy abstracts and January 12, 2001 for e-mail abstracts. E-mail submissions are strongly encouraged. Contributed papers will be presented as platform presentations or poster presentations. Authors should indicate their session preference and preferred presentation format i.e., poster or platform. For uniformity, all the abstracts must be in the following format: the title of the talk should be in all capital letters followed by the names and addresses of the authors. The presenter's name should be underlined. Abstracts are limited to 200 words. E-mail abstracts should be as text in the body of the e-mail or an attached Microsoft Word file. Send abstracts to Dr. Lucille Giannuzzi, Program Chair, 407 275-4354, lag-at-mail.ucf.edu. Authors will be contacted by their session chair in mid-January 2001 concerning their abstract's status.
Registration You may register on line by clicking on the following link:
General Meeting Information You may find more information by following the "AVS Info; Chapters; Florida Chapter" links at: www.vacuum.org or the "Local Affiliate Societies" link at www.microscopy.org or contact Fred Stevie, 407-371-7626, stevie-at-lucent.com or Jo Ann Moore, jamoore-at-com1.med.usf.edu 1-813-974-9446
In order to receive email updates and future information about the meeting please forward your email address to Pete Ries pjries-at-lucent.com.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Postdoctoral Position in Transmission Electron Microscopy - Rice University
Rice University, Department of Mechanical Engineering and Materials Science, located in Houston, TX, is seeking candidates for a postdoctoral position in transmission electron microscopy of multicomponent oxide thin films. Applicants should have extensive and demonstrated experience in several areas of TEM and a strong background and interest in materials problem solving. Preference will be given to candidates with experience in high-resolution imaging and electron energy-loss spectroscopy as well as conventional diffraction contrast imaging. Facilities at Rice include a JEOL 2010, field-emission SEM and high-resolution X-ray diffractometers, as well as state-of-the-art sample preparation. The project will be carried out in close collaboration with the University of Houston using a state-of-the-art field-emission TEM (JEOL 2010F), with annular dark-field detector, Oxford link EDS, Gatan GIF and STEM capabilities. Ability and interest to take part in an upgrade of existing facilities to high resolution scanning TEM capability is expected. The position is available immediately. Duration about 1-2 years, salary is commensurate with qualifications. Candidates with a Ph.D. in Materials Science or Physics will be given preferred consideration. Interested candidates should send a curriculum vitae, publication list and the names of at least three references with their contact addresses to:
Prof. Susanne Stemmer Rice University Department of Mechanical Engineering and Materials Science MS 321 6100 Main Street Houston, TX 77005-1892 stemmer-at-rice.edu
Applicants must have proof of legal authorization to work in the United States. Rice University is an Affirmative Action/Equal Opportunity Employer.
Unfortunately Agfa have recently stopped supplying their Scienta film that we used in some of our TEMs so we are now changing to Kodak 4489 or SO163 as the Agfa runs out.
Ron
On Fri, 15 Dec 2000 10:21:27 -0500 Heidi Smith {SMITHH-at-em.agr.ca} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Seasons greetings to all! } } I am currently in the process of ordering Kodak electron microscope film 4489 (3.25 x 4 in.), however, it seems that I can no longer order the film from Kodak Canada Inc. Kodak is still producing the film but only in the U.S. I guess. This may not seem problematic but the price of the film from the Canadian distributers is at least two times as much (almost three times in one case) as the price paid the last time it was ordered. } } I believe that AGFA produces a similar type of film but I do not know the product name or who to contact. If anyone has information on distributers, of either Kodak or the AGFA film, with reasonable prices I would appreciate hearing from you. Alternatively, if anyone has another favorite TEM film that they use and would like to share that information I would be happy to hear from you. } } Please send me the information offline at the following address: } smithh-at-em.agr.ca } } Thank you in advance! } Heidi Smith } Microscopy Technician } Agriculture and Agri-Food Canada } Kentville, NS, Canada } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
There are now several copies of the book available on different servers, so access should not be a problem. The book is available at:
{http://loner.phys.lsu.edu/Users/brent/Stereology.pdf} (Thanks to Brent Neal at Louisiana State Univ. in Baton Rouge. This was the original host machine, but there was a general power failure just as I was posting the original note - another proof of Murphy's Law - and it took a while to get things back up; it is fine now)
{http://www.abdn.ac.uk/~ort056/stereology.pdf} (Thanks to Jenny Gregory {j.gregory-at-abdn.ac.uk} in Aberdeen, Scotland)
{http://128.101.243.155/Stereology.pdf} (Thanks to Mike Herron {herro001-at-umn.edu} at the University of Minnesota)
{http://www.biotech.ufl.edu/~emcl/Stereology.pdf} (Thanks to Greg Erdos {gwe-at-biotech.ufl.edu} at the University of Florida in Gainesville)
It should be available very soon from {http://www.practical-stereology.org} (Thanks to Greg Strout {gstrout-at-ou.edu} at the University of Oklahoma)
A few more sites are in process, and everything will be linked from the original main pages at {http://members.aol.com/drjohnruss/links.htm} {http://members.aol.com/ImagProcTK/update.htm}
Many thanks to all of the others out there who offered help or advice.
Hi everyone; just before everyone disappears for the weekend.... I will be cutting some cryosections of infected wheat heads (on monday, of course), and wanted to fix briefly, then put into a cryoprotectant to keep the cells from rupturing when I section (for light microscopy). I know I have successfully used sucrose in the past, but I can't seem to find the concentration. (4% rings a very distant bell...). Can anyone out there tell me what works for them? (or if you have other cryoprotectants that you put your plant samples into before sectioning, I'd be interested in those, too!)
thanks, as always, in advance shea
Dr. S.Shea Miller Agriculture & AgriFood Canada Eastern Cereal and Oilseed Research Centre Rm. 2068 Neatby Building Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 E-mail: millers-at-em.agr.ca
All this talk about what y'all charge your users is no different than what prompted Congress to pass the Sherman Anti-Trust Act:
to protect Interstate commerce from price-fixing.
The discussion and setting of rates by communication between vendors isn't proper to my legally untrained eye.
Though it's OK to publish your rates, if that's what you really charge.
Now, it also turns out that what you charge users is nowhere near what Amenex Associates as a private, for-profit company has to charge to stay in business:
http://207.103.140.16/webpage/amnxfees.htm
Chuck Garber might want to chime in as to why independent lab owners get steamed up about all this.
Best regards, George Langford amenex-at-amenex.com
I was contacted today by a person who needs to locate some EM images of veterinary specimens. They have an immediate need for images, probably SEM's, of organs of the digestive system.
For further information, please contact them directly, not the listserv, at the following:
Jill Kahn 952-852-7358 kahn-at-collemcvoy.com
I'm just the messenger,
John john.chandler-at-colostate.edu Colorado State University
Thank you to evryone who responded to the free EDS detector. The two that I had are now on their way to the University of North Carolina.
I still have one EDS detector that was removed working from a PHI Auger microprobe. I also have some line voltage stabilizers that were removed from a JEOL 840 SEM. Keep in mind the stabilizers are HEAVY and the shipping costs are proportional.
Please contact me offline for details.
Thank You,
Earl Weltmer Scanservice Corp. SEM Maintenance (714) 573-9158
I can see your point of concern regarding this issue. However, if I understand the context within these folks are discussing fees, there is a different reference basis. They are talking about user fees in an academic environment. These fees are for services to members of their academic community--and in fact, for members of their respective institutions. This is quite different from for-profit enterprises' sphere of interest.
If the universities engage in external fee-based work, your point would tend to have merit. But what these folks are trying to do is to spread their cost center's burden over those who are using it, within their small community of users.
If the situation is other than this, of course, that is a different issue.
gary g.
At 01:29 PM 12/15/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm somewhat amazed by how many varied responses there are on rotary shadowing technique, and how few seem to agree with what works for us. So, here is yet another response.
We shadow biological molecules from the connective tissue matrix, usually ranging in size from 16 to 300 kd. Many are linear, some are globular. We spray the molecules in solution with 70% glycerol. The other 30% is 100 microgram/ml of protein, preferably in a volatile buffer such as 1% acetic acid or 0.1M ammonium bicarbonate, pH 7.8. Other buffers can be used but salt crystals can be a big problem. Spraying is accomplished using an air brush, and we spray onto freshly cleaved mica discs cut from sheets using a hole punch.
The sample is dried in vacuum, though we often accumulate 20 samples so some drying occurs outside vacuum. We are careful to pre-pump the vacuum chamber and heat the guns thoroughly to out-gas, then vent the jar with nitrogen, introduce the samples, and pump again. Our system uses a turbo-molecular pump. We can vary the angle of the rotary stage from outside the chamber, so we tilt the specimen away from the gun and outgas thoroughly again so that little vacuum loss is seen during evaporation. This is really important. Gas is introduced to the rods whenever the system reaches atmospheric pressure. It is also important to keep the system clean, as outgassing a dirty gun takes much longer than outgassing a clean gun. We try to complete evaporation without entering the 10 -5 range, and we begin in the 10-7 range. We evaporate at a slow rate, usually taking 3 to 5 minutes to complete a run, which also seems to improve resolution. Final film resolution is very proportional to vacuum, the better the vacuum the smaller the grain size. We use a quartz monitor for controlling the amount of Pt-C coming from the electron beam gun, but we also use a folded piece of filter paper placed 90 degrees relative to the source, and monitor the color which should be dark gray (not black, not brown). We evaporate at 6 degrees relative to sample as the sample rotates. Following Pt-C deposition, we then tilt the sample to 90 degrees relative to a resistance carbon source and evaporate a backing film of carbon onto the mica. The thickness of the film is monitored with a piece of folded filter paper placed 90 degrees relative to the carbon source, and the correct amount is just visible tan color (not gray) on the filter paper. Our film thickness monitor is not sensitive enough to monitor carbon deposition. We find this carbon film absolutely necessary for sample stability, perhaps because we use so little PT-C. However, too much carbon will certainly affect final resolution, loosing edge detail. Finally, the replicas are exposed to the vapors of 1% acetic acid in a petri dish for about a half-hour prior to floating in distilled water (the acid is very useful in helping the replica to release from the mica (so that they float off as one intact film ). We use high-transmission 600 mesh grids to support the films.
For many years we evaporated Pt wire from carbon rods using a resistance source. We wound 2.3 cm of wire around a cylindrical jig which was the same diameter as the sharpened carbon rods (about 1mm). Prior to coiling the wire, we passed it through a alcohol burner flame till it was orange, which made it more malleable and perhaps cleaned it a bit. The coil was transferred to the resistance source, spanning the intersection of two rods (therefore the site of most resistance and primary heating) held together with moderate spring tension. The carbon rods with accompanying Pt coil was at 6 degrees and about 11 cm away from the mica discs. Using a welders plate (Fullam #12511), we observed the metal as current was increased through the rods and just after the wire melted, the current was turned up just a bit more and left there until the Pt was seen to evaporate completely. It was necessary to observe this through the welders plate to get a good shadow and know when to turn the current down to avoid too much carbon. We evaporated a backing film of pure carbon from a second source oriented at 90 degrees from a distance of 11 cm so that the rods just started to spark, counted to 1.5 seconds, at which time the amount of carbon was probably about right (as judged by filter paper color and a bit of luck).
Sorry for the novel,
Doug
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
Stainless steel funnel poses also some danger - taking it with bare hand after the fill (no such problem with the plastic ones). The funnel I use has a crack which makes it more safe from an internal stress point of view.
Gary Gaugler wrote: ========================================================== I can see your point of concern regarding this issue. However, if I understand the context within these folks are discussing fees, there is a different reference basis. They are talking about user fees in an academic environment. These fees are for services to members of their academic community--and in fact, for members of their respective institutions. This is quite different from for-profit enterprises' sphere of interest.
