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Hi I dont know if this technique is the best but it certainly works for us. Depending on what part of the beetle you want to examine, we stick and glue with silver dag, an entomological pin (for the small beetles) or a normal pin (for larger specimens)to an area of the specimen which is not going to be observed. From here we place the specimen and pin in a stub like vice. This allows you to gold coat the beetle and place in the SEM chamber with minimal handling. Ok yes the pin can be seen under the SEM. The idea is the mount the specimen in such away that the pin will not obscure in anyway the views that are wanted. ie if dorsal and front view are wanted then the pin would be placed in the ventral side at an angle less than 90 degrees sloping backwards towards the posterior end. This will give a greater angle to work with when observing the frontal position.
I know this is a brief explanation, however if you want to ask any questions please ring me on 02 9320 6198
Hope this helps
Sue Lindsay
SEM Lab Australian Museum
I would like to know if there is any new technique about SEM and beetles, what is the best way to mount the beetle.
Keep care and be of good cheer.
Regards
Vratislav Richard Eugene Maria John Baptiste of Bejsak (Bayshark)-Collorado-Mansfeld
Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
Temporally home address: 32 Girrawheen Ave. Kiama NSW 2533 Australia e-mail: 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.
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BEGIN:VCARD VERSION:2.1 N:Bejsak-Collorado-Mansfeld;Vratislav Richard;Eugene Maria John Baptist FN:Vratislav Richard Eugene Maria John Baptist Bejsak-Collorado-Mansfeld ORG:Bayshark Research Laboratory TITLE:director NOTE;ENCODING=QUOTED-PRINTABLE:Marketing and Coaching=0D=0A=0D=0ATenebrionidae Orbis and higher taxonomy TEL;WORK;VOICE:(+61 2) 9319 6380 TEL;CELL;VOICE:(+61 414) 540 465 ADR;WORK:;;29 Edward Street;Darlington, SYDNEY;;NSW 2008;Australie LABEL;WORK;ENCODING=QUOTED-PRINTABLE:29 Edward Street=0D=0ADarlington, SYDNEY NSW 2008=0D=0AAustralie ADR;HOME;ENCODING=QUOTED-PRINTABLE:;;(temporaly address):=0D=0A32 Girrawheen Ave;KIAMA;;NSW 2533;AUSTRALIA LABEL;HOME;ENCODING=QUOTED-PRINTABLE:(temporaly address):=0D=0A32 Girrawheen Ave=0D=0AKIAMA NSW 2533=0D=0AAUSTRAL= IA URL: URL:http://www.coleoptera.org EMAIL;INTERNET:ricardo-at-login.cz EMAIL;PREF;INTERNET:vratislav-at-bigfoot.com EMAIL;INTERNET:ricardo-at-ans.com.au REV:19981231T063007Z END:VCARD --IMA.Boundary.638414519--
} Hi } I dont know if this technique is the best but it certainly works for us. } Depending on what part of the beetle you want to examine, we stick and glue } with silver dag, an entomological pin (for the small beetles) or a normal } pin (for larger specimens)to an area of the specimen which is not going to } be observed. From here we place the specimen and pin in a stub like vice. } This allows you to gold coat the beetle and place in the SEM chamber with } minimal handling. } Ok yes the pin can be seen under the SEM.
Paint the pin with dag before mounting and if you get your contrast right the pin dissapear into the bagground!!!
Just a usefull hint. Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
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I highly reccomend MOC (Microscopical Optical Consulting) to service your microtomes. This is a small, independant company in Cottage Valley, NY. They do all of our's every year - Reichart, Leica, Sorvall, etc. Their # is 914-268-6450. Good luck! Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
A Research Fellow is required in the Department of Materials, University of Leeds, for a fixed period of two years, to work on an electron microscope study of the mechanism of graphitisation in steels. Applicants should have, or be near completing, a PhD in Metallurgy, Materials Science or a related discipline. Experience in transmission electron microscopy would be advantageous.
Salary: Research IA within the range stlg15,735- stlg19,197 p.a. according to qualifications and relevant experience.
Application forms and further particulars may be obtained from Professor D V Edmonds, Department of Materials, University of Leeds, Leeds, LS2 9JT, UK. tel: 0113 233 2348, fax: 0113 233 2384.
Job ref: 062-066-004-027.
Closing date: 29 January 1999.
Towards Equal Opportunities
Professor David V Edmonds Department of Materials School of Process, Environmental and Materials Engineering University of Leeds Leeds LS2 9JT United Kingdom
-- Jane M. Woodruff Polysciences Inc Phone: 215-343-6484 400 Valley Rd. 800-523-2575 Warrington, PA 18976 Fax:215-343-0214 E-mail jwoodruf-at-polysciences.com
Project MICRO is MSA's middle school educational outreach program. In his capacity as MSA webmaster Nestor gave MICRO a wonderful Christmas present; a major expansion of its web page. The new URL is http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html There is a LOT of microscopy education info there, including the quotations that listserver readers contributed last year. Please visit the site, and send me your comments.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I am a vendor, and we developed and sell "Entomounts", which are basically specimen mounts with the entomology pins already in them. They are provided as a convenience, and are so reasonably priced that I don't care much one way or the other whether we sell a bunch of them or not {grin} .
Happy New Year! Steven Slap
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
I have a position open for an EM TECHNICIAN, SENIOR, open. The job entails negative staining and thin sectioning of clinical specimens for viral examination. The salary is $26K plus good health benefits. Send resume to me below, or email for questions.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We have a researcher who is interested in imaging some molecules (proteins, primarily) in an aqeuous medium in order to determine if any conformational changes are occurring. Originally, this work was being done using x-ray microscopy; however, since there are only a very few such microscopes in the world, I suggested cryo TEM.
We are now looking for central facilities where high resolution cryo-TEM work might be done.
Thank you.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I fear that my copy of Microscopy News (I think that is correct) which had a procedure for the preparation of snowflakes my have hit the recycling bin without my knowledge or consent.
Can anyone forward a copy of the procedure to me? I would greatly appreciate it.
Thanks.
Alan Stone ASTON Metallurgical Services as-at-mcs.com
Maria Lucia Ribeiro Caldas wrote: ============================================= Does anyone have a protocol for the resin SPI-PON 812 (Sigma). ============================================== This product originally came from our firm, SPI Supplies. It is our SPI-Pon 812 epoxy resin, it is one of the components of our "Epon replacement" kit, and is described on our website given below.
We have not yet put up on our website the "Use Instructions" for this product, but if you send me your FAX number, I will make sure that a complete set is FAXed off to you right away. SPI-Pon 812 resin is "plug in" compatible with all known protocols based on the original Epon 812 protocols , so you could continue using whatever protocol(s) you have found successful in the past.
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: www.2spi.com ############################ ==================================================
------------------------------------------------------------------- FOR THOSE PLANNING TO SUBMIT A PAPER TO THE MICROSCOPY AND MICROANALYSIS MEETING.
This year information about the authors of Papers submitted to the M&M '99 meeting in Portland, OR should be submitted electronically. Unfortunately, the URL for submission was inadvertently left out of the "Call for Papers" booklet. The URL is:
The abstract itself still should be mailed to Microscopy & Microanalysis "99 Meeting Management (as indicated in the Call for Papers).
Additional information about the meeting and a link to the data entry page are available at:
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If you need a copy of the "Call for Papers" contact M&M '99 meeting management toll free at 877-672-6271 or toll call at 708-361-6045. Fax number: 708-361-6166.
Hello everybody, I would like to get some information on TEM diffraction pattern analysis. Specifically;
1) What software is available for analysis of diffraction patterns (both ring patterns and spot patterns)? What kind of accuracy can be obtained - are we getting close to the accuracy of X-ray diffractometry yet, or are there fundamental reasons such as lens aberrations, smaller Bragg angles, and accuracy of measurement which mean that we'll never get there?
2) What are the typical procedures people use for, say, measuring camera length or identifying unknown phases using diffraction?
I will make a summary of replies and distribute it to anyone who is interested.
Many thanks in advance, and a Happy New Year to all,
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK e-mail richard.beanland-at-gecm.com
Make a 1% solution of Formvar in methylene chloride (chloroform will work too). Chill the solution and some glass slides (leave them outside with a cover to keep the snow off). Dip a toothpick in the Formvar solution and place drops on a slide, then catch snowflakes on a piece of black velvet or something similar. With a toothpick wetted with the solution, touch a snowflake ever so slightly and it will cling so you can transfer it to a drop on the slide. Cover the slide and let the solvent evaporate (this happens very quickly). Take the slide inside and the snowflake will melt, leaving the Formvar replica. (To preserve the most delicate structure, leave the slide outside longer to let the water sublimate).
This is a great treatment for cabin fever. Happy New Year.
I've been using a variation of the "ento pin" for years that provides a little more flexibility than a rigid pin. Cut a long, thin triangular piece of thick aluminum foil (like that in a weighing dish - in a pinch you can use aluminum or copper tape), bend the base at a 90 degree angle, and stick it to the stub with carbon paint or your favorite conductive adhesive. Mount the insect on the point with conductive adhesive and coat. After coating, re-paint the stub surface and pin with carbon paint to darken the background. The mount is flexible enough to make fine adjustments to the positioning
of the insect (to get an exactly lateral view, or to hide the pin or whatever) and can be bent 90 degrees in any direction to get dorsal, ventral or other views. This works a lot better than trying to tilt the stage, as in most scopes you lose the ability to move in the X or Y direction at high tilts. Plus the background remains darker if you don't have to tilt.
Hope this helps
JME
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Lucy, I'm not familiar with that specific mounting media but if it is for permanent slides (similar to Cargill melt mounts), then place the container (if glass) on a hot plate and slowly heat it up until the crystals go back into solution. You may need to repeat this if recrystallization occurs between uses. Hope this is helpful. Mike Bucker Consolidated Labs of Va Feed Microscopy Unit
} } } maria lucia ribeiro caldas {caldasml-at-amcham.com.br} 01/04 6:29 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all
I have an old 100g bottle of Ultramount II (Ladd) which is completely crystallized. I would like to know if there any solution for this.
maria lucia ribeiro caldas wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Dear all } } I have an old } 100g bottle of } Ultramount II } (Ladd) which is } completely } crystallized. I } would like to } know if there } any solution } for this. } } Thanks } } Lucy
Dear Lucy,
It would depend. If there is some solvent left ther might be a chance to save it. If you wish to contact me directly and give me a liittle more detail I can advise you further. On your other post, SPI-812 is a replacement for the old Epon 812 as is our LX-112 and similar products sold be other supply houses. SPI-812 I believe probably is a trademark product from SPI, but I could supply you with the protocols from our product if you wish.
Thanks,
Dr. Charles Duvic Chemist -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
Is anyone using or has anyone used Improvision's OpenLab software? We have had it in for a demo and are curious as to how it performs in "real life", how is the tech support, etc. Any comments (positive or negative) will be appreciated.
I have acquired 2 sets of human blood smears - one stained with Wright's stain and one with cresyl violet to show reticulocytes. I would like to incorporate them into my histology class student slide sets. I have coverslipped the smears but there is still a little stained area outside some of the coverslipped area. Are there any safety issues with stained smears or can one assume that any potential viral material would be killed by the staining step. I am unfamilair with the exact staining procedure but thought that there is ethanol in the stain. any comments from knowledge histotechs. Thanks in advance. Tom Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Date sent: Tue, 5 Jan 1999 09:55:29 -0500 (EST) } From: Tamara Howard {howard-at-cshl.org} To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} We have been using openlab here for about 1 year now. It is being used for both time lapse imaging and low light GFP imaging. We are happy with the software. We have had no real problems. The only problems that we did run into were generally users (my self included) getting a little confused. The whole package is very complex and complete, depending on which modules you have and does require a little time to learn. I did find the automator, a module for automating repetitive or long protracted tasks, could become slightly cluttered, again more dependant on how the user was laying out the tasks. We did run into a couple of problems, or tasks that made us scratch our heads. When we phoned their UK tech support line, there was always someone able to help and they resolved all the problems that we had. When the line was busy they always phoned back. These remarks all relate to the earlier version of open lab, Open lab 2 only arrived with us just before christmas so I have not had long to play with it, but so far it appears to be as good as before. They also appear to be setting up internet user groups and an improved web site.
If you need any more info please give me a call.
Peter
Peter McHardy Technology Services Manager, Beatson Institute, Glasgow University, Garscube Estate, Bearsden, Glasgow G61 1BD Tel 0141 330 4818 Fax 0141 330 4127 http://www.beatson.gla.ac.uk/pmh
I am trying to embed L. dispar spermathecae for TEM, however I'm having = a terrible time dissecting / fixing the organs without losing the sperm = contained within them. Does anyone have any suggestions? ( I have = tried fixing the organs before dissecting them from the insect - this has not worked well )
Thanks,
Laura K. Garvey University of Connecticut Dept. of Molecular and Cell Biology=20 U-131, Beach Hall Storrs, CT 06269=20
DEPARTMENTS OF PHYSICS & ASTRONOMY AND CHEMISTRY=20
POST-DOCTORAL RESEARCH ASSISTANT =20
RA1A =A315,537 - =A323,651=20
A post-doctoral position is available for up to 24 months to work on an EPSRC funded project, "The use of XANES and ELNES for the characterisation of stabilised zirconia". The project is a collaboration between Glasgow University, The Queen's University, Belfast, MEL Chemicals Ltd and Johnson Matthey Ltd. The part of the project associated with this post involves modelling the near edge fine structure present on the edges observed in x-ray absorption spectroscopy and electron energy loss spectroscopy. Experience with first principles band structure calculations is essential and a background in the theoretical interpretation of spectroscopic techniques such as ELNES and XANES would be highly desirable, as would a knowledge of many-body physics. The post will be based in Glasgow but will involve extended periods at The Queens University working with Professor Finnis and the Atomic Simulations Group. =20
Further information is available at http://www.ssp.gla.ac.uk/ or from Professor Alan Craven, Department of Physics and Astronomy, University of Glasgow, Glasgow G12 8QQ. (Tel 0141 330 5892, FAX 0141 330 4464, a.craven-at-physics.gla.ac.uk) to whom applications, including a CV and the names of two referees, should be sent. Closing date - 12 January 1999.
------------------------------------------------------------------------ Dr Dave McComb Lecturer in Materials Chemistry Department of Chemistry University of Glasgow Glasgow G12 8QQ UK
} } Hello everybody, } I would like to get some information on TEM diffraction pattern } analysis. Specifically; } } 1) What software is available for analysis of diffraction patterns (both ring } patterns and spot patterns)?
One package is Desktop Microscopist. I have some FORTRAN code (written for a UNIX platform) which is helpful for indexing spot patterns to a known structure (If you are interested, you may contact me). I am sure there are others, too. A place to look would be the Sincris site at
http://www.lmcp.jussieu.fr/sincris/logiciel/
} What kind of accuracy can be obtained - are we } getting close to the accuracy of X-ray diffractometry yet, or are there } fundamental reasons such as lens aberrations, smaller Bragg angles, and } accuracy of measurement which mean that we'll never get there?
Because of lens hysteresis, it's not possible on a standard TEM to calibrate the camera constant L*(lambda) to an accuracy of greater than about 2 percent. A way to get more accuracy is to use an internal standard. For a good discussion of the problems, see Hirsch, Howie, Nicholson, Pashley and Whelan, Electron Microscopy of Thin Crystals, end of chapter 1.
In spite of the camera length inaccuracy, the RELATIVE spacings for two spots isn't affected. Therefore, it is potentially limited only by measurement inaccuracies (how precisely can we locate the center of the spot), and by the relrods (see Hirsch et al).
I believe that a spherical-type distortion of the pattern occurs if you use the beam convergence (condenser lens) to compensate for poor focus of the diffraction pattern. I've never read a good discussion of this (anyone know of one?) Also, ring patterns can be distorted from circular by astigmatism effects (in any post-specimen lens). These optical factors would also limit the accuracy of relative d-spacings, though to some extent one may be able to correct for them.
One place where electrons have a significant advantage over x-rays is with respect to noise. The interaction of electrons with matter is strong, so in very short experimental times good statistics can be obtained from miniscule sample volumse. Electron diffraction therefore has potential for structure refinements (e.g. recent work by Jansen and Zandbergen, L. D. Marks et al for surface diffraction). Also, the photographic film or CCD is a 2D detector (rare in XRD), so you can win big time in reducing noise. This can be used in studies of amorphous materials, via circumferential averaging of the patterns. However, strictly speaking, both dynamical and inelastic effects have to be considered in quantitatively interpreting the data.
} } 2) What are the typical procedures people use for, say, measuring camera length } or identifying unknown phases using diffraction?
Again, you can measure camera length as accurately as you want. Turn the scope off and on and it may differ by a couple of percent.
} } I will make a summary of replies and distribute it to anyone who is interested.
Thanks, I would be interested in hearing.
Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
mailto:wharton.sinkler-at-anlw.anl.gov
} } Many thanks in advance, and a Happy New Year to all, } } Richard Beanland } GMMT Ltd., } Caswell, } Towcester, } Northants NN12 8EQ } UK } e-mail richard.beanland-at-gecm.com } } }
Workshop on Quantitative Image Analysis May 20-22 and May 24-26, 1999, North Carolina State University, Raleigh, North Carolina, USA June 14-16, 1999, Danish Technological Institute, Taastrup, Denmark
This highly regarded hands-on course taught by Dr. John Russ and other expert faculty has been presented annually for more than 15 years. It deals with all phases of quantitative and computer-assisted imaging from acquisition and processing through measurement and stereological interpretation. Attendees receive The Image Processing Handbook plus a CD-ROM containing images, algorithms (Photoshop-compatible for Mac and Windows) and an extensive tutorial. The course is appropriate for professionals scientists, technicians and administrators using these techniques for research. Attendees typically come from materials science, geology, biological and medical sciences, pharmaceuticals, food science, industrial quality control, remote sensing, and other disciplines.
For detailed information and registration contact Cindy Allen, Dept. of Continuing and Professional Education, N. C. State University, Raleigh, NC 27695-7401, 919-515-8171, fax 919-515-7614, email: cindy_allen-at-ncsu.edu
On-line information is available at http://members.aol.com/IPCourse/
Dear listservers,=20 This topic was discussed awhile ago. Hopefully, some new developments = are now here to solve the problem easily. We are looking for a dedicated = software package for scheduling, in this case, instrument use = incorporated into our webpage. We have several microscopes, = workstations, microtomes, etc. and would like to have calenders or = something similar for each, so users could sign up in advance using the = web. It should be possible to assign different levels of privilege to = each user. Once an entry is made it could only be changed by an = administrator other than the original user. We like would like to then = to automatically transfer this information into a billing database, = either Access or FileMaker Pro based. Is something like what I have = described commercially available?
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
I don't have the URL immediately at hand, but you might try the Filemaker website. They have additional templates besides those shipping with Filemaker. there are also links to Filemaker consultants who have free templates or who may be able to advise you regarding feasibility.
Regards, Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Wed, 6 Jan 1999, hank p adams 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 listservers, } This topic was discussed awhile ago. Hopefully, some new developments are now here to solve the problem easily. We are looking for a dedicated software package for scheduling, in this case, instrument use incorporated into our webpage. We have several microscopes, workstations, microtomes, etc. and would like to have calenders or something similar for each, so users could sign up in advance using the web. It should be possible to assign different levels of privilege to each user. Once an entry is made it could only be changed by an administrator other than the original user. We like would like to then to automatically transfer this information into a billing database, either Access or FileMaker Pro based. Is something like what I have described commercially available? } } Hank Adams } Cell Biology } Integrated Microscopy Core } Baylor College of Medicine } One Baylor Plaza } Houston, Tx 77030 } } }
} I would like to get some information on TEM diffraction } pattern } analysis. Specifically; } } 1) What software is available for analysis of diffraction patterns (both ring } patterns and spot patterns)?
There is an operation in the SPIDER image processing program which will refine a lattice and determine the background-subtracted intensities. I have written a procedure which subtracts the circu- larly symmetric background, which improves the subsequent linear background subtraction. I have also written a routine (both in SPIDER and as a stand-alone) to determine the center and radius of a ring from up to 20 points. I can send you the source code for the stand-alone version.
} What kind of accuracy can be obtained - are we } getting close to the accuracy of X-ray diffractometry yet, or are there } fundamental reasons such as lens aberrations, smaller Bragg angles, and } accuracy of measurement which mean that we'll never get there? } When I selected 20 points from each of 13 rings on a gold pattern (5 from each quadrant), I got r/d* = 402.79+-0.49 pixels. The range of values was 401.89 to 403.71. The precision was close to 0.1%. Furthermore, there seemed to be no pattern of systematic variation, except that the larger rings, which were less intense and more diffuse, gave somewhat larger errors.
} 2) What are the typical procedures people use for, say, measuring camera } length } The best procedure, if it can be done, is to evaporate gold, or another standard, onto the crystal whose lattice constants are to be measured. This way one gets the lattice points and the standard on the same negative. I scan my negatives on a Perkin-Elmer micro- densitometer using a 10 mu x 10 mu window. For lattice constant measurement, I do not interpolate the file, but for intensity measure- ments, I reduce by a factor of 5 by averaging a 5 x 5 array for each pixel in the reduced image. This reduction does not produce errors in the background-subtracted intensities.
} I will make a summary of replies and distribute it to anyone who is } interested. } Thanx. I am on both listservers, so only those responses to you directly, rather than to a list, would be required.
} Many thanks in advance, and a Happy New Year to all, } And to you. Yours, Bill Tivol
We've had abit of a clean up here at RMIT and have some goodies to give away. We have: Two boxes Tungsten filaments, Item no. A050 purchased from AGAR these were used in an old ETEC. One H.V. supply for the ETEC, I've been told that when the ETEC was decommissioned it still worked!!!
Yes! they are old items, but perhaps someone has a creative use for them. If you want them you can contact me on the details below. Email has the best chance of getting me.
Richard, One major source of inaccuracy in the measurement of electron diffraction patterns lies in the rather large variation of camera length with the specimen height. If you use a double-tilt stage, the specimen height changes with tilt (only one tilt axis is eucentric). It is possible to calibrate the camera length against objective current when the specimen is in focus, but, as pointed out by Wharton Sinkler, switching the microscope off and on will undo your work. The height change with tilt can be obviated by using a rotation-tilt stage instead, but it can drive you nuts, especially if the crystal is not at the centre of the grid.
As was suggested, the best method is to use an internal standard as this compensates for both camera-length changes and projector astigmatism. It also enables the operator to estimate d-spacings on screen in order to decide whether the crystal is in a sensible orientation, and one can very quickly tell whether a pattern is rectangular or oblique.
I hope this helps, Eric ---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
} } Hi all. } } We are currently setting up a cryo-vitrification unit for TEM sample } analysis. Research into what is the best } cryogen to use for vitrification of our sample type gave the answer of } liquid ethane. } } Does anybody have any advice as to what materials to use or to NOT use } with liquid ethane (for material } incompatibility reasons - chemically and physically (i.e. extreme } temperatures))? Is copper okay with } liquid ethane? I've vaguely read somewhere that some groups use a copper } coil (in liquid nitrogen) to condense } their ethane. Any other confirmations? } } A small flexible length of tubing will also be needed to join the copper } (?) tubing to the cylinder regulator } (so we can move the coil in and out of the liquid N2 easily). Apparently } natural rubber is NOT good with ethane. } } Any other pearls of wisdom out there? } } Thanks in advance. I'll summarise my replies. } } Terri } } ------------------------------------- } Ms Terri Soar, PhD student, } University of South Australia } Email: terri-at-drage.com.au {mailto:Terri-at-drage.com.au} } ------------------------------------- }
Electron microscopy specialist needed immediately. Prepare ultra thin sections and photograph them using JEOL1200EX. Darkroom, Adobe Illustrator/Photoshop. Salary commensurate with experience.
Mail, e-mail, or FAX resume and two letters of recommendation to:
Dr. Peter Sterling 123 Anatomy-Chemistry BLDG Department of Neuroscience University of Pennsylvania Philadelphia, PA 19104-6058
I have used Formvar for over 50 years now and never known exactly what it is. In the back of my mind I seem to recall hearing that it is polyvinylformal, but I'm not certain that is correct. I have just consulted two polymer scientists in our department, and neither one of them knows what it is, nor could they find a chemical formula for it in their reference books.
If anyone knows what it is, I'd like to know too.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Philippe-Andr=E9 Buffat wrote: } =20 } I would be strongly interested in softwares able to handle digital } diffraction patterns. In particular a soft to flatten the background is } needed. From my experience, I can see on negatives faint rings or spots= , } but under the same condition it is impossible to see them on a computer } screen without increasing the contrast but then the range between black= and } white covers only a small part of densities in the whole pattern and th= e } observable area between the white center and the black periphery become= s } unacceptably small. The use of color coding of the intensities improves } only a little bit the visible range. An other soft to determine } automatically the spot position or the ring position by a refinement me= thod } or deconvolution of gaussian/lorentzian (or else) curves would also be } welcome.
Dear Philippe, In my old paper (Ultramicroscopy (1982) 9:117-130) I discuss=20 a technique of subtracting circularly-symmetric background, and give=20 a reference to an even older paper by Fraser et al. (Appl. Cryst.=20 (1977) 10:64- ). I have since written a procedure using the opera- tions in the image-processing program SPIDER (I'm sure the operations are also part of other programs), and I'd be happy to send you a copy. Briefly, the procedure refines a lattice, masks out the spots, replaces them with a local average background, makes a 2-D rotational average image, and subtracts this from the original pattern. The remaining=20 background is linear enough so that the usual form of background sub- traction will work. This process cannot be used for ring patterns, although a modification could work. Not only does this circularly- symmetric subtraction improve the visibility on a screen, it also increases the accuracy with which the intensities can be measured. As one can surmise, SPIDER has an operation to determine intensities and refine a lattice. It can also find the center and radius of a ring, and I have a stand-alone version of this. This program works by one choosing from 3 to 20 points on the ring, using the first 3 to make an initial guess, then least-squares-fitting the points (if there are more than 3). I'd be happy to send you the=20 FORTRAN code for the stand-alone ring program. Yours, Bill Tivol
Wharton, Nice summary, I would like to add a practical hint to those who may be new to ED. You can use an extrinsic standard such as a gold film. The improtant point is maintaining the same conditions for the standard and sample. To accomplish this the lens currents should be duplicated. An easy way to accomplish this is to focus the sample with the Z position while maintaining the lens currents from the standard. When using extrinsic standards the potential for a Z position mismatch and subsequent change in the objective lens current is the most likely error in camera length. Russ
-----Original Message----- } From: Wharton Sinkler [mailto:wharton.sinkler-at-anlw.anl.gov] Sent: Tuesday, January 05, 1999 2:07 PM To: Richard Beanland +44 1327 356363; Microscopy Listserver; lemas Listserver
} } Hello everybody, } I would like to get some information on TEM diffraction pattern } analysis. Specifically; } } 1) What software is available for analysis of diffraction patterns (both ring } patterns and spot patterns)?
One package is Desktop Microscopist. I have some FORTRAN code (written for a UNIX platform) which is helpful for indexing spot patterns to a known structure (If you are interested, you may contact me). I am sure there are others, too. A place to look would be the Sincris site at
http://www.lmcp.jussieu.fr/sincris/logiciel/
} What kind of accuracy can be obtained - are we } getting close to the accuracy of X-ray diffractometry yet, or are there } fundamental reasons such as lens aberrations, smaller Bragg angles, and } accuracy of measurement which mean that we'll never get there?
Because of lens hysteresis, it's not possible on a standard TEM to calibrate the camera constant L*(lambda) to an accuracy of greater than about 2 percent. A way to get more accuracy is to use an internal standard. For a good discussion of the problems, see Hirsch, Howie, Nicholson, Pashley and Whelan, Electron Microscopy of Thin Crystals, end of chapter 1.
In spite of the camera length inaccuracy, the RELATIVE spacings for two spots isn't affected. Therefore, it is potentially limited only by measurement inaccuracies (how precisely can we locate the center of the spot), and by the relrods (see Hirsch et al).
I believe that a spherical-type distortion of the pattern occurs if you use the beam convergence (condenser lens) to compensate for poor focus of the diffraction pattern. I've never read a good discussion of this (anyone know of one?) Also, ring patterns can be distorted from circular by astigmatism effects (in any post-specimen lens). These optical factors would also limit the accuracy of relative d-spacings, though to some extent one may be able to correct for them.
One place where electrons have a significant advantage over x-rays is with respect to noise. The interaction of electrons with matter is strong, so in very short experimental times good statistics can be obtained from miniscule sample volumse. Electron diffraction therefore has potential for structure refinements (e.g. recent work by Jansen and Zandbergen, L. D. Marks et al for surface diffraction). Also, the photographic film or CCD is a 2D detector (rare in XRD), so you can win big time in reducing noise. This can be used in studies of amorphous materials, via circumferential averaging of the patterns. However, strictly speaking, both dynamical and inelastic effects have to be considered in quantitatively interpreting the data.
} } 2) What are the typical procedures people use for, say, measuring camera length } or identifying unknown phases using diffraction?
Again, you can measure camera length as accurately as you want. Turn the scope off and on and it may differ by a couple of percent.
} } I will make a summary of replies and distribute it to anyone who is interested.
Thanks, I would be interested in hearing.
Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
mailto:wharton.sinkler-at-anlw.anl.gov
} } Many thanks in advance, and a Happy New Year to all, } } Richard Beanland } GMMT Ltd., } Caswell, } Towcester, } Northants NN12 8EQ } UK } e-mail richard.beanland-at-gecm.com } } }
You wrote: } } } Does anybody have any advice as to what materials to use or to NOT use } } with liquid ethane (for material } } incompatibility reasons - chemically and physically (i.e. extreme } } temperatures))? Is copper okay with } } liquid ethane? I've vaguely read somewhere that some groups use a copper } } coil (in liquid nitrogen) to condense } } their ethane. Any other confirmations? } } Both Cu and Al are compatable with LEt. The best scheme might be to pass the N2 vapor at ~80 K through the tube to liquify, but not freeze, the Et. We use an Al cup cooled by LN2, and there are problems with the Et solidifying.
} } A small flexible length of tubing will also be needed to join the copper } } (?) tubing to the cylinder regulator } } (so we can move the coil in and out of the liquid N2 easily). Apparently } } natural rubber is NOT good with ethane.
Tygon is also not good. Teflon tubing retains its strength and flexibility at 77 K, so I reccommend it. Good luck. Yours, Bill Tivol
I have done some more research on the matter of the composition of Formvar. In the book 'Techniques for Electron Microscopy' D. H. Kay, Ed., Blackwell Scientific, 1965 I find a statement indicating that Formvar is Polyvinyl Formal (p. 60) In the book 'Polymer Chemistry' by M. P. Stevens, Oxford Univ. Press, 1990, p.302, I find that the reaction of vinyl alcohol with butyl aldehyde produces a polymer called polyvinyl butyral. By analogy, if vinyl alcohol were reacted with formaldehyde (HCHO) one might assume it would produce polyvinyl formal. If this is so, AND IT IS ONLY A GUESS, then by analogy the chemical formula for the repeating unit in the polymer chain might be:
CH2 / \ -[CH2-CH CH]- | | O O \ / CH2
I hope this formula survives the process of being transmitted across the internet. This word processer is not ideal for writing organic chemical formulas.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Wil Bigelow wrote: } } I have used Formvar for over 50 years now and never known exactly what it is. In the back of my mind I seem to recall hearing that it is } polyvinylformal, but I'm not certain that is correct. I have just } consulted two polymer scientists in our department, and neither one of them knows what it is, nor could they find a chemical formula for it in their reference books.
Will et al:
According to the free sample, yes, I said free sample, I got from Monsanto, Formvar is a "polymer from polyvinyl alcohol and formaldehyde as as copolymer with polyvinyl acetate". If that is not enough information you could call Monsanto in St. Louis. I believe that it was originally developed to coat copper wire. Note that are several different types of Formvar. I think the type us EM folks use is 15/95 but I could be wrong.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Wilbur C. Bigelow wrote: =================================================== I have used Formvar for over 50 years now and never known exactly what it is . In the back of my mind I seem to recall hearing that it is polyvinylformal, but I'm not certain that is correct. I have just consulted two polymer scientists in our department, and neither one of them knows what it is, nor could they find a chemical formula for it in their reference books.
If anyone knows what it is, I'd like to know too. ================================================= You are correct, it is generically, polyvinyl formal, and the term "Formvar" is a trade name, originally registered (if my memory is correct) by Monsanto Chemical Company in St. Louis.
Disclaimer: SPI is a supplier of Formvar resin for use in electron microscopy.
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: www.2spi.com ############################ ==================================================
You will find all the information in my book "low Temperature Microscopy and Analysis" Plenum Press New York 1992
Patrick Echlin University of CambridheOn Wed, 6 Jan 1999, Mail Delivery
Subsystem wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } Hi all. } } } } We are currently setting up a cryo-vitrification unit for TEM sample } } analysis. Research into what is the best } } cryogen to use for vitrification of our sample type gave the answer of } } liquid ethane. } } } } Does anybody have any advice as to what materials to use or to NOT use } } with liquid ethane (for material } } incompatibility reasons - chemically and physically (i.e. extreme } } temperatures))? Is copper okay with } } liquid ethane? I've vaguely read somewhere that some groups use a copper } } coil (in liquid nitrogen) to condense } } their ethane. Any other confirmations? } } } } A small flexible length of tubing will also be needed to join the copper } } (?) tubing to the cylinder regulator } } (so we can move the coil in and out of the liquid N2 easily). Apparently } } natural rubber is NOT good with ethane. } } } } Any other pearls of wisdom out there? } } } } Thanks in advance. I'll summarise my replies. } } } } Terri } } } } ------------------------------------- } } Ms Terri Soar, PhD student, } } University of South Australia } } Email: terri-at-drage.com.au {mailto:Terri-at-drage.com.au} } } ------------------------------------- } } } } } }
We use Tygon brand tubing connected to a copper coil when liquifying gaseous ethane. The tubing is connected to the ethane tank regulator at one end and the copper coil at the other. The copper coil is placed in a beaker in a styrofoam box in a fume hood. The ethane is turned on and liquid nitrogen is poured around the coil. As the copper and ethane start to get cold, the ethane will start to liquify and collect in the beaker. It should take approximately 2-3 minutes to start to liquify and 5-7 minutes to collect 2-300 ml.
You need to be extremely careful with liquid ethane. Besides the obvious (it's extremely cold and will produce severe burns rather quickly), it is volatile if it comes in contact with liquid nitrogen. So when you have a beaker of liquid ethane immersed in a box of liquid nitrogen, the potential for injury cannot be overstated. Hand (and forearm) as well as eye protection are essential.
We've been using liquid ethane for years and I even have photos of the setup in our lab. If you'd like, I could send you a copy of these. Feel free to write or call.
Hope this helps....
Russ
Russell W. Desnoyer Senior Research Technologist Cleveland Clinic Foundation Department of Molecular Cardiology 9500 Euclid Avenue Cleveland, Ohio 44195 Ph: (216) 444-4673 Fax: (216) 445-6062 E-mail: desnoyr-at-cesmtp.ccf.org
I need some info on binary alloys. Typically Aluminium, Titanium and Carbon Nitrides, Borides, etc. These are typically grown as thin films using cathodic arc deposition. What I need is some general info: What research has been done in the past? What research is currently happening? Where is the research is likely to go? What applications do these materials have as thin films and as bulk samples?
I have come across this several times, and there is no simple solution that I am aware of. I believe the reason has a lot to do with how the eye/brain interprets images. We do a good job of excluding noise, and we can often see patterns when it is very difficult to quantitatively measure them on a computer. In a sense we do a type of Maximum Entropy analysis -- we have a prior model of what is in the image and find features that fit this model, for instance weak spots or rings. (Of course, this also means that sometimes we find things that are not there.)
The best method that I am aware of is to combine a rank filter (good at reducing shot noise), some sort of high-pass filter to remove only the low frequencies (reducing the background) and pasting togethor images at different contrast levels to prevent the high intensity regions from dominating. At least for a picture this often works, although you have to play a lot with the kernel sizes. Quantitation is very hard. You have to set up a model (Maximum Entropy, Maximum Likelihood, Least-Squares) and perform a numerical fit. Sometimes Least-Squares works; I have never tried Maximum Entropy which should do better.
Philippe-Andr=E9 Buffat wrote: } =20 } I would be strongly interested in softwares able to handle digital } diffraction patterns. In particular a soft to flatten the background is } needed. From my experience, I can see on negatives faint rings or spots, } but under the same condition it is impossible to see them on a computer } screen without increasing the contrast but then the range between black a= nd } white covers only a small part of densities in the whole pattern and the } observable area between the white center and the black periphery becomes } unacceptably small. The use of color coding of the intensities improves } only a little bit the visible range. An other soft to determine } automatically the spot position or the ring position by a refinement meth= od } or deconvolution of gaussian/lorentzian (or else) curves would also be } welcome.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Formvar is indeed used to coat copper wire. My wife works for an overhead transformer manufacturing firm and they buy the stuff by the barrels for coating the wire (not just copper). It is a different grade and formulation, otherwise I would've been tempted to never buy the stuff again after purchasing a 55 gal drum of it...
On Wed, 06 Jan 1999 14:53:02 -0800 Geoff McAuliffe {mcauliff-at-UMDNJ.EDU} wrote: .. I got from } Monsanto, Formvar is a "polymer from polyvinyl alcohol and formaldehyde } as as copolymer with polyvinyl acetate". If that is not enough } information you could call Monsanto in St. Louis. I believe that it was } originally developed to coat copper wire. Note that are several } different types of Formvar.
I have some filters with particles of differing size but fixed density distributed over the surface. I would like to quantify and compare the "uniformity" of the particle mass distribution on each filter by SEM. My software estimates the volume of each particle using the min and max diameters and assuming an oblate spheroid. Is there a statistically sound recipe for using the observed variation in Vf - where Vf is the particle volume per field measured over many randomly selected fields - to quantify the uniformity of the sample's mass distribution and compare one sample to another? If two samples have different particle loadings, does one use different field areas on the two samples to get the same average Vf for both? The recipe should also include some confidence factor related to the fraction of filter area analyzed.
Thanks for your suggestions.
Bob Willis ManTech Environmental Technology, Inc Research Triangle Park, NC
For background and previous research, try "Phase Diagrams of Binary Titanium Alloys", edited by J. L. Murray. There may also be similar books (published by ASM) for aluminium and carbon. If not, the phase diagram compilation by T. B. Massalski would be a place to start, as well as "Journal of Phase Equilibria" (formerly "Bulletin of Alloy Phase Diagrams" ).
Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
mailto:wharton.sinkler-at-anlw.anl.gov
---------- } From: "George Theodossiou" {GEORGE-at-bunyip.ph.rmit.edu.au} } To: microscopy-at-sparc5.microscopy.com } Subject: Binary Alloys } Date: Thu, Jan 7, 1999, 10:43 AM }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One of our researchers has mouse embryos that express gfp-linked material. These embryos were preserved and maintained in PBS plus freshly made 4% p formaldehyde and 0.5% Tween 20. Initially, his control samples were pink in color without fluorescence. After 3 months, his control samples have started to turn green under fluorescent light. In so far as these are controls for his gfp expression samples, this fluorescence is not helpful.
Any idea what is causing this change in color? I recommended he should store his samples long-term in PBS plus 0.5% formaldehyde in PBS. Anyone have any better suggestions?
Also, those of you who work with gfp.....Could he treat his embryos, say with ammonium chloride, to try to get rid of his unwanted background fluorescence? Can he do an equivalent treatment of his controls and experimentals without damaging the gfp fluorescence he has previously observed?
thanks in advance
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-8759 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting remote access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
You should look up the proceedings from the International Conference on Metallurgical Coatings and Thin Films for the past several years. The proceedings are two volumes and the papers are full papers. One volume is printed in Thin Solid Films and the other in Surface and Coatings Technology. You can find out more about this year's conference at the following web site: http://www.vacuum.org/icmctf/icmctf.html
Incidentally, I am a session chair for the "Microstructural, Microanalytical and Imaging Characterization" session in the "Coating and Thin Film Characterization" symposium.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: George Theodossiou To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hi all
I need some info on binary alloys. Typically Aluminium, Titanium and Carbon Nitrides, Borides, etc. These are typically grown as thin films using cathodic arc deposition. What I need is some general info: What research has been done in the past? What research is currently happening? Where is the research is likely to go? What applications do these materials have as thin films and as bulk samples?
i've been reading principals of heat treatment of steel by krauss and in the references of chapter 5 reference 4 lists a paper by S.Chattopadhyay and C.M. Sellars titled Quantitative measurements of pearlite spheroidization,Metallography,vol10,1977 pg 89-105
does anyone have this paper? if you do could you please fax it to me at 716-684-9433
or could you tell me what Metallography is ,is it a magizne?
Sounds like you may want to check with John Russ about many of these issues. I thought he was right there in your backyard. But I will offer a couple thoughts.
I think you would be better use the same field size for all of your measurements. Besides, Vf should have the same mean value no matter how large the field. The larger the field you measure, the less variation in field-specific measurements between fields. Supposing I find a standard deviation of 4% for some measurement for a single field (determined by measureing multiple fields). Then, if I measured a field 5 times as wide or measured 25 fields and averaged the results, the standard deviation on that measurement would be sqrt(25) less or 0.8%. Either way, 25 times more area was analyzed. So, you could make some adjustments to match up your measurements even if you did use different areas.
Regarding measuring the variation in Vf and compare samples - our work involves the measuring of void distributions. We are routinely measuring 20 frames per sample and calculating the variation in Af, but that is to have a measure of confidence in our Af measurements. It gives us some insight into the nature of our samples. A lower variability normally translates into a smaller average particle size, but it can also indicate some things about uniformity of distribution for the smae feature size between samples.
As to the statistical test for determining when the variation is significantly different between two populations with identical means, I will leave that to those that are better versed in statistics than I am at this moment. There must be one out there.
Hoping this helps. Warren
At 08:51 AM 1/7/99 -0600, you wrote: } I have some filters with particles of differing size but fixed density } distributed over the surface. I would like to quantify and compare the } "uniformity" of the particle mass distribution on each filter by SEM. My } software estimates the volume of each particle using the min and max } diameters and assuming an oblate spheroid. Is there a statistically } sound recipe for using the observed variation in Vf - where Vf is the } particle volume per field measured over many randomly selected fields - } to quantify the uniformity of the sample's mass distribution and compare } one sample to another? If two samples have different particle loadings, } does one use different field areas on the two samples to get the same } average Vf for both? The recipe should also include some confidence } factor related to the fraction of filter area analyzed. } } Bob Willis } ManTech Environmental Technology, Inc } Research Triangle Park, NC
For some reason, Philippe-Andre Buffat's posting to the listserver = never showed up in my mail. I wonder if this is common--getting partial = conversations.
Seeing faint features on TEM negatives and not on the digitized images = sounds as if it has more to do with limitations of the digitization process used = than it does with visualization. 8 bits probably just isn't enough integers to = preserve the faint image detail and keep the black and white extremes at the = same time. If the data is not preserved in the digital images, no amount of fancy = filtering is going to recover it. I'd suggest a careful look at what's going on = in the digitization process you're using. If your scanner allows spreading = the film density data over 12 or 14 bits instead of the usual 8, you may be able = to extract the fine detail information by postprocessing high-bit images = to level backgrounds, adjust tones, and filter. The unsharp mask filter in = Photoshop sometimes does a good job at enhancing faint details that are otherwise = lost or blurred in the scanning process.
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA=20
---------- From: L. D. Marks Sent: Thursday, January 7, 1999 12:59 PM To: Microscopy List Subject: Re: TEM; diffraction pattern analysis
The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com On-Line Help = http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html = -----------------------------------------------------------------------.=
I have come across this several times, and there is no simple solution that I am aware of. I believe the reason has a lot to do with how the eye/brain interprets images. We do a good job of excluding noise, and we can often see patterns when it is very difficult to quantitatively measure them on a computer. In a sense we do a type of Maximum = Entropy analysis -- we have a prior model of what is in the image and find features that fit this model, for instance weak spots or rings. (Of course, this also means that sometimes we find things that are not there.)
The best method that I am aware of is to combine a rank filter (good at reducing shot noise), some sort of high-pass filter to remove only the low frequencies (reducing the background) and pasting togethor images at different contrast levels to prevent the high intensity regions from dominating. At least for a picture this often works, although you have to play a lot with the kernel sizes. Quantitation is very hard. You have to set up a model (Maximum Entropy, Maximum Likelihood, Least-Squares) and perform a numerical fit. Sometimes Least-Squares works; I have never tried Maximum Entropy which should do better.
Philippe-Andr=E9 Buffat wrote: } =20 } I would be strongly interested in softwares able to handle digital } diffraction patterns. In particular a soft to flatten the background is } needed. From my experience, I can see on negatives faint rings or spots, } but under the same condition it is impossible to see them on a computer } screen without increasing the contrast but then the range between black and } white covers only a small part of densities in the whole pattern and the } observable area between the white center and the black periphery becomes } unacceptably small. The use of color coding of the intensities improves } only a little bit the visible range. An other soft to determine } automatically the spot position or the ring position by a refinement method } or deconvolution of gaussian/lorentzian (or else) curves would also = be } welcome.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Greetings, I need assistance with flat embedding of rat cerebellum (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically we are worried about tissue curling during the dehydration. My first inclination is argarose embed (before epon), but I would take any expert advise. Thank you,
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
A postdoctoral position is available in the Interface Physics Group at the University of Illinois at Chicago (UIC). Research in the Interface Physics Group focuses on the use atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. Current research programs involve ceramics, high-Tc superconductors and optoelectronic/high-power semiconducting materials and devices. The experimental facilities to perform this research are comprehensive: a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detector (Z-contrast), Gatan Imaging Filter, and Noran EDS; a VG HB501A Field-Emission dedicated STEM with EDS, EELS and Auger spectrometers; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; a JEOL 6320 =46ield-Emission SEM with EDS and Cathodoluminescence; a JEOL JXA733 microprobe; and a Topometrix AFM/STM. In addition to the electron microscopes, specimen preparation facilities include a Gatan Duo-mill, =46ischione precision ion-mill, SouthBay plasma cleaner and Leica Ultramicrotome. The Interface Physics Group has a Silicon Graphics R10000 Power Indigo workstation with a Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The physics department has additional workstations and access to the UIC Convex Exemplar Supercomputer and the National Center for Supercomputing Applications at UIUC. =20
This position is a joint postdoctoral appointment with Professor Susanne Stemmer in the Department of Physics at UIC. Research performed by the successful candidate for this position will involve the investigation of grain boundaries and defect structures in ionic and mixed ionic/electronic conducting oxide ceramics. The aim of the program is to incorporate experimental results into comprehensive atomic scale models for ionic/electronic transport in these materials.=20 It is anticipated that this position will involve a significant amount of industrial collaboration.
Candidates should be recent Ph.D. graduates in physics, metallurgy, or materials science with a background in the relevent materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume, names, addresses of three references and a publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience. UIC is an equal opportunity employer.
} Date: Thu, 7 Jan 1999 15:06:29 -0500 (EST) } From: MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu} } To: microscopy-at-sparc5.microscopy.com } Subject: flat embedding of vibratome 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 } -----------------------------------------------------------------------. } } } Greetings, } I need assistance with flat embedding of rat cerebellum } (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically } we are worried about tissue curling during the dehydration. My first } inclination is argarose embed (before epon), but I would take any expert } advise. } Thank you, } } Mike D } } We do this all the time. Sections don't curl. We originally tried to put them into flat molds standing up on their edges, but they would fall over sometimes. We now cut the pointed end off a BEEM capsule; snap the cap on the other end and wrap the junction with a sliver of Parafilm to prevent leaking; put a drop of resin in the lid (now upside down, with the Beem capsule sticking upwards); and lift the thick section into the lid with a wooden applicator stick broken into a flat wedged end with the tip then bent up into the shape of a hoe then fill the capsule with resin. Do this on a light box and under a dissecting scope.
If you need to keep straight which side is which, you can trim the tiny thick section with a razor blade into a funny shape that you will recognize. We use a trapezoid-like shape/state of Nevada shape: __ | \ |___\
That way you can keep the side of interest outward, e.g., confocal scope-selected areas: Miller SE, Levenson RM, Aldridge C, Hester S, Kenan DJ, Howell DN. 1997. Identification of focal viral infections by confocal microscopy for subsequent ultrastructural analysis. Ultrastruc. Pathol. 21:183-193.
Your sections are wider; thus, you will have to embed in a larger mouthed container if you want sections parallel with the face. But the principle is the same. If the sections have enough room to "swim" in the resin, you can gently stretch them out with the applicator stick, i.e., they won't premanently curl even though they will be flimsy as they float around in the resin.
Good luck.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Mike D- we routinely embed flat sections after the immuno proceedures, do any post fixation (OsO4), followed by dehydration, infiltration with resin, then to embed we place the 100 micron section between two glass slides * one of which has been subbed with a gelatin material, the other whioch has been subbed with Trenmittel (we purchase ours from EMS) the trenmittel is something like teflon, it is a release agent. remove all exess resin from around the section, then clamp the two slides together (clothes pins or large paper clips) polymerize as usual, then using a knife blade pry the two glass slides apart, this takes a little practice. finally reembed by filling a gelatin capsule with resin, let it thicken a little, then invert it over thetissue section which remained on the subbed slide, polymerize overnight. to free the section/block; hold slide over an alcohol burner flame (use pliers) then after 6-8 seconds snap off the block with the aid of haemostats or pliers. it really can work quite well. but practice first it is a little tricky -Mike
On Thu, 7 Jan 1999, MICHAEL DELANNOY 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 need assistance with flat embedding of rat cerebellum } (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically } we are worried about tissue curling during the dehydration. My first } inclination is argarose embed (before epon), but I would take any expert } advise. } Thank you, } } Mike D } } }
In a message dated 1/7/99 2:07:07 PM, wesaia-at-iastate.edu wrote:
} Sounds like you may want to check with John Russ about many of these } issues. I thought he was right there in your backyard. But I will offer a } couple thoughts. } } I think you would be better use the same field size for all of your } measurements. Besides, Vf should have the same mean value no matter how } large the field.
Guess I'll throw in my $.02 worth since my name has been taken in vain.... ;-)
Field size does matter, alas. He is trying to measure particle size which means he needs to ignore those which touch the edge of the image field, and this creates several problems: a) large particles are more likely to touch the edge than small ones; b) larger fields will have proportionately fewer edge- touching particles. It can get complicated but the idea is to determine the number of edge-touching features and from that and the size distribution of those measured, adjust the effective area of the image so that the number per unit area is corrected for edge effects. Contact me directly via email if I can be of help
I am new to microscopy - and am trying to figure out what is the best general purpose mountant. I read that Euparal is good and stable for many years, but I can't find any except in Australia. I just want to make some basic slides for now, nothing exotic. I would appreciate any advice from anyone. Thanks.
David W. Bass Appalachian State University Boone, North Carolina dwbass-at-appstate.campuscw.net
Hello All, I have had 21 replies to my original request for information. Many thanks to everybody who responded, and apologies to anybody I misrepresent in this summary!
The state of the art in measurement accuracy was claimed to be close to that of X-ray diffraction by several people. [Of course, it depends what kind of X-ray diffraction you're talking about - I don't personally believe it will ever be possible to get accuracies as good as that of double (triple, etc.) crystal X-ray diffraction from spot patterns, although CBED/HOLZ line analysis comes close.] Jouk Jansen mentioned his paper in Acta Cryst of Jan 1998 on analysis of diffraction patterns. 2% was mentioned as the best day-to-day reproducibility one could hope for without taking special precautions such as evaporating gold onto your sample to include a standard pattern on the same negative as the pattern you want to measure. Lens hysteresis, astigmatism and pincushion/barrel distortion due to poor focusing may make it even worse. Eric Lachowski mentioned the huge effect that being away from eucentric height can have [I came across this myself a little while ago - I was horrified to find that a 50% change of diffraction pattern spacing was possible, even though the image size only changed by 5%, when tilting a sample.] An accuracy of 0.1% in measurement was about the limit, using computer-aided measurements of digital images. The distortion of diffraction patterns due to reciprocal lattice spiking was mentioned by quite a few people as potentially being the limiting factor in measurement. Most people seem to use evaporated gold as a standard for measuring camera length. There seems to be quite a wide variety of measurement methods out there, ranging from the good old fashioned way (i.e. lupe and graticule) to quite sophisticated analysis of digitized images. The software people use seems to fall into two categories; packages used to measure the position of spots and/or rings, and packages which simulate diffraction patterns which can be used for comparison with the real thing. Measurement packages included Gatan's Digital micrograph and NIH-Image. A few people have put a lot of work into producing packages which automatically make measurements: EDP, by Jaap Brink (http://ncmi.bioch.bcm.tmc.edu/~brink), which is free. MacLispix by Dave Bright (http://www-sims.nist.gov/MLx/doc/home.nclk), also free but runs only on a power Mac (sob). Bill Tivol has written plug-ins for the SPIDER image processing package that subtracts circular backgrounds, measure the centre and radius of a ring, and obtain background-subtracted intensities for a spot pattern. Corneliu Sarbu mentioned the free package PATTERN, running on PCs, which can be used as an aid to interpretation of spot patterns (see Microscopy Today 98-9 (Nov 1998)). Analysis packages: Carine, Crystal Designer, CRISP, Desktop microscopist and of course EMS were mentioned as packages which are used to simulate diffraction patterns for a particular crystal and zone axis. Wharton Sinkler has written a FORTRAN program to aid indexing of patterns to a known structure (http://www.lmcp.jussieu.fr/sincris/logiciel/).
Many thanks again to all those who replied.
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK e-mail richard.beanland-at-gecm.com
As far as sectioning, we tend to look for the Tg of the various components. As long as all of the components in your system have Tg well above room temperature, then sectioning at room temperature will result in relatively small deformations. In your case, PVC has a Tg of about 85 C and room temperature sectioning should be OK. In the case of PVB ( I assume B = Butadiene ?) the Tg is bellow RT and therefore it is better to use cryo sectioning for this material. Typically we would section this at about -100C .
As far as staining, osmium should stain the butadiene , but so will RuO4. I am not familiar with a stain for PVC, but you could try etching techniques (e.g. plasma etching). Check Linda Sawyers book on Polymer microscopy .
I hope this helps.
Jordi Marti ---------------------------------- You wrote: What is eveyones favorite stain for enhancing contrast during polymer TEM? OS4 or otherwise? The polymer materials are PVC and PVB.
Also, does anyone out there have some advice on a recommended temperature for ultramicrotomy of the above polymers? We DO have a cryo-ultramicrotome.
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Mike, You shouldn't have much trouble with curling during fixation and dehydration. The problem comes with getting absolutely flat sections for polymerization. Your sections are a bit thicker than I used and we were usually able to cut down the area of interest further but try this procedure. I think it should work fine. I have used it to embed vibratomed x-sections of rat brain which had been subjected to pre-embedding ICC reactions. In this case, since the amount of reaction diminished as you cut further into the section, it was critical to have absolutely flat material for serial sectioning.
Try embedding in droplets of resin on a 22x22mm plastic coverslip (available through most of the EM supply companies). Cover with another coverslip and weight with metal washers or nuts. The weighting is important as it really pressed the sections down insuring that they are very flat and squeezes out extra resin. After 24 hr. polymerization, cut edges of cover slips and they can then easily be separated. Cut the end off of gelatin or beam capsules to give a tube. Place the capsules over areas of interest on the sections, add one small drop of resin and polymerize a number of hours to secure capsules to the sections. Then fill up the capsules and finish polymerization.
When ready to section, blocks are easily broken or cut off of the coverslips. There will be very little extra resin to cut through before getting into the tissue making sectioning fairly easy once you are properly lined up.