If the universities engage in external fee-based work, your point would tend to have merit. But what these folks are trying to do is to spread their cost center's burden over those who are using it, within their small community of users. ============================================================= I am obviously not a lawyer, however, one should always keep in mind that Congress never granted any kind of exemptions to the anti-trust laws to tax- exempt organizations. So George Langford is absolutely correct, none of us should be discussing (with competitors) how we set our pricing.
Those of us who follow such things know that the penalties can be quite severe for those who are found to be guilty of violating the anti-trust laws
Gary Gaugler wrote: ========================================================== I can see your point of concern regarding this issue. However, if I understand the context within these folks are discussing fees, there is a different reference basis. They are talking about user fees in an academic environment. These fees are for services to members of their academic community--and in fact, for members of their respective institutions. This is quite different from for-profit enterprises' sphere of interest.
If the universities engage in external fee-based work, your point would tend to have merit. But what these folks are trying to do is to spread their cost center's burden over those who are using it, within their small community of users. ============================================================= I am obviously not a lawyer, however, one should always keep in mind that Congress never granted any kind of exemptions to the anti-trust laws to tax- exempt organizations. So George Langford is absolutely correct, none of us should be discussing (with competitors) how we set our pricing.
Those of us who follow such things know that the penalties can be quite severe for those who are found to be guilty of violating the anti-trust laws
Last Friday I got my first quantitative analytical results out of the JSM-840A which I have been converting to a 1 x eds, 3 x wds JXA version over the past year.
This has been a big fun project, I've learned heaps, and I couldn't have done it without this list and the help of the many listers who have given freely of advice and suggestions, and more.
So thanks, Nestor, and all you good people out there. Please drop in if you ever find yourselves in Auckland, New Zealand, it would be great to be able to replace the mental images which I have of people with factually-based ones!
cheers and happy Christmas
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I do not agree with your analysis of this situation nor with your conclusion. Drawing on a somewhat fading memory of prior law classes (yes, I too am not a lawyer), I recollect that antitrust is covered by the Sherman Antitrust Act.
The purpose of this Act and those is various states is to prevent trusts from creating restraints on trade or commerce and thus, reducing competition. The Sherman Antitrust Act was designed to maintain economic liberty, and to [try to] eliminate restraints on trade and competition.
The Act applies to all transactions and businesses involved in interstate commerce. If the activities are local, the act applies to transactions affecting interstate commerce. Therefore, as I understand universities, they are not engaged in interstate commerce. Neither are they engaged in commerce, per se. Thus, they do not engage in transactions affecting interstate commerce. Their fundamental basis of action is to recover costs of ownership of university owned equipment and facilities as applied to users of such items at the university. I do not sense a wholesale effort by universities to engage in external commerce. Note also that many of the managers are not recovering the cost of their salaries. These are sunk costs by their respective universities.
It therefore seems to me that antitrust does not apply to universities. They are not engaged in commerce, they do not affect interstate commerce, they do not effect restraints on trade and commerce, and they are not in competition with one another. And I think they would agree that they are not in competition with private industry.
The law works in two ways: it specifies actions which are allowed and may specify actions which are not allowed. Lawyers exist because so much of the law is open to interpretation. I've just provided my interpretation of the law. Since I am not a lawyer, there is no charge for this.
gary g.
At 08:38 AM 12/16/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I sincerely thank those of you who provided me with information and support regarding the TEM user fee. I really appreciate it.
Regarding what "Garber, Charles A." wrote:
} .... So George Langford is absolutely correct, none of us } should be discussing (with competitors) how we set our pricing.
I don't think this is relevant to what we have done. What we have discussed is mostly about maintenance cost recovery, not making profit. Furthermore, we are not competitors, and we mostly provide service for research and teaching within our own university or institution. There is no competition among us.
"It therefore seems to me that antitrust does not apply to universities. They are not engaged in commerce, they do not affect interstate commerce, they do not effect restraints on trade and commerce, and they are not in competition with one another. And I think they would agree that they are not in competition with private industry."
Hmmm. This logic is quite interestinng. Then even though Universities provide a service in competition with private enterprise they are not competing. The logic in all of this escapes me. Hmmm.
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Garber, Charles A." {cgarber-at-2spi.com} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, December 16, 2000 2:48 PM
I am not friendly with American law ether, but our medical school stated as "non-profit organization". So, to me it mean, that University is not involved in trading and thereafter is not a subject for anti-trust law.
Our hospital, for instance, charge people for the service, but they did not make a profit, same as when we charge people for using our facilities. I don't see any problem here as long as we did not make profit on it.
Merry Christmas! Happy New Year and New Millennium for all ListServer readers! I wish to all of us, that our microscopes will work better in New Millennium and we will have enough users to recover at least 70% of maintaining cost.
Sergey.
} Date: Sat, 16 Dec 2000 14:48:49 -0800 } From: Gary Gaugler {gary-at-gaugler.com} } Subject: Re: User fee discussion } X-Sender: gaugler-at-pop.calweb.com (Unverified) } To: "Garber, Charles A." {cgarber-at-2spi.com} } Cc: MSA listserver {Microscopy-at-sparc5.microscopy.com} } X-Mailer: QUALCOMM Windows Eudora Version 5.0 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
Chuck Garber's reply to the User Fee discussion was truncated, due to a subtle effect of the Email system here. In fairness I am reposting it for him so that you can see the rest of his comments.
For those of you that are curious, the message was truncated because of a line in the original text which started with a "." followed by a blank/empty line. This caused the Email system to believe the message was completed.
============================================================= I am obviously not a lawyer, however, one should always keep in mind that Congress never granted any kind of exemptions to the anti-trust laws to tax- exempt organizations. So George Langford is absolutely correct, none of us should be discussing (with competitors) how we set our pricing.
Those of us who follow such things know that the penalties can be quite severe for those who are found to be guilty of violating the anti-trust laws.
So to my (also) legally untrained eye, for universities to be discussing how they set their pricing is no different than if independent laboratories (like Amenex and Structure Probe) started a parallel discussion on how we should be establishing our pricing. Anyone in academia who has ever written a proposal for funding in recent years knows that universities are in direct competition with each other in ways that are no different from the way independent laboratories compete with each other. George alluded to another issue, that is, the case when universities start offering their services to private companies. I am not aware of anyone having gone to jail for this, but it does raise enormous issues of ethical values, and the impact on students, and their own views of what is right and what is wrong, when the university's facilities are so commercialized. All of us should have some concern about such matters.
Chuck
Disclaimer: Structure Probe, Inc. is an independent for-profit tax-paying laboratory offering electron microscopy services to clients, often times in direct competition with universities who claim they "don't compete with private companies".
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. /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: www.2spi.com ############################ ==================================================
Agfa Scientia TEM film was discontinued earlier this year. I do not know of a manufacturer other than Kodak that is selling a "dedicated" TEM in North America.
George Laing National Graphic Supply
Heidi Smith wrote:
} I believe that AGFA produces a similar type of film but I do not } know the product name or who to contact. If anyone has } information on distributers, of either Kodak or the AGFA film, } with reasonable prices I would appreciate hearing from you. } Alternatively, if anyone has another favorite TEM film that they } use and would like to share that information I would be happy to } hear from you. } }
I'd like to add my 0.004$ or R0.02 to the LN2 discussion:
An alternative to both the exploding plastic funnel and freezing finger steel funnel is to make one out of very thick brown paper (held together with staples rather than sticky tape).
I also staple a tissue paper filter in the funnel mouth to try to remove any ice from the LN2.
Regards Alison Tuling
Engineer Industrial Metal and Minerals Research Institute Department Materials science and Metallurgical engineering University of Pretoria South Africa +27 11 420 4556
It is interesting that this discussion comes up every 12 to 18 months or so. I also note a subtle difference in the way the question was posed, and see that the responses have, perhaps without intending, responded to both aspects of the original question. First, it would appear that asking for rates, particularly at potentially competitive facilities, is at least a gray area, if not a violation of the anti-trust laws. Second, asking assistance in how to determine how to set rates seems to be less problematic. It is within that context that anti-trust laws would appear to be less applicable. For individuals who have not had to deal with the issues of making a facility self-supporting, it can seem a daunting task. Thus, getting advice from other facility managers who have done the task aids the requester to determine what costs must be included, how to determine other factors, etc, and not miss any important/essential costs. It is important for all of us to be aware of the differences in the way questions are posed, especially as regards to this type of issue. This has been an enlightening discussion all around.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc
NB: These are personal opinions only, and in no way reflect any other person or organization.
On Sat, 16 Dec 2000 21:25:04 -0800, Earl Weltmer 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 } -----------------------------------------------------------------------. } } } Gary Gaugler wrote: } } "It therefore seems to me that antitrust does not apply to } universities. They are not engaged in commerce, they do } not affect interstate commerce, they do not effect } restraints on trade and commerce, and they are not } in competition with one another. And I think they would } agree that they are not in competition with private industry." } } Hmmm. This logic is quite interestinng. Then even though Universities } provide a service } in competition with private enterprise they are not competing. The logic in } all of this escapes me. } Hmmm. } } ----- Original Message ----- } } From: "Gary Gaugler" {gary-at-gaugler.com} } To: "Garber, Charles A." {cgarber-at-2spi.com} } Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} } Sent: Saturday, December 16, 2000 2:48 PM } Subject: Re: User fee discussion } } } } ------------------------------------------------------------------------ } } 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 do not agree with your analysis of this situation nor with } } your conclusion. Drawing on a somewhat fading memory } } of prior law classes (yes, I too am not a lawyer), I recollect } } that antitrust is covered by the Sherman Antitrust Act. } } } } The purpose of this Act and those is various states is to } } prevent trusts from creating restraints on trade or commerce } } and thus, reducing competition. The Sherman Antitrust Act } } was designed to maintain economic liberty, and to [try to] eliminate } } restraints on trade and competition. } } } } The Act applies to all transactions and businesses involved in } } interstate commerce. If the activities are local, the act applies } } to transactions affecting interstate commerce. Therefore, as } } I understand universities, they are not engaged in interstate } } commerce. Neither are they engaged in commerce, per se. } } Thus, they do not engage in transactions affecting interstate } } commerce. Their fundamental basis of action is to recover } } costs of ownership of university owned equipment and facilities } } as applied to users of such items at the university. I do not } } sense a wholesale effort by universities to engage in external } } commerce. Note also that many of the managers are not } } recovering the cost of their salaries. These are sunk costs } } by their respective universities. } } } } It therefore seems to me that antitrust does not apply to } } universities. They are not engaged in commerce, they do } } not affect interstate commerce, they do not effect } } restraints on trade and commerce, and they are not } } in competition with one another. And I think they would } } agree that they are not in competition with private industry. } } } } The law works in two ways: it specifies actions which are } } allowed and may specify actions which are not allowed. } } Lawyers exist because so much of the law is open to } } interpretation. I've just provided my interpretation of the } } law. Since I am not a lawyer, there is no charge for this. } } } } gary g. } } } } } } } } } } } } At 08:38 AM 12/16/00, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } Gary Gaugler wrote: } } } ========================================================== } } } I can see your point of concern regarding this issue. However, if I } } } understand the context within these folks are discussing fees, there is a } } } different reference basis. They are talking about user fees in an } academic } } } environment. These fees are for services to members of their academic } } } community--and in fact, for members of their respective institutions. } This } } } is quite different from for-profit enterprises' sphere of interest. } } } } } } If the universities engage in external fee-based work, your point would } tend } } } to have merit. But what these folks are trying to do is to spread their } } } cost center's burden over those who are using it, within their small } } } community of users. } } } ============================================================= } } } I am obviously not a lawyer, however, one should always keep in mind that } } } Congress never granted any kind of exemptions to the anti-trust laws to } tax- } } } exempt organizations. So George Langford is absolutely correct, none of } us } } } should be discussing (with competitors) how we set our pricing. } } } } } } Those of us who follow such things know that the penalties can be quite } } } severe for those who are found to be guilty of violating the anti-trust } laws } } } } } }
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
} Colleagues } } Chuck Garber's reply to the User Fee discussion was } truncated, due to a subtle effect of the Email system here. } In fairness I am reposting it for him so that you can } see the rest of his comments. } -------------------------------------------------- } } ============================================================= } I am obviously not a lawyer, however, one should always keep in mind that } Congress never granted any kind of exemptions to the anti-trust laws to tax- } exempt organizations. So George Langford is absolutely correct, none of us } should be discussing (with competitors) how we set our pricing.