An alternative method is to embed between teflon-coated glass slides which are then also weighted. I have found this a bit more cumbersome than using the plastic coverslips but it may work better for thicker specimen material.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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Greetings, I need assistance with flat embedding of rat cerebellum (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically we are worried about tissue curling during the dehydration. My first inclination is argarose embed (before epon), but I would take any expert advise. Thank you,
Mike D
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Hi, everybody, I am looking for where I can buy an interference microscopy, which can be used to evaluate the thickness of thin film on a glass substrate by comparing the fringe at the edge of the thin film. I like to know also that if there is any kind of glass binder (adhesive material) which can be used inside a vacuum chamber, can last several hours in vacuum before working and can be used as air-tight sealing. Thanks a lot. Zgliu
Liu Zugang Departamento de Fisica Universidade de Aveiro 3810 Aveiro Portugal Fax:+351-34-24965 Email:zugang-at-fis.ua.pt
I would just like to correct one item in Richard's summary: CRISP and (more relevantly) ELD are programs which extract data from experimental images and diffraction patterns (rings and spots), respectively. They are not used to simulate diffraction patterns.
Disclaimer: Calidris sells CRISP and ELD, and we have a vested interest in making sure that microscopists receive accurate information about the programs.
I will be happy to send details to anyone who is interested.
Best wishes for the New Year,
George Farrants.
Richard wrote: } Analysis packages: Carine, Crystal Designer, CRISP, Desktop microscopist and of } course EMS were mentioned as packages which are used to simulate diffraction } patterns for a particular crystal and zone axis.
Does anyone have an answer to those questions? Working with a Variable Pressure SEM; 1- What can we see with it on polymers? 2- Where can I find pictures of polymers on a VP-SEM? 3- Is it possible to see spherolites in polymers without any pre-treatment (etching...) 4- Is it possible to see the "Skin effect" on injected parts in polymers and measure its thickness ?
We have tried to cleave rock salt (1cm cube, purchased from a microscopy supplier) for use as a substrate. We cleaved it with a razor blade, and had hoped to get a near-atomically smooth surface so we could deposit an aluminum film on it. However, the cleaved surfaces appear to be far rougher this, on the order of tens of microns.
Are there any special procedures that we should follow to get a near-atomically smooth surface?
Thank you all for your suggestions.
Mick Thomas
Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
} From: "Ursel Bangert" {USCHI-at-fs2.phy.umist.ac.uk} Organization: Umist To: MTLRMDB-at-LUCS-MAC.NOVELL.LEEDS.AC.UK
Good Morning All,
I am looking for suggestions on dispersing agglomerated crystals. Does anyone have a recommendation on what "dispersing agents" are available for dispersing sub-micron ( {100 nm) crystals?
I have only used ultrasonication of particles in acetone/water so far and that works well for micron sized crystals.
Thank you,
Mohan Kalyanaraman
Sr. Staff Material Scientist Mobil Technology Company PO Box 480 Paulsboro, NJ 08066 mohan_kalyanaraman-at-email.mobil.com
Does anyone have an answer to those questions? Working with a Variable Pressure SEM; 1- What can we see with it on polymers? 2- Where can I find pictures of polymers on a VP-SEM? 3- Is it possible to see spherolites in polymers without any pre-treatment (etching...) 4- Is it possible to see the "Skin effect" on injected parts in polymers and measure its thickness ? 5- Does anyone work on polymers for quality issues?
Does anyone have any tips/protocol/adivce for the preparation of human chromosomes for SEM. I'm just starting the project and I could use anything to get started... Thanks!
Rob Reff } From the Lab of: Professor William J. Perreault Lawrence University
} What is everyones favorite stain for enhancing contrast during polymer } TEM? OS04 or otherwise? The polymer materials are PVC and PVB.
} From the literature, ruthenium trioxide seems to be the most popular. We have used chlorosulphonic acid, but this mainly works for polyolefins, and I think it would chew up PVB (is that poly vinyl butyral?).
} Also, does anyone out there have some advice on a recommended temperature } for ultramicrotomy of the above polymers? We DO have a cryo-ultramicrotome.
The following article is superb. However, the author is out of reprints, so it might be better to use some form of library loan, if you don't take the journal.
*} TI: Reflections on the use of microtomy for materials science specimen preparation AU: Plummer_HK NA: FORD MOTOR CO,RES LAB,MAIL DROP 3028,SRL,DEARBORN,MI,48121 JN: MICROSCOPY AND MICROANALYSIS, 1997, Vol.3, No.3, pp.239-260 IS: 1431-9276 DT: Review
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
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.
------=_NextPart_000_012E_01BE3BBB.A789C660 Content-Type: text/x-vcard; name="Vratislav Richard Eugene Maria John Baptist Bejsak-Collorado-Mansfeld.vcf" Content-Transfer-Encoding: quoted-printable Content-Disposition: attachment; filename="Vratislav Richard Eugene Maria John Baptist Bejsak-Collorado-Mansfeld.vcf"
BEGIN:VCARD VERSION:2.1 N:Bejsak-Collorado-Mansfeld;Vratislav Richard;Eugene Maria John Baptist FN:Vratislav Richard Eugene Maria John Baptist Bejsak-Collorado-Mansfeld ORG:Bayshark Research Laboratory TITLE:director NOTE;ENCODING=3DQUOTED-PRINTABLE:Marketing and = Coaching=3D0D=3D0A=3D0D=3D0ATenebrionidae Orbis and higher taxonomy TEL;WORK;VOICE:(+61 2) 9319 6380 TEL;CELL;VOICE:(+61 414) 540 465 ADR;WORK:;;29 Edward Street;Darlington, SYDNEY;;NSW 2008;Australie LABEL;WORK;ENCODING=3DQUOTED-PRINTABLE:29 Edward = Street=3D0D=3D0ADarlington, SYDNEY NSW 2008=3D0D=3D0AAustralie ADR;HOME;ENCODING=3DQUOTED-PRINTABLE:;;(temporaly address):=3D0D=3D0A32 = Girrawheen Ave;KIAMA;;NSW 2533;AUSTRALIA LABEL;HOME;ENCODING=3DQUOTED-PRINTABLE:(temporaly address):=3D0D=3D0A32 = Girrawheen Ave=3D0D=3D0AKIAMA NSW 2533=3D0D=3D0AAUSTRAL=3D IA URL: URL:http://www.coleoptera.org EMAIL;INTERNET:ricardo-at-login.cz EMAIL;PREF;INTERNET:vratislav-at-bigfoot.com EMAIL;INTERNET:ricardo-at-ans.com.au REV:19990108T233435Z END:VCARD
I am trying to gather information regarding Scanning Transmission = Electron Microscopy. I currently use an Amray 1645 SEM which has the = capacity for STEM work, but I have never used it in this mode. Any = comments or literature citings on this subject would be greatly = appreciated.
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3509.100"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I am trying to gather information = regarding=20 Scanning Transmission Electron Microscopy. I currently use an Amray 1645 = SEM=20 which has the capacity for STEM work, but I have never used it in this = mode. Any=20 comments or literature citings on this subject would be greatly=20 appreciated. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Thank you for your help, = {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Chris = Holp {/FONT} {/DIV} {/BODY} {/HTML}
A technical difficulty in my lab is coming on the table: How to increase resolution of digitized SEM images, especially for low magnification ( {50X). The pixel size of SEM image (50X)in my machine (LEO-435VP) is 4.5 um. In other word, no matter how good image software performs, the measurement error is at least 4.5 um. We are going to use SEM as a routine measurement tool under 100X, what is the disadvantage? Does any one have excellent idea to solve this problem, in terms of : Increase number of pixels and save as compressed .jpg to reduce file size? Stage mapping? Software solution? What else?
Mick: Maybe the salt could work, but why not use freshly cleaved mica, its not sensitive to humidity, dead easy to cleave and cheaper. I must declare that ProSciTech (and several other EM suppliers) stock mica. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 09, 1999 5:13 AM, Mick Thomas [SMTP:mgt3-at-msc.cornell.edu] wrote: } Fellow microscopists, } } We have tried to cleave rock salt (1cm cube, purchased } from a microscopy } supplier) for use as a substrate. We cleaved it with a } razor blade, and } had hoped to get a near-atomically smooth surface so we } could deposit an } aluminum film on it. However, the cleaved surfaces appear } to be far } rougher this, on the order of tens of microns. } } Are there any special procedures that we should follow to } get a } near-atomically smooth surface? } } Thank you all for your suggestions. } } Mick Thomas } } } } } Mick Thomas } UHV-STEM Laboratory } E-1 Clark Hall } Cornell University } Ithaca, NY 14853 } } Phone: 607-255-0650 } Fax: 607-255-7658 } e-mail: mgt3-at-msc.cornell.edu
Chris: Just a couple of the more obvious differences between SEM and STEM (as an attachment to a TEM), using secondary mode - 1. STEM has generally higher kV - greater soft specimen penetration, charging is worse, but on "hard specimen" better resolution. 2. Working distance is low in STEM, greater resolution, poor depths of field. 3. STEM has small sample access and often limited tilt/ rotate facilities. Suitable specimens can give great images, but with more difficulties.
In STEM mode, which is also possible with some SEMs (including yours). The important differences are: The specimen is a section and this is penetrated by the beam. The detector (photo multiplier) is below the specimen. The penetration envelope is not formed because a relative thin section is used and the beam diameter is the most important determinant of image resolution. So when in a conventional SEM resolution in soft (biological) specimen is limited to say 6nm, you could resolve, say 2nm in STEM because the resolution is largely determined by the beam diameter. Better still, in low contrast specimens contrast can be increased at will - until electronic noise takes over. Maybe the best use of STEM is in image analysis of soft specimens. X-ray scattering from the penetration envelope in an SEM at around 15kV will result in spatial X-ray resolution of about 20 micrometer, which is often pretty near useless. In STEM the spatial X-ray resolution is only a fraction thereof. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 09, 1999 11:56 AM, Chris [SMTP:cholp-at-ncweb.com] wrote: } I am trying to gather information regarding Scanning } Transmission Electron Microscopy. I currently use an } Amray 1645 SEM which has the capacity for STEM work, but } I have never used it in this mode. Any comments or } literature citings on this subject would be greatly } appreciated. } } Thank you for your help, } } Chris Holp } { { File: ATT00001.html } }
Jena (name also of city), was pre WW2 the Zeiss centre. Zeiss Jena and Zeiss Oberkochen run as two separate companies in East and West Germany. Zeiss Oberkochen later took over the Jena works. Its now known simply as Zeiss. Zeiss should know about these optics, but I doubt that they can or would supply these. Your chances are in the secondhand market. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 09, 1999 9:35 AM, ricardo [SMTP:ricardo-at-ans.com.au] wrote: } Dear colleagues } } I am looking for objectives and oculars or other } accessories for microscope } called Technival 2 from old East Germany company JENA.. } } Any help? } } Keep care and be of good cheer. } } Regards } } Vratislav Richard Eugene Maria John Baptiste } of Bejsak (Bayshark)-Collorado-Mansfeld } } Coleoptera - Australia, Tenebrionidae of World } (incl. Lagriinae, Alleculinae) } } Temporally home address: } 32 Girrawheen Ave. } Kiama NSW 2533 } Australia } e-mail: vratislav-at-bigfoot.com } ricardo-at-ans.com.au } (before Ricardo-at-compuserve.com } and ricardo-at-login.cz ) } } http://www.coleoptera.org } phone : 0414 540 465 (Australia) } +61 414 540 465 (International) } } 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. } } } { { File: Vratislav Richard Eugene Maria John Baptist } Bejsak-Collorado-Mansfeld.vcf } }
Liu: I expect that several of the major microscope manufacturers still make double beam interference microscopes (Certainly Leitz used to). All of these would have scary price tags. Perhaps you can find one second-hand or an alternative method for measuring film thickness.
The properties you seek for sealing air/ glass under vacuum are those of Apiezon T. This item is in our online (page M2) and I must declare an obvious interest. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Friday, January 08, 1999 8:17 PM, Liu Zugang [SMTP:zugang-at-ideiafix.fis.ua.pt] wrote: Hi, everybody, } I am looking for where I can buy an interference } microscopy, which } can be used to evaluate the thickness of thin film on a } glass } substrate by comparing the fringe at the edge of the thin } film. } I like to know also that if there is any kind of glass } binder } (adhesive material) which can be used inside a vacuum } chamber, can } last several hours in vacuum before working and can be } used as } air-tight sealing. } Thanks a lot. } Zgliu } } Liu Zugang } Departamento de Fisica } Universidade de Aveiro } 3810 Aveiro } Portugal } Fax:+351-34-24965 } Email:zugang-at-fis.ua.pt
In a message dated 1/9/99 1:47:00 AM, zhiyuw-at-worldnet.att.net wrote:
} A technical difficulty in my lab is coming on the table: How to increase } resolution of digitized SEM images, especially for low magnification } ( {50X). The pixel size of SEM image (50X)in my machine (LEO-435VP) is 4.5 } um. In other word, no matter how good image software performs, the } measurement error is at least 4.5 um. } ... } Does any one have excellent idea to solve this problem, in terms of : } Increase number of pixels and save as compressed .jpg to reduce file size? } Stage mapping? } Software solution? } What else? } If your beam size and actual imaging resolution is sub-micron, then the 4.5 micron limit is arising from the timing of your ADC, in other words how many samples it takes along each scan line, and from the spacing of the lines. If you can alter than then you can acquire images with more pixels. But DON'T compress them with jpeg or any lossy compression scheme or you will lose the benefits - these methods cause brightness and location shifts for features and will not get the accuracy you want.
If you can't fiddle the acquisition, your other choice is to acquire a series of higher magnification images and stitch them together as a mosaic. This isn't always easy to do, since stage mechanisms aren't very precise and if the sample isn't flat and horizontal you will have magnification that varies from side to side and makes fitting impossible.
On the other hand, what is it that you need to measure that can't be sampled at higher magnification - do you really need One Big Picture?
a Happy and prosperous new year to all! I am a new subscriber and I work at a local branch of the Min. of Health as a veterinary hygiene inspector in Italy. I am working on a substitution of malachite green in aquaculture, but I would like to know more about its nature, use and toxity, how dose it act? I've recently learned that it's used as a dye in staining certain cell tissues; surely somebody has posed himself the problem and has even found some explanation. Can somebody please give me some information or wher I can get it (ex. Web Sites)? Excusing me for having drifted you away from your prevalent work I anticipate my thanks to those how will answer and a good luck to evrybody! regards
emidio fazzini
(...up here from downunder!)
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Jim Darley wrote: ============================================== Maybe the salt could work, but why not use freshly cleaved mica, its not sensitive to humidity, dead easy to cleave and cheaper. I must declare that ProSciTech (and several other EM suppliers) stock mica. =============================================== Jim is of course correct in that salt is sensitive to moisture, and it is that very characteristic that causes freshly cleaved NaCl to be the substrate of choice for some researchers. When studies of epitaxial effects are being done, especially at elevated temperatures, it can be problematic to remove the thin film coating from the substrate, but in the case of NaCl, it can be readily dissolved in water. And even when mica could be used, in many instances, NaCl is also used in parallel since the unit cell dimensions both in size and symmetry are quite a bit different. And they give effects that can be quite different as well.
The impact of the differences in unit cell geometry and unit cell dimensions can also result in a significant difference in annealing effects of small crystals on the surfaces of these substrates. However, from the standpoint of smoothness, for example, if one was making carbon support films, mica would be better (if not also easier and cheaper) than NaCl.
An "old" reference from the literature that shows some of this can be found at Die Makro. Chemie, 113, 246 (1968), "Polyoxymethylene Single Crystals. II. The Effect of Substrate of Annealing Behavior".
Chuck
Disclaimer: SPI is a supplier of both mica and fine single crystal NaCl as are also some of the other main suppliers of consumables to the microscopy and microanalysis market.
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Hello: Can anyone comment on the difference in the volume or resolution of a point EDS analysis at 1kv or even 200V vs. the more standard 10 or 20kV. If we were to use a FE-SEM at 200V could we expect a significant increase in resolution of our EDS for a bulk sample compared our convention SEMs. Any comments are greatly appreciated. Steve
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
Sure Stephen, lower kV equals better spatial X-ray resolution. But. . . . But what X-rays would you be able to excite with these low voltages? Look at a table of X-ray energies and remember that the very definition of the X-ray energy lines is the minimum voltage required to produce those X-rays. Generally 1.8 x that energy is required to obtain maximum fluorescence. I expect that you would like to analyse light elements (biological samples), and these are in most cases unsatisfactory in SEM because of the poor spatial resolution. For these TEM or STEM with EDS are the answer. (Made a posting on STEM yesterday with more info) Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
} } } Hello: Can anyone comment on the difference in the volume } or resolution of } a point EDS analysis at 1kv or even 200V vs. the more } standard 10 or 20kV. } If we were to use a FE-SEM at 200V could we expect a } significant increase in } resolution of our EDS for a bulk sample compared our } convention SEMs. Any } comments are greatly appreciated. Steve } } } ------------------------------ } Stephen McCartney } Research Associate } Virginia Tech } Materials Institute } 2108 Hahn Hall } Blacksburg, VA 24061-0344 } USA } } TEL: 540-231-9765 } FAX: 540-231-8517 } ------------------------------ }
} Hi Mark, I must be getting real old as I remember my gransfather used to } make chairs out of plant material. Large plants I believe. They wood cut the } larger stems into structural members and glue the parts together into many } different shapes. I sure these were not as functional or beautiful as the } bent steel and plastic used today. Maybe you could find one of these } antiques for your purpose. } Russ
Hi Russ,
your suggestion has stirred some racial memories in my mind. I have a vague recollection of seeing such ancient furniture in scratchy black & white movies (assuming they were not plastic/steel replicas of the cellulose originals).
I will enquire as to the availability of chairs fashioned from our forefather's favourite material, and also purhaps from adobe brick, bamboo, or rock. 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.
Hi everybody who interested in the problem of 3-D biological tissue architectures reconstruction. This problem (especially 3-D epithelia structure) is interesting for me too. So I would greatly appreciate a copy of your papers. Also I am inviting you to visit our homepage on same subject. URL is followed: http://members.tripod.com/~Gensav/index.htm Sincerely yours Gennady A. Savostyanov
Dr. Gennady A. Savostyanov E-mail: savost-at-ief.spb.su savost-at-hotmail.com Sechenov Institute for Evolutional Physiology and Biochemistru Russian Academy of Science 44 M. Thorez, 194223 St. Petersburg, Russia
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} } } Hank Adams wrote ........... Dear listservers, We are looking for a dedicated software package for scheduling, in this case, instrument use incorporated into our webpage. We have several microscopes, workstations, microtomes, etc. and would like to have calenders or something similar for each, so users could sign up in advance using the web. It should be possible to assign different levels of privilege to each user. Once an entry is made it could only be changed by an administrator other than the original user. We like would like to then to automatically transfer this information into a billing database, either Access or FileMaker Pro based. Is something like what I have described commercially available
We have just such a system written in house. It has run very smoothly now for 5 years with constant upgrades. You can buy it if you like it.
Just go to the website in my signature.
The introductory material explains how we work.
Click on Booking System, Access to Images and use login {guest} password {guest} to try out the system.
If you are interested get back to me.
Mel Dickson ***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Hi: I am beginning a project on glass and I will be using a contract microscope. Can you explain with diagrams how the phase contrast scope works. Thank you in advance, dsm
High Steve! As far as I remember, the optimum (in sense of intensity) ratio between energy of electrons and excitation energy of X-ray line is from 2 to 3. At that the spatial X-ray resolution is not very high. It can be improved by diminishing this ratio. But the intensity of the line drops very strongly during decreasing of electrons energy to excitation energy of the line. But I think there are not so many X-ray lines interesting for you on EDS spectrum in the range up to 200 eV :-)). Regards. Victor Sidorenko, ANTRON Co. Ltd., Moscow, Russia.
} Hello: Can anyone comment on the difference in the volume or resolution of } a point EDS analysis at 1kv or even 200V vs. the more standard 10 or 20kV. } If we were to use a FE-SEM at 200V could we expect a significant increase in } resolution of our EDS for a bulk sample compared our convention SEMs. Any } comments are greatly appreciated. Steve } } } ------------------------------ } Stephen McCartney } Research Associate } Virginia Tech } Materials Institute } 2108 Hahn Hall } Blacksburg, VA 24061-0344 } USA } } TEL: 540-231-9765 } FAX: 540-231-8517 } ------------------------------ } } }
Hi Zhiyu Wang - Lets hope its me who is confused: I don't care. If a monitor is 100mm across and is represented by 4um pixel (100 divided by 0.004=) 2500 would be required for a single line. If one pixel was missing I would just forget about that, although the percentage error would be constant, regardless of magnification. What I would worry about is the large variation in magnification readings, which is possible because of the SEM's great depths of field and tilt angles. Calibrated latex spheres have been used for decades in TEM. Now larger calibrated spheres are available and these can be routinely and economically applied to SEM and light microscopy specimen to provide a reliable size comparison. Disclaimer: ProSciTech supplies latex particles (page "S2" online) and thus has a vested interest. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 09, 1999 4:21 PM, Zhiyu Wang [SMTP:zhiyuw-at-worldnet.att.net] wrote:
} Hi, All: } } A technical difficulty in my lab is coming on the table: } How to increase } resolution of digitized SEM images, especially for low } magnification } ( {50X). The pixel size of SEM image (50X)in my machine } (LEO-435VP) is 4.5 } um. In other word, no matter how good image software } performs, the } measurement error is at least 4.5 um. } We are going to use SEM as a routine measurement tool } under 100X, what is } the disadvantage? } Does any one have excellent idea to solve this problem, in } terms of : } Increase number of pixels and save as compressed .jpg to } reduce file size? } Stage mapping? } Software solution? } What else? } } Thank you for help } } Zhiyu Wang } } } }
Conn's Biological Stains, 9th edition edited by r.d. Lillie, page 248. under Diaminotriphenylmethanes. It is used as a vital dye so I woul 'assume' that it is not very toxic but there was no mention of toxicity.
-- Begin original message --
} From: Emidio FAZZINI {emifax-at-hotmail.com} } Date: Sat, 09 Jan 1999 09:56:24 -0600 } Subject: malachite green in aquaculture } 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 } -----------------------------------------------------------------------. } } } } Hi everybody, } } a Happy and prosperous new year to all! } I am a new subscriber and I work at a local branch of the Min. of Health } as a veterinary hygiene inspector in Italy. I am working on a } substitution of malachite green in aquaculture, but I would like to know } more about its nature, use and toxity, how dose it act? } I've recently learned that it's used as a dye in staining certain cell } tissues; surely somebody has posed himself the problem and has even } found some explanation. } Can somebody please give me some information or wher I can get it (ex. } Web Sites)? } Excusing me for having drifted you away from your prevalent work I } anticipate my thanks to those how will answer and a good luck to } evrybody! } regards } } } emidio fazzini } } (...up here from downunder!) } } } ______________________________________________________ } Get Your Private, Free Email at http://www.hotmail.com } } } }
-- End original message -- best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
Depending on the size of the spherulites, you can see them readily with polarized light microscopy, with no sample preparation. Also, by using a first order red plate, you can determine the sign of the spherulite.
If you can cross-section the part, you can readily see the skin effect and measure its thickness, again using light microscopy. While EM certainly has a place in the polymer lab, there is a tremendous amount to be learned from light microscopy, with much less sample prep and investment in equipment. I'd really suggest that you start here first.
There are a number of different alternatives, but first you need to make some choices regarding the level of measurement. For very delicate measurements (1/10th - 1/400th of a wave, or on the order of 10-50 nm), you will need to use a multiple beam interferometer with a monochromatic light source. If you just need more standard thicknesses (70 nm or higher), you can go with a regular dual beam interferometer, using a white light source or even just a good narrow band filter.
For general purposes, there are several add-on interferometers which can be used to replace a typical objective (most commonly, the 10x objective). The one I have used most is a small Michelson which used to be available through Hacker in this country. (I will fax you an information sheet).
Regarding multiple-beam/add ons: talk to Nikon. As I remember, in addition to a Michelson, they also make a Tolanksy. The challenge with the Tolansky is getting a reference mirror which has a reflectivity matched to the reflectivity of your sample. I haven't worked with the Nikon system for some years, but I seem to remember that they had a small turret, providing you with a choice of several different R values.
Re: larger systems:
Your choices in Europe tend to be much greater than ours in the US. If you can get your hands on a Reichert Polyvar Met on the used equipment market, they had a very respectable and easily used interferometer accessory. Reichert is now part of the Leica family, but I don't know if they have picked up this neat little attachment. Other, older microscopes which I have used include the Interphako from Jena (now Zeiss) and the Pluta, from PZO. The Interphako was amazingly inexpensive in comparison with its capabilities: in half shade mode it could readily measure to 10 nm. Finally, Leitz used to make the Linnik, the flagship of interferometers. I understand that these are no longer available, but, again, you can probably find one on the used-equipment market.
I am sure that I have missed someone (my apologies), but these comments should give you a starting point.
One more issue: Thin films are transparent and most of these (exception: Linnik and Interphako) work in reflected light. By mounting your sample on a front surfaced mirror, you can overcome this problem.
Phase contrast microscopes have two components: a ring which is placed in the condenser and a special phase-changing plate which is mounted in the objective. They can be viewed in the back focal plane of the objective either by removing the eyepiece and looking down the tube into the back of the objective or replacing the eyepiece with the centering telescope provided with the phase kit. Make sure that the microscope is set up for phase first. That is, make sure that the right ring is rotated or moved into position in the condenser and that you are using the matching phase objective.
In the back focal plane of the objective (BFPo), you will see the smoky phase ring from the objective's plate overlaying the bright ring in the condenser.
First, the principles:
1. a. The basic underlying concept behind phase contrast comes from Nature's kind provision for a fairly predictable delay in the light passing through biological and similar structures. These structures cause the light passing through them to lag by a quarter of a wavelength behind the light which passes through the surrounding material.
b. The microscope image is formed by the interference between light passing through the specimen and light passing through the background. For improved contrast (brighter brights and darker darks), the ideal situation would be for the light from the specimen to either be completely in step or to lag by half a wavelength. The intensity of the light in the image directly proportional to the SQUARE of the amplitude of the wave which results when the two waves are "added together". This is a bit difficult to explain without a drawing, but here's the gist:
If the waves are perfectly in step, the resulting,additive wave has an amplitude twice that of the original components and the light in the image is 2 squared or 4 times brighter.
If the waves are half a wavelength out of step by half a wavelength, then they meet peak to trough and cancel each other. That part of the image will have "zero intensity" and be dark.
All we need to do is design a microscope which controls the light from specimen and background to meet these conditions.
Now, for the purpose of each:
2. The function of the ring in the condenser is to carefully define what will be known as the "background light" and place it very specifically in the smokey area of the phase plate mounted in the objective. The smokey has a channel cut in it so that the background light has to go through less glass
3. When a portion of that light interacts with the sample it (a) scatters, to fill the whole back focal plane of the objective and (b) undergoes approximately a quarter of a wave shift.
4. When that "specimen light" reaches the phase plate, most of it goes through the thicker, non smokey part of the phase plate. The phase plate is designed so that the thicker section adds another quarter of a wave lag to the specimen light. (quarter wave lag from specimen) + (quarter wave lag from phase plate) = half wave lag required for destructive interference and improved contrast.
5. But why is the phase plate smokey? Because the light from the background is MUCH brighter than the light passing through the specimen. That is, its amplitude is much greater. For the two waves to destructively interfer, their amplitudes must match. The manufacturers coat the cut in the phase plate with neutral density material so that the background intensity is brought into range with the light coming from the specimen. You can see the effect in the phase image: the background is cut to about 15% its original intensity, resulting in a soft, pearly gray.
Finally: how can you fine-tune a phase contrast image?
Since the underlying principle is based on a difference in refractive index between the sample and its mounting, one possibility is to change the mountant. For your glass, for instance, you might find that there are terrible haloes around the glass particles when they are mounted in air. Try this test: mount some test samples (all the same refractive index) in (a) water, ri 1.33 (b) glycerin, ri 1.47 and (c) immersion oil, ri 1.55. You will see the image get crisper and cleaner as you move toward the immersion oil, which is a much better match for glass (typically on the order of ri 1.5152).
Secondly, this whole process is wavelength dependent, yet we never specified WHICH wavelength. Look in your phase kit for a green filter (if it is an interference filter, it will appear yellow and mirrored). Place this filter over the light port. It defines the wavelength for which your phase kit was built, usually 548 nm.