What rubbish!! Of course we can discuss what standards we use to set fees. Businesses do this all the time.
} Those of us who follow such things know that the penalties can be quite } severe for those who are found to be guilty of violating the anti-trust laws.
Everytime someone gets on this list (and other lists) and asks about setting prices for users of lab services, Mr Langford and/or Dr. Garber starts making veiled threats about anti-trust violations. Isn't it interesting that no actual statues are cited, just vague references and generalizations, and that none of the citers are legally trained?
} So to my (also) legally untrained eye, for universities to be discussing how } they set their pricing is no different than if independent laboratories } (like Amenex and Structure Probe) started a parallel discussion on how we } should be establishing our pricing. Anyone in academia who has ever written } a proposal for funding in recent years knows that universities are in direct } competition with each other in ways that are no different from the way } independent laboratories compete with each other.
Not true of NIH funding of individual investigators.
} George alluded to another } issue, that is, the case when universities start offering their services to } private companies.
If you will read the original posting, that is NOT what was being talked about.
} I am not aware of anyone having gone to jail for this,
I am sure that if anyone had even been formally charged, you or Mr. Langford would be sure to let us know!
} } but it does raise enormous issues of ethical values, and the impact on } students, and their own views of what is right and what is wrong, when the } university's facilities are so commercialized. All of us should have some } concern about such matters.
Ethical behavior is a two-way street.
} Chuck } } Disclaimer: Structure Probe, Inc. is an independent for-profit tax-paying } laboratory offering electron microscopy services to clients, often times in } direct competition with universities who claim they "don't compete with } private companies". } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } Structure Probe, Inc. /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: www.2spi.com } ############################ } ==================================================
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Regarding the contact info that Scott mentioned awhile back, we happen to sell what is probably that film dryer. You can see it at
http://www.tedpella.com/photo_html/photo11.htm
Tom
"Walck, Scott D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } There is a company called California Stainless that made film dryers that } some of the EM supply houses sold. I bought one from them a number of years } ago. Sorry that I don't have the contact info, but I'm sure that this is } just what you want. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } -----Original Message----- } From: Donald Lovett [mailto:lovett-at-tcnj.edu] } Sent: Monday, November 20, 2000 9:51 AM } To: Microscopy.lst } Subject: Film dryers } } } --------------------------------------------------------------- } --------- } 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/MicroscopyListserv er/FAQ.html } } } } --------------------------------------------------------------- } --------. } } } } Years ago I worked in a lab that had a small forced (and filtered) film } dryer (with heater). It was just large enough to hold two racks of TEM } negatives. I cannot find one in any of the catalogues. I can } only find } film dryers for 35 mm roll film. Could anyone please } recommend a source } of a similar dryer? (Vendors, please feel free to respond directly to } me). } } Thanks, } } Don } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } P.O. Box 7718 fax: (609) 637-5118 } The College of New Jersey } Ewing, NJ 08628-0718 } } } }
} It is interesting that this discussion comes up every 12 to 18 months or so.
Yes. Some listmember asks for a little guidance and get several replys. Then two people (the same two people, if my memory serves me) start yelling "anti-trust violation". No facts, no applicable case law, just the suggestion of lawlessness.
} I also note a subtle difference in the way the question was posed, and see } that the responses have, perhaps without intending, responded to both } aspects of the original question. First, it would appear that asking for } rates, particularly at potentially competitive facilities, is at least a } gray area, if not a violation of the anti-trust laws.
How? Please tell us exactly how asking what some other lab charges for XYZ is illegal.
} Second, asking } assistance in how to determine how to set rates seems to be less } problematic. It is within that context that anti-trust laws would appear to } be less applicable. For individuals who have not had to deal with the } issues of making a facility self-supporting, it can seem a daunting task. } Thus, getting advice from other facility managers who have done the task } aids the requester to determine what costs must be included, how to } determine other factors, etc, and not miss any important/essential costs. } It is important for all of us to be aware of the differences in the way } questions are posed, especially as regards to this type of issue. This has } been an enlightening discussion all around. } } Roger Moretz, Ph.D. } Dept of Toxicology } Boehringer Ingelheim Pharmaceuticals, Inc } } NB: These are personal opinions only, and in no way reflect any other person } or organization. } } On Sat, 16 Dec 2000 21:25:04 -0800, Earl Weltmer wrote: } } } Gary Gaugler wrote: } } } } "It therefore seems to me that antitrust does not apply to } } universities. They are not engaged in commerce, they do } } not affect interstate commerce, they do not effect } } restraints on trade and commerce, and they are not } } in competition with one another. And I think they would } } agree that they are not in competition with private industry." } } } } Hmmm. This logic is quite interestinng. Then even though Universities } } provide a service } } in competition with private enterprise they are not competing. The logic } in } } all of this escapes me. } } Hmmm. } } } } ----- Original Message ----- } } } From: "Gary Gaugler" {gary-at-gaugler.com} } } To: "Garber, Charles A." {cgarber-at-2spi.com} } } Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} } } Sent: Saturday, December 16, 2000 2:48 PM } } Subject: Re: User fee discussion } } } } } } } I do not agree with your analysis of this situation nor with } } } your conclusion. Drawing on a somewhat fading memory } } } of prior law classes (yes, I too am not a lawyer), I recollect } } } that antitrust is covered by the Sherman Antitrust Act. } } } } } } The purpose of this Act and those is various states is to } } } prevent trusts from creating restraints on trade or commerce } } } and thus, reducing competition. The Sherman Antitrust Act } } } was designed to maintain economic liberty, and to [try to] eliminate } } } restraints on trade and competition. } } } } } } The Act applies to all transactions and businesses involved in } } } interstate commerce. If the activities are local, the act applies } } } to transactions affecting interstate commerce. Therefore, as } } } I understand universities, they are not engaged in interstate } } } commerce. Neither are they engaged in commerce, per se. } } } Thus, they do not engage in transactions affecting interstate } } } commerce. Their fundamental basis of action is to recover } } } costs of ownership of university owned equipment and facilities } } } as applied to users of such items at the university. I do not } } } sense a wholesale effort by universities to engage in external } } } commerce. Note also that many of the managers are not } } } recovering the cost of their salaries. These are sunk costs } } } by their respective universities. } } } } } } It therefore seems to me that antitrust does not apply to } } } universities. They are not engaged in commerce, they do } } } not affect interstate commerce, they do not effect } } } restraints on trade and commerce, and they are not } } } in competition with one another. And I think they would } } } agree that they are not in competition with private industry. } } } } } } The law works in two ways: it specifies actions which are } } } allowed and may specify actions which are not allowed. } } } Lawyers exist because so much of the law is open to } } } interpretation. I've just provided my interpretation of the } } } law. Since I am not a lawyer, there is no charge for this. } } } } } } gary g. } } } } } } } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } Gary Gaugler wrote: } } } } ========================================================== } } } } I can see your point of concern regarding this issue. However, if I } } } } understand the context within these folks are discussing fees, there } is a } } } } different reference basis. They are talking about user fees in an } } academic } } } } environment. These fees are for services to members of their academic } } } } community--and in fact, for members of their respective institutions. } } This } } } } is quite different from for-profit enterprises' sphere of interest. } } } } } } } } If the universities engage in external fee-based work, your point } would } } tend } } } } to have merit. But what these folks are trying to do is to spread } their } } } } cost center's burden over those who are using it, within their small } } } } community of users. } } } } ============================================================= } } } } I am obviously not a lawyer, however, one should always keep in mind } that } } } } Congress never granted any kind of exemptions to the anti-trust laws } to } } tax-exempt organizations. So George Langford is absolutely correct, none } of us } } } } should be discussing (with competitors) how we set our pricing. } } } } } } } } Those of us who follow such things know that the penalties can be } quite } } } } severe for those who are found to be guilty of violating the } anti-trust } } laws } } _______________________________________________________ } Send a cool gift with your E-Card } http://www.bluemountain.com/giftcenter/
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Ah yes, this reminds me of a similar Close Encounter with liq N2 that I had: I removed a stainless steel rack from a Dewar, the type used for holding 5"X5" boxes as a cell freezer. Heavy, and dripping liq N2, I placed it down on one of those gray round rubber-topped step stools found in most labs. It was quite exciting when the rubber exploded with a CRACK and sent shards of frozen rubber in every direction. Luckily, I was alone and never 'fessed up to the missing rubber top on the step stool.............. Marilyn
Best wishes for an environmentally conscious, socially responsible, low stress, non-addictive, gender neutral, winter solstice holiday, practiced within the most joyous traditions of the religious persuasion of your choice, but with due respect for the religious persuasion of others who choose to practice their own religion, as well as those who choose not to practice a religion at all.
Additionally, (topical bit here!) a fiscally successful, personally fulfilling, and medically uncomplicated recognition of the generally accepted calendar year 2001, but not without due respect for the alternative calendars of choice of other cultures whose contributions have helped make our society great, without regard to the race, creed, color, religious, or sexual preferences of the wishes.
May the deity of our choice bless us each and every one but should you not be a believer (as above) then please accept our collective wish as fellow sentient (?) beings for the full development of your inner self.
Yours, Chris ==================================================
(Disclaimer: This greeting is subject to confirmation, clarification or withdrawal. It implies no promise by the sendeer actually to implement any of the expressed wishes for her/himself or others and no responsibility for any unintended emotional stress these greetings may or may not bring to those not caught up in the holiday spirit.)
Hope y'all manage to get a life this Christmas ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
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Reply to: RE: Fwd: Re: MRS follow-up
Dear Experts of Quasicrystals,
Recently I received two e-mails from the MRS QC meeting organizer to this entire group of recipients I am addressing, concerning the binary QCs. Since the binary QCs are interesting, I would like to make some more discussions.
In the literature, many binary QCs have been observed, including the Al-, Cd-, Ti-, and Cr-based binary systems (to see Pr. Guyot's review, Rep. Prog. Phys., 1991, 54, p. 1373, Table 1). The Ta- and Ga-based systems found later are not listed there. Some of these constituents may be substituted by other elements, not only for the Cd-Cu system (Bendersky et al., Scripta metall., 1987, 21, 531) but also for other alloy systems. Since the substitution seems to be relatively easier in binary alloys, probably there are some more chances to explore new QC alloys by the substitution method (little changes in composition maybe needed), Among those QCs observed so far, some may still remain unclear if they can form from the liquid directly or not.
The interesting thing to us is that why some QCs can form from the liquid directly. However, even if a QC forms directly from the liquid, it doesn't mean that it must be a stable phase (for example, we recently observed that the Mg-Zn-RE IQCs are not thermodynamically stable). Therefore, if a QC is not thermodynamically stable, it should be more accurately renamed as an "as-cast" QC instead of a "stable" QC.
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id QAA02924 for dist-Microscopy; Mon, 18 Dec 2000 16:51:47 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id QAA02921 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 18 Dec 2000 16:51:17 -0600 (CST) Received: from pdx.edu (tuttle.oit.pdx.edu [131.252.120.29]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id QAA02914 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 18 Dec 2000 16:51:02 -0600 (CST) Received: from [131.252.192.235] (host-192-235.dhcp.pdx.edu [131.252.192.235]) by pdx.edu (8.11.1/8.9.3) with ESMTP id eBIMnnu22753 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 18 Dec 2000 14:49:49 -0800 (PST) Message-Id: {200012182249.eBIMnnu22753-at-pdx.edu} X-Mailer: Microsoft Outlook Express Macintosh Edition - 4.5 (0410)
Hi everyone,
thanks for the deluge of TEM film holders - we have plenty now thanks.