For more background (plus important diagrams), may we suggest "Optimizing Light Microscopy for Biological and Clinical Laboratories"? Even though it is biologically oriented, you will find a great deal of sound, basic information which will help you in your microscopy. Details are available on our website:
{ {http://www.MME-Microscopy.com/education} . Also, a reminder that the ACS course on Applied Microscopy for Chemists is just around the corner. Details are also available on the website.
I have been requested to forward this announcement to this group.
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
-----Original Message----- } From: Mancini [SMTP:mancini-at-bcm.tmc.edu] Sent: Sunday, January 10, 1999 6:24 PM To: CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU
Hello !
The female threads , which hold the cone screw that secures the specimen holder in our Ultracut E are partially stripped. One option is for our machine shop to rethread them. This would involve removing the heater and thermocouple wires that are at the top of the bridge. I was wandering if any of you had the same problem and how you went about solving it.
I also checked about replacing the whole bridge, but this option is quite expensive and there is a question about part availability.
I read an interview with a photojournalist who does all digital photography who saves her work on a Jaz drive "in the field"(PhotoMetro interview between ADColeman&Maggie Hallahan). She commented that "...the Jaz only lasts a few months and after that I have to archive everything to CD." Is this true? It's rather alarming as Iomega claims that the storage life of the cartridges(under *ideal* conditions) is 10 years. Someone is planning to supply our microscopy lab with a Jaz drive for image storage, but if it's not archival, I don't think it's worth the investment. Does anyone have experience with long term(at least a year) image storage on Jaz disks?
I am presently looking at the potential purchase of a basic SEM, probably variable pressure for a local museum. Are there any one of you using SEM for public shows? If yes may I have more information on what is exactly done.
The other alternative (more likely) is presentation of a "microzoo" similar to Oceanographic institute of Monaco. Small organisms (plancton, benthos, etc.) are presented via a fully motorized stereoscopic microscope. Any personnal experience to share on the subject? Ciao! -- L.Paul Bedard, ing. Ph.D. DocuScience inc.
Dear Stephen, I have successfully used lower kV electrons to reduce the volume of material excited for x-ray spectroscopy and a good Monte Carlo x-ray program, such as David Joy's, will help you to determine the volume excited. At 5 kV I was able to analyse 0.5 micron layers in a cross-sectioned semi-conductor. However, you must limit your analyses to those elements whose x-rays are excited by this energy of electrons, generally the energy of the x-rays is half or less than the energy of the electrons going in. I don't believe there is much, if any, resolution advantage to lower electron energy. At 200V, there will be very few x-rays excited that an EDX system can detect. You wrote: } Hello: Can anyone comment on the difference in the volume or resolution of } a point EDS analysis at 1kv or even 200V vs. the more standard 10 or 20kV. } If we were to use a FE-SEM at 200V could we expect a significant increase in } resolution of our EDS for a bulk sample compared our convention SEMs. Any } comments are greatly appreciated. Steve } } } ------------------------------ } Stephen McCartney Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
We embed vibratome sections of neural tissue that has been DAB treated between two pieces of Aclar. If a minimum of resin is placed between the sheets, the sections won't move much as a weight is applied to flatten the sections. There are different grades of Aclar, and I had to go directly to Allied Signal to get the one I prefer. The Aclar peels easily away from the polymerized resin, leaving one with a "section" that can be viewed with a LM for subsequent trimming. I super glue it to a blank beem capsule for thin sectioning. Best of luck, Randy -- Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own. http:\\www.uiowa.edu\~cemrf
Are there any references to descriptors for various morphology of grown crystals? We see several morphologies here, but I feel that I am only coming up with vegetable analogies - cauliflower, cabbage, etc..
While they sometimes capture the spirit of the picture, they seem technically inadequate. Are there technical terms that would be more appropriate to describe crystal clusters? Ideally, is there any reference (website etc.) where the terms are defined along with pictures?
If there is interest, I will post a summary. And thanks to those that offered leads on dispersing methods
Mohan Kalyanaraman
Sr. Staff Material Scientist Mobil Technology Company PO Box 480 Paulsboro, NJ 08066 609-224-3989 609-224-3608 (fax) mohan_kalyanaraman-at-email.mobil.com
I'm looking to see if someone knows anyone that prints cell images on slides--I need some black and white reference images of cellular material on a standard slide that can be used to calibrate some equipment. There are the usual problems with bio material that I really don't want to have to address since this is going to serve as a calibration standard. They will be viewed at 4x magnification, so they don't even need to be especially excellent reproductions. The biggest key (besides looking like cells) is that the printed image is *thin* (microns?), as I am trying to accurately determine the thickness of the coverslip/permount conglomeration that gets mounted on top of it.
Can anyone spit a quick Sudan IV protocol for lipid staining (for en bloc or embedded tissues-plant) to my email (or the net if anyone else is curious)?? Much obliged!! Tracey
Tracey Pepper Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
{html} Can anyone spit a quick Sudan IV protocol for lipid staining (for {i} en bloc {/i} or embedded tissues-plant) to my email (or the net if anyone else is curious)?? {br} {x-tab} {/x-tab} Much obliged!! {br} {x-tab} {/x-tab} Tracey {br} {br} {br} {br} {div} Tracey Pepper {/div} {div} Bessey Microscopy Facility {/div} {div} Iowa State University {/div} {div} ph: 515-294-3872 {/div} {div} fax: 515.294.1337 {/div} {/html}
I basically agree with what John says below, only stitching may be your only choice, if you really need a large field of view AND a high resolution.
I suppose, the resolution of 4.5 um comes from the fact, that you digitize a large field of view into a given number of pixels (for example, a field of view of about 9 mm digitized into 2Kx2K would give you such a resolution). The problem is, that going to 4K x 4K resolution may give you a better resolution but more likely is not. The reason is this:
For a digital image acquisition (and I believe, the 435 is digital), you divide your normal x and y sweep into the required number of voltage levels. The total sweep is normally of the order of a few Volts. If you divide that by a few thousand pixels, you end up with a few millivolts per pixel. Any noise of that level will essentially prohibit to keep the beam stationary to the level you would require. This is normally true independent of actual magnification as the digitization is done before the scan amplifier, which translates the x and y sweep into the actual voltages required to activate the scan coils.
So, to stay with the example above (9 mm field of view): if you need to digitize that to, let's say 1 um, you would need a resolution of 9000 x 9000 pixels. As I explained above, you probably would not be able to achive that without image stitching. Also, it would give you an 81 MB image (at 8 bits per pixel) or 162 MB (at 16 bits per pixel). I'm not sure you want to deal with that! John is absolutely right regarding lossy compression: Why spend a lot of money and effort to achieve high resolution to throw it all away through compression? Lossless compression may reduce the files by a factor of 2 or so.
We do have software and hardware for image acquisition and processing, especially but not limited to LEO microscopes, and we migh be able to help you with some of your challenges. If you need further info, please contact me through email.
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: info-at-soft-imaging.com
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. } ******************************************************* } ---------- } From: } "DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com[SMTP:"DrJohnRuss-at-aol.com"-at-sparc5. } microscopy.com] } Sent: Saturday, January 9, 1999 6:23 AM } To: zhiyuw-at-worldnet.att.net; Microscopy-at-sparc5.microscopy.com } Subject: Re: Resolution of digital SEM image } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I did not read the article, but perhaps the author was saying that when the jaz disk fills up, she archives to CD to clear out the jaz disk. This may be true, because the relative cost of a few CD's is nothing when compared to a jaz disk (a few dollars vs. over $100US). In this manner, she would only need to purchase one or two jaz disks. I have been archiving images on jaz disks for well over a year now and have had no problems with the file longevity.
Hope this helps,
David A. Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (800) 221-1975x2108
"Pauline C. Yu" {splene-at-pw.usda.gov} on 01/11/99 11:49:06 AM
Please respond to "Pauline C. Yu" {splene-at-pw.usda.gov}
To: Microscopy list {Microscopy-at-Sparc5.Microscopy.Com} cc: (bcc: David Bell/NA/Millipore)
I read an interview with a photojournalist who does all digital photography who saves her work on a Jaz drive "in the field"(PhotoMetro interview between ADColeman&Maggie Hallahan). She commented that "...the Jaz only lasts a few months and after that I have to archive everything to CD." Is this true? It's rather alarming as Iomega claims that the storage life of the cartridges(under *ideal* conditions) is 10 years. Someone is planning to supply our microscopy lab with a Jaz drive for image storage, but if it's not archival, I don't think it's worth the investment. Does anyone have experience with long term(at least a year) image storage on Jaz disks?
I have not used a Jaz drive, but if it is like the Zip (basically a floppy disk inside), I would not consider it an "archive" media. On the other hand, unless the enviroment was really poor, I would expect a much greater life than you mentioned. Also, the equipment capable of retreiving the data is limited. Will you have a working Jaz drive in 10 years? Another down side to the Jaz is the cost of storage compared to a CD-R (650 MB for {$1.50). I would suggest a CD-R archive for cost, longevity, and an increased probability that it can be read a few years down the road. It remains to be seen if the manufacturers are correct, but most rate the CD-R for 100 years or more storage.
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I read an interview with a photojournalist who does all digital photography who saves her work on a Jaz drive "in the field"(PhotoMetro interview between ADColeman&Maggie Hallahan). She commented that "...the Jaz only lasts a few months and after that I have to archive everything to CD." Is this true? It's rather alarming as Iomega claims that the storage life of the cartridges(under *ideal* conditions) is 10 years. Someone is planning to supply our microscopy lab with a Jaz drive for image storage, but if it's not archival, I don't think it's worth the investment. Does anyone have experience with long term(at least a year) image storage on Jaz disks?
by post.its.mcw.edu (8.8.8/8.8.8) with ESMTP id OAA25356 for {microscopy-at-sparc5.microscopy.com} ; Mon, 11 Jan 1999 14:48:35 -0600 (CST) Message-ID: {369A633B.6E10F13A-at-mcw.edu}
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Hi, dear all -
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max size, 2 Afterglow type, 150x135mm CRTs, 1 non-afterglow type120x90mm
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EM Facility Department of Microbiology Medical College of Wisconsin
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I forget the source, but a few months ago I recall a discussion on the web of CD longevity. It had been the conventional wisdom that CDs had a shelf life on the order of 30 years. But people were discovering that CDs 5 years old or so were becoming defective through delamination or other processes, just with age and not necessarily with handling, so there was some skepticism about the 30yr shelf life figure.
Like all media types, there is no question that after 10 years or so you might expect that the ability to read specific disks or cards or whatever would be difficult due to dramatic changes in technology (who has 8-track tapes any longer?) and the natural migration to new technologies that causes older technology to disappear. So any discussion of shelf life for electronic storage is probably a moot point (unlike film, which could be good almost indefinitely with proper handling). I fully expect to have to, sooner or later, transfer all of my archived data over to some new type of storage. Just the way life is...
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
Jim J Darley wrote: } } In STEM mode, which is also possible with some SEMs } (including yours). The important differences are: } The specimen is a section and this is penetrated by the } beam. The detector (photo multiplier) is below the } specimen.
One can also put a parallel EELS detector under the specimen and collect position-tagged spectra. This gives a very high-resolu- tion compositional image.
} Better still, in low contrast specimens contrast can be } increased at will - until electronic noise takes over. } Maybe the best use of STEM is in image analysis of soft } specimens.
Looking at the low-loss region of an EELS spectrum can dif- ferentiate among lipid, protein and nucleic acid, as reported a few years ago by Rich Leapman at MSA. This could be ideal for cryo- specimens or other unstained biological specimens. Yours, Bill Tivol
I am trying to locate Ms. Lisa Hartnell. We were colleagues a few years ago at RMC. After departing from RMC, Lisa worked for a while in Paul Webster's lab at Yale.
If anyone knows of Lisa's whereabouts or has an e-mail address for her, please contact me off-list. Or feel free to forward this message to her.
Thanks for your help!
Bob **************************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Research Microscopy Products *****************************************
It might be useful to contact the Royal Microscopical Society. I know that in the past they have displayed material on open access on a SEM with restricted access to controls. I haven't personally seen it, but read about it in the "RMS Proceedings". I think a manual SEM was used with covers over certain controls. A similar effect could be achieved using a simple PC control interface.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: "pbedard-at-saglac.qc.ca"-at-sparc5.microscopy.com {"pbedard-at-saglac.qc.ca"-at-sparc5.microscopy.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Hello all,
I am presently looking at the potential purchase of a basic SEM, probably variable pressure for a local museum. Are there any one of you using SEM for public shows? If yes may I have more information on what is exactly done.
The other alternative (more likely) is presentation of a "microzoo" similar to Oceanographic institute of Monaco. Small organisms (plancton, benthos, etc.) are presented via a fully motorized stereoscopic microscope. Any personnal experience to share on the subject? Ciao! -- L.Paul Bedard, ing. Ph.D. DocuScience inc.
I agree with Larry. If you want to "Archive" images there is only one stable medium - Conventional Film.
Jaz & Zip disks are fine for temporary storage of images. I have been using Jaz disks for a few years without any failures, but due to cost transfer data to CD-R's. The Jaz is only used for rapid access to the images. As pointed out the Jaz/Zip technology will probably be superseded soon ( 1GB drives are gone already ! ) and in a few years it will be difficult to read data if your drive fails. This was a major factor along with cost ) 5 years ago when we decided on a CD-R storage system, over Magneto-Optical which was the current favourite then.
CD-R's are not in themselves a good long-term archive medium. We have experienced a number of failures of CD-R's given to customers over the five years. These failures are usually attributed to environmental factors since the CD-R's are extremely sensitive to bad handling. Our archive has not experienced any failures in this time and the CD-R's are carefully stored in drawers. We always recommend that customers take their own copy so that a second copy exists. Any images lost over the five years were due to hard disk failures prior to storage. CD-R's are so cheap now that it is probably worth writing two disks at a time and storing them separately. At least five years on all the archive can still be read on any computer.
I suppose DVD ( 5.2 GB ) will be the next way to go as soon as industry settle on a universal standard ?
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Larry Allard {l2a-at-ornl.gov} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I confirm that my address is right. Please add it to the mailing list.
Thank you. --------------------------------------------------- Dr. J. Ricote Laboratoire du Physique et l'Etat Condens=E9 (LPEC) Facult=E9 des Sciences Universit=E9 du Maine-Le Mans Avenue Olivier Messiaen BP 535 72085 Le Mans cedex FRANCE
The 5-year life of CDs is for older technology CDs. Current CDs are nominally rated at 70-200 years, depending on the manufacturer. Magnetic media, regardless of how it is packaged (floppy, winchester, Zip, Jaz, SyQuest, whatever), all fall prey to the same problems that all magnetic media fall to -- archival for a decade or two, less than that under "real world" use.
For a good discussion of the CD longevity debate, see http://www.cd-info.com/CDIC/Industry/news/media-chronology.html.
For a good discussion of how these "100 year" lifetime determinations are made, see
I think that one has to remember that the environmental conditions are exquisitely important when discussing longevity. There *is no* single number that one can look to unless one knows how the media is stored and used.
For an example of how this plays for photographic media, look at http://www.kodak.com/cluster/global/en/consumer/education/imageStability.shtml
While the above is for photographic prints, not CDs or magnetic media, the importance of environmental issues are analogous.
billo
On Mon, 11 Jan 1999, Larry Allard 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 } -----------------------------------------------------------------------. } } } Woody and all: } } Just a couple of additional cents... } } I forget the source, but a few months ago I recall a discussion on the web } of CD longevity. It had been the conventional wisdom that CDs had a shelf } life on the order of 30 years. But people were discovering that CDs 5 } years old or so were becoming defective through delamination or other } processes, just with age and not necessarily with handling, so there was } some skepticism about the 30yr shelf life figure. } } Like all media types, there is no question that after 10 years or so you } might expect that the ability to read specific disks or cards or whatever } would be difficult due to dramatic changes in technology (who has 8-track } tapes any longer?) and the natural migration to new technologies that } causes older technology to disappear. So any discussion of shelf life for } electronic storage is probably a moot point (unlike film, which could be } good almost indefinitely with proper handling). I fully expect to have to, } sooner or later, transfer all of my archived data over to some new type of } storage. Just the way life is... } } Larry } } } } } } ------------------------------------------------------------------------ } } 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 Pauline, } } } } I have not used a Jaz drive, but if it is like the Zip (basically a floppy } } disk } } inside), I would not consider it an "archive" media. On the other hand, } } unless } } the enviroment was really poor, I would expect a much greater life than you } } mentioned. Also, the equipment capable of retreiving the data is limited. } } Will } } you have a working Jaz drive in 10 years? Another down side to the Jaz is } } the } } cost of storage compared to a CD-R (650 MB for {$1.50). I would suggest a } } CD-R } } archive for cost, longevity, and an increased probability that it can be } } read a } } few years down the road. It remains to be seen if the manufacturers are } } correct, but most rate the CD-R for 100 years or more storage. } } } } Woody } } } } ______________________________ Reply Separator } } _________________________________ } } Subject: Jaz disk archivalness? } } Author: "Pauline C. Yu" [SMTP:splene-at-pw.usda.gov] at CORP } } Date: 1/11/99 10:49 AM } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I read an interview with a photojournalist who does all digital } } photography who saves her work on a Jaz drive "in the field"(PhotoMetro } } interview between ADColeman&Maggie Hallahan). She commented that "...the } } Jaz only lasts a few months and after that I have to archive everything to } } CD." } } Is this true? It's rather alarming as Iomega claims that the storage } } life of the cartridges(under *ideal* conditions) is 10 years. Someone is } } planning to supply our microscopy lab with a Jaz drive for image storage, } } but if it's not archival, I don't think it's worth the investment. } } Does anyone have experience with long term(at least a year) image storage } } on Jaz disks? } } } } thanx } } Pauline Yu } } Microscopist Technician } } USDA-ARS-WRRC } } - - -- --- ----- -------- ------------- --------------------- } } } 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 } } 423-574-4981 } 423-574-4913 Fax } l2a-at-ornl.gov } } }
They do all sorts of materials printed on glass. We routinely use their image analysis calibration slide and stage micrometers as part of our on-site workshops.
Caveat: MME has no commercial interest in this product.
Suggest you see the Particle Atlas, available from McCrone, now in a very convenient CD Rom format which is quick and easy to search. Try contacting
Dina Mattes at McCrone Associates, Westmont, IL 800-622-8122.
There are also good descriptions of all sorts of crystal families and habits the old reference, Chamot (pronounced Cha-mo) and Mason's Handbook of Chemical Microscopy. I think McCrone Assoc. has also brought this back into print. It's a great reference because it shows you how to control crystal growth of various types under the microscope. Drs. Chamot and Mason reigned over the chemical microscopy facility at Cornell U. for nearly 100 years.... Their book contains lots of good "benchtop wisdom".
Many thanks to all who responded to my question. Below is a summary of the responses to cleaving rock salt:
1) Irradiate the salt (with a Co60 source, for example) to set up point defects in the crystal, making it easier to cleave.
2) Ensure you are using a high quality salt crystal.
3) Use a sharp, brand new razor blade for each cleave.
3) After cleaving, take a piece of lens cleaning paper, place it on a smooth surface, and put a little water on it. Using tweezers, pick up the salt and rub it in a circular motion in the puddle of water.
4) Consider using alternate substrates, depending on their suitability to the particular experiment on hand. Suggestions included mica, Barium sulfate, and Highly Ordered Pyrolytic Graphite (HOPG).
5) Be realistic in your expectations; even a very well cleaved salt crystal will have some cleavage steps.
Thanks again, these suggestions have been very helpful to me.
There was a wonderful microscopy exhibits at both the American Museum of Natural History (NY) and at the Smithsonian in the early-mid 80's. Cecil Fox, then with Armed Forces Institute of Pathology, helped organize both. The Smithsonian exhibit not only had a major display of old and new equipment but a working SEM. I have lost track of Dr. Fox ... last I heard he was still in the Silver Spring area. I have left a message for him and will send you info when available.
The other alternative is to talk to our illustrious listmaster, Nestor. He has a tremendous amount of experience with telemicroscopy which should be easily transferable to this type of situation.
Quoted from "Staining Procedures", 3rd ed. Biological Stain Commission pg 212: Sudan IV to show: suberized walls, cuticle, fat or oil globules
fix: either none or any botanical fixative embedding: paraffin or freehand
Preparation of staining and mounting solutions: Make a saturated solution of Sudan IV in 95% EtOH Add an equal volume of glycerin and filter if stain precipitates out on the sections, dilute further as necessary with a mixture of equal parts glycerine and EtOH Keep in a dropper bottle
Staining schedule: 1) Bring sections into 30% EtOH 2) Mount sections on a slide in a few drops of the staining and mounting solution 3) Seal edges of coverslip with "vas-par"*
Results: Cuticle, cutinized and suberized walls, and fat globules--orange.
*equal parts melted paraffin and vasaline; apply with a metal rod about 1/8th inch in diameter, bent into an L shape with a 6" handle and a 1" leg: warm applicator over a burner/hot plate, etc., then melt and pick up a few drops of vas-par and apply to coverslip edges. pg 239
Reference: Rawlins, T.E. 1933. "Phytopathological and Botanical Research Methods." John Wiley & Sons, NY.
(Pardon me while I go wipe off my keyboard.)
Phil
} Can anyone spit a quick Sudan IV protocol for lipid staining (for en bloc or } embedded tissues-plant) to my email (or the net if anyone else is curious)?? } Much obliged!! } Tracey } } } } Tracey Pepper } Bessey Microscopy Facility } Iowa State University } ph: 515-294-3872 } fax: 515.294.1337
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net
Thanks for the informative links. I was not aware of the extensive "debate" that has gone on regarding CD lifetime expectancy.
It is interesting that models suggest a lifetime of CD-R of 217 years. I think it is safe to assert that no one presently alive will be around to test this hypothesis. And if they were, it is probably safer to assert that there would be no device still in existance that could read such an ancient recording...
Larry
The 5-year life of CDs is for older technology CDs. Current } CDs are nominally rated at 70-200 years, depending on the } manufacturer. Magnetic media, regardless of how it is } packaged (floppy, winchester, Zip, Jaz, SyQuest, whatever), } all fall prey to the same problems that all magnetic media } fall to -- archival for a decade or two, less than that } under "real world" use. } } For a good discussion of the CD longevity debate, see } http://www.cd-info.com/CDIC/Industry/news/media-chronology.html. } } For a good discussion of how these "100 year" lifetime } determinations are made, see } } http://www.cd-info.com/CDIC/Technology/CD-R/Media/Kodak.html. } } I think that one has to remember that the environmental } conditions are exquisitely important when discussing } longevity. There *is no* single number that one can } look to unless one knows how the media is stored and used. } } For an example of how this plays for photographic media, } look at } http://www.kodak.com/cluster/global/en/consumer/education/imageStability.shtml } } While the above is for photographic prints, not CDs or magnetic } media, the importance of environmental issues are analogous. } } billo } } } On Mon, 11 Jan 1999, Larry Allard 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 } } -----------------------------------------------------------------------. } } } } } } Woody and all: } } } } Just a couple of additional cents... } } } } I forget the source, but a few months ago I recall a discussion on the web } } of CD longevity. It had been the conventional wisdom that CDs had a shelf } } life on the order of 30 years. But people were discovering that CDs 5 } } years old or so were becoming defective through delamination or other } } processes, just with age and not necessarily with handling, so there was } } some skepticism about the 30yr shelf life figure. } } } } Like all media types, there is no question that after 10 years or so you } } might expect that the ability to read specific disks or cards or whatever } } would be difficult due to dramatic changes in technology (who has 8-track } } tapes any longer?) and the natural migration to new technologies that } } causes older technology to disappear. So any discussion of shelf life for } } electronic storage is probably a moot point (unlike film, which could be } } good almost indefinitely with proper handling). I fully expect to have to, } } sooner or later, transfer all of my archived data over to some new type of } } storage. Just the way life is... } } } } Larry } } } } } } } } } } } ------------------------------------------------------------------------ } } } 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 Pauline, } } } } } } I have not used a Jaz drive, but if it is like the Zip (basically a floppy } } } disk } } } inside), I would not consider it an "archive" media. On the other hand, } } } unless } } } the enviroment was really poor, I would expect a much greater life than you } } } mentioned. Also, the equipment capable of retreiving the data is limited. } } } Will } } } you have a working Jaz drive in 10 years? Another down side to the Jaz is } } } the } } } cost of storage compared to a CD-R (650 MB for {$1.50). I would suggest a } } } CD-R } } } archive for cost, longevity, and an increased probability that it can be } } } read a } } } few years down the road. It remains to be seen if the manufacturers are } } } correct, but most rate the CD-R for 100 years or more storage. } } } } } } Woody } } } } } } ______________________________ Reply Separator } } } _________________________________ } } } Subject: Jaz disk archivalness? } } } Author: "Pauline C. Yu" [SMTP:splene-at-pw.usda.gov] at CORP } } } Date: 1/11/99 10:49 AM } } } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } I read an interview with a photojournalist who does all digital } } } photography who saves her work on a Jaz drive "in the field"(PhotoMetro } } } interview between ADColeman&Maggie Hallahan). She commented that "...the } } } Jaz only lasts a few months and after that I have to archive everything to } } } CD." } } } Is this true? It's rather alarming as Iomega claims that the storage } } } life of the cartridges(under *ideal* conditions) is 10 years. Someone is } } } planning to supply our microscopy lab with a Jaz drive for image storage, } } } but if it's not archival, I don't think it's worth the investment. } } } Does anyone have experience with long term(at least a year) image storage } } } on Jaz disks? } } } } } } thanx } } } Pauline Yu } } } Microscopist Technician } } } USDA-ARS-WRRC } } } - - -- --- ----- -------- ------------- --------------------- } } } } } } 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 } } } } 423-574-4981 } } 423-574-4913 Fax } } l2a-at-ornl.gov } } } } } }
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
Sorry Zhiyu Wang, I still have trouble with that concept. A single pixel in a line of 2500 pixel represents an error any microscopist could ignore. The pixel size may be 4.5um - on the charge coupled device; on the specimen, depending on how small the area examined (higher power less field) that pixel may represent rather more. What matters is the observed (enlarged) image and here, if the image width is 100mm, that single pixel represents 100 over 2500= 0.04mm or an 0.04% error. If only part of the image is used, like with a photographic negative the error does not change dramatically. One would not calculate a magnification by ignoring that only half of the enlarged image is used.