Yet another plea - last week we noticed we were suddenly getting nasty streaks on our SEM photos from light getting to our film holder. Close examination revealed that the plastic casing has cracked through.
In light of the fact that lots of people are going digital with their SEM images, does anyone have an old 550 Polaroid film holder that they are willing to sell to us? If not, we are willing to sell off our 30 packs of polaplan film that goes in this type of holder.
cheers Liz McKenzie
******************************************************* Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
I must protest the blatant and insensitive hemispherism implicit in this posting.
Merry er....just be happy, OK?
Sally Stowe (Canberra, Australia, where its 30 degrees and rising).
Chris Jeffree wrote:
---
Dear All
Best wishes for an environmentally conscious, socially responsible, low stress, non-addictive, gender neutral, winter solstice holiday, practiced within the most joyous traditions of the religious persuasion of your choice, but with due respect for the religious persuasion of others who choose to practice their own religion, as well as those who choose not to practice a religion at all. ........... etc
Folks, this is the year 2000. Anti trust laws dont mean, nor do diddly squat. It is all rhetoric spun off to make it look like things are peachy keen. In reality it is pure B_ _ _S_ _T. Same for when the gurus talk about how great the economy is. Look into your own wallet and assess if you are REALLY doing better than you were 10-20 year ago. The answer is NOT. Lou Solebello
From root Mon Dec 18 20:28:14 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id TAC00250 for dist-Microscopy; Mon, 18 Dec 2000 19:38:18 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id SAA03116 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 18 Dec 2000 18:52:01 -0600 (CST) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id SAA03109 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 18 Dec 2000 18:51:51 -0600 (CST) X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {v03130301b66461281365-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Folks, this is the year 2000. Anti trust laws dont mean, nor do diddly squat. It is all rhetoric spun off to make it look like things are peachy keen. In reality it is pure B_ _ _S_ _T. Same for when the gurus talk about how great the economy is. Look into your own wallet and assess if you are REALLY doing better than you were 10-20 year ago. The answer is NOT. Lou Solebello
To anyone who has experience in attempting to view and determine emulsions bead size using a SEM and determining average particle size of a past wax/paint sealant.
Please respond to Thomas Van Doozer tvd-at-gvtc.com Thank you.
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Zaluzec-at-sparc5.microscopy.com wrote:
} Colleagues } } Chuck Garber's reply to the User Fee discussion was } truncated, due to a subtle effect of the Email system here. } In fairness I am reposting it for him so that you can } see the rest of his comments. } -------------------------------------------------- } } ============================================================= } I am obviously not a lawyer, however, one should always keep in mind that } Congress never granted any kind of exemptions to the anti-trust laws to tax- } exempt organizations. So George Langford is absolutely correct, none of us } should be discussing (with competitors) how we set our pricing.
What rubbish!! Of course we can discuss what standards we use to set fees. Businesses do this all the time.
} Those of us who follow such things know that the penalties can be quite } severe for those who are found to be guilty of violating the anti-trust laws.
Everytime someone gets on this list (and other lists) and asks about setting prices for users of lab services, Mr Langford and/or Dr. Garber starts making veiled threats about anti-trust violations. Isn't it interesting that no actual statues are cited, just vague references and generalizations, and that none of the citers are legally trained?
} So to my (also) legally untrained eye, for universities to be discussing how } they set their pricing is no different than if independent laboratories } (like Amenex and Structure Probe) started a parallel discussion on how we } should be establishing our pricing. Anyone in academia who has ever written } a proposal for funding in recent years knows that universities are in direct } competition with each other in ways that are no different from the way } independent laboratories compete with each other.
Not true of NIH funding of individual investigators.
} George alluded to another } issue, that is, the case when universities start offering their services to } private companies.
If you will read the original posting, that is NOT what was being talked about.
} I am not aware of anyone having gone to jail for this,
I am sure that if anyone had even been formally charged, you or Mr. Langford would be sure to let us know!
} } but it does raise enormous issues of ethical values, and the impact on } students, and their own views of what is right and what is wrong, when the } university's facilities are so commercialized. All of us should have some } concern about such matters.
Ethical behavior is a two-way street.
} Chuck } } Disclaimer: Structure Probe, Inc. is an independent for-profit tax-paying } laboratory offering electron microscopy services to clients, often times in } direct competition with universities who claim they "don't compete with } private companies". } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } Structure Probe, Inc. /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: www.2spi.com } ############################ } ==================================================
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Hi guys, Anybody who can give me tips on interpretation of thin film micrographs. Or where I can get info on the same subject on the net. Thanks for all your help.
Mr. O. O. ILORI DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING OBAFEMI AWOLOWO UNIVERSITY, ILE-IFE, OSUN STATE NIGERIA.
We are trying to take the low-power photomicrographs for negative images to identify the connections past the interstitial subnucleus in the Nucleus tractus solitarius by neuronal tracer. Any guidance on this would be very much appreciated Thank you in advance.
Hyung-Tae Kim
Postdoctoral Fellow Laryngeal and Speech Section, MNB, NINDS Bldg. 10 Rm 5D38 10 Center Drive, MSC 1416 Bethesda, MD 20892-1416
At 3:27 PM -0500 12/15/00, Shea Miller wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Best wishes for an environmentally conscious, socially responsible, low stress, non-addictive, gender neutral, winter solstice holiday, practiced within the most joyous traditions of the religious persuasion of your choice, but with due respect for the religious persuasion of others who choose to practice their own religion, as well as those who choose not to practice a religion at all.
Additionally, (topical bit here!) a fiscally successful, personally fulfilling, and medically uncomplicated recognition of the generally accepted calendar year 2001, but not without due respect for the alternative calendars of choice of other cultures whose contributions have helped make our society great, without regard to the race, creed, color, religious, or sexual preferences of the wishes.
May the deity of our choice bless us each and every one but should you not be a believer (as above) then please accept our collective wish as fellow sentient (?) beings for the full development of your inner self.
Yours, Chris ==================================================
(Disclaimer: This greeting is subject to confirmation, clarification or withdrawal. It implies no promise by the sendeer actually to implement any of the expressed wishes for her/himself or others and no responsibility for any unintended emotional stress these greetings may or may not bring to those not caught up in the holiday spirit.)
Hope y'all manage to get a life this Christmas ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
Thanks for your all your replies on this topic. Some liked it, and some didn't but there was a substantial number of spoiled ballots, so the jury is still out. Best wishes to you all anyway! Chris ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ University of Oregon EPMA/SEM Technician ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
The Department of Geological Sciences at the University of Oregon invites applications for an electron beam specialist in a modern microanalytical facility. The facility houses a four-spectrometer Cameca SX-50 microprobe and a JEOL JSM-6300 SEM. It serves faculty and students in the Department of Geological Sciences, Chemistry, Physics and the Materials Science Institute. Primary responsibilities include instrument maintenance, instruction and research collaboration. Additional responsibilities include maintenance and operation of support and sample preparation equipment as well as optical microscopes and a network of computers used for file archival and data/image analysis.
We seek an individual with expertise and experience in WDS and EDS analytical techniques, SEM imaging and instrument maintenance. Additional desired skills include experience with network computers (UNIX/Windows), applied computer-aided microscopy and instruction in instrument use. An advanced degree in petrology, mineralogy or analytical chemistry is desirable.
This is a full-time, annually renewable position. Salary and title will be commensurate with experience and education. Applicants should send curriculum vitae, a statement of experience and interests, and the names, addresses, phone numbers and email addresses of at least three referees to the EPMA/SEM Search Committee, Department of Geological Sciences, 1272 University of Oregon, Eugene OR 97403-1272. We will begin reviewing completed applications February 1, 2001, and will continue until the position is filled.
The University of Oregon is an equal opportunity/affirmative action institution committed to cultural diversity and compliance with the Americans with Disabilities Act.
A PDF of the text is available ... we would genuinely appreciate this announcement's distribution. http://epmalab.uoregon.edu/epma_position.pdf
Feel free to visit the facility's wwwsite for more information regarding the Dep't of Geological Sciences, the University of Oregon, and the community of Eugene. http://epmalab.uoregon.edu/ ...
to ask any specific questions ... about the facility: mailto:epmalab-at-darkwing.uoregon.edu about the position: Jack Rice mailto:jrice-at-darkwing.uoregon.edu
... and a special thanx to my peers ... http://epmalab.uoregon.edu/images/zircon-xmas.jpg
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
We at university-based facilities are not competing with commercial labs, nor are we out to steal any business away from you. Believe me, most of us have our hands full trying to keep up with the needs/demands of our in-house clients. We are here to provide service at reasonable cost to the research faculty, students, and staff. I know that my facility is expected to cover its non-personnel operating costs (service contracts, supplies, maintenance contracts) and NOT turn a profit. That is against NIH guidelines (take a look at the tapes/transcripts of the M&M sessions devoted to facility management). Since most if not all of my clients are federally funded, I am not allowed to charge them more than a fee calculated to cover my expenses for the procedure involved. My institution maintains a separate EM facility for clinical applications, in part because of the billing differences. I think that the original question posted was a plea for help in establishing de novo an approximation of what is a reasonable price....high enough to cover operating costs and keep from operating too far "in the red", but not so high as to be unreasonable or out of the range for people operating within grant budgets.
It is not unreasonable for people to want to be able to get work done without having to send it out or schlepp it half-way across town. It has become too costly for individuals to maintain this type of equipment in their own labs, and many people need access only on a sporadic basis. That is why central core facilities get set up.
Chill out guys.
Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
} Dear Colleagues: } } We are trying to take the low-power photomicrographs for negative images to } identify the connections past the interstitial subnucleus in the Nucleus } tractus solitarius by neuronal tracer. } Any guidance on this would be very much appreciated } Thank you in advance. } } } Hyung-Tae Kim } } Postdoctoral Fellow } Laryngeal and Speech Section, MNB, NINDS } Bldg. 10 Rm 5D38 } 10 Center Drive, MSC 1416 } Bethesda, MD 20892-1416 } } email : kimth-at-ninds.nih.gov } Phone : 301-402-0157 } Fax : 301-480-0803
Could you be more specific about the problems you are encountering? What is keeping you from getting useful images?
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I am looking for a used control unit for Reichert Ultracut Ultramicrotome. This is a separate box sits right next to the main microtome and some key functions of the microtome can be controlled from it. If anyone has one for sale, I will be very interested in buying it. Please contact me directly, not to the listserve.
Thank you,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab and Fluorescence Imaging Facility Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
Help! I'm looking for objective lenses for a Zeiss STEMI SV8 stereomicroscope.
The Zeiss part numbers include: 47 50 73 S 50 mm. 47 50 61 S 100 mm. 47 50 62 PLS 100 mm. 47 50 63 DS 100 mm. 47 50 72 Photo S 100 mm. 47 50 74 S 150 mm. and 47 50 71 S 200 mm.
Please respond to me directly at nessonm-at-ucs.orst.edu with prices and condition. DON'T just hit the REPLY button and bother everyone else in the newsgroup.
TIA, M. Nesson _______________________________________________________________________ Michael Nesson, Ph.D. Department of Biochemistry & Biophysics 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 (541)737-5245 FAX:(541)737-0481 nessonm-at-ucs.orst.edu
How might I find out if there is anything we can do with our old RCA EMU3G TEM other than simply scrapping it? How could I list it where someone who might need it for parts would see it? It was working until the day we stopped teaching the course here in 1990, and with the arrival of our new SEM in January the old TEM has to go to make room for it.