If you worry about magnification use a build in calibration, like latex spheres as earlier suggested. With care an SEM may be accurate to 5%, though some people have claimed 1%. In either case those pixel will not be your problem - the way I see it. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Tuesday, January 12, 1999 5:37 PM, Zhiyu Wang [SMTP:zhiyuw-at-worldnet.att.net] wrote: } Hi, Jim: } } Thank you for responding my message. Your calculation is } right, but the } question is that we do not want to ignore the smallest } unit on large scale } of measurement. Say 1/2500 is a small number in } persentage, but its } absolute value is 4 um, my sample is not covered by total } 2500 pixels, } only part of them instead. If I measure multipal samples, } the error should } be +/- 4 um, ie. 8 um, close to 0.01 mm. This is too } rough for quality } control of machinary (in semoconductor industry) } } Basiclly I do not believe SEM is a good machine for } dimension measurement } as many factors involve on imaging system. Therefore, its } high electron } optical resolution and high depth of view attract me and } my boss to do } something different. I am collect information around the } world. I } appreciate your help and we can share some interest idea } later. } } Thanks, } } Zhiyu Wang } ---------- } } From: Jim J Darley {jim-at-proscitech.com.au} } } To: 'Zhiyu Wang' {zhiyuw-at-worldnet.att.net} ; } 'Microscopy-at-sparc5.microscopy.com' } {Microscopy-at-Sparc5.Microscopy.Com} } } Subject: RE: Resolution of digital SEM image } } Date: Monday, January 11, 1999 4:28 AM } } } } } } --------------------------------------------------------- } } --------------- } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } } ml } } ---------------------} } } } } Hi Zhiyu Wang - } } Lets hope its me who is confused: I don't care. } } If a monitor is 100mm across and is represented by 4um } } pixel } } (100 divided by 0.004=) 2500 would be required for a } } single } } line. If one pixel was missing I would just forget about } } } } that, although the percentage error would be constant, } } regardless of magnification. } } What I would worry about is the large variation in } } magnification readings, which is possible because of the } } } } SEM's great depths of field and tilt angles. } } Calibrated latex spheres have been used for decades in } } TEM. } } Now larger calibrated spheres are available and these } } can } } be routinely and economically applied to SEM and light } } microscopy specimen to provide a reliable size } } comparison. } } Disclaimer: ProSciTech supplies latex particles (page } } "S2" } } online) and thus has a vested interest. } } Cheers } } Jim Darley } } ProSciTech } } Microscopy } } PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } ********************** www.proscitech.com.au } } ***** } } } } } } } } On Saturday, January 09, 1999 4:21 PM, Zhiyu Wang } } [SMTP:zhiyuw-at-worldnet.att.net] wrote: } } } } } Hi, All: } } } } } } A technical difficulty in my lab is coming on the } } } table: } } } How to increase } } } resolution of digitized SEM images, especially for } } } low } } } magnification } } } ( {50X). The pixel size of SEM image (50X)in my } } } machine } } } (LEO-435VP) is 4.5 } } } um. In other word, no matter how good image } } } software } } } performs, the } } } measurement error is at least 4.5 um. } } } We are going to use SEM as a routine measurement tool } } } under 100X, what is } } } the disadvantage? } } } Does any one have excellent idea to solve this } } } problem, } } in } } } terms of : } } } Increase number of pixels and save as compressed .jpg } } } to } } } reduce file size? } } } Stage mapping? } } } Software solution? } } } What else? } } } } } } Thank you for help } } } } } } Zhiyu Wang } } } } } } } } } } } } } }
} Bill: } } Thanks for the informative links. I was not aware of the extensive } "debate" that has gone on regarding CD lifetime expectancy. } } It is interesting that models suggest a lifetime of CD-R of 217 years. I } think it is safe to assert that no one presently alive will be around to } test this hypothesis. And if they were, it is probably safer to assert } that there would be no device still in existance that could read such an } ancient recording... } } Larry }
I'm not sure how good these models are, though. It's a little like mutagenesis tests on bacteria. Yeah, it's a measure of carcinogenicity, but it's a loose one. I'm much happier using these tests as a relative scale rather than an absolute one.
Hello: I am currently working on mechanical behavior / microstructure correlations in single colony near apha Ti Alloys. I am currently preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but have run into the problem of a very uneven alpha phase morphology, the alpha phase has island formation - we are using a cold stage to minimize the hydride formation at the interface. I am using 3 micron/6micron diamond paste during the dimpling process - the foil itself otherwise is fairly clean in terms of the dislocation content. Could anyone in the microscopy land have some suggestions in terms of what the problem may be?
thanks - if you want you can respond directly to suri.3-at-osu.edu
Pauline Yu wrote: ============================================ I read an interview with a photojournalist who does all digital photography who saves her work on a Jaz drive "in the field"(PhotoMetro interview between ADColeman&Maggie Hallahan). She commented that "...the Jaz only lasts a few months and after that I have to archive everything to CD." Is this true? It's rather alarming as Iomega claims that the storage life of the cartridges(under *ideal* conditions) is 10 years. Someone is planning to supply our microscopy lab with a Jaz drive for image storage, but if it's not archival, I don't think it's worth the investment. Does anyone have experience with long term(at least a year) image storage on Jaz disks? ============================================== When the 1 GIG Iomega drives came out some months ago, I thought then they were the greatest thing since sliced bread. I have used one on my home desktop repeatedly without problems. And ditto for some number of the office desktops (except for one).
But another one that I dedicated for use with my laptop for when I travel has been another story. During the warranty period, when I could get through on the Iomega lines, they did keep replacing the entire drive, which apparently did not survive the bouncing around during travel. The drive itself was packed in my checked luggage but well packed. But the survival rate was about three months. After the warranty period was up, they stopped replacing them.
In any case, my experience seemed to be consistent with that of the photojournalist: If you carry them around, you have problems. But so far as I can tell, after very heavy use of the 1 GIG JAZ drives, if they are permanently installed, we have not had any particular problems. In talking with their customer service people, apparently, something goes wrong with the drive if carried around and when that happens, it does do damage to the replaceable disc when you next try to use it.
Chuck
Disclaimer: In this instance, I have no financial interest in this product, just a satisfied user who wished Iomega had more customer service lines.
=================================================== 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: www.2spi.com ############################ ==================================================
"In Situ Hydride Formation in Zirconium and Titanium during Ion Milling" Graham J.C. Carpenter et al, JMSA Vol.1 No. 4 pp 175-184 1995.
=46rom the paper you will see that a key factor in eliminating hydride formation is having a clean sample free from hydrocarbon contamination. =
This contamination could be remnants from the dimpling process or other pre-thinning steps or it could be caused by the back streaming of diffusi= on pump oil in your ion mill. As titanium has a high chemical affinity for hydrogen, you may want to look over your preparation steps and try to eliminate any areas of possible contamination. If you are still having a=
problem, a quick cleaning of both the specimen and the specimen holder in= a Plasma Cleaner should take care of it.
If you have an interest, I can send you a copy of the above referenced paper. I also have papers on plasma cleaning which may be of interest.
NOTE: South Bay Technology does manufacture the PC150 Plasma Cleaner and therefore I have a vested interest in promoting its use.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Satyarth Suri } Hello: I am currently working on mechanical behavior / microstructure correlations in single colony near apha Ti Alloys. I am currently preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but have run into the problem of a very uneven alpha phase morphology, the alpha phase has island formation - we are using a cold stage to minimize the hydride formation at the interface. I am using 3 micron/6micron diamond paste during the dimpling process - the foil itself otherwise is fairly clean in terms of the dislocation content. Could anyone in the microscopy land have some suggestions in terms of what the problem may be?
thanks - if you want you can respond directly to suri.3-at-osu.edu
Pauline, We have had a horrible time with the Jaz drives (hooked to an SGI archiving confocal images). Apparently when they get near full capacity they screw up. I've sent three unreadable disks to an image retrieval company, with no luck. The told me that when the disk is near full and you try to add another file it will put data back at the begining of the disk, erasing file allocation tables, basically making the disk unreadable. We have several defective disks and I have been forced to the zip drive.
Hello, My lab is currently looking for a used carbon coater for our TEM lab. Our Emitech has passed away and is in England trying to be repaired, but it looks grim. I also have a Denton coater that will not pump down. So I am looking into other options.
If your are selling such a machine or might know someone who is, please write back or drop a line to our website
I need some advice as to the best protocol for fixing fresh plant leaves for routine TEM. My only experience has been with animal tissue. Thanks, MG Engle
We just got a G3 Mac and an Epson digital projector-I have seen some really nice digital slide shows and am excited by the increase in quality and decrease in effort making slides. I expect, however, to spend some effort finding out the best way to put together a slide show, and am wondering what advice or experience might be out there. I would like to show ~10MB images without any computer stuff showing at the same time, but could consider compression (eg., jpeg) if there is no loss of quality. Wondering whether to get into a slide presentation program or just make a stack and open them sequentially (I have 190+ MB of memory). There is also a neat IR pointer that seems to bounce off the screen and feed into software in the projector that allows you to move the cursor around and click/double click with the IR pointer. Too bad they didn't include a laser pointer in this device!....Many thanks for any help or advice!...Tom Reese
Reply to: RE: Ti Alloys - TEM foil preparation Pure titanium has been jet electropolished here at Argonne National = Laboratory for over 10 years. A South Bay 550-B single jet instrument is = used because it has in-situ viewing of the specimen during the process to = speed up the determination of proper electropolishing conditions.
Conditions: -20 degrees C. 70 volts, 35 mA, (one side of 3 mm disc)
Note: After polishing about half way thru the specimen,=
it is cleaned, dried, and a dot of = Microshield stop- off lacquer is placed on the polished "side = one" dimple. The inverted specimen is then remounted on = the polish- ing instrument and thinned to perforation = using the = sensitive optical termination system. It = should be = possible to make several foils per day in = this manner.
Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il, 60439
E-mail: bkestel-at-anl.gov South Bay Technology wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
{HTML} {HEAD} {/HEAD} {BODY} {PRE WIDTH=3D"132"} Reply to: RE: Ti Alloys - TEM foil preparation
{/PRE} {FONT = FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} Pure titanium has = been jet electropolished here at Argonne = National Laboratory for over 10 years. = A South Bay 550-B single jet instrument is = used because it has in-situ viewing of the = specimen during the process to speed up = the determination of proper electropolishing = conditions. {BR} {BR} Electrolyte: = 30 ml. perchloric acid {BR} = 295 ml. methanol {BR} = 175 ml. butyl cellosolve {BR} {BR} = Conditions: -20 degrees C. {BR} = 70 volts, = 35 mA, (one side of 3 mm disc) {BR} {BR} = Note: After polishing about = half way thru the specimen, {BR} = it is cleaned, dried, = and a dot of Microshield stop- {BR} = off lacquer is placed = on the polished "side one" dimple. {BR} = The inverted = specimen is then remounted on the polish- {BR} = ing instrument = and thinned to perforation using the {BR} = sensitive optical = termination system. It should be {BR} = possible to make = several foils per day in this manner. {BR} {BR} = Bernard Kestel {BR} Materials = Science Division {BR} Argonne National = Laboratory {BR} Argonne, Il, 60439 {BR} {BR} = E-mail: bkestel-at-anl.gov {BR} South = Bay Technology wrote: {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#= 000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR} I also have papers on plasma cleaning which = may be of interest. {BR} > {BR} >NOTE: South = Bay Technology does manufacture the PC150 = Plasma Cleaner and {BR} >therefore I have = a vested interest in promoting its use. {BR} > {BR} >Best = regards- {BR} > {BR} >David = {BR} >Writing at 9:38:23 = AM on 1/12/99 {BR} > = {BR} >***************************************************************** {BR} >********** {BR} >************************ {BR} > {BR} >David = Henriks = TEL: {BR} >800-728-2233 (toll = free in the USA) {BR} >South Bay Technology, = Inc. +1-949-492-2600 {BR} >1120 = Via Callejon = FAX: +1-949-492-1499 {BR} >San Clemente, = CA 92673 USA e-mail: {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} henriks-at-southbaytech.com {/U} {= /FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} > {BR} >***************************************************************** {BR} >********** {BR} >************************ {BR} > {BR} > = >>>>> Please visit us = at {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.= southbaytech.com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} <<<<< {BR} > {BR} >Manufacturers = of precision sample preparation equipment = and supplies for {BR} >metallography, crystallography = and electron microscopy. {BR} > {BR} >Message = text written by Satyarth Suri {BR} >> {BR} >Hello: {BR} >I = am currently working on mechanical behavior = / microstructure {BR} >correlations in single = colony near apha Ti Alloys. I am currently {BR} >preparing = the foils using a dual ion mill (6kV, 12deg, = 1mA), but {BR} >have run into the problem = of a very uneven alpha phase morphology, {BR} >the = alpha phase has island formation - we are = using a cold stage {BR} >to minimize the = hydride formation at the interface. I am = using {BR} >3 micron/6micron diamond paste = during the dimpling process - the {BR} >foil = itself otherwise is fairly clean in terms = of the dislocation {BR} >content. Could = anyone in the microscopy land have some = suggestions {BR} >in terms of what the problem = may be? {BR} > {BR} >thanks - if you want = you can respond directly to {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D1 COLOR=3D"#0000FF"} {U} suri.3-at-osu.edu {/U} {/FONT} {FONT FACE=3D"= Geneva" = SIZE=3D1 COLOR=3D"#000000"} {BR} > {BR} >-satyarth {BR} > {BR} >< {BR} > {BR} > {BR} > {BR} >RFC822 = header {BR} >----------------------------------- {BR} > {BR} > = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id MAA28629; = Tue, 12 Jan 1999 12:21:30 -0600 {BR} > Received: = from Sparc5.Microscopy.Com (sparc5.microscopy.com = {BR} >[206.69.208.10]) by dns2.anl.gov = (8.9.1a/8.6.11) with SMTP id MAA02315; Tue, = 12 Jan 1999 {BR} >12:22:10 -0600 (CST) {BR} > = Received: (from {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = daemon-at-localhost {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) {BR} >id LAA19027 for dist-Microscopy; = Tue, 12 Jan 1999 11:48:51 -0600 {BR} > Received: = from no_more_spam.com (Sparc5 [206.69.208.10]) = by {BR} >Sparc5.Microscopy.Com (8.6.11/8.6.11) =
Hi Guys, I'm looking for a shareware program for the mac that will allow me to batch process 12 bit images to 8 bit images. Thanks,
--Ciprian ________________________________________________________________ Ciprian A. Almonte Phone: (412) 648-9796 Center for Biologic Imaging FAX: (412) 648-8330 University of Pittsburgh URL:http://sbic6.sbic.pitt.edu Pittsburgh, PA 15261 USA mailto:calmonte-at-pitt.edu ________________________________________________________________
American Chemican Society "Applied Optical Microscopy for Chemists" March 4-5-7 in Beautiful Orlando, Florida Three days of total immersion, hands-on experience with all phases of optical microscopy, including one whole day of Polarized Light (both qualitative and quantitative) Strongly supported with the latest in equipment from the major manufacturers for HANDS-ON labs Special Saturday evening session on video imaging and image analysis
For further details, including registration information, visit the MME website: {http://www.MME-Microscopy.com/education}
Looking forward to seeing you there!
Barbara Foster, Course Coordinator
Microscopy/Microscopy Education ...Educating microscopists for greater productivity. 125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
This is a multi-part message in MIME format. --------------6C36AB32CC464C8C85112D45 Content-Type: text/plain; charset=iso-8859-9 Content-Transfer-Encoding: 7bit
Dear all microscopists;
I am a young electron microscopist who is trying to organise an EM lab in Turkey.
I had a chance to study with a wonderful electron microscopist and also a great teacher, Dr. Robert J.Kayton(the president of PNEMS) in Portland, OR for 1 year. I learned the technical aspects of electron microscopy from him. I have never had much more enjoyable experience than that. Now, I am back and have been asked to organise an EM lab.
All I have in this lab is an old TEM(Carl Zeiss EM 9) and an old ultramicrotome. As you see, I need so many things. I really want to rebuilt a nice EM lab and to apply my experience I have learned in U.S. It will be very hard for me since it is not easy to find financial support for that in my country.
Would you like to help me? I would like to ask you to send me any equipments, appliances, dark room stuffs, books etc. which you don't use anymore in your EM lab. I will prepare a chart for being able to thank all of you on the front door of my lab and write the names of persons, companies, facilities etc. which help me. I also would like to bring small presents from Turkey for you, dear helpfull electron microscopists, when I come to the Microscopy&Microanalysis'1999,in Portland,OR.
I will greatly appreciate any kind of help.
Thanks in advance,
Ranan Gulhan AKTAS, M. D. Trakya University, Faculty of Medicine Pathology Department 22030 Edirne, TURKEY Tel: +90 284 235 44 68 Fax: +90 284 235 76 52 e-mail: ranaoz-at-turk.net
Unless you have a compelling reason to use ion milling for this preparation, I recommend using jet electropolishing, possibly with just a final touch-up in the ion mill. Although both electropolishing and ion milling commonly produce sample preparation artifacts, the artifacts from ion-milling reactive metals such as Ti and Zr are not well understood and can be harder to eliminate. Despite Carpenter et. als work (see D. Henriks posting) implicating hydrocarbon contamination as a source of hydriding during ion milling, my personal experience ion milling Zr samples in 'oil-free' systems indicates that water vapor is the main culprit in the hydridng. Surface irregularities on ion milled Zr/Ti samples (surface terracing, for example) usually have other causes, possibly related to air leaks in the ion mill or oxygen in the feed gas. Another source of irregular surfaces on ion milled samples is embedded polishing compound. Cubic boron nitride (CBN) sometimes gives better results in dimpling metals than diamond, and even the diamond polishing compounds from different suppliers can give different results. Conversely, brief ion milling of electropolished samples--especially at low incidence angles and very low kV--can improve the surface finish and mill away irregularities caused by second-phase particles. Often the 'trick' to eliminating milling artifacts is minimizing time spent in the ion mill.
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA
---------- From: South Bay Technology Sent: Tuesday, January 12, 1999 6:00 PM To: Satyarth Suri; Microscopy ListServer Subject: Ti Alloys - TEM foil preparation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Dear Satyarth:
An excellent paper on the subject is:
"In Situ Hydride Formation in Zirconium and Titanium during Ion Milling" Graham J.C. Carpenter et al, JMSA Vol.1 No. 4 pp 175-184 1995.
From the paper you will see that a key factor in eliminating hydride formation is having a clean sample free from hydrocarbon contamination. This contamination could be remnants from the dimpling process or other pre-thinning steps or it could be caused by the back streaming of diffusion pump oil in your ion mill. As titanium has a high chemical affinity for hydrogen, you may want to look over your preparation steps and try to eliminate any areas of possible contamination. If you are still having a problem, a quick cleaning of both the specimen and the specimen holder in a Plasma Cleaner should take care of it.
If you have an interest, I can send you a copy of the above referenced paper. I also have papers on plasma cleaning which may be of interest.
NOTE: South Bay Technology does manufacture the PC150 Plasma Cleaner and therefore I have a vested interest in promoting its use.
David Henriks TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Satyarth Suri } Hello: I am currently working on mechanical behavior / microstructure correlations in single colony near apha Ti Alloys. I am currently preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but have run into the problem of a very uneven alpha phase morphology, the alpha phase has island formation - we are using a cold stage to minimize the hydride formation at the interface. I am using 3 micron/6micron diamond paste during the dimpling process - the foil itself otherwise is fairly clean in terms of the dislocation content. Could anyone in the microscopy land have some suggestions in terms of what the problem may be?
thanks - if you want you can respond directly to suri.3-at-osu.edu
I am compiling for the MSA Education Committee a listing of laboratories involved in outreach and/or remote access programs with local schools or just across campus. Please contact me with information about your respective programs. This list will be mounted on the MSA webpage.
I will be compiling lists for scanning electron microscopes, light microscopes(standard and confocal), scanning probes, confocal, and transmission microscopes. I'ld like to know what school levels your program is tartgeted to, what instruments you are using, a web site if you have one, and your email so I can follow up on the information.
Please contact me directly at sbarlow-at-sunstroke.sdsu.edu (NOT THE LISTSERVER LIST) and I will see the information is compiled and posted.
thanks in advance
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
I agree with Chuck. When I took Dr. McKenzie's Digital Microscopy course, he informed us that these things couldn't be banged around. We've had a 1 GB (permanently installed) for over a year and a 2 GB "portable" drive (that never gets moved) for almost a year, and have never had any trouble with either.
I don't know about over filling disks. My fattest one still has 170 MB left and is perfectly fine. For really important files/pictures, I keep 2 backups on separate disks. We have about 20 of these things. We also use Zips as a more portable medium. Go iomega.
No commercial interest; just a satisfied customer.
Sara
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} Dear All, } } Is anyone aware of an electropolishing solution for Mg alloys that (i) } thins very slowly, and (ii) works at room temperature? } } thanks. } Sincerely yours
D. H. Ping
------------------------------------------------------------------------- D. H. Ping, Dr Materials Physics Division National Research Institute for Metals (NRIM) 1-2-1 Sengen, Tsukuba 305-0047, Japan Phone: +81-298-59-2717 FAX:+81-298-59-2701 E-mail: Ping-at-tamamori.nrim.go.jp WWW: http://inaba.nrim.go.jp/apfim/ Home phone: +81-298-59-0918
Can anyone out there tell me a bit about endpoint detection systems for Reactive Ion Etchers? What different types there are, how they work, what situations they are optimal for, and {shudder} approximate price ranges? I was told to write up a summary within the week regarding the possibility of either attaching one to our existing RIE (PlasmaTherm Batchtop) or getting another. (Wow! They actually came to me and offered to buy stuff, instead of me having to beg!!! Either I'm doing stuff really right, or really wrong!)
Much thanks in advance,
Marisa Ahmad R&D Specialist mahmad-at-semiconductor.com
"It's not how hard you fall, it's how high you bounce."
Much as I hate to say it, Microsoft PowerPoint does a pretty good job with digital slide presentation. You definitely do *not* want to simply make a stack of slides and open them independently...it will seem clumsy and will be distracting. PowerPoint opens slides instantly, and you can do a lot of tricks to presumably enhance the production (although sometimes the tricks are somewhat distracting themselves). The best thing is to have a PowerBook with XGA (1024 x 768) resolution, and a projector with the same XGA resolution, to get the best projection results. We have been using digital projection almost exclusively for over a year now with this setup, and it works extremely well. The best thing is that you can be making slides just about up to the minute of your talk...a great capability for those of us who tend to procrastinate just a teeny bit... ;-).
Larry
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Hi Tom, Firstable congratulation on you new G3, awesome machine. I recommend Graphic converter for slide show. It's pretty easy to set up and it allow you to fade down the images. If you need help seeting it up let me know. } } We just got a G3 Mac and an Epson digital projector-I have seen some really } nice digital slide shows and am excited by the increase in quality and } decrease in effort making slides. I expect, however, to spend some effort } finding out the best way to put together a slide show, and am wondering } what advice or experience might be out there. I would like to show ~10MB } images without any computer stuff showing at the same time, but could } consider compression (eg., jpeg) if there is no loss of quality. Wondering } whether to get into a slide presentation program or just make a stack and } open them sequentially (I have 190+ MB of memory). There is also a neat IR } pointer that seems to bounce off the screen and feed into software in the } projector that allows you to move the cursor around and click/double click } with the IR pointer. Too bad they didn't include a laser pointer in this } device!....Many thanks for any help or advice!...Tom Reese
--Ciprian
_____________________________________________________________ Ciprian A. Almonte Phone: (412) 648-9796 University of Pittsburgh FAX: (412) 648-8330 Center for Biologic Imaging http://sbic6.sbic.pitt.edu Pittsburgh, PA 15261 USA mailto:calmonte-at-pitt.edu _____________________________________________________________
The problem with overfilling Jaz disks sounds like more of a software problem than a problem with the Jaz. I have filled a number of Jaz disks to capacity and simply get a message telling me it's too full. This has worked with both PC's and Unix systems.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Sara Miller {saram-at-duke.edu} To: Garber, Charles A. {cgarber-at-2spi.com} Cc: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
At 10.46 -0500 99-01-12, Satyarth Suri wrote: } } Hello: } I am currently working on mechanical behavior / microstructure } correlations in single colony near apha Ti Alloys. I am currently } preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but } have run into the problem of a very uneven alpha phase morphology, } the alpha phase has island formation - we are using a cold stage } to minimize the hydride formation at the interface. I am using } 3 micron/6micron diamond paste during the dimpling process - the } foil itself otherwise is fairly clean in terms of the dislocation } content. Could anyone in the microscopy land have some suggestions } in terms of what the problem may be?
Why can't you electropolish the specimens? For my PhD work we prepared a lot of (commercial purity alpha) Ti specimens using electropolishing with minimal hydride formation. This has been continued by another PhD student who also did work on Ti at the University of Birmingham (England).
We used a non-acid electrolyte developed by B.J. Kestel of Argonne National Lab. He recommends using it in a South Bay Technology single jet electropolisher but we made it work with a Struers Twin jet Tenupol, I guess you could probably make it work with a variety of other jet electropolishing devices. I can give further details of our procedure if anyone wants.
Hope this helps
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren PLEASE NOTE NEW CONTACT DETAILS: Department of Experimental Physics Chalmers University of Technology SE-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 =46AX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
I am polishing a low carbon steel that is coated with zinc. The situation i'm experiencing is when i try to etch the base material with 2%nital for grain size determination i seam to get some effect from the zinc in that the area near the surface does not etch. Is it because of polishing practice dragging that zinc onto the surface?
even if you do use something like 'Powerpoint' for the final slide presentation, it might be worth seeing if you can get hold a program such as 'Thumbsplus'. It's shareware for the PC but I don't know if it's available for the Mac. I have found 'Thumbsplus' is much quicker for simple image management and filing images because of it's use of thumbnail images and it can even produce a basic slide show.
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------------------------------------------------------------------------- ----------------- Tom:
Much as I hate to say it, Microsoft PowerPoint does a pretty good job with digital slide presentation. You definitely do *not* want to simply make a stack of slides and open them independently...it will seem clumsy and will be distracting. PowerPoint opens slides instantly, and you can do a lot of tricks to presumably enhance the production (although sometimes the tricks are somewhat distracting themselves). The best thing is to have a PowerBook with XGA (1024 x 768) resolution, and a projector with the same XGA resolution, to get the best projection results. We have been using digital projection almost exclusively for over a year now with this setup, and it works extremely well. The best thing is that you can be making slides just about up to the minute of your talk...a great capability for those of us who tend to procrastinate just a teeny bit... ;-).
Larry } } We just got a G3 Mac and an Epson digital projector-I have seen some really nice digital slide shows and am excited by the } increase in quality and decrease in effort making slides. I expect, however, to spend some effort finding out the best way to } put together a slide show, and am wondering what advice or experience might be out there. I would like to show ~10MB } images without any computer stuff showing at the same time, but could consider compression (eg., jpeg) if there is no loss of } quality. Wondering whether to get into a slide presentation program or just make a stack and open them sequentially (I have 190+ } MB of memory). There is also a neat IR pointer that seems to bounce off the screen and feed into software in the projector that } allows you to move the cursor around and click/double click with the IR pointer. Too bad they didn't include a laser pointer in this } device!....Many thanks for any help or advice!...Tom Reese
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
In message {3306033D.951200C9-at-mauromedia.com} , Michael Draper {quex-at-mauromedia.com} writes } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE: Ti Alloys - TEM foil preparation Re: Electropolishing of Ti alloys = Due to interest in this subject, I will list some additional = electrolytes I have used on Ti alloys. (3mm, jet polishing).
Ti-8 w/% Al 13% HCL -60 degrees C Ti-811 87% methanol 70 to 90 volts Ti-6Al-4V 25-35 mA.
Ti-13 Sn 130 ml HCL -50 degrees C 670 ml methanol 150 volts 100 ml butyl 60 mA.
Ti-3V 60 ml perchloric acid -50 degrees C Ti-20 Zr 590 ml methanol 50 volts Ti-14 Al 350 ml butyl cellosolve 10-15 mA
Ti-8 Al 60 ml perchloric acid -15 t0 -20 C (rolled) 460 ml ethyl alcohol, 95% 180 volts 280 ml butyl alcohol 40 mA. 100 ml butyl cellosolve = Ti 5.3 g. lithium chloride -40 degrees C. nanocrystals, 11.16 g. magnesium 150 volts compacted chloride 30 mA 500 ml. methanol 100 ml. butyl cellosolve (Note: the two salts above must be added to the combined solvents one = "powder" at a time, while stirring. The two dry salts react violently if = mixed together. This electrolyte has nearly as wide an application as = perchloric acid mixtures and is known as B K-2). I suspect that B K-2 will cause the least hydride problem. It was = developed to eliminate hydrides in vanadium TEM foils. The voltages given = would need to be reduced about 20% for a twin jet system due to lower = electrical resistance of that configuration. My work was done on a South = Bay vertical single jet unit. I have no vested interest in the above equipment, but have enjoyed = the ease with which good polishing conditions can be obtained and = reproduced since 1975 or so. Good luck! = Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
E-mail: {bkestel-at-anl.gov} Thomas, Larry wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
by mail.unixg.ubc.ca with esmtp (Exim 1.92 #2) for Microscopy-at-Sparc5.Microscopy.Com id 100TFD-0006xh-00; Wed, 13 Jan 1999 08:34:12 -0800 X-Sender: ech-at-pop.unixg.ubc.ca Message-Id: {v0300780000e8124c7823-at-[137.82.136.14]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: Microscopy-at-Sparc5.Microscopy.Com
Following the discussion about filling Jaz discs:
We collect images from a Bio-Rad MRC 600 confocal to Jaz disc on an IBM computer and transfer to a Jaz drive on a Mac G3 for making projections and plates with NIH Image and Photoshop. We used to transfer by ethernet but it was too slow and taking a disc out of one drive and putting into another drive is by far the quickest approach. Using the Mac G3 to work on the images allows another researcher to collect their images on the confocal and doesn't hold them up while data is being transfered.