Thanks for any assistance.
Sincerely, Lee Hadden Professor of Biology Department of Biology Wingate University Wingate, NC 28174
Can anybody tell me how to prepare TEM samples for biomaterials such as coral? Thanks. Feng ************************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
Philips delivered their first commercial electron microcope in October 1949 (bought by 'Statens Serum Institute' in Denmark for research into the polio virus)
Source: FEI Company - Scope 11 - December 1999 - page 3
Decraen Erwin
Belgian Building Research Institute (BBRI) Division of Materials Av. P. Holoffe 21 B - 1342 Limelette
-----Original Message----- From: Ekstrom, Harry [SMTP:harry.ekstrom-at-honeywell.com] Sent: Wednesday, December 20, 2000 4:57 AM To: '"Geinfam-at-aol.com"-at-sparc5.microscopy.com'; Microscopy-at-sparc5.microscopy.com Subject: RE: Electron microscope first demonstration
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-----Original Message----- } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, December 19, 2000 4:36 PM To: Microscopy-at-sparc5.microscopy.com Subject: Electron microscope first demonstration
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I have a date of Tues 07 April 1931 for the first two-stage (magnification of an electron image further magnified by a second lens) image of a platinum grid being obtained by Ruska.
Ref. T. Mulvey, Dept. Physics, The University of Aston, Birmingham. - An article based upon the opening address of the fifth European congress on electron microscopy, EMCON 72 held in Manchester 5-12 September 1972.
S.C. Hyman Chief Technician The Electron Microscope Laboratory Faculty of Medicine and Biological Sciences Adrian Building University of Leicester University Road Leicester LE1 7RH
Tel. (0116) 252 3370
-----Original Message----- } From: Ekstrom, Harry [mailto:harry.ekstrom-at-honeywell.com] Sent: 20 December 2000 03:57 To: '"Geinfam-at-aol.com"-at-sparc5.microscopy.com'; Microscopy-at-sparc5.microscopy.com
My source quotes M.Knoll first demonstrated the SEM in 1935 but no specific date.
} Can anyone tell me the exact date in the "30's when the electron microscope } was first demonstrated?
I have a little brochure in front of me called "The First North American Electron Microscope", prepared by U.M. Franklin, G.C. Weatherly and G.T. Simon, in 1978. In a section about the early European design efforts, they state that Ruska had "constructed the pioneer two-stage transmission model in 1933", but don't give any further information on it.
Frank Thomas Geological Survey of Canada Atlantic Dartmouth, Nova Scotia Canada
Hello! This is to inform members and other interested individuals that the newsletter for the Southeastern Microscopy Society is now online at http://www.biotech.ufl.edu/sems/ . This issue contains information about our annual meeting to be held May 23-25, 2001 at Clemson University. The issue also includes information and forms for joining the Society. Thanks John P. Shields, PhD Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080 FAX 706-542-4271 jshields-at-cb.uga.edu
Eragonite or polyp? Hard or soft? Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, December 20, 2000 6:35 PM, Feng Wu [SMTP:fwu-at-gbumail.bgu.ac.il] 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 } -----------------------------------------------------------------------. } } } Can anybody tell me how to prepare TEM samples for biomaterials such as } coral? } Thanks. } Feng } ************************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } ************************************************ }
In 1935? That was a TEM, not an SEM. Knoll was Ernst Ruska's PhD supervisor and later collaborator. They designed and built the first TEMs. The very first model had no specimen chamber and just proved that the lens system could work. So the first model, which I seem to remember was before 1935 was really not a microscope. Von Ardenne described the operating principle of an SEM in the late 30th, but I believe none was build until the early 60th at Cambridge University, UK. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, December 20, 2000 1:57 PM, Ekstrom, Harry [SMTP:harry.ekstrom-at-honeywell.com] wrote: } } } My source quotes M.Knoll first demonstrated the SEM in 1935 but no specific } date. } } Harry Ekstrom } Materials Laboratory } } (602) 231-2744 } e-mail: harry.ekstrom-at-honeywell.com } } } -----Original Message----- } } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com } [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] } Sent: Tuesday, December 19, 2000 4:36 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Electron microscope first demonstration } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Can anyone tell me the exact date in the "30's when the electron microscope } was first demonstrated? } } geinfam-at-aol.com }
I think Ernst Ruska is generally credited for building the protootype of today's microscope as his PhD project. He and Max Knoll published a description in 1932 (Knoll M, Ruska E. The electron microscope. Z Physik 78:318-39, 1932.).
However, others contributed to the design and knowledge of electron behavior and lenses, including de Broglie on wave theory (Phil Mag 47:446-58, 1924) and Busch on lenses (Ann Physik 15 (Ser 5):145-66, 1932). Also Rudenberg received patents describing and EM in 1932, and Bruche and Johanson built the first electrostatic EM that could enlarge images of self-luminous objects in 1932.
The first commercial EM was developed by von Borries and Ruska and produced in Germany by Siemens and Halske in 1939, and the first commercial EM in the US was the RCA developed by Hillier, Vance, and some others.
This is probably more than you wanted to know, but "the" electron microscope was really a series of developments, and today's microscopes benefitted from additional contributions and refinements of others.
Hope this helps. Sara Miller
On Tue, 19 Dec 2000, Ekstrom, Harry wrote:
} Date: Tue, 19 Dec 2000 20:56:52 -0700 } From: Ekstrom, Harry {harry.ekstrom-at-honeywell.com} } To: "'\"Geinfam-at-aol.com\"-at-sparc5.microscopy.com'" {"Geinfam-at-aol.com"-at-sparc5.microscopy.com} , } Microscopy-at-sparc5.microscopy.com } Subject: RE: Electron microscope first demonstration } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } My source quotes M.Knoll first demonstrated the SEM in 1935 but no specific } date. } } Harry Ekstrom } Materials Laboratory } } (602) 231-2744 } e-mail: harry.ekstrom-at-honeywell.com } } } -----Original Message----- } } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com } [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] } Sent: Tuesday, December 19, 2000 4:36 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Electron microscope first demonstration } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Can anyone tell me the exact date in the "30's when the electron microscope } was first demonstrated? } } geinfam-at-aol.com } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
An excellent, brief history of EM development, written by Ernst Ruska himself (as the text of his Nobel Prize address) appears in Reviews of Modern Physics Vol. 59 No. 3 Part I p.p. 627-638 (1987). It includes some interesting photos, micrographs, and drawings
To answer the question, in this article Ruska states that he and Max Knoll first demonstrated magnified electron images of a specimen on 7 April 1931. ........................................................................... ...................................................... Jeffrey A. Fortner Argonne National Laboratory CMT/205 9700 S. Cass Avenue Argonne, IL 60439-4837
(630) 252-5594 (voice) (630) 252-4771 (fax)
} ---------- } From: Jim at ProSciTech } Reply To: jim-at-proscitech.com } Sent: Wednesday, December 20, 2000 7:53 AM } To: 'Ekstrom, Harry'; '"Geinfam-at-aol.com"-at-sparc5.microscopy.com'; } Microscopy-at-sparc5.microscopy.com } Subject: RE: Electron microscope first demonstration } } ------------------------------------------------------------------------ } 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 1935? That was a TEM, not an SEM. Knoll was Ernst Ruska's PhD } supervisor and } later collaborator. They designed and built the first TEMs. } The very first model had no specimen chamber and just proved that the lens } } system could work. So the first model, which I seem to remember was before } 1935 } was really not a microscope. } Von Ardenne described the operating principle of an SEM in the late 30th, } but I } believe none was build until the early 60th at Cambridge University, UK. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, December 20, 2000 1:57 PM, Ekstrom, Harry } [SMTP:harry.ekstrom-at-honeywell.com] wrote: } } } } } } My source quotes M.Knoll first demonstrated the SEM in 1935 but no } specific } } date. } } } } Harry Ekstrom } } Materials Laboratory } } } } (602) 231-2744 } } e-mail: harry.ekstrom-at-honeywell.com } } } } } } -----Original Message----- } } } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com } } [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] } } Sent: Tuesday, December 19, 2000 4:36 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Electron microscope first demonstration } } } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } Can anyone tell me the exact date in the "30's when the electron } microscope } } was first demonstrated? } } } } geinfam-at-aol.com } } } }
I could not find an exact date ether but Burton and Kohl "The Electron Microscope," Reinhold publishing corp., 1942 in a chapter on History of the Electron Microscope says the following (p149) " In 1932 both types of electron microscopes were described and the first images were shown. E. Bruche and H. Hohannson produced the first electron images of an oxide cathode with an aperture lens system utilizing 300-volt electron beams. M. Knoll and E. Ruska developed the first magnetic electron microscope utilizing 60,000-volt electron beams. The theory of electron lenses was largely developed by J. Picht, O. Shcherzer and W. Henneberg." In the Acknowledgments at the start of the book Dr. Kohl (one of the authors) is said to have given a demonstration of the oxide cathodes experiments: "Such an electron microscope was demonstrated before the seminar of the McLennan Laboratory in April, 1934; this probably was the first such demonstration in Canada." according to Burton. This puts the non-scanning types back a few years before the SEM date given below. Burton and Kohl don't provide any better dates than these, that I could find. Hope this might help you in your search. Jim.
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } "Ekstrom, Harry" {harry.ekstrom-at-honeywell.com} 12/19/00 07:56PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
My source quotes M.Knoll first demonstrated the SEM in 1935 but no specific date.
4. National Sales Equipment 800-545-0540 83 Eastman St, Easton, MA 02334
5. International Equipment Trading 708-913-0777 960 Woodlands Pkwy., Vernon Hills, IL 60061 You might try one of these (some addresses and phone numbers may be out of date), or ytou can probably find other companies that deal with used equipment on the Internet.
Good luckn and Happy Holidays
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Can anybody tell me how to prepare TEM samples for Single crystal? Thanks for your interested
Dear Erdem, It depends on the crystal and other parameters. Are you doing selected-area (SAED), convergent beam (CBED), or reflection (RHEED) diffraction? Is your specimen radiation resistant--such as a mineral--or labile--such as an organic? Will it stay solid in the vacuum, or will it need to be kept at LN2 temperature (as does anthracene)? What do you need to measure, intensities or lattice constants? What voltage will you use? The preparation can be very simple--e.g., MgO, which is made by burning Mg and collecting the smoke on a formvar-coated grid--or complicated--e.g., growing copper perchlorophthalocyanine epitaxially on freshly-cleaved alkali halide from a vapor or isolating a protein and finding crystallization conditions that produce well-ordered platy single crystals. Let me know some details, and I'll see if I can help. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Many thanks for the awful warnings, confessions and suggestions. We use two ordinary 9cm plastic funnels which have cracked and one 20cm funnel labelled KENUTUF, supplied with the mic to top up the Xray analysis dewar, which still seems as good as new after at least six years.
I love the Origami solution but to replace the little funnels I'll go with the stainless type for durability and add an insulated handle.
On the bung popping dewar front, the answer is not tin hats and a shout of 'incoming' but to provide a small vent. Even pressurised dewars need a relief valve.
Try Dr. Joseph L. Scott at The College of William and Mary (Williamsburg, VA)... jlscot-at-facstaff.wm.edu His research includes TEM of coralline algae.
Patty Clow
~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Patricia A. Clow, Ph.D. Staff Scientist I / Microscopy Genzyme Corporation One Mountain Road PO Box 9322 Framingham, MA 01701-9322 phone: 508-270-2053 fax: 508-872-9080 email: patricia.clow-at-genzyme.com
-----Original Message----- } From: Feng Wu [mailto:fwu-at-gbumail.bgu.ac.il] Sent: Wednesday, December 20, 2000 3:35 AM To: Microscopy-at-sparc5.microscopy.com
Can anybody tell me how to prepare TEM samples for biomaterials such as coral? Thanks. Feng ************************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
The current discussion of user fees appears to have strong, contrary opinions, so it seems worthwhile to take a look at what is posted at the US Department of Justice web site.