We once lost 1GB of data because the user filled up the Jaz disc completely. Now we wouldn't lose that data because as I understand it: When you transfer to a Mac, there is a Mac header put onto the disc. If the disc is full, this header overwrites the PC directory tree. The IBM doesn't recognise the disc and the Mac thinks it is empty. The solution (thanks to Eric Jervis) is to put the Jaz disc back into the IBM. Look for hidden files, delete them, and use Norton Utilities to rebuild the tree directory. We have a rule now which is "Try to leave 10% free on a disc," then this problem never arises.
We use Jaz and Zips regularly as temporary storage and for quick transfers between computers but we always archive to CD. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
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Hey Ciprian,
I'm not aware of a shareware tool, however, commercially IPLab (http://www.iplab.com/) can do it, and I believe so can ImportAccess (http://www.desacc.com/). Good luck.
Brian C. Tryon MCP-Hahnemann School of Medicine Ann Preston Hall, Box 551 3300 Henry Avenue Philadelphia, PA 19129 USA
----------------------------------------- "Quantifying is a committing task." - Cruz-Orive, 1994.
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." - Richard Feynman --------------------------------------------------------------
I travel quite a bit with my Jaz, and have not had the same experience mentioned earlier with drive problems. For what it's worth, when traveling I carry 2 carry-ons, one for my laptop, the other (a cool bag from Kensington) carries any drives, media, and accessories. I rarely pack my drives in checked luggage (my paranoia). One reason I carry on all drives (besides brutish bag-manglers, boy, do I have stories) is because I believe this avoids some of the temperature fluctuations that you might experience with checked bags. I've been to some cold regions and noticed luggage items quite cold when upacked. Typically, I'm working on something just prior to catching a flight so going from warm to cold couldn't help drive performance. Likewise, I typically take some dessicant and moisture-proof bags with me, quickly wrap the drive in, throw in a bag of dessicant and I believe this helps avoid condensation on the drive and when I get to my destination, I can get right to work without worry of letting the drive reach room temp or needing a papertowel to drive off the casing.
So, I've had Jaz drives (I currently have 3) since they came out, never a problem with the drive. I do routinely check the disk with disk utility programs, and have occasionally (once every 2 months with routine weekend checks of mission critical data) found some errors but nothing that could not be repaired.
I do store image data (triplicates) on Jaz and CD-ROM, but my main archival tool is CD-ROM, I've had excellent performance from a Pinnacle drive and Toast software and would strongly recommend to others to try CD-ROM storage. Again, I "burn" duplicates of all data stored on Jaz, one copy goes in the safe, the other is kept within easy reach, and the Jaz is used for day to day data working. When not being used, all Jaz are stored in their containers and filed in a clean unit. Most of the computers on my network get a weekend quick cleaning to keep dust and garbage from interrupting the work flow. Hope it helps,
Brian C. Tryon MCP-Hahnemann School of Medicine Ann Preston Hall, Box 551 3300 Henry Avenue Philadelphia, PA 19129 USA
----------------------------------------- "Quantifying is a committing task." - Cruz-Orive, 1994.
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." - Richard Feynman --------------------------------------------------------------
My colleague here at Polaroid has recently purchased an old microscope. He is interested in finding out as much information about it as possible. The markings on the microscope are the following: Waldemar Straufs Berlin, S.W. 68 N0. 3692 Note that the S and F is written in a strange script and may not be translated correctly. It is a black stand microscope with "brass accents" and a brass optical tube. It has a condenser which slides out, a polarizing stage, a mirror for capturing the light, a pivot arm in the stand. There are three objectives on the nose piece with the following markings: J. Thamm A.G. Berlin N.W. 6, one is an oil immersion. It came with three eye pieces as well. Pls. let us know if you have any information on this microscope. You can contact my colleague directly, Russ Gaudiana Gaudiar-at-Polaroid.com
We have a graduate student who is looking at concrete & the weathering of the steel rebar inside of it. He needs to section the steel rebar. We have never worked with steel, we are kinda biological oriented. So I'm asking for help. What's the best way for him to get thin sections of steel? Any & all suggestions gratefully accepted, I will pass them on to him.
Steeling myself for your replies,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
I am one of those people who moved from the bench to management (in USDA), and just can't seem to give up my roots (mycology/plant pathology) so keep plugging along with my self-funded research. I have been using glass knives for LM sections, and as part of a recent trade picked up a new EdgeCraft Standard Ultrathin 2.7mm diamond knife. Turns out that I am pretty happy with my glass knife sections, and would be better off with something else or some money instead of the diamond knife. The knife sells for about $2000, but I would be open to offers of about half that, or trades (or trade + cash) involving a better sterio microscope(s) - I am pretty happy with my B&L SZ5, but keep dreaming of a Wild - other areas of interest are older Leitz microscopes (170 mm Ortholux, Dialux, etc.), as well as objectives and components. If you want more information on the diamond knife (model, angle, etc.) - I can provide that. My objective is to find a home for something that I don't need, and in the process keep up the support for my own work.
RE: Ti Alloys - TEM foil preparation 1/13/99 3:47 PM
Bernard Kestel wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America = = } electrolytes I have used on Ti alloys. (3mm, jet polishing). } } Ti-8 w/% Al 13% HCL -60 degrees C } Ti-811 87% methanol 70 to 90 volts } Ti-6Al-4V 25-35 mA. } } Ti-13 Sn 130 ml HCL -50 degrees C } 670 ml methanol 150 volts } 100 ml butyl 60 mA. } } Ti-3V 60 ml perchloric acid -50 degrees C } Ti-20 Zr 590 ml methanol 50 volts } Ti-14 Al 350 ml butyl cellosolve 10-15 mA } } Ti-8 Al 60 ml perchloric acid -15 t0 -20 C } (rolled) 460 ml ethyl alcohol, 95% 180 volts } 280 ml butyl alcohol 40 mA. } 100 ml butyl cellosolve } Ti 5.3 g. = lithium = } chloride -40 degrees C. } nanocrystals, 11.16 g. magnesium 150 volts } compacted chloride 30 mA } 500 ml. methanol } 100 ml. butyl cellosolve } (Note: the two salts above must be added to the combined solvents = one = } "powder" at a time, while stirring. The two dry salts react violently if = mixed = } together. This electrolyte has nearly as wide an application as = perchloric acid = } mixtures and is known as B K-2). } I suspect that B K-2 will cause the least hydride problem. It was = } developed to eliminate hydrides in vanadium TEM foils. The voltages = given would = } need to be reduced about 20% for a twin jet system due to lower = electrical = } resistance of that configuration. My work was done on a South Bay = vertical single jet unit. } I have no vested interest in the above equipment, but have enjoyed = the = } ease with which good polishing conditions can be obtained and reproduced = since = } 1975 or so. Good luck! } Bernard Kestel } Materials Science Division } Argonne National Laboratory } Argonne, Il., 60439 } } E-mail: {bkestel-at-anl.gov} } Thomas, Larry 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 } } -----------------------------------------------------------------------. } } } } } } Satyrith- } } } } Unless you have a compelling reason to use ion milling for this = preparation, I } } recommend using jet electropolishing, possibly with just a final touch-= up = } in the } } ion mill. Although both electropolishing and ion milling commonly = produce } } sample preparation artifacts, the artifacts from ion-milling reactive = metals } } such as Ti and Zr are not well understood and can be harder to eliminate.=
} } Despite Carpenter et. als work (see D. Henriks posting) implicating = hydrocarbon } } contamination as a source of hydriding during ion milling, my personal } } experience ion milling Zr samples in 'oil-free' systems indicates that = water } } vapor is the main culprit in the hydridng. Surface irregularities on = ion = } milled } } Zr/Ti samples (surface terracing, for example) usually have other causes,=
} } possibly related to air leaks in the ion mill or oxygen in the feed gas. } } Another source of irregular surfaces on ion milled samples is embedded = } polishing } } compound. Cubic boron nitride (CBN) sometimes gives better results in = dimpling } } metals than diamond, and even the diamond polishing compounds from = different } } suppliers can give different results. Conversely, brief ion milling of } } electropolished samples--especially at low incidence angles and very low = } kV--can } } improve the surface finish and mill away irregularities caused by second-= phase } } particles. Often the 'trick' to eliminating milling artifacts is = minimizing } } time spent in the ion mill. } } } } Larry Thomas } } Mechanical and Materials Engineering } } Washington State University } } Pullman, WA } } } } } } ---------- } } From: South Bay Technology } } Sent: Tuesday, January 12, 1999 6:00 PM } } To: Satyarth Suri; Microscopy ListServer } } Subject: Ti Alloys - TEM foil preparation } } } } ------------------------------------------------------------------------=
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America = } To = } Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.= html } } -----------------------------------------------------------------------.=
} } } } } } Dear Satyarth: } } } } An excellent paper on the subject is: } } } } "In Situ Hydride Formation in Zirconium and Titanium during Ion Milling"=
} } Graham J.C. Carpenter et al, JMSA Vol.1 No. 4 pp 175-184 1995. } } } } From the paper you will see that a key factor in eliminating hydride } } formation is having a clean sample free from hydrocarbon contamination. = } } This contamination could be remnants from the dimpling process or other } } pre-thinning steps or it could be caused by the back streaming of } } diffusion } } pump oil in your ion mill. As titanium has a high chemical affinity = for } } hydrogen, you may want to look over your preparation steps and try to } } eliminate any areas of possible contamination. If you are still having } } a } } problem, a quick cleaning of both the specimen and the specimen holder } } in a } } Plasma Cleaner should take care of it. } } } } If you have an interest, I can send you a copy of the above referenced } } paper. I also have papers on plasma cleaning which may be of interest. } } } } NOTE: South Bay Technology does manufacture the PC150 Plasma Cleaner = and } } therefore I have a vested interest in promoting its use. } } } } Best regards- } } } } David } Writing at 9:38:23 AM on 1/12/99 } } } = } } *************************************************************** } ** } } ********** } } ************************ } } } } David Henriks TEL: = } } 800-728-2233 (toll free in the USA) } } South Bay Technology, Inc. +1-949-492-2600 } } 1120 Via Callejon FAX: } } +1-949-492-1499 } } San Clemente, CA 92673 USA e-mail: } } henriks-at-southbaytech.com } } } } = } } *************************************************************** } ** } } ********** } } ************************ } } } } } } } } } Please visit us at http://www.southbaytech.com { { { { { } } } } Manufacturers of precision sample preparation equipment and supplies = for } } metallography, crystallography and electron microscopy. } } } } Message text written by Satyarth Suri } } } } } Hello: } } I am currently working on mechanical behavior / microstructure } } correlations in single colony near apha Ti Alloys. I am currently } } preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but } } have run into the problem of a very uneven alpha phase morphology, } } the alpha phase has island formation - we are using a cold stage } } to minimize the hydride formation at the interface. I am using } } 3 micron/6micron diamond paste during the dimpling process - the } } foil itself otherwise is fairly clean in terms of the dislocation } } content. Could anyone in the microscopy land have some suggestions } } in terms of what the problem may be? } } } } thanks - if you want you can respond directly to suri.3-at-osu.edu } } } } -satyarth } } } } { } } } } } } } } } } RFC822 header } } ----------------------------------- } } } } Received: from dns2.anl.gov (dns2.anl.gov [146.139.254.3]) by = } } horus.et.anl.gov (8.6.11/8.6.11) with ESMTP id SAA02218; Tue, 12 Jan = 1999 18:17:43 -0600 } } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com = } } [206.69.208.10]) by dns2.anl.gov (8.9.1a/8.6.11) with SMTP id SAA19261; = Tue, 12 Jan 1999 = } } 18:18:22 -0600 (CST) } } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = } (8.6.11/8.6.11) } id RAA22399 for dist-Microscopy; Tue, 12 Jan 1999 17:41:= 01 -0600 } } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id RAA22384 for = } } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 12 Jan 1999 17:40:30 -= 0600 } } Received: from Sparc5AAEM (aaem.amc.anl.gov [146.139.72.3]) by = } } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA22377 for = } } {microscopy-at-sparc5.microscopy.com} ; Tue, 12 Jan 1999 17:40:17 -0600 } } Received: from pnl.gov (relay.pnl.gov [130.20.128.34]) by Sparc5AAEM = } } (8.6.11/8.6.11) with ESMTP id RAA14848 for {MICROSCOPY-at-aaem.amc.anl.gov} ;= Tue, 12 Jan = } 1999 } 17:52:22 -0600 } } Received: from pnlmse1.pnl.gov by pnl.gov (PMDF V5.1-12 #28154) } } with ESMTP id {01J6GJ2FP8R4970ZC9-at-pnl.gov} for MICROSCOPY-at-aaem.amc.anl.= gov; } } Tue, 12 Jan 1999 15:54:41 PST } } Received: by PNLMSE1.pnl.gov with Internet Mail Service (5.5.2232.9) } } id {CR51M2B5} ; Tue, 12 Jan 1999 15:54:42 -0800 } } Content-return: allowed } } Date: Tue, 12 Jan 1999 15:54:36 -0800 } } From: "Thomas, Larry" {Larry.Thomas-at-pnl.gov} } } Subject: RE: Ti Alloys - TEM foil preparation } } To: Satyarth Suri {suri-at-mse.eng.ohio-state.edu} , } } Microscopy ListServer {MICROSCOPY-at-aaem.amc.anl.gov} , } } "'South Bay Technology'" {Henriks-at-CompuServe.COM} } } Message-id: {7A04D1887189D2118EEB00A024BF29DA41CB17-at-pnlmse2.pnl.gov} } } X-Mailer: Internet Mail Service (5.5.2232.9) } } Errors-to: Microscopy-request-at-sparc5.microscopy.com } } Content-Type: text } } Content-Length: 5172 } } Status: } } } } } RFC822 header } ----------------------------------- } } Received: from dns2.anl.gov (dns2.anl.gov [146.139.254.3]) by = } horus.et.anl.gov (8.6.11/8.6.11) with ESMTP id KAA07881; Wed, 13 Jan 1999 = 10:30:15 -0600 } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com = } [206.69.208.10]) by dns2.anl.gov (8.9.1a/8.6.11) with SMTP id KAA11490; = Wed, 13 Jan 1999 = } 10:30:55 -0600 (CST) } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) = } id JAA28860 for dist-Microscopy; Wed, 13 Jan 1999 09:57:58 -0600 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id JAA28855 for = } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 13 Jan 1999 09:57:27 -= 0600 } Received: from Sparc5AAEM (aaem.amc.anl.gov [146.139.72.3]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id JAA28848 for = } {microscopy-at-sparc5.microscopy.com} ; Wed, 13 Jan 1999 09:57:14 -0600 } Received: from horus.et.anl.gov (horus.et.anl.gov [146.139.240.27]) by = } Sparc5AAEM (8.6.11/8.6.11) with ESMTP id KAA15405 for {MICROSCOPY-at-aaem.= amc.anl.gov} ; = } Wed, 13 Jan 1999 10:09:20 -0600 } Received: from 146.139.240.184 (et212mac184.et.anl.gov [146.139.240.184])= by = } horus.et.anl.gov (8.6.11/8.6.11) with SMTP id KAA07653; Wed, 13 Jan 1999 = } 10:11:07 -0600 } Message-Id: {199901131611.KAA07653-at-horus.et.anl.gov} } Date: 13 Jan 99 10:11:49 -0500 } From: Bernard Kestel {kestel-at-anl.gov} } Subject: RE: Ti Alloys - TEM foil preparation } To: "'South Bay Technology'" {Henriks-at-CompuServe.COM} , } "Thomas, Larry" {Larry.Thomas-at-pnl.gov} , } Microscopy ListServer {MICROSCOPY-at-aaem.amc.anl.gov} , } Satyarth Suri {suri-at-mse.eng.ohio-state.edu} } X-Mailer: QuickMail Pro 1.5.4 (Mac) } X-Priority: 3 } MIME-Version: 1.0 } Reply-To: Bernard Kestel {kestel-at-anl.gov} } Content-Transfer-Encoding: quoted-printable } Errors-to: Microscopy-request-at-sparc5.microscopy.com } Content-Type: text/plain; charset=3D"US-Ascii" } Content-Length: 10127 } Status: = } --====56524855544856555650===1 Content-Type: text/html; charset="US-Ascii" Content-Transfer-Encoding: quoted-printable
{HTML} {HEAD} {/HEAD} {BODY} {PRE WIDTH=3D"132"} RE: Ti Alloys - TEM foil preparation 1/13/99 3:47 PM
{/PRE} {FONT = FACE=3D"Geneva" SIZE=3D2 COLOR=3D"#000000"} {BR} Bernard Kestel wrote: {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR} Reply to: RE: Ti Alloys - TEM = foil preparation {BR} > Re: Electropolishing = of Ti alloys {BR} > Due to interest = in this subject, I will list some additional = {BR} >electrolytes I have used on = Ti alloys. (3mm, jet polishing). {BR} > {BR} > = Ti-8 w/% Al 13% HCL = -60 degrees C {BR} > Ti-811 = 87% methanol = 70 to 90 volts {BR} > Ti-6Al-4V = 25-35 = mA. {BR} > {BR} > Ti-13 Sn = 130 ml HCL -50 degrees = C {BR} > = 670 ml methanol 150 volts {BR} > = 100 ml butyl = 60 mA. {BR} > {BR} > = Ti-3V 60 ml perchloric = acid -50 degrees C {BR} > Ti-20 = Zr 590 ml methanol = 50 volts {BR} > Ti-14 Al = 350 ml butyl cellosolve 10-15 mA {BR} > {BR} > = Ti-8 Al 60 ml perchloric = acid -15 t0 -20 C {BR} > (rolled) = 460 ml ethyl alcohol, 95% = 180 volts {BR} > = 280 ml butyl alcohol = 40 mA. {BR} > = 100 ml butyl cellosolve {BR} > = Ti = 5.3 g. lithium {BR} >chloride = -40 degrees C. {BR} > nanocrystals, = 11.16 g. magnesium 150 = volts {BR} > compacted = chloride 30 mA {BR} > = 500 ml. methanol {BR} > = 100 ml. butyl = cellosolve {BR} > (Note: the two salts = above must be added to the combined solvents = one {BR} >"powder" at a time, = while stirring. The two dry salts react = violently if mixed {BR} >together. This = electrolyte has nearly as wide an application = as perchloric acid {BR} >mixtures and is = known as B K-2). {BR} > I suspect that = B K-2 will cause the least hydride problem. = It was {BR} >developed to eliminate hydrides = in vanadium TEM foils. The voltages given = would {BR} >need to be reduced about 20% = for a twin jet system due to lower electrical = {BR} >resistance of that configuration. = My work was done on a South Bay vertical = single jet unit. {BR} > I have no vested = interest in the above equipment, but have = enjoyed the {BR} >ease with which good = polishing conditions can be obtained and = reproduced since {BR} >1975 or so. Good = luck! {BR} > Bernard Kestel {BR} > = Materials Science Division {BR} > = Argonne National Laboratory {BR} > = Argonne, Il., 60439 {BR} > {BR} > = E-mail: < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = bkestel-at-anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} >Thomas, Larry = wrote: {BR} >>-------------------------------------------------------------------= ----- {BR} >>The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America >To {BR} >Subscribe/Unsubscribe = -- Send Email to {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {= U} ListServer-at-MSA.Microscopy.Com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >>On-Line Help = {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.msa.= microscopy.com/MicroscopyListserver/FAQ.html {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >>-------------------------------------------------------------------= ----. {BR} >> {BR} >> {BR} >>Satyrith- {BR} >> {BR} >>Unless = you have a compelling reason to use ion = milling for this preparation, I {BR} >>recommend = using jet electropolishing, possibly with = just a final touch-up {BR} >in the {BR} >>ion = mill. Although both electropolishing and = ion milling commonly produce {BR} >>sample = preparation artifacts, the artifacts from = ion-milling reactive metals {BR} >>such = as Ti and Zr are not well understood and = can be harder to eliminate. {BR} >>Despite = Carpenter et. als work (see D. Henriks posting) = implicating hydrocarbon {BR} >>contamination = as a source of hydriding during ion milling, = my personal {BR} >>experience ion milling = Zr samples in 'oil-free' systems indicates = that water {BR} >>vapor is the main culprit = in the hydridng. Surface irregularities = on ion {BR} >milled {BR} >>Zr/Ti samples = (surface terracing, for example) usually = have other causes, {BR} >>possibly related = to air leaks in the ion mill or oxygen in = the feed gas. {BR} >>Another source of = irregular surfaces on ion milled samples = is embedded {BR} >polishing {BR} >>compound. = Cubic boron nitride (CBN) sometimes gives = better results in dimpling {BR} >>metals = than diamond, and even the diamond polishing = compounds from different {BR} >>suppliers = can give different results. Conversely, = brief ion milling of {BR} >>electropolished = samples--especially at low incidence angles = and very low {BR} >kV--can {BR} >>improve = the surface finish and mill away irregularities = caused by second-phase {BR} >>particles. = Often the 'trick' to eliminating milling = artifacts is minimizing {BR} >>time spent = in the ion mill. {BR} >> {BR} >>Larry = Thomas {BR} >>Mechanical and Materials = Engineering {BR} >>Washington State University {BR} >>Pullman, = WA {BR} >> {BR} >> {BR} >> ---------- {BR} >> From: = South Bay Technology {BR} >> Sent: Tuesday, = January 12, 1999 6:00 PM {BR} >> To: = Satyarth Suri; Microscopy ListServer {BR} >> Subject: = Ti Alloys - TEM foil preparation {BR} >> {BR} >> ------------------------------------------------------------------= ------ {BR} >> The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America > To {BR} >Subscribe/Unsubscribe = -- Send Email to {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {= U} ListServer-at-MSA.Microscopy.Com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> On-Line Help = {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.msa.= microscopy.com/MicroscopyListserver/FAQ.html {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> ------------------------------------------------------------------= -----. {BR} >> {BR} >> {BR} >> Dear = Satyarth: {BR} >> {BR} >> An excellent = paper on the subject is: {BR} >> {BR} >> "In = Situ Hydride Formation in Zirconium and = Titanium during Ion Milling" {BR} >> Graham = J.C. Carpenter et al, JMSA Vol.1 No. 4 pp = 175-184 1995. {BR} >> {BR} >> From = the paper you will see that a key factor = in eliminating hydride {BR} >> formation = is having a clean sample free from hydrocarbon = contamination. {BR} >> This contamination = could be remnants from the dimpling process = or other {BR} >> pre-thinning steps or = it could be caused by the back streaming = of {BR} >>diffusion {BR} >> pump oil = in your ion mill. As titanium has a high = chemical affinity for {BR} >> hydrogen, = you may want to look over your preparation = steps and try to {BR} >> eliminate any = areas of possible contamination. If you = are still having {BR} >>a {BR} >> problem, = a quick cleaning of both the specimen and = the specimen holder {BR} >>in a {BR} >> Plasma = Cleaner should take care of it. {BR} >> {BR} >> If = you have an interest, I can send you a copy = of the above referenced {BR} >> paper. = I also have papers on plasma cleaning which = may be of interest. {BR} >> {BR} >> NOTE: = South Bay Technology does manufacture the = PC150 Plasma Cleaner and {BR} >> therefore = I have a vested interest in promoting its = use. {BR} >> {BR} >> Best regards- {BR} >> {BR} >> David = > Writing = at 9:38:23 AM on 1/12/99 {BR} >> = > {BR} >>*************************************************************** {BR} =
>** {BR} >>********** {BR} >> ************************ {BR} >> {BR} >> David = Henriks = TEL: {BR} >> 800-728-2233 = (toll free in the USA) {BR} >> South = Bay Technology, Inc. = +1-949-492-2600 {BR} >> 1120 Via = Callejon = FAX: {BR} >>+1-949-492-1499 {BR} >> San = Clemente, CA 92673 USA e-mail: {BR} >> {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} henriks-at-southbaytech.com {/U} {= /FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> {BR} >> {BR} >>*************************************************************** {BR} =
>** {BR} >>********** {BR} >> ************************ {BR} >> {BR} >> = >>>>> Please visit us = at {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.= southbaytech.com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} <<<<< {BR} >> {BR} >> Manufacturers = of precision sample preparation equipment = and supplies for {BR} >> metallography, = crystallography and electron microscopy. {BR} >> {BR} >> Message = text written by Satyarth Suri {BR} >> > {BR} >> Hello: {BR} >> I = am currently working on mechanical behavior = / microstructure {BR} >> correlations = in single colony near apha Ti Alloys. I = am currently {BR} >> preparing the foils = using a dual ion mill (6kV, 12deg, 1mA), = but {BR} >> have run into the problem = of a very uneven alpha phase morphology, {BR} >> the = alpha phase has island formation - we are = using a cold stage {BR} >> to minimize = the hydride formation at the interface. = I am using {BR} >> 3 micron/6micron diamond = paste during the dimpling process - the {BR} >> foil = itself otherwise is fairly clean in terms = of the dislocation {BR} >> content. = Could anyone in the microscopy land have = some suggestions {BR} >> in terms of = what the problem may be? {BR} >> {BR} >> thanks = - if you want you can respond directly to = {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} suri.3-at-osu.edu {/= U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> {BR} >> -satyarth {BR} >> {BR} >> < {BR} >> {BR} >> {BR} >> {BR} >> {BR} >>RFC822 = header {BR} >>----------------------------------- {BR} >> {BR} >> = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >>horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id SAA02218; = Tue, 12 Jan 1999 18:17:43 -0600 {BR} >> = Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com = {BR} >>[206.69.208.10]) by dns2.anl.gov = (8.9.1a/8.6.11) with SMTP id SAA19261; Tue, = 12 Jan 1999 {BR} >>18:18:22 -0600 (CST) {BR} >> = Received: (from {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = daemon-at-localhost {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ) by Sparc5.Microscopy.Com = {BR} >(8.6.11/8.6.11) >id RAA22399 for = dist-Microscopy; Tue, 12 Jan 1999 17:41:01 = -0600 {BR} >> Received: from no_more_spam.com = (Sparc5 [206.69.208.10]) by {BR} >>Sparc5.Microscopy.Com = (8.6.11/8.6.11) with SMTP id RAA22384 for = {BR} >>" {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = MicroscopyFilteredEmail-at-msa.microscopy.com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} "; Tue, 12 Jan 1999 = 17:40:30 -0600 {BR} >> Received: from = Sparc5AAEM (aaem.amc.anl.gov [146.139.72.3]) = by {BR} >>Sparc5.Microscopy.Com (8.6.11/8.6.11) = with ESMTP id RAA22377 for {BR} >>< {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} microscopy-at-sparc5.microscopy.= com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >; Tue, 12 Jan 1999 = 17:40:17 -0600 {BR} >> Received: from = pnl.gov (relay.pnl.gov [130.20.128.34]) = by Sparc5AAEM {BR} >>(8.6.11/8.6.11) = with ESMTP id RAA14848 for < {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D1 COLOR=3D"#0000FF"} {U} MICROSCOPY-at-aaem.amc.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >; Tue, 12 Jan {BR} >1999 = >17:52:22 -0600 {BR} >> Received: = from pnlmse1.pnl.gov by pnl.gov (PMDF V5.1-12 = #28154) {BR} >> with ESMTP id < {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} 01J6GJ2FP8R4970ZC9-at-pnl.gov {/= U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > for {/FONT} {FONT FACE=3D"= Geneva" = SIZE=3D1 COLOR=3D"#0000FF"} {U} MICROSCOPY-at-aaem.amc.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ; {BR} >> Tue, 12 = Jan 1999 15:54:41 PST {BR} >> Received: = by PNLMSE1.pnl.gov with Internet Mail Service = (5.5.2232.9) {BR} >> id <CR51M2B5>; = Tue, 12 Jan 1999 15:54:42 -0800 {BR} >> = Content-return: allowed {BR} >> Date: = Tue, 12 Jan 1999 15:54:36 -0800 {BR} >> = } From: "Thomas, Larry" < {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} Larry.Thomas-at-pnl.gov {/U} {/= FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} >> Subject: = RE: Ti Alloys - TEM foil preparation {BR} >> = To: Satyarth Suri < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#= 0000FF"} {U} suri-at-mse.eng.ohio-state.edu {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >, {BR} >> = Microscopy ListServer < {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D1 COLOR=3D"#0000FF"} {U} MICROSCOPY-at-aaem.amc.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >, {BR} >> = "'South Bay Technology'" < {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} Henriks-at-CompuServe.COM {/U} {/= FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} >> Message-id: = < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = 7A04D1887189D2118EEB00A024BF29DA41CB17-at-pnlmse2.pnl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} >> X-Mailer: = Internet Mail Service (5.5.2232.9) {BR} >> = Errors-to: {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = Microscopy-request-at-sparc5.microscopy.com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> Content-Type: = text {BR} >> Content-Length: 5172 {BR} >> = Status: > {BR} > {BR} > {BR} > {BR} >RFC822 = header {BR} >----------------------------------- {BR} > {BR} > = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id KAA07881; 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by mtiwmhc02.worldnet.att.net (InterMail v03.02.07 118 124) with ESMTP id {19990114001451.EPZL10402-at-worldnet.att.net} ; Thu, 14 Jan 1999 00:14:51 +0000 Message-ID: {369D37CA.56F575DD-at-worldnet.att.net}
Dear Paula,
The success of thin sections will depend on the blade type and size which will then be dictated on the type of saw you have. Slow speed diamond saws have a blade capacity of 5" Diameter which can limit the sample size to be cut and take a long time to cut.. High speed saws can accomodate larger blades and perform cuts much faster (5 minutes or less) with the same accuracy as a slow speed saw. Feed pressure can affect the blade during the cut on either saw. If the blade is too thin, too much feed pressure will cause the blade to wander form the original cut line affecting the thickness of the section. This can be remedied by using a large flange or more rigid, thicker blade.