The following is taken "http://www.usdoj.gov/atr/foia/divisionmanual/ch2.htm".
Trusts, etc., in restraint of trade illegal; penalty
Every contract, combination in the form of trust or otherwise, or conspiracy, in restraint of trade or commerce among the several States, or with foreign nations, is declared to be illegal. Every person who shall make any contract or engage in any combination or conspiracy hereby declared to be illegal shall be deemed guilty of a felony, and, on conviction thereof, shall be punished by fine not exceeding $10,000,000 if a corporation, or, if any other person, $350,000, or by imprisonment not exceeding three years, or by both said punishments, in the discretion of the court.
§ 2 Sherman Act, 15 U.S.C. § 2
Monopolizing trade a felony; penalty
Every person who shall monopolize, or attempt to monopolize, or combine or conspire with any other person or persons, to monopolize any part of the trade or commerce among the several States, or with foreign nations, shall be deemed guilty of a felony, and, on conviction thereof, shall be punished by fine not exceeding $10,000,000 if a corporation, or, if any other person, $350,000, or by imprisonment not exceeding three years, or by both said punishments, in the discretion of the court.
(There are many more sections, but I believe these two summarize the main issues of the Act.) { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { {
Based on a common sense reading of the above, it seems to me that "restraint of trade" and the "attempt to monopolize" are the key factors that that Sherman Antitrust Act is designed to prevent, however, the issue of "intent" is not obvious.
I would think that a discussion by EM facility managers about how they charge for the use of their facilities and services, where the main emphasis seems to be how to cover the expenses of running the facilities, does not appear to be an intentional "restraint of trade" or an "attempt to monopolize" their trade.
I would agree that any suggestion of price fixing or how to undercut the rates of private companies would certainly violate the Sherman Antitrust Act, but I hardly think anyone can argue that any such comments have been made in the current discussion.
However, the question remains that if the well intentioned discussions between managers of nonprofit and/or university supported facilities on how to charge to cover their operating costs results in wide spread rates that actually do undercut private companies, is that illegal? I will refrain from presenting my opinion, since I believe that only someone with the appropriate legal background can give an authoritative answer to this question. And considering that every lawsuit has lawyers representing the opposing sides, the answer may only be clear if there is already a precedent on record where a similar case has actually gone through the courts.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Personnel salaries ARE part of the operating costs of running any type of laboratory facility. In addition, the cost of an electron microscope is also part of the operating expense of operating a lab. To suggest that only the non personnel costs such as service contracts, supplies etc. should be counted as operating costs really distorts the true picture.
I came from a university lab that originally was small and able to meet all of its operating expenses including salaries. Eventually visionaries saw the need for a larger more grand facility that would include some instruments with an annual service contract around 35K, other contract costs were in the range of 9K to 20K per year. The facility was built and the rates were kept low for academic users (about $25/hr.) and rates for non university users were scaled depending upon association with the facility. It didn't matter that it required about 1400 hours of beam time on one instrument just to pay the service contract because the facility received 8-10 million dollars per year in support from NSF. Principle instruments were obtained in grants and other subsidies not related to operating expenses. Eventually the NSF contract expired and now with aging equipment technology is passing them by.
The point of this is that nearly all university facilities will eventually have to be accountable for all of their expenses and adjust their rates to cover the costs. But universities still will have an advantage in that they usually don't have to purchase the principle equipment from operating expenses. How NSF granted instrumentation can be used is described in NSF Important Rule 91.
When university labs have to account for all of their operating expenses, they will become competitors of private non-subsidized labs. If they offer instruments rates less than private lab, they are cheating.
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
I get a rare opportunity to renovate some space to move our EM suite and have to turn in the ideal lab concept very soon. I know there is a book on this very thing. Anyone recall the title and author??
I am now in charge of a Phillips XL30 ESEM and have a couple of questions. What are you doing to prevent contamination of the specimen chamber. I had a user wanting to view human tissue today unfixed and am reluctant to do so. Anyone have a solution. Mine was to fix and process for standard SEM but that may not be an option for some of the in-situ studies or preserved museum specimens they don't want to wash or damage. Thanks
} Small stainless steel funnels are used to fill many LN2 } anticontamination devices on electron microscopes. I have found that a } Java jacket (designed for holding hot coffee cups) makes a great } reuseable jacket for picking up these funnels when they are cold.
I want to cheaply mount an inexpensive video camera onto the optical microscope of, yes, you guessed it, a JEOL 840. Economically, you might say.
Somebody been there, or near there?
I'm not very knowledgeable about optics, do I need to get
1 a lensless video camera and take off the microscope eyepiece,
2 a lensless video camera and retain the microcope eyepiece,
3 a video camera with a lens and take off the microscope eyepiece,
4 a video camera with a lens and retain the microscope eyepiece,
or
5 none of the above?
I'm sure I can devise the neccessary spacer/adaptor
tia
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
One volume is "Design of the Electron Microscope Laboratory", by Ronald Alderson. It is part of the Glauert series, "Practical Methods in Electron Microscopy". The edition we have is from 1975 and I don't know if there is a newer one.
Good luck, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Scott Whittaker [mailto:Whittaker.scott-at-nmnh.si.edu] Sent: Wednesday, December 20, 2000 3:08 PM To: Microscopy-at-sparc5.microscopy.com
I get a rare opportunity to renovate some space to move our EM suite and have to turn in the ideal lab concept very soon. I know there is a book on this very thing. Anyone recall the title and author??
I think the point we are missing here is geinfam-at-aol.com asked when the first EM microscope was first "demonstrated". My source states the theoretical "description" of the SEM was brought to us by H. Stintzing in 1929. However the first "demonstration" of an SEM was in 1935 by M. Knoll. In 1932, the first TEM was "constructed" by Knoll and Ruska. I wonder how they all felt about the electoral college.
----- Original Message ----- } From: Fortner, Jeffrey A. {fortner-at-cmt.anl.gov} To: 'Microscopy Listserver' {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, December 21, 2000 3:32 AM
I worked out the real, full-costs-recovery cost of our emlab a few years ago, and came to the conclusion that we would have to charge around £1250 a day to our users for access to a TEM. The charge for basic access to a top-grade analytical FEG TEM at a commercial rate could easily be in the order of £3000 per day. The task for most of us, as you indicate, is to keep the running costs down to a level which the University can tolerate. Most University facilities I know of in the UK are free of the requirement to earn the replacement cost of the equipment, and have to earn only limited staff costs. I am aware that there is ever-increasing pressure on Universities to earn more of the costs of their operations. The latest rumour is that electricity and water charges are just around the corner, but that one is at least a decade old now. I'll believe it when they install the meters. Maybe it'll come, maybe it won't. But in the past 15 years of operating a facility I swear that the user attitudes to charging don't reflect any recognition of this. Internal users still lobby for access to be free, or at least to appear free at the point of use. We therefore make just a few k per annum towards our costs by selling a little slack time to any local industries that have a need for our services. Who exactly are we cheating by doing this? An alternative way of looking at it is that we are offering the public some access to a facility they paid for in the first place, at a rate which is no more than is necessary to keep the machines running. I personally don't know of any commercial operation anywhere within hundreds of miles of Edinburgh that we are in competition with. Our operation is strictly self-limiting. once we have made the few k required to cover our costs the focus is back on internal user support. If there is any real industrial EM business out there any competent commercial operation would have no difficulty whatever in surviving whatever level of pseudo-commercial activity Universities are involved in. If there is any real evidence to the contrary I would like to hear it (and would be amazed) We're no threat, so please don't keep shooting us! Chris
} The point of this is that nearly all university facilities will eventually
} have to be accountable for all of their expenses and adjust their rates to } cover the costs. But universities still will have an advantage in that } they usually don't have to purchase the principle equipment from operating } expenses. How NSF granted instrumentation can be used is described in NSF } Important Rule 91. } } When university labs have to account for all of their operating expenses, } they will become competitors of private non-subsidized labs. If they offer } instruments rates less than private lab, they are cheating. } } John Humenansky/Staff Scientist } Physical Electronics, Inc. (PHI) } 6509 Flying Cloud Drive } Eden Prairie, MN 55344 } 952-828-6387 } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
Can anybody suggest how to measure thickness of the conductive coating for EM specimen? If it is transparent, ellipsometer could be used. Since metallic or carbon coating is not transparent, I could not use that method. Then do I have only way to measure directly, like measuring thickness on the cross-section or on the surface by using EM or SPM? Is there any other way?
I have the following problem during my TEM observation. I am looking at titania (TiO2), rutile, particles which are placed on top of a carbon coated cooper grid. Those particles are ca. 90 nm long and ca. 15 nm wide. At 200 kV the particles are very unstable under the electron beam. I think charging occurs. The attempt to coat the sample with carbon did show some success, but even after very long sputtering times (15 min. at 25 A) most particles twinkle.
If anybody has an idea for me how to avoid the twinkling, preferentially without sputtering any carbon on top of the sample, I would be very thankful.
Sincerely, Joon
************************************************************ Joon Hwan CHOI, Ph.D. Post-doctoral Researcher Materials Department University of California at Santa Barbara (UCSB) Santa Barbara, CA 93106
Ernst Ruska received in 1986 the Nobel Prize in Physics "for his fundamental work on electron optics and for the design of the first electon microscope".
You can read more on the Official Web Site of the Nobel Foundation: http://www.nobel.se/physics/laureates/1986/index.html
Cheers,
Paul ====================== Paul Baggethun Alcoa Technical Center Alcoa Center, PA 15069 USA ======================
-----Original Message----- } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, December 19, 2000 6:36 PM To: Microscopy-at-sparc5.microscopy.com
I think the twinkling problem is more electron beam heating of your sample as opposed to charging. If the samples were charging, they would be moving on the carbon support film. Carbon coating the samples will help. Are you using holey carbon grids or continuous support films. The latter should help somewhat, but you have about 200A of carbon that adds to the thickness. You could also try a cold stage to see if that could help. You could also try a higher voltage machine if one is available to you. Higher voltages allow the electrons to pass through your sample with less interaction and therefore less heating occurs.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Joon Hwan CHOI [mailto:jchoi-at-engineering.ucsb.edu] Sent: Wednesday, December 20, 2000 9:32 PM To: Microscopy-at-sparc5.microscopy.com Subject: TEM specimen preparation of titania particles
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
I have the following problem during my TEM observation. I am looking at titania (TiO2), rutile, particles which are placed on top of a carbon coated cooper grid. Those particles are ca. 90 nm long and ca. 15 nm wide. At 200 kV the particles are very unstable under the electron beam. I think charging occurs. The attempt to coat the sample with carbon did show some success, but even after very long sputtering times (15 min. at 25 A) most particles twinkle.
If anybody has an idea for me how to avoid the twinkling, preferentially without sputtering any carbon on top of the sample, I would be very thankful.
Sincerely, Joon
************************************************************ Joon Hwan CHOI, Ph.D. Post-doctoral Researcher Materials Department University of California at Santa Barbara (UCSB) Santa Barbara, CA 93106
About the firts TEM, the question is answered. But about the SEM, a few precisions.
Some interresting papers gives the essential informations about its history. But we must keep in mind that the early SEM was a "reflexion microscope" designed in analogy with the optical microscope. And the development were made in paralell with the TEM. So the design looks like a modified TEM, and not like what we know today as a SEM. It was also the case with the first Stereoscan.
The papers :
M. Von Ardenne Zur Geschichte der Rasterelektronenmikroskopie und der Elektronenmikrosonde. Optik 50 (1978) No 3, 177-188
C.W. Oatley The early history of the scanning electron microscope J. Appl. Phys. 53(2) Feb 1982
The first papers of Von Ardenne were from 1937 and 39 (Zeitschrift fur Physik), and he had built an instrument and published some photography. The intrument was destroyed by the bombardement at the end of WW II.
An other interresting paper is :
V.K. Zworykin, J. Hillier and R.L.Snder A Scanning Electron Microscope ASTM bulletin August 1942
After a lot of nice experiments (with FE guns!) they conclued that it was too expensive, with a poor signal and to less resolution.
J. Faerber IPCMS-GSI 23, rue de Loess 67037 Strasbourg CEDEX France
We fix human cells for safety reasons but sometimes look at them wet (hard work!). Chris Gilpin argues that fixation helps specimens resist beam damage and that most distortion occurs during drying rather than fixation.
Dave
On Wed, 20 Dec 2000 16:15:13 -0500 Scott Whittaker {Whittaker.scott-at-nmnh.si.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am now in charge of a Phillips XL30 ESEM and have a couple of questions. What are you doing to prevent contamination of the specimen chamber. I had a user wanting to view human tissue today unfixed and am reluctant to do so. Anyone have a solution. Mine was to fix and process for standard SEM but that may not be an option for some of the in-situ studies or preserved museum specimens they don't want to wash or damage. Thanks } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
At 4:08 PM -0500 12/20/00, Scott Whittaker wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
******************* Hi Scott, I just pulled the volume off my shelf. It is part of the series edited by Audrey Glauert: Practical Mehods in Electron Microscopy Volume 4 "Design of the Electron Microscope Laboratory" written by Ronald H. Alderson original copyright 1975. There may be a more recent edition, but this one certainly covers the main considerations.
I envy your opportunity to "start from scratch" many of us end up retrofitting spaces designed for other purposes.Have fun.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
By analogy, any combination (even among nonprofits) for the purposes of setting rates must be illegal if the well publicized divvying up of applicants to graduate school was an illegal restraint of trade.
Private for-profit industry was not involved in that particular conspiracy in any way.
On the other hand, the mere listing of categories of expense and the discussion of how best to allocate those costs between users and providers seem appropriate. That's essentially the same as a discussion of the principles of accounting or the geometry of the Ewald sphere.
When specific rates are mentioned, that's what looks like price fixing.
Best regards, George Langford, Sc.D. amenex-at-amenex.com
I disagree that posting rates implies core facilities are attempting price-fixing. Surely commercial entities look at the published costs of their competitors all the time. When one airline drops its rates, the others usually follow within 24 hrs. Price-fixing only results if you make your decisions in concert with your competitors. Knowing how much other core facilities charge has several benefits. First, the core knows what level of fees allow investigators to afford to use a technology. If I charged enough to fully reimburse the true expenses of my core, most of my researchers would simply switch to alternative techniques since they have limited grant dollars. Secondly, it allows core directors to demonstrate to administrators what portion of operating expenses other cores attempt to recover from user fees. I always tell my administrators that I am to make a profit that rivals our university library! Lots of cores say they try to recover operating expenses but unlike industry, we never account for building costs, electricity and heat, maintenance, etc. Researchers pay for those types of things with their indirect costs that accompany virtually all grants. This makes a comparison of commercial vs university fees impossible to judge. Indirect costs also help many universities pay for the "matching portion" that most federal grant agencies require as a part of matching equipment grants. Whether I use the flow cytometer on my campus or not, my indirect costs help pay for it. In addition, as a tax-paying citizen of the state of Missouri, my tax dollars help subsidize our University. We pay these taxes, in part, because we want our students to have access to the cutting edge technologies that will make them competitive citizens. What good would having all these fun toys be if they were too expensive to allow our undergrads and grads some time on them. These subsidies have no equivalent with private industry.
Anyone wanting to see what MU charges for confocal and other types of LM, should surf over to http://www.biotech.missouri.edu/mcc/prices.html . The costs for our EM core are at http://www.biotech.missouri.edu/mbp/cores/ . I am responsible for posting the LM core prices but Randy Tindall is responsible for the EM core prices being posted so please direct all lawsuits regarding them at him (Sorry, Randy). See you in court.
Merry Christmas and Happy Holidays to all. Tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear Scott I had the same opportunity to design for an EM lab when I moved to the University of Nebraska -- new facility, new space. Fortunately the lab spaces offered included one that I checked to be virtually ideal. This was in the basement of a building that had 3 floors above ground plus the basement and that was intended to be suitable for adding another 3 floors on top. The footings and the (1 foot thick reinforced concrete) wall surrounding the lab were really massive and confer the best stability I've heard of -- floor vibration velocities { 0.5micrometers per second (at frequencies 1 to } 50 Hz) and amplitudes { 0.5 micrometers (peak to peak) and AC magnetic fields {0.5 mG (before EM equipment installation). I got the architectural and construction team to provide several important provisions for use of the space for EM: 1. laminar air inlets to rooms (positioned so as not to affect the TEM columns!), 2. near-floor level return air vent in the TEM rooms (to satisfy needs to remove SF6 since it is denser than air), 3. acoustic lining of air inlet ducts, 4. temperature regulation to +/- 1 C with a maximum rate 1 C / hour, 5. humidity regulation in the range 40 to 60% relative humidity year round 6. water chillers in a mechanical facilities room adjacent to, but outside, the EM facility.
The above aspects overlap and go beyond the necessarily somewhat outdated recommendations in Anderson's book.
There were still issues that required care in order for the TEM to meet the image drift specifications I required of the manufacturers. These measures included adjustment of the air flow in the HREM room and reduction of the temperature difference between the TEM water temperature and the room air temperature. The TEM water chiller also required to be equipped with the precision electronic temperature regulation option. Under these conditions, there is no need to install curtains or otherwise to curtail air circulation during high resolution work.
I am very happy that our facilities design and management staff, construction and maintenance people, and microscope manufacturer's representatives could all be persuaded to achieve the desired end result -- continued, essentially trouble-free, stable operations of our lab in all conditions of weather and use of the rest of the building. It is definitely worth taking great care over the major and minor details before you start!
If I can provide other information. please let me know. Best wishes Brian Robertson
*********************************************************** Assoc. Prof. Brian W. Robertson Department of Mechanical Engineering and Center for Materials Research and Analysis University of Nebraska-Lincoln, N124 WSEC, 17th & Vine Sts., Lincoln, NE 68588-0656, USA ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 **
HI Ritchie, here is my cheap fix ( {1K$US) As you mention in 1-4 the eye piece is the trick. If you have std 23 mm eye pieces (For larger slip in eye pieces you can make a bushing) you can buy a relay lens from Edmund Scientific & others (23 mm x c-mount). Then buy a color security camera w/c-mount & connect to your video recording device. I use a Snappy TM interface to my laptop. I can throw all this in my laptop tote sack & aquire pretty decent images at almost any optical u-scope I walk up to. ball park on economics, optics $500, camera $200-300, Snappy {$100US For higher res. images there has been a lot of buzz on this list server about interfacing the Cool Pic TM digital camera. ck the archives. Don't forget to recalibrate you magnification.
If you have thread on eye pieces... I'd like to know what you find out.
disclaimer: no financial interest in companies or products mentioned.
Happy holidays, Bruce Brinson Rice U.
Ritchie Sims wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi again } } I want to cheaply mount an inexpensive video camera onto the optical } microscope of, yes, you guessed it, a JEOL 840. Economically, you } might say. } } Somebody been there, or near there? } } I'm not very knowledgeable about optics, do I need to get } } 1 a lensless video camera and take off the microscope eyepiece, } } 2 a lensless video camera and retain the microcope eyepiece, } } 3 a video camera with a lens and take off the microscope eyepiece, } } 4 a video camera with a lens and retain the microscope eyepiece, } } or } } 5 none of the above? } } I'm sure I can devise the neccessary spacer/adaptor } } tia } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Tell you what, folks. Next time we marauding academic pirates are in need of guidance on fees, let's just ask for links to web pages where other labs already have their rates published. Maybe that will make everybody happy and serve the same purpose as our usual blatant conspiracy does. That way we can take over the whole world legally!
Yeah, that's the ticket.....
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Has some a recept to clean the ceramic holder from a LaB6 filament. It is shorted (300 kohm at rohm temp., 0 at working temp.) with the filament. The tip is OK. Is it a way to remove the evaported coating (LaB6, decomposed LaB6 or carbon ?) on the ceramic, without dammaging the tip. Mechanically seems to bee inpracticable, it's too small (10 mm diameter, 5 mm heigth between the ceramic holder and the graphite blocs), and I have no "microbillage" (I don't know in english). But chemicaly ? I don't know the manufacturer of the tip, it comes from a LEED optic.
Thanks
J. Faerber IPCMS-GSI 23, rue de Loess 67037 Strasbourg CEDEX France
Your reply proves my point. If you were a business your personnel salaries and instrument costs would have to be factored into the ACTUAL operating costs. The instrument rates that you charge for outside clients are based on YOUR operating costs which do not include salaries or instruments and can therefore be much lower than what a business MUST charge to recover their costs and make a profit. Many universities are not in competition to private business because they are funded internally and exist primarily for education and research. Too often however the economics of operating a university facility collide with the reduction in institutional funding and then outside users are sought which is the unfair competition aspect unless the university rates for outside users are the same as what the businesses are charging for similar equipment.
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
} We pay these taxes, in part, because we want our } students to have access to the cutting edge technologies that will } make them competitive citizens. What good would having all these fun } toys be if they were too expensive to allow our undergrads and grads } some time on them.
I would also argue that students who never have a chance to see or use an electron microscope during their training are less likely to seek out those services later when they go to work in the private sector. I strongly suspect that the availability of EM in Universities is an important factor in creating a market for private sector EM providers.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
} } } One point that seems to have been missed is that the various } university labs do not compete with each other but rather need to recover } costs within their institution. This is a cost recovery and } redistribution process from within a single fiscal entity. (All grants } etc. are university funds and no longer belong to the granting agency or } the PI.) So I don't get this restraint of trade nonsense. We do not } sell anything within our institution. We provide a service that must be } financially supported in one way or another and it is most appropriate to } have the users of our facility provide all or part of those costs. So I } figure we can discuss anything we damn well please among professionals } who provide these services. } } At 10:01 AM 12/21/2000 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
I think all opinions have been aired and nothing new has been said for awhile. I'm going to suggest that the thread be left to ferment for another year when I'm sure the question will get asked once again.
It's a good bet that it will work. If it doesn't, try changing the distance between the microscope and the camera. Also, if you cannot fill the video chip completely with the image, or if it is over filled or distorted, try using the video lens with an eyepiece preceding it before you consider purchasing any additional optics. The alignment of the eyepiece and video lens from side to side, as well as the distance separation them are critical. A small deviation can lead to large changes in the image appearance and quality. There is a sweet spot, so patience prevails. Again, this is not the optimal way to interface a video camera and a microscope, but it is cost effective and performs well in many non-critical applications. If the video lens has an adjustable focus, it may be possible to use it without the eyepiece, and it is worth a try if all else fails.
Best of luck
John
John Turner, Ph.D. Director, Advanced Chemical Imaging Facility Cleveland State University Cleveland, Ohio 44115
I beleive the first high resolution SEM was built by Oliver Wells as a PhD project in 1959. It was enabled by the Everhart-Thornley secondary electron detector. My understanding is that the theory of how an SEM should work was developed in the '30s but specimen current and other available signals couldn't be amplified sufficiently or cleanly enough to be useable. Most of them still can't be used for high resolution work.