CBN is the only choice for cutting steels on a low speed saw without destroying the blade or sample, but can also be used on a high speed saw. Aluminum Oxide blades cut the most effieciently on a high speed saw and cost much less. DO NOT attempt cutting with diamond blades, they will only become loaded with steel and this will prohibit the sample from being cut. Even frequent dressing will not enhance the performance. Aluminum Oxide blades are better suited for steels and Allied HighTech has a wide selection of blades for any saw you may have.
I know of two labs at LBL that have a high speed saw and can get you this information should you require it. In additon, if you have any application questions, please feel free to contact me at 800-675-1118 to further discuss your interest.
Good Luck,
Gary Liechty
Paula Sicurello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hey there boarders, } } We have a graduate student who is looking at concrete & the } weathering of the steel rebar inside of it. He needs to section the steel } rebar. We have never worked with steel, we are kinda biological oriented. } So I'm asking for help. } What's the best way for him to get thin sections of steel? Any & } all suggestions gratefully accepted, I will pass them on to him. } } Steeling myself for your replies, } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207 } http://biology.berkeley.edu/EML
-- Gary Liechty Product Application Specialist
Allied High Tech Products, Inc. 2376 E. Pacifica Place Rancho Dominguez, CA 90220 800-675-1118 310-635-2466 310-762-6808 Fax
Equipment and Consumable Products for Materialographic, SEM and TEM Sample Prepration
Try Graphic converter, it's a shareware that can read and translate almost every format. It can be able to solve your problem. You can find it on every info-mac mirror.
Eric LEROY Dr. Laboratoire de Chimie Metallurgique des Terres Rares UPR 209 - CNRS Groupe des Laboratoires de Thiais 2-8, rue Henri Dunant 94320 THIAIS cedex
A few years ago I got hold of a beta version of a SEM Macintosh simulation program, SuperSEM. It was a Macintosh program that was developed with Supercard software. It was misplaced during the process of upgrading my computers. Does anyone know if it exists?
Thanks in advance, Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
Colin raises a good point of media being orphaned that we feel the pinch of already. Our HP 1300T MO drive (~650 MB each side) failed on us a while back. It will no longer accept the cartridges - it keeps kicking them out. We had already offloaded much of the data to CD but we have a few cartridges that I would still like to retrieve. However, we appear to be the only ones on campus that ever purchased the HP drive.
Is there anyone out there with a working HP MO drive (gathering dust or not) that could help us retrieve the last few cartidges?
At 07:10 AM 1/12/99 +0000, you wrote: {snip} } As pointed out the Jaz/Zip technology will probably be superseded } soon ( 1GB drives are gone already ! ) and in a few years it will be } difficult to read data if your drive fails. This was a major factor } along with cost ) 5 years ago when we decided on a CD-R storage system, } over Magneto-Optical which was the current favourite then. {snip} } Colin Reid, } Electron Microscope Unit, } Trinity College Dublin,
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
Please remember that 3/4ths of your 10 MB image won't be doing you any good during the slide show. If your projector supports true 24-bit color (as opposed to 16-bit or 8-bit) and 1024x768 pixels, you will only be displaying 2.25 MB of image. You might as well match your image size to the projector as you make up your presentation. It would help file size and the speed of transition between slides, but these fast computers are handling that pretty well.
I concur with the others. Most imaging programs have a slide show capability built in. They may not have all the fancy transitional effects, but they should do the show. Even my shareware LViewPro on the PC had that.
At 05:29 PM 1/12/99 -0400, you wrote: } } We just got a G3 Mac and an Epson digital projector-I have seen some really } nice digital slide shows and am excited by the increase in quality and } decrease in effort making slides. I expect, however, to spend some effort } finding out the best way to put together a slide show, and am wondering } what advice or experience might be out there. I would like to show ~10MB } images without any computer stuff showing at the same time, but could } consider compression (eg., jpeg) if there is no loss of quality. Wondering } whether to get into a slide presentation program or just make a stack and } open them sequentially (I have 190+ MB of memory). There is also a neat IR } pointer that seems to bounce off the screen and feed into software in the } projector that allows you to move the cursor around and click/double click } with the IR pointer. Too bad they didn't include a laser pointer in this } device!....Many thanks for any help or advice!...Tom Reese } ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
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Reply to: RE: Ti Alloys - TEM foil preparation Re: Electropolishing of Ti alloys = Due to interest in this subject, I will list some additional = electrolytes I have used on Ti alloys. (3mm, jet polishing).
Ti-8 w/% Al 13% HCL -60 degrees C Ti-811 87% methanol 70 to 90 volts Ti-6Al-4V 25-35 mA.
Ti-13 Sn 130 ml HCL -50 degrees C 670 ml methanol 150 volts 100 ml butyl 60 mA.
Ti-3V 60 ml perchloric acid -50 degrees C Ti-20 Zr 590 ml methanol 50 volts Ti-14 Al 350 ml butyl cellosolve 10-15 mA
Ti-8 Al 60 ml perchloric acid -15 t0 -20 C (rolled) 460 ml ethyl alcohol, 95% 180 volts 280 ml butyl alcohol 40 mA. 100 ml butyl cellosolve = Ti 5.3 g. lithium chloride -40 degrees C. nanocrystals, 11.16 g. magnesium 150 volts compacted chloride 30 mA 500 ml. methanol 100 ml. butyl cellosolve (Note: the two salts above must be added to the combined solvents one = "powder" at a time, while stirring. The two dry salts react violently if = mixed together. This electrolyte has nearly as wide an application as = perchloric acid mixtures and is known as B K-2). I suspect that B K-2 will cause the least hydride problem. It was = developed to eliminate hydrides in vanadium TEM foils. The voltages given = would need to be reduced about 20% for a twin jet system due to lower = electrical resistance of that configuration. My work was done on a South = Bay vertical single jet unit. I have no vested interest in the above equipment, but have enjoyed = the ease with which good polishing conditions can be obtained and = reproduced since 1975 or so. Good luck! = Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
E-mail: {bkestel-at-anl.gov} Thomas, Larry wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE: Ti Alloys - TEM foil preparation Re: Electropolishing of Ti alloys = Due to interest in this subject, I will list some additional = electrolytes I have used on Ti alloys. (3mm, jet polishing).
Ti-8 w/% Al 13% HCL -60 degrees C Ti-811 87% methanol 70 to 90 volts Ti-6Al-4V 25-35 mA.
Ti-13 Sn 130 ml HCL -50 degrees C 670 ml methanol 150 volts 100 ml butyl 60 mA.
Ti-3V 60 ml perchloric acid -50 degrees C Ti-20 Zr 590 ml methanol 50 volts Ti-14 Al 350 ml butyl cellosolve 10-15 mA
Ti-8 Al 60 ml perchloric acid -15 t0 -20 C (rolled) 460 ml ethyl alcohol, 95% 180 volts 280 ml butyl alcohol 40 mA. 100 ml butyl cellosolve = Ti 5.3 g. lithium chloride -40 degrees C. nanocrystals, 11.16 g. magnesium 150 volts compacted chloride 30 mA 500 ml. methanol 100 ml. butyl cellosolve (Note: the two salts above must be added to the combined solvents one = "powder" at a time, while stirring. The two dry salts react violently if = mixed together. This electrolyte has nearly as wide an application as = perchloric acid mixtures and is known as B K-2). I suspect that B K-2 will cause the least hydride problem. It was = developed to eliminate hydrides in vanadium TEM foils. The voltages given = would need to be reduced about 20% for a twin jet system due to lower = electrical resistance of that configuration. My work was done on a South = Bay vertical single jet unit. I have no vested interest in the above equipment, but have enjoyed = the ease with which good polishing conditions can be obtained and = reproduced since 1975 or so. Good luck! = Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
E-mail: {bkestel-at-anl.gov} Thomas, Larry wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
by hawk.csrv.uidaho.edu (8.8.6 (PHNE_14041)/) with ESMTP id IAA24952 for {Microscopy-at-Sparc5.Microscopy.Com} ; Thu, 14 Jan 1999 08:45:37 -0800 (PST) Received: from OAK/SpoolDir by oak.csrv.uidaho.edu (Mercury 1.43); 14 Jan 99 08:47:23 -0800 (PST) Received: from SpoolDir by OAK (Mercury 1.43); 14 Jan 99 08:46:51 -0800 (PST)
I am looking for a used, but functional, EDX specimen holder for a JEOL 1200 EX TEM. If anyone has one for sell, please contact me at: jfb-at-uidaho.edu (208)885-6656
Thank you.
Franklin Bailey Holm Research Center University of Idaho Moscow, ID 83844-2204
A pair of postdocs in Plant Pathology came in to the facility to ask about a cold stage for a light microscope, which no one at this university seems to have. They want to look at cryostat sections of wood, and need digital images for image analysis, and the sections must remain frozen. I have here a light microscope fitted with a digital camera hooked to the Mac with NIH Image. I have liquid nitrogen, styrofoam, and a machine shop. I also have high humidity. Can anyone tell me how to kludge together a cold enough stage that we can get images of these frozen sections while preventing frost? I have some general ideas, but I'd appreciate any specific plans or ideas.
The idea is to look at the number of vacuoles filled with gas vs the number filled with water. Up to now they have had images taken with an SEM with a cold stage in Canada, and would prefer to do them locally, at lower mag, and cheaper.
Thanks in advance for any advice!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
A colleage from an industrial SEM lab contacted me about a problem with contamination on the final aperture on an variable pressure SEM. This SEM was recently purchased by one of their branch plants to examine semiconductor products. Although it is a variable pressure SEM, they have only operated it in the high vac mode like a standard SEM.
The problem they are experiencing is that the final aperture presumably gets so dirty they have to change the aperture every two weeks. Unfortunately the manufacturer has not been able to shed any light on this situation.
If the vacuum system is alright, I suggested that the problem might be with the samples. Supposedly the branch lab operators are following the same sample prep protocol established in the local SEM lab.
Any other ideas out there?
Thanks in advance, Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
Products like Dazzle can digitize the video over to MPG files which should be transportable. We bought a unit for about $200 and are still coming up to speed with it. If you have a CD-R, you should be able to cut the movies onto a CD which they could read over there.
Disclaimer: We have no interest in Dazzle other than its the unit we bought. There are many other products on the market which can also accomplish the task.
Warren S.
At 12:15 PM 1/14/99 -0500, you wrote: } I have in-situ results currently stored on VHS tape that I need to present } in Europe. } } Any recipes available? } } Thanks. } David E. Luzzi } Professor } Department of Materials Science } University of Pennsylvania
To all We have a client trying to convert from glass plates to sheet film. Can anyone help us to obtain film adapters for the Siemens 1A camera? Thank you. Peter Stolzenberg
I'm just posting to ask if anyone has developed a fixation protocol for Drosophila embryos suitable for use in TEM studies. Is it necessary to remove the chorion and vitelline membranes as in light microscopy procedures. Also, I'm wondering if the use of methanol is strictly taboo, or can it be tolerated if the exposure time is short?
Does anyone have a good protocol or reference for virus isolation = from food other than shellfish? We are trying to isolate and concentrate = Norwalk virus from some foods implicated in a poisoning outbreak in = Detroit. The foods in question are salami, garbanzo beans and some kind of = cheese (I think provolone). We need to concentrate to at least 1 million = particles/ml in order to use our prep for negative staining and TEM = observation. Any suggestions would be greatly appreciated!
Sincerely, Peggy Casey Michigan Dept. of Community Health Lansing, Mich. phone: 517-335-8102 e-mail: caseym-at-state.mi.us
I have been told to search for "M6", an adhesive which was used in the somewhat distant past for bonding samples for cross-sectioning. ..I cannnot find a company which has even heard of this. Any suggestions as to where I can find it?
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3392 [x4] `___/ //\./=/~\====\ (704)509-6622 [Hm] \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
} Hello, } } A colleague is interested in examining rat kidney tubules at the TEM level. } The tubules are approximately 40 um in length. Unfortunately, once they } are placed in fixative in a 1 ml centrifuge tube, the tubules are no longer } visible which creates processing problems. We are going to try placing } the tubules in agar as a way of transporting the tubules through the } processing procedure so as not to lose them in transit. } } Also, the few cells (not tubules) we did see appeared to be washed out. We } used the following solutions: } } 2% glutaraldehyde in sodium cacodylate buffer } 2% osmium tetroxide } alcohol } propylene oxide } polybed } } Any suggestions would be greatly appreciated. } } Thank you, } } Ginger Baker } EM Lab Manager } Oklahoma State University College of Osteopathic Medicine } lizard-at-osu-com.okstate.edu } 918-561-8232
Through the years I have approached, evaluated and re-evaluated "hole sizing" from SEM micrographs for various technologies: biology, electron optics, polymers, chemistry and microelectronics. The paramount objectives are always the same: 1) spatial resolution(of digitizing hardware and software) 2) repeatability 3) SEM background contrast (gray scales) differentiation 4) Maximizing Automation 5) Minimizing user judgement (auto-thresholding, etc.) 6) Manual extraction of artifacts 7) Stereological considerations to mention a few. Also such techniques were attempted to improve the above such as: Image projection, edge detection, pre- and post- image or frame(or pixel) averaging and processing. Well once again this re-evaluation in light of current advances in hard and soft digital technologies is required. I thought I would see if commercial and non-commercial members of this forum would share any current technological information with me. Hopefully after the first 5 or 10 responses, to avoid driving subscribers crazy , you could respond directly to my email. I will gather and redistribute this data in any fashion requested at a later date to avoid congesting the mailservor. Thank you for your interest. Jeff Day/ 'JD' DBA Texas Industrial Mesquite, Texas Email: WA5EKH-at-juno.com
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
} } } I have been told to search for "M6", an adhesive which was used in the } somewhat distant past for bonding samples for cross-sectioning. ..I } cannnot find a company which has even heard of this. Any suggestions as } to where I can find it?
Great stuff! Does work well. M6 is a short abbreviation for M-Bond 610 which was originally desighned for strain gages. Can be purchased from "MM" Micro Measurement Division at Raleigh North Carolina USA. Phone: (919) 365 3800 Fax: (919) 365 3945
Standard disclaimer.
Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
Tim (TJ) LaFave Jr. wrote: ================================================= I have been told to search for "M6", an adhesive which was used in the somewhat distant past for bonding samples for cross-sectioning. ..I cannnot find a company which has even heard of this. Any suggestions as to where I can find it? ================================================== Could it be that you are looking for M-Bond(TM) 610? It is all described on our website below, if that is what you had in mind. Maybe there was a predecessor called M6 but in any case, M-Bond 610 is perhaps what you had in mind? The product is manufactured by a division of Vishay Technology.
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: www.2spi.com ############################ ==================================================
} I have in-situ results currently stored on VHS tape that I need to present } in Europe. } } Any recipes available? } } Thanks. } David E. Luzzi } Professor } Department of Materials Science } University of Pennsylvania } 3231 Walnut Street } Philadelphia, PA 19104-6272 } } 215-898-8366 } 215-573-2128 - fax } luzzi-at-lrsm.upenn.edu {mailto:luzzi-at-lrsm.upenn.edu} } } Hi David,
Here in Europe we use different standards in different countries, however, we also have multi-standard machines that will play the 3 commonest standards (NTSC, PAL and SECAM). I suggest that before you go to too much trouble check if your hosts can play NTSC. VHS is the commonest format here anyway. Regards, Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
I did my first bifringant crystals with Polaroid light to night. My experiment in marginal at best. The swift is not too swift and my polarizing setup was a Grey photo polarize and the resolver was a broken pair of brown sun glasses. Probably the worst choice I could have come up with.
But it was a hoot.
I am trying a little slower drying solution and hope for larger salt crystals.
I have gone through my stock of Siemens spares, and have found 23 cut film inserts to fit the Elmiskop 1a plate holders. They are second-hand, but appear to be in very good condition. If you are interested please Email me directly.
Regards,
Bob ********************************************** Bob Phillips, MicroServiS Electron Microscopy Services, 11 Grafton Close, St. Ives, Huntingdon, Cambs. PE17 6DL United Kingdom. Email: microservis-at-dial.pipex.com ****************************************** -----Original Message----- } From: "PESTOEM-at-aol.com"-at-Sparc5.Microscopy.Com {"PESTOEM-at-aol.com"-at-Sparc5.Microscopy.Com} To: Microscopy-at-Sparc5.Microscopy.Com {Microscopy-at-Sparc5.Microscopy.Com} Cc: uelzen-at-erols.com {uelzen-at-erols.com}
O.k., all you good microscopists, I'm looking for a source for either a Uranium Glass Block or a suitable replacement for visualizing/teaching the light path on a light microscope. The block is placed in the microscope stage and the illumination path way can be viewed inside the block - particularly as the condensor is focused and as either the stage aperature or field aperature are varied.
I know this can be visualize with a dilute milk solution but I really would prefer something a little more stable (and non-perishable).
Thanks!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meeting point for developers and users working in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
If you pulse-spiin the tubules in an Eppendorf centrifuge and then osmicate (I would use 1% OSO4 in 0.1M sodium cacodylate, 30 min to 1 hr, room temp), then spin again, you should be able to see them. At that point you can embed them in a small drop of 2% agarose and continue the processing. Good luck.
On Thu, 14 Jan 1999, Ginger R Baker 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, } } } } A colleague is interested in examining rat kidney tubules at the TEM } level. } } The tubules are approximately 40 um in length. Unfortunately, once they } } are placed in fixative in a 1 ml centrifuge tube, the tubules are no } longer } } visible which creates processing problems. We are going to try placing } } the tubules in agar as a way of transporting the tubules through the } } processing procedure so as not to lose them in transit. } } } } Also, the few cells (not tubules) we did see appeared to be washed out. } We } } used the following solutions: } } } } 2% glutaraldehyde in sodium cacodylate buffer } } 2% osmium tetroxide } } alcohol } } propylene oxide } } polybed } } } } Any suggestions would be greatly appreciated. } } } } Thank you, } } } } Ginger Baker } } EM Lab Manager } } Oklahoma State University College of Osteopathic Medicine } } lizard-at-osu-com.okstate.edu } } 918-561-8232 } } } }
I am a new faculty member in a small department of veterinary pathology. I am interested in obtaining the equipment necessary to produce good quality ultrathin sections for TEM examination at our University's central EM facility. I am particularly interested in ultratomes (1 or 2), vacuum oven, embedding molds, and other necessities. Equipment from laboratories that are down-sizing would be considered ideal. Please respond directly to my e-mail address: sfrasca-at-canr1.cag.uconn.edu
Richard: Here is a summary I had saved of a previous uranyl glass thread. Hope it helps. Tom
--------------------------------------
Newport Industrial Glass, Inc. 1631 Monrovia Ave. Costa Mesa, CA 92627 Tel: 714-642-9980 Fax: 714-645-6800 Contact person: Bill Larsen (you can tell him I sent you). Sold as a 6.5x6.5" sheet (you can specify 1 mm height), so you may want to form a "consortium" to have Newport pre-cut a sheet to slide size (nominal extra cost, but your lab only needs one or two slides). If there is a lot of interest, my company may start selling single slides.
As for references and the Shading Correction equation: please see my article in the 11/94 issue of Journal of NIH Research 6(11): 80 (usual Internet disclaimer: yes, that is an ad from my company on the facing page). Also look at Jericevic et al (1989) Methods in Cell Biology 30:47-83.
MutliSpeck beads: The Molecular Probes pre-mounted slide kits should be ideal for DAPI and Fluorescein. I believe they were optimized for Rhodamine, but should still work ok for Texas Red. If your problem is with mounting, Mol. Probes now sells the beads in solution, so you can 'sprinkle' some on your specimens. If you have a different problem with the current MultiSpeck's, Mol. Probes may be able to work something out for you.
Sorry, but I usually buy my reference material from Mol. Probes and don't keep close track of other slide manufacturers.
Sincerely,
Dr. George McNamara Universal Imaging Corporation George_M-at-Image1.com
} Dear confocalers,
} I read through the archives for information on uranyl glass test slides, } but couldn't figure out if there is a definitive answer to the question, } Does anyone still manufacture uranyl glass, and where could I get some?
You can obtain micro slide sized pieces of uranyl glass from:
Newport Industrial Glass, Inc. 1631 Monrovia Avenue Costa Mesa, CA 92627 (714) 645 - 1500 (714) 645 - 6800 [FAX]
They have this glass (glass #3750) in large stock pieces, so it must be cut down to the size of a micro slide. The will grind it down to whatever thickness you desire, as well. We ordered two micro slide sized peices in July of 1993. The cost was $86.
Good luck!
Cheers, Bill Bug
************************* * Bill Bug * * Dept. of Biology * * Swarthmore College * *************************} O.k., all you good microscopists, I'm looking for a source for either a Uranium Glass } Block or a suitable replacement for visualizing/teaching the light path on } a light } microscope. The block is placed in the microscope stage and the } illumination path way } can be viewed inside the block - particularly as the condensor is focused } and as either } the stage aperature or field aperature are varied. } } I know this can be visualize with a dilute milk solution but I } really would prefer } something a little more stable (and non-perishable). } } Thanks! } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax) --============_-1295708639==_ma============ Content-Type: text/enriched; charset="us-ascii"
Richard: Here is a summary I had saved of a previous uranyl glass thread. Hope it helps. Tom
--------------------------------------
} From: Not Specified { {Kris_Kavanau-at-dmcmail.ucsf.edu} Attn: George M
Return-Path: Kris_Kavanau-at-dmcmail.ucsf.edu To: Microscopy-at-aaem.amc.anl.gov (M)
Organization: University of California, SF Division of Molecular Cytometry Date: Fri, 03 Feb 1995 10:03:40 PST
OFFICE MEMO Uranyl Glass/FM Stds. Date: 2/3/95
Dear Microscopists,
Does anyone have any uranyl glass, or know where it might be obtained? I have been told that it is no longer manufactured commercially. It might be an excellent "generic" fluorescence microscopy control. Are there any commercially available, pre-mounted fluorescence standards besides "MultiSpeck" from Molecular Probes? They are very convenient, but they are not ideal for our applications as DAPI, fluorescein, and Texas Red specific controls. Unfortunately, Flow Cytometry Standards Co. no longer makes pre-mounted standards.
I have been managing the UCSF core flow and image cytometry facility ("Lab for Cell Analysis") for 2 years, but I had no real QC for our 2 occasionally used fluorescence microscopes. Now I need to establish QC protocols for 6 additional multi-user, computerized fluorescence (one scanning confocal) microscopes in the "National Molecular Cytogenetics Resource." I was surprised that so few standards (and journal references) seem to be available.
Any suggestions or comments would be greatly appreciated. Thank you very much. Kris Kavanau; kavanau-at-dmc.ucsf.edu
To: Microscopy-at-AAEM.amc.anl.gov
} From: George M { {George_M-at-image1.com}
Reply to: RE} Uranyl Glass/FM Stds.
Hi Kris,
Uranium glass slides can be purchased from:
Newport Industrial Glass, Inc.
1631 Monrovia Ave.
Costa Mesa, CA 92627
Tel: 714-642-9980
Fax: 714-645-6800
Contact person: Bill Larsen (you can tell him I sent you). Sold as a 6.5x6.5" sheet (you can specify 1 mm height), so you may want to form a "consortium" to have Newport pre-cut a sheet to slide size (nominal extra cost, but your lab only needs one or two slides). If there is a lot of interest, my company may start selling single slides.
As for references and the Shading Correction equation: please see my article in the 11/94 issue of Journal of NIH Research 6(11): 80 (usual Internet disclaimer: yes, that is an ad from my company on the facing page). Also look at Jericevic et al (1989) Methods in Cell Biology 30:47-83.
MutliSpeck beads: The Molecular Probes pre-mounted slide kits should be ideal for DAPI and Fluorescein. I believe they were optimized for Rhodamine, but should still work ok for Texas Red. If your problem is with mounting, Mol. Probes now sells the beads in solution, so you can 'sprinkle' some on your specimens. If you have a different problem with the current MultiSpeck's, Mol. Probes may be able to work something out for you.
Sorry, but I usually buy my reference material from Mol. Probes and don't keep close track of other slide manufacturers.
} I read through the archives for information on uranyl glass test slides, but couldn't figure out if there is a definitive answer to the question, Does anyone still manufacture uranyl glass, and where could I get some?
} Thanks,
} Chi-Bin Chien
} chien-at-jeeves.ucsd.edu
Chi-Bin Chien,
You can obtain micro slide sized pieces of uranyl glass from:
Newport Industrial Glass, Inc.
1631 Monrovia Avenue
Costa Mesa, CA 92627
(714) 645 - 1500
(714) 645 - 6800 [FAX]
They have this glass (glass #3750) in large stock pieces, so it must be cut down to the size of a micro slide. The will grind it down to whatever thickness you
desire, as well. We ordered two micro slide sized peices in July of 1993. The cost was $86.
Good luck!
Cheers,
Bill Bug
*************************
* Bill Bug *
* Dept. of Biology *
* Swarthmore College *
*************************} O.k., all you good microscopists, I'm looking for a source for either a Uranium Glass
} Block or a suitable replacement for visualizing/teaching the light path on a light
} microscope. The block is placed in the microscope stage and the illumination path way
} can be viewed inside the block - particularly as the condensor is focused and as either
} the stage aperature or field aperature are varied.
}
} I know this can be visualize with a dilute milk solution but I really would prefer
} something a little more stable (and non-perishable).
by zahn.acpub.duke.edu (8.8.5/Duke-4.7.0) with ESMTP id KAA20344; Fri, 15 Jan 1999 10:36:52 -0500 (EST) Received: (from saram-at-localhost) by godzilla4.acpub.duke.edu (8.8.5/Duke-4.7.0) id KAA15860; Fri, 15 Jan 1999 10:36:49 -0500 (EST)
On Thu, 14 Jan 1999, Margaret Casey wrote:
} Date: Thu, 14 Jan 1999 16:02:26 -0500 } From: Margaret Casey {CaseyM-at-state.mi.us} } To: microscopy-at-sparc5.microscopy.com } Subject: TEM:virus isolation from foods for negative staining } } ------------------------------------------------------------------------ } 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 Fellow Listers, } } Does anyone have a good protocol or reference for virus isolation from food other than shellfish? We are trying to isolate and concentrate Norwalk virus from some foods implicated in a poisoning outbreak in Detroit. The foods in question are salami, garbanzo beans and some kind of cheese (I think provolone). We need to concentrate to at least 1 million particles/ml in order to use our prep for negative staining and TEM observation. Any suggestions would be greatly appreciated! } } Sincerely, } Peggy Casey } Michigan Dept. of Community Health } Lansing, Mich. } phone: 517-335-8102 } e-mail: caseym-at-state.mi.us } The best way to detect these kinds of viruses in dilute concentrations is PCR. In this case, viral numbers are likely to be very low, and getting enough to see by negative staining, unless you aggregate them with antiserum, may be difficult. If you want to proceed anyway, see concentration methods in Hayat and Miller, Negative Staining, McGraw-Hill.