Ken Converse owner Quality Images third party SEM service Delta, PA
Jim at ProSciTech 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 } -----------------------------------------------------------------------. } } In 1935? That was a TEM, not an SEM. Knoll was Ernst Ruska's PhD supervisor and } later collaborator. They designed and built the first TEMs. } The very first model had no specimen chamber and just proved that the lens } system could work. So the first model, which I seem to remember was before 1935 } was really not a microscope. } Von Ardenne described the operating principle of an SEM in the late 30th, but I } believe none was build until the early 60th at Cambridge University, UK. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, December 20, 2000 1:57 PM, Ekstrom, Harry } [SMTP:harry.ekstrom-at-honeywell.com] wrote: } } } } } } My source quotes M.Knoll first demonstrated the SEM in 1935 but no specific } } date. } } } } Harry Ekstrom } } Materials Laboratory } } } } (602) 231-2744 } } e-mail: harry.ekstrom-at-honeywell.com } } } } } } -----Original Message----- } } } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com } } [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] } } Sent: Tuesday, December 19, 2000 4:36 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Electron microscope first demonstration } } } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } Can anyone tell me the exact date in the "30's when the electron microscope } } was first demonstrated? } } } } geinfam-at-aol.com } }
Is there somebody who know how to calculate fine structures near L23 edges by using ICXANES, while this code is restricted to spectra involving core levels of s symmetry, i.e. K, L1...?
} Are you going to fill us in on the answer, or do you even know it? } ... } Inquiring minds want to know.
I thought I had ... it is a mystery to me how some posts, for example this facility supervisor vacancy's announcement, get cross-posted everywhere, and why the subsequent discussion of this image missed this list :o)
http://epmalab.uoregon.edu/images/zircon-xmas.jpg
It is an image of e-beam induced CL as acquired via RGB filters. The blue is somewhat typical of igneous (or primary) quartz ... the red halo is evidence of radiation damage from a nearby zircon inclusion. Other images from the same rock actually show us the "smoking gun". We investigated a number of sandstones, and otherwize quartz bearing rocks from this environment, and another type of radiation induced CL we observed was red indistinct rims on qtz grains in sandstone. We imagined radioactive fluids percolating through the rocks.
My turn for True Confessions or surviving liquid nitrogen incidents.
Many years ago in another time warp (70's) a "green" technician (no, not environmental green) was dutifully following his supervisor's orders to fill EDX detector Dewars with liquid nitrogen. The staff had used a heavy-duty plastic Nalgene pitcher, wrapped with lab tape, to accomplish this chore. Thus, the technician followed protocol and used the same pitcher to collect the cryogenic fluid. And it worked well for many, many weeks. However, one day as he was filling the wrapped container, there was an enormous explosion (or implosion?). The technician was left holding the handle of the pitcher with the rest of the pitcher vanished into hundreds of small plastic pieces and liquid nitrogen splattered everywhere. Holy mackerel! But for the grace of God, not one piece of plastic or drop of liquid nitrogen hit the SURPRISED technician!
However, to this day that technician remembers his mother's admonitions about wearing clean underwear, you never know when an eighteen wheeler will hit you.
Bruce F. Ingber, Biologist USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 (504) 286-4270 phone (504) 286-4419 fax bingber-at-commserver.srrc.usda.gov
=================================================== Fernando D. Balducci Laboratorio de Microscopia Electrónica Facultad de Ingeniería - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
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Hi, all Thanks for your attention to my question on coral TEM specimen. The coral is hard, but not dense. A lot of platelets arranged in radiation way were observed in optical microscope. Can anybody give me some suggestion to prepare it? Thanks again. ************************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
fax 972-7-6472944 tel 972-7-6461473(o) -6281432(h) email: fwu-at-bgumail.bgu.ac.il ************************************************ ----- Original Message ----- } From: Feng Wu {fwu-at-bgumail.bgu.ac.il} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, December 20, 2000 10:34 AM
Dear Dr. Feng Wu,
I have used the following procedure to fix specimen of Leptogorgia virgulata, an octocoral (sea whip) found off the eastern coast of the USA. When fixed according to this procedure, whole tissue specimen, as well as the harvested sclerites by themselves, can be prepared for viewing in either SEM or TEM. I hope you find this information helpful.
Regards,
Elizabeth Bray Plant Chemist SCE&G Central Laboratory 2102 N. Lake Dr. Columbia, SC 29212 USA
Fixing and Embedding Protocol for Octocorals and Their Sclerites ================================================================
Reagent Preparation: 1. Purchase the following: 16% Paraformaldehyde (PFA) 50% Gluteraldehyde (GA) NaCacodylate, reagent grade [Sodium cacodylate (CH3)2AsO2Na · 3H2O, MW = 214.02] LR White acrylic embedding medium
2. 0.1 M NaCacodylate (CH3)2AsO2Na · 3H2O, MW = 214.02 21.4 g NaCacodylate diluted to 1000 mL with DW. Adjust the pH to 7.4 with 4% NaOH or 1:10 HCl.
3. Fixative: 2% PFA, 0.15% GA, 0.05 M NaCacodylate in filtered Sea Water: 6.3 mL of 16% PFA 0.15 mL of 50% GA 25.0 mL of 0.1 M NaCacodylate, pH = 7.4 18.55 mL of Millipore filtered Sea Water Total Volume of fixative = 50 mL
Procedure: 1. Fix the specimen in PFA/GA/NaCaco fixative for 1 hour. 2. Rinse in 0.05 M NaCaco Buffer 3 x 10 minutes. 3. Dehydrate in a series of EtOH (ethyl alcohol) washes of ever increasing strength: 50% EtOH 10 minutes. 75% EtOH 10 minutes. 95% EtOH 15 minutes. 100% EtOH 15 minutes x 2, done under vacuum.
4. Three (3) changes of LR White, all done under vacuum. The first two changes for at least 30 minutes, and the last change overnight. The last change should be done in the BEEM capsules in which the specimen will be cured.
NOTE: Be sure that when you fill the capsules with the LR White for the final time, there are no air bubbles in them. This will cause the level of embedding medium to become lower in the capsule as it sits overnight under vacuum, and when you cap them for curing in the oven, the air inside will lead to incomplete curing making it difficult, if not impossible, to properly section the hard coral material. To ensure that there are no bubbles present, allow the capsules to sit under vacuum for 30 minutes to 1 hour, go back and top off any capsules that need it, then reestablish the vacuum and allow them to sit overnight. Before capping, ensure that the level of embedding medium is at the top of the capsule. If it is not, carefully add sufficient medium to restore the proper level and proceed with capping.
5. Cap to ensure the exclusion of air and cure overnight in a 60o C oven.
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You may be referring to Vol. 4 of Practical Methods in Electron Microscopy, A. Glauert, ed., North Holland Pub., 1975. The book is a monograph by Ronald H Alderson on "Design of the Electron Microscope Laboratory". It has a lot of good information, but may be getting a bit dated for the ultimate level of environment for today's most sophisticated and sensitive instruments.
hope this helps...
Larry
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Hi, I am looking for a protocol for embedding plant tissues in LR White resin. I am working with endosperm and plant embryo of seeds of Ilex paraguariensis (maté). The endosperm contains a large amount of proteins and lipids. Thank's.
Dr. Rinaldo Pires dos Santos Dept. of Botany - UFRGS Brazil E-mail: rinaldop-at-uol.com.br Tel: + 55 51 4964053 (home) + 55 51 3167634 (UFRGS)
Thank you for your kind replies. The equipment that you mentioned seems too expensive for me to use, especially when I need just a couple of times thickness measurement for the calibration of my sputter coater. Is there other way to measure the thickness? Tobias suggested to use ellipsometry technique because the metallic or carbon coating transmits light when it is very thin. Have any you ever tried this before?
Best wishes,
Jondo Yun Kyungnam University, Korea jdyun-at-hanma.kyungnam.ac.kr
There are special machines called "thickness monitor" on the market. You have to go to your preferable EM supplier and ask them about that. It's very usual piece of equipment. It measures directly the thickness of the deposited material on the sensor. Sensor is mounted inside the vacuum chamber close to your sample. It is pretty sensitive (down to the 0.1 nm). The cost - couple of thousand backs. If you have further questions - I'll be happy to help you.
Have a good holiday season.
Sergey.
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
=============================================================== Jondo, Keep in mind that unless you deposit a really thick coating, carbon or metal coats do transmit light so ellipsometry could work. I have no personal experience with this, I just wanted to make the point. Good luck. TB
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I am currently involved with a cryo substitution project with human red blood cells. I have replaced the non crystalline ice with 2% Osmium in 100% acetone and have OK morphology, but the contrast is a bit low. I am thinking about trying to follow the Osmium step with uranyl acetate, before embedding in plastic. Does any one out there know if 1%uranly acetate will be OK in 100% acetone? Thanks, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
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Dear Timothy, We also worked with acetone and I have found that if one uses 4% OsO4 in acetone the contrast should be rather good.
Happy New Year!
Yours, Sasha
On Thu, 28 Dec 2000, Timothy Schneider wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Out There: } } I am currently involved with a cryo substitution project with human red } blood cells. I have replaced the non crystalline ice with 2% Osmium in 100% } acetone and have OK morphology, but the contrast is a bit low. I am } thinking about trying to follow the Osmium step with uranyl acetate, before } embedding in plastic. Does any one out there know if 1%uranly acetate will } be OK in 100% acetone? Thanks, Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu } } }
Can you deposit your coating onto a sample of polymerized TEM embedding media, then cut the sample at 90 degrees on a microtome, then measure the thickness using a TEM?
Good luck,
Doug
On Thu, 28 Dec 2000 15:38:21 +0900 =?ks_c_5601-1987?B?Sm9uZG8gWXVuIMCxwbi1tQ==?= {jdyun-at-kyungnam.ac.kr} wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- Send Email } to ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Sergey, Tobias and other listers: } } Thank you for your kind replies. } The equipment that you mentioned seems too expensive for } me to use, especially when I need just a couple of times } thickness measurement for the calibration of my sputter } coater. Is there other way to measure the thickness? } Tobias suggested to use ellipsometry technique because the } metallic or carbon coating transmits light when it is very } thin. Have any you ever tried this before? } } Best wishes, } } Jondo Yun } Kyungnam University, Korea } jdyun-at-hanma.kyungnam.ac.kr } } ==================================================================== } Hello Jondo } } It looks like nobody answer your question. } } There are special machines called "thickness monitor" on } the market. You have to go to your preferable EM supplier } and ask them about that. It's very usual piece of } equipment. It measures directly the thickness of the } deposited material on the sensor. Sensor is mounted inside } the vacuum chamber close to your sample. It is pretty } sensitive (down to the 0.1 nm). The cost - couple of } thousand backs. If you have further questions - I'll be } happy to help you. } } Have a good holiday season. } } Sergey. } } Sergey Ryazantsev, Ph.D. } Electron Microscopy } Department of Biological Chemistry, School of Medicine } University of California, Los Angeles } Box 951737 } Los Angeles, CA 90095-1737 } } (310) 825-1144 (office) } Pager: (310) 845-0248 } FAX: (310) 206-5272 (departmental) } mailto:sryazant-at-ucla.edu } } =============================================================== } Jondo, } Keep in mind that unless you deposit a really thick } coating, carbon or metal coats do transmit light so } ellipsometry could work. I have no personal experience with } this, I just wanted to make the point. Good luck. TB } } } } Jondo,?------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email } to ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } Dear Listers: } } } Can anybody suggest how to measure thickness of the } conductive } coating for EM specimen? If it is transparent, } ellipsometer could be } used. Since metallic or carbon } coating is not transparent, I could } not use that method. } Then do I have only way to measure directly, } like } measuring thickness on the cross-section or on the surface } by } using EM or SPM? Is there any other way? } } } Jondo Yun } } jdyun-at-kyungnam.ac.kr } } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 } Tucker Hall / / / / \ \ \ } Biological Sciences /_ / __ /__ \ \ \__ } University of Missouri / / / \ \ } \ Columbia, MO USA / / / } \ \ \ 65211-7400 / / ___ / \ } \__/ \ ____ voice: 573-882-0173 fax: } 573-882-0123 } } }
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
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