Under separate cover, I will send you the name of an expert who does PCR on Norwalk. She is on the faculty at UNC and the CDC.
SM
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
How fast can you work and how long will the sections stay frozen? We l= ooked at frozen samples on a light microscope by covering the scope in a glov= e bag (~$15 from scientific supply houses) and doing all our work inside the = bag. We started by flooding the bag with dry nitrogen for a couple of hours = to drive out all humidity. We were able to freeze our samples in the bag,=
transfer them to the stage and back to the LN2 when we thought they wer= e starting to melt. You might be able to transfer the frozen sections in= to the bag and keep them frozen with some LN2 and quickly transfer them to the= stage of the microscope. This is a very kludgy setup, but it worked for us a= nd was dirt cheap.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-3537 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com =
since a couple of weeks I have to face a problem with our LEO EM912 which I could not solve up to now: I can't correct for the astigmatism in diffraction mode any more. If I use the "Image Stig xy" buttons they influence the appearence but the possible range of this potentiometers is not large enough for the correction. In addition the zero-beam moves very slowly across the screen (approx. 0.5mm/min on the final screen). I did a thorough alignment of all the settings of the microscope and the problem persists. We had service here and were told that it is probably some dirt particle in a part of the column which we can't clean. The only possibility would be to try to "burn" the particle in low-mag mode with high illumination aperture and "playing" with the illumination tilt in diffraction mode or the illumination shift in spot mode. And in fact, this helps for a short time, but the problem reappears latest after half an our again.
We have no problems with astigmatism in the image mode.
Does anybody out there has an idea what we could do, check, exchange ... to help solving the problem?
Greetings,
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
Lou - } } A few years ago I got hold of a beta version of a SEM Macintosh simulation } program, SuperSEM. It was a Macintosh program that was developed with } Supercard software. It was misplaced during the process of upgrading my } computers. Does anyone know if it exists?
Might you be thinking of Brian Griffen's "Virtual SEM"? It's a great simulation, and you'll find it listed in the CD-ROM section of the bibliography on Project MICRO's web page (URL below).
Caroline }
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Reply to: RE: Ti Alloys - TEM foil preparation Re: Electropolishing of Ti alloys Due to interest in this subject, I will list some additional = electrolytes I have used on Ti alloys. (3mm, jet polishing).
Ti-8 w/% Al 13% HCL -60 degrees C Ti-811 87% methanol 70 to 90 volts Ti-6Al-4V 25-35 mA.
Ti-13 Sn 130 ml HCL -50 degrees C 670 ml methanol 150 volts 100 ml butyl 60 mA.
Ti-3V 60 ml perchloric acid -50 degrees C Ti-20 Zr 590 ml methanol 50 volts Ti-14 Al 350 ml butyl cellosolve 10-15 mA
Ti-8 Al 60 ml perchloric acid -15 t0 -20 C (rolled) 460 ml ethyl alcohol, 95% 180 volts 280 ml butyl alcohol 40 mA. 100 ml butyl cellosolve Ti 5.3 g. = lithium chloride -40 degrees C. nanocrystals, 11.16 g. magnesium 150 volts compacted chloride 30 mA 500 ml. methanol 100 ml. butyl cellosolve (Note: the two salts above must be added to the combined solvents one = "powder" at a time, while stirring. The two dry salts react violently if = mixed together. This electrolyte has nearly as wide an application as = perchloric acid mixtures and is known as B K-2). I suspect that B K-2 will cause the least hydride problem. It was = developed to eliminate hydrides in vanadium TEM foils. The voltages given = would need to be reduced about 20% for a twin jet system due to lower = electrical resistance of that configuration. My work was done on a South = Bay vertical single jet unit. I have no vested interest in the above equipment, but have enjoyed = the ease with which good polishing conditions can be obtained and = reproduced since 1975 or so. Good luck! Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
E-mail: {bkestel-at-anl.gov} Thomas, Larry 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 Satyarth: } } An excellent paper on the subject is: } } "In Situ Hydride Formation in Zirconium and Titanium during Ion Milling" } Graham J.C. Carpenter et al, JMSA Vol.1 No. 4 pp 175-184 1995. } } From the paper you will see that a key factor in eliminating hydride } formation is having a clean sample free from hydrocarbon contamination. } = This contamination could be remnants from the dimpling process or other } pre-thinning steps or it could be caused by the back streaming of } diffusion } pump oil in your ion mill. As titanium has a high chemical affinity for } hydrogen, you may want to look over your preparation steps and try to } eliminate any areas of possible contamination. If you are still having } a } problem, a quick cleaning of both the specimen and the specimen holder } in a } Plasma Cleaner should take care of it. } } If you have an interest, I can send you a copy of the above referenced } paper. I also have papers on plasma cleaning which may be of interest. } } NOTE: South Bay Technology does manufacture the PC150 Plasma Cleaner and } therefore I have a vested interest in promoting its use. } } Best regards- } } David } Writing at 9:38:23 AM on 1/12/99 } } } ******************************= *********************************** } ********** } ************************ } } David Henriks TEL: } 800-= 728-2233 (toll free in the USA) } South Bay Technology, Inc. +1-949-492-2600 } 1120 Via Callejon FAX: } +1-949-492-1499 } San Clemente, CA 92673 USA e-mail: } henriks-at-southbaytech.com } } } ***************************************************************** } ********** } ************************ } } } } } } } Please visit us at http://www.southbaytech.com { { { { { } } Manufacturers of precision sample preparation equipment and supplies for } metallography, crystallography and electron microscopy. } } Message text written by Satyarth Suri } } } Hello: } I am currently working on mechanical behavior / microstructure } correlations in single colony near apha Ti Alloys. I am currently } preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but } have run into the problem of a very uneven alpha phase morphology, } the alpha phase has island formation - we are using a cold stage } to minimize the hydride formation at the interface. I am using } 3 micron/6micron diamond paste during the dimpling process - the } foil itself otherwise is fairly clean in terms of the dislocation } content. Could anyone in the microscopy land have some suggestions } in terms of what the problem may be? } } thanks - if you want you can respond directly to suri.3-at-osu.edu } } -satyarth } } { } } } } } RFC822 header } ----------------------------------- } } Received: from dns2.anl.gov (dns2.anl.gov [146.139.254.3]) by } horus.et.= anl.gov (8.6.11/8.6.11) with ESMTP id SAA02218; Tue, 12 Jan 1999 18:17:43 -= 0600 } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com } [206.69.208.= 10]) by dns2.anl.gov (8.9.1a/8.6.11) with SMTP id SAA19261; Tue, 12 Jan = 1999 } 18:18:22 -0600 (CST) } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) } id RAA22399 for dist-Microscopy; Tue, 12 Jan 1999 17:41:01 -0600 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by } Sparc5.= Microscopy.Com (8.6.11/8.6.11) with SMTP id RAA22384 for } "= MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 12 Jan 1999 17:40:30 -= 0600 } Received: from Sparc5AAEM (aaem.amc.anl.gov [146.139.72.3]) by } Sparc5.= Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA22377 for } {microscopy-at-= sparc5.microscopy.com} ; Tue, 12 Jan 1999 17:40:17 -0600 } Received: from pnl.gov (relay.pnl.gov [130.20.128.34]) by Sparc5AAEM } (8.= 6.11/8.6.11) with ESMTP id RAA14848 for {MICROSCOPY-at-aaem.amc.anl.gov} ; Tue,= 12 Jan 1999 } 17:52:22 -0600 } Received: from pnlmse1.pnl.gov by pnl.gov (PMDF V5.1-12 #28154) } with ESMTP id {01J6GJ2FP8R4970ZC9-at-pnl.gov} for MICROSCOPY-at-aaem.amc.anl.= gov; } Tue, 12 Jan 1999 15:54:41 PST } Received: by PNLMSE1.pnl.gov with Internet Mail Service (5.5.2232.9) } id {CR51M2B5} ; Tue, 12 Jan 1999 15:54:42 -0800 } Content-return: allowed } Date: Tue, 12 Jan 1999 15:54:36 -0800 } From: "Thomas, Larry" {Larry.Thomas-at-pnl.gov} } Subject: RE: Ti Alloys - TEM foil preparation } To: Satyarth Suri {suri-at-mse.eng.ohio-state.edu} , } Microscopy ListServer {MICROSCOPY-at-aaem.amc.anl.gov} , } "'South Bay Technology'" {Henriks-at-CompuServe.COM} } Message-id: {7A04D1887189D2118EEB00A024BF29DA41CB17-at-pnlmse2.pnl.gov} } X-Mailer: Internet Mail Service (5.5.2232.9) } Errors-to: Microscopy-request-at-sparc5.microscopy.com } Content-Type: text } Content-Length: 5172 } Status: }
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by pilsener.ucs.ualberta.ca (8.9.1a/8.9.1) with ESMTP id MAA10247; Fri, 15 Jan 1999 12:46:33 -0700 (MST) Received: from localhost (mingchen-at-localhost) by gpu2.srv.ualberta.ca (8.8.5/8.8.5) with SMTP id MAA67494; Fri, 15 Jan 1999 12:46:33 -0700
Hi Mary,
I have done a lot of fixation for plant material for more than 25 yrs. I think 2% paraformaldehyde-2.5% glutaraldehyde fixative in 0.2 M cacodylate or phosphate buffer (pH 7.2) is the best.
Good luck.
Ming
On Tue, 12 Jan 1999, Mary Gail Engle 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 need some advice as to the best protocol for fixing fresh plant leaves } for routine TEM. My only experience has been with animal tissue. } Thanks, } MG Engle } } }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * #1074B Dentistry Pharmacy Building * * University Of Alberta. * * Edmonton, Alberta, Canada T6G 2N8 * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Lou Ross wrote: } } } } A colleage from an industrial SEM lab contacted me about a problem with } contamination on the final aperture on an variable pressure SEM. This SEM } was recently purchased by one of their branch plants to examine } semiconductor products. Although it is a variable pressure SEM, they have } only operated it in the high vac mode like a standard SEM. } } The problem they are experiencing is that the final aperture presumably } gets so dirty they have to change the aperture every two weeks. } Unfortunately the manufacturer has not been able to shed any light on this } situation. } } If the vacuum system is alright, I suggested that the problem might be with } the samples. Supposedly the branch lab operators are following the same } sample prep protocol established in the local SEM lab. } } Any other ideas out there? } } Thanks in advance, } Lou Ross } Senior Electron Microscope Specialist } Room 101 } Department of Geological Sciences } University of Missouri } Columbia, MO 65211 } (573) 882-4777 } (573) 882=5458 fax } www.missouri.edu/~geosclmr/ebaf.html
Dear Lou Ross,
Based on your message concerning the contaminated aperture, my feeling is that the problem probably rests somewhere other than the aperture itself. But since there is the slightest possibility that there might be a problem with the aperture perhaps we can assist you. Ladd produces the vast majority of apertures/microholes used in the United States and the aperture you have may very will be a Ladd aperture. To make sure there is no residual contamination on an aperture we have a post production protocol that is designed to eliminate any contamination that results during production. In fact we have some apertures/microholes used in fluid and gas control that are required to be absolutely contamination free. We have a protocol to ensure that. It is also important that an aperture be shipped or stored in a glass or non-contaminating vial and that the aperture surface should not touch tissue or even lint free cloth. We would be glad to examine the aperture to see if the contamination was a result of packaging, handling or the production process. Please contact us if you if you wish us to do that.
John Arnott Chairman
-- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
My institution needs to purchase a new cryostat in the next several months. I would appreciate any comments from cryostat users about their satisfation/ dissatisfaction with particular makes and models of relatively new cryostats.
We do not anticipate particularly heavy usage of this equipment, but the tissues that will need to be sectioned range from soft tissues such as brain and heart all the way to mouse and rat bone. Some investigators would like to collect sections in excess of 30 microns for confocal microscopy projects, and others would like to serial section whole organs from transgenic mice. Are disposable knives sufficient for this range of usage?
Also, cryostat vendors are welcome to email me directly. Surprisingly, it has been like pulling teeth to get some companies to talk to a ready and willing customer.
Thanks for your help,
Dr. Steve Fields Oklahoma Medical Research Foundation 825 N.E. 13th Street Oklahoma City, OK 73104
Good Morning, Lou! There are several reasons why the final aperture may be getting dirty on a variable pressure SEM, and why this may be more noticeable on semiconductor samples. Several causes contribute to contamination within SEM chambers. If this system has been used for any other analysis other than semiconductor, at low vac modes, residues from prior analysis may be lining the chamber walls - these are difficult to remove even by baking the chamber out. Additionally, this system probably has oil-cooled pumps - the deleterious effect of backstreaming oils into chamber which "gums" onto the final aperture is well established, but can be diminished with a strategically located dry N2 purge port in the sample exchange chamber. Mounting media is also of concern - since most conductive paints outgas, these vapors can also contaminate the F.A. quickly, especially if the material is not exceedingly well dried. Simple cleanliness may also be the culprit - by handling a metal sample fixture with bare hands, oils and sweats are deposited on the fixture. These deposits love to attach to the F.A. (and cold tip of EDS detectors too!) As most semiconductor analysis requires fairly high magnification, and often lower KeV, these problems are exacerbated. I've delt with these and many other problems concerning the optimal performance of SEM's with semiconductor materials, and would be pleased to look over his system for performance improvements. Let me know if I may be of assistance!
In a message dated 1/14/99 8:41:46 PM Mid-Atlantic Standard Time, RossLM-at-missouri.edu writes:
{ { A colleage from an industrial SEM lab contacted me about a problem with contamination on the final aperture on an variable pressure SEM. This SEM was recently purchased by one of their branch plants to examine semiconductor products. Although it is a variable pressure SEM, they have only operated it in the high vac mode like a standard SEM.
The problem they are experiencing is that the final aperture presumably gets so dirty they have to change the aperture every two weeks. Unfortunately the manufacturer has not been able to shed any light on this situation.
If the vacuum system is alright, I suggested that the problem might be with the samples. Supposedly the branch lab operators are following the same sample prep protocol established in the local SEM lab.
Any other ideas out there?
Thanks in advance, Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html } }
I am asking for help. We bought a JSM 5410 SEM (3.5 nm at 30kv 8mm working distance) and JEOL 2010 microscopes (without FEG) here. Both microscopes has almost finished installation. For JEOL 2010 I checked our quotation written by JEOL that they should provide working pressure of 6x10-6 Pa or better at specimen as read by the ion pump. But now we got best vacuum is 1.5x10-5 Pa. Is there anyone who has JEOL 2010 TEM and can tell me your best working value ASAP? Thank you very much in advance.
As for JSM 5410, is there anybody who has JSM 5000 series SEM and can send me your resoultion shooting photos by JEOL engineer (resolution 3.5nm) to let me have a look. I will pay the Fedex fee and send you back by Fedex express. Thank you very much.
Sincerely yours,
Weilie Zhou (Ph.D)
************************************************ Advanced Materials Research Institute University of New Orleans Science Building 2021 New Orleans, LA 70148 Tel:(504) 280-5570 Fax:(504) 280-3185 ************************************************
I have given the following advice to several users of variable pressure SEMs who have dirty microscopes and contamination, and they all said it cured the problem.
Don't leave it in high Vacuum mode all the time. Some variable pressure SEMs are very dirty due to backstreaming at high vacuum. They are designed for low vacuum not high vacuum. By placing the system into low vacuum mode, the chamber is placed into viscous flow vacuum dynamics which stops backstreaming and purges out contaminants. Try leaving it in low vacuum mode overnight for a month and check the results.
Let me know if this helps.
Ronald Vane XEI Scientific
Disclaimer: XEI is in the anti-contamination business. } } -----Original Message----- } From: Lou Ross {RossLM-at-missouri.edu} } To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com} } Date: Thursday, January 14, 1999 1:48 PM } ----------------------------------------------------------------------. } } } } } } A colleage from an industrial SEM lab contacted me about a problem with } } contamination on the final aperture on an variable pressure SEM. This SEM } } was recently purchased by one of their branch plants to examine } } semiconductor products. Although it is a variable pressure SEM, they have } } only operated it in the high vac mode like a standard SEM. } } } } The problem they are experiencing is that the final aperture presumably } } gets so dirty they have to change the aperture every two weeks. } } Unfortunately the manufacturer has not been able to shed any light on this } } situation. } } } } If the vacuum system is alright, I suggested that the problem might be with } } the samples. Supposedly the branch lab operators are following the same } } sample prep protocol established in the local SEM lab. } } } } Any other ideas out there? } } } } Thanks in advance, } } Lou Ross } } Senior Electron Microscope Specialist } } Room 101 } } Department of Geological Sciences } } University of Missouri } } Columbia, MO 65211 } } (573) 882-4777 } } (573) 882=5458 fax } } www.missouri.edu/~geosclmr/ebaf.html } } } } }
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Happy New Year all,
A question for the New Year: what are EM laboratories using for pure water = these days? Is it possible to use deionized water, or even the distilled = water from Rite Aid or Arrowhead? In the past I had a still which was fed = from a deionized water supply. We had no problems at all with this.
If the preferred purification process turns out to be deionized water, = which quality is best - Type 1 (HPLC etc.) or will lower quality suffice? = Also has anyone had problems with resin in the water?
Regards,
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/apw.htm
microscopy-at-Sparc5.Microscopy.Com, tina-at-pbrc.hawaii.edu MMDF-Warning: Parse error in original version of preceding line at rfdata.net
Another quick cheap fix is dress warm and do the work in a walk in freezer. The glove bag would work well to keep your breath from freezing on the scope.
When I was working at Ag Engineering we had and etymology and vet student that spent the summer in a walk in cooler monitor tick behavior from freezing to 55 degrees F.
Gordon
Gordon Couger gcouger-at-couger.com Owner PRAG-L PRactical AGriculture List www.couger.com/prag-l Stillwater, OK 405 624-2855 GMT -6:00
How fast can you work and how long will the sections stay frozen? We looked at frozen samples on a light microscope by covering the scope in a glove bag (~$15 from scientific supply houses) and doing all our work inside the bag. We started by flooding the bag with dry nitrogen for a couple of hours to drive out all humidity. We were able to freeze our samples in the bag, transfer them to the stage and back to the LN2 when we thought they were starting to melt. You might be able to transfer the frozen sections into the bag and keep them frozen with some LN2 and quickly transfer them to the stage of the microscope. This is a very kludgy setup, but it worked for us and was dirt cheap.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-3537 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com
} From Microscopy-request-at-sparc5.microscopy.com Thu Jan 14 14:44:05 1999 } } Date: Thu, 14 Jan 1999 09:11:29 -1000 (HST) } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } To: Microscopy Listserver {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Cold stage for LM } } Happy New Year to you all } } .......... I have liquid nitrogen, styrofoam, and a machine } shop. I also have high humidity. Can anyone tell me how to kludge } together a cold enough stage that we can get images of these } frozen sections while preventing frost? I have some general ideas, but } I'd appreciate any specific plans or ideas.............. } } } Aloha, } Tina } }
Tina, We have done some work with a cold stage for cathodoluminescence (CL) studies of minerals, ceramics, etc. A simple vacuum chamber (mechanical pump - pressures of 30 to 100 millitorr) is used for the CL work and the chamber sits on the stage of the light microscope. For cold stage work we use a simple system consisting of a chillable plate that is cooled in liquid nitrogen outside the chamber and then inserted and pumped down. Temperatures low enough to dramatically change the CL behaviour are achieved for about 10 - 12 minutes and there is minimum condensation. The vacuum helps to provide thermal insulation for the chillable plate and also minimizes the amount of water vapor that can condense. The plate itself is designed to minimize thermal transfer to the other parts of the vacuum chamber.
Don Marshall
(Claimer: RELION is in the business of CL instrumentation for light microscopes and chillable sample plates are one of the items we offer.)
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
A three month (June, July, and August) position is open for a microscopy oriented technician at the Marine Biological Laboratory, Woods Hole, MA. We would like to attract someone with some knowledge of biological preparative techniques and experience in laser scanning confocal microscopy, TEM, SEM, and/or LM.
The technician will assist in the Central Microscopy Facility. The technician's duties will be to check out incoming investigators in the usage of our equipment and then to supervise its continuing usage and to perform contract work for investigators. This may include fixation, embedding, sectioning, scope use, darkroom work, etc. The technician will also provide routine maintenance.
This is a short term and scientifically rewarding position. Salary will be in the $7 to $10/hour range. Housing may be available to rent through MBL.
For more information, including a more detailed position description, please contact Louis Kerr at: MBL, 7 MBL Street, Woods Hole, MA 02543. Telephone, 508-289-7273; or at Email: lkerr-at-mbl.edu. Please apply to: Human Resources, MBL, 7 MBL Street, Woods Hole, MA 02543. or resume-at-mbl.edu.
An Equal Opportunity/Affirmative Action Employer/ Non-smoking workplace.
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
} SUMMER MICROSCOPY TECHNICIAN POSITION AVAILABLE } } A three month (June, July, and August) position is open for a microscopy } oriented technician at the Marine Biological Laboratory, Woods Hole, MA....
} ...This is a short term and scientifically rewarding position. Salary will be } in the $7 to $10/hour range. Housing may be available to rent through MBL... } } ...For more information, including a more detailed position description, } please contact Louis Kerr at: MBL, 7 MBL Street, Woods Hole, MA 02543. } Telephone, 508-289-7273; or at Email: lkerr-at-mbl.edu. } Please apply to: Human Resources, MBL, 7 MBL Street, } Woods Hole, MA 02543. or resume-at-mbl.edu.
I once had a senior student (at UC Berkeley) who took this job. He was recruited into a top MD/PhD program and met his wife, all in a summer of hard work at MBL. Apply, folks!
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Perhaps a little off-topic, but for those of you who archive microscopy images to recordable CD-ROM, does anyone know of a strategy for placing a "software lock" on the CD-ROM which would require a "key" to access, view, or copy the CD-ROM contents? I'd rather not compress or encrypt images and use a password to access but to have a security option initiated upon mounting of the CD-ROM.
Thanks very much for any info,
Brian C. Tryon MCP-Hahnemann School of Medicine Ann Preston Hall, Box 551 3300 Henry Avenue Philadelphia, PA 19129 USA
----------------------------------------- "Quantifying is a committing task." - Cruz-Orive, 1994.
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." - Richard Feynman --------------------------------------------------------------
I am a school counselor at the elementary level. Our leadership team is spending some of our grant money on microscopes for a Science lab which we are developing. I would like to know basic information on purchasing microscopes for the elementary student. We hope to purchase five-six scopes depending on the price. Can you help with this information? Please e-mail me at diorado-at-stc.net Thanks! Diane
Tina: I am afraid I will not provide detailed plans, just a couple of hints. I expect that the easiest way would be to built a suitable, insulated enclosure with a minimal opening for the objective lens. On top of that small opening a resistance heater loop may be required. Somewhere you would have an inlet for dry industrial nitrogen gas and with a small sealed chamber and the only opening near the lens, no air and therefore no moisture can enter. Gas flow could be quite low - if the outlet hole is not too large. Gasflow would need to be activated well before freezing. Temperature control and nitrogen flow are other problems, but avoiding frost tends to be the greatest challenge. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
Hi all
Here is a 'WISH' list of bits that we want for our labs, if anyone has these bits that they want to giveaway, trade, sell, we look forward to hearing from you.
Spares for a Gatan model 600 ion beam thinner
Spares for JEOL 35-CF Spares for JEOL 100-CX
EDXS for a JEOL 35-CF with with electronics. It doesn't have to have excellent energy resolution. EELS for a JEOL 2010 or JEOL 100-CX Four quadrant Back Scatter detector (UHV Compatible)
An oven Any specimen preparation gear for physical and biological specimens Microtome and accessories
Enlargers working or broken Timers for enlargers Print dryer
Printers - B&W and color
Dessicators (vacuum)
Chemical storage cupboard
I know some of these are extravagant, but I have to ask. The Gods just might smile upon me today.
Thank you George G. Theodossiou Dept Applied Physics RMIT Email: George-at-bunyip.ph.rmit.edu.au ph:+61 3 9925 3394 fax:+61 3 9925 5290
Hi all Best wishes for the new year to all. We have a small problem that needs fixing and would like to get a few = other ideas on the matter before the user spends money on fixing it. The user has an ISI SEM which has the typical back streaming problems. = This oil is contaminating the ED detector window and therefore causing = absorption of the light x-rays. Other than cleaning the window regularly, which carries the danger of = blowing the window and detector, we were looking at methods to stop this = contamination. We are fitting a foreline trap to the Rotary pump, but = experience shows that you still get oil back streaming. We could try a = Ln2 cold finger but this is a hassle to keep toped up and can only be = fitted above the diff pump which makes engineering a problem. A SEM clean gas leak system would also help but they are very short on = cash. ( surprise surprise) They also need to work at high mag and so = whilst the SEM is in use the SEM clean system could not be effective.=20 The only other idea we have is to fit a peltier cold finger to the SEM = chamber in the hope of attracting most of the oil to the cold finger = and thus not contaminating the detector. This we though could be = controlled to have a fairly low temp whilst the SEM is in use and = frequent sample changes are required, then turn up the power for when = the SEM is not in use to be more efficient.
Has any one fitted a peltier to their EM before ? What experiences have = you had with them and what size and power would work.=20 All ideas welcome.=20 Thanks =20 Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
Hello fellow listservers, I've recently discovered a problem with a batch of a particular type of cold setting epoxy resin. I have been using this batch of resin on an infrequent basis for about 2 years (and is probably about 4 years old). When I analysed some of the cured resin by microanalysis last week appreciable bromine (Br) was detected as well as the usual chlorine content (Br} Cl). The 'introduction' of the Br is a recent thing because I also analysed some resin (from the same batch) prepared approximately one year ago. The analysis of this showed no Br with Cl the only detectable element present (with the Be window of the EDS detector in place that is). Is it possible therefore for the resin to in some way deteriorate with time and for Br compounds to form? If so, what are the chemical reactions going on here. I'm very certain that the Br is not a contaminant. Also, I should point out that if this is a chemical degradation of the resin then the physical characteristics of the cured resin are normal. I would appreciate any thoughts on the matter. Regards
Martin Roe Macaulay Land Use Research Institute Aberdeen AB15 8QH Scotland United Kingdom
Greetings All, } } I have been reading this list for well over a year and am confident that this is the place to come if one has a question regarding microscopy...so here goes. } } The Chemical Hygiene Officer of the company I work for has decided that all laboratory personnel must wear full eye protection when in a lab. } This presents a problem to me, because of my near sightedness, I prefer to look through the optical/IR microscope without glasses and this results in a side splash safety hazard. Does anyone have any suggestions on how to fully protect myself from side impact and satisfy the safety requirements and view my microscopy images? Thank you all in advance and if anyone wants a summary of responses, I will be more than happy to oblige. } } } } } Deborah Hills-Haney } Research Analytical Services/NMR Lab } International Flavors and Fragrances R&D } 1515 US Highway 36 } Union Beach, NJ 07735 } } Phone: (732) 335-2663 } Fax: (732) 335-2591
______________________________________________________ Get Your Private, Free Email at http://www.hotmail.com
} Hello fellow listservers, } I've recently discovered a problem with a batch of a particular } type of cold setting epoxy resin. I have been using this batch of } resin on an infrequent basis for about 2 years (and is probably } about 4 years old). When I analysed some of the cured resin by } microanalysis last week appreciable bromine (Br) was detected as well } as the usual chlorine content (Br} Cl). The 'introduction' of the Br } is a recent thing because I also analysed some resin (from the same } batch) prepared approximately one year ago. The analysis of this } showed no Br with Cl the only detectable element present (with the Be } window of the EDS detector in place that is). Is it possible } therefore for the resin to in some way deteriorate with time and for } Br compounds to form? If so, what are the chemical reactions going on } here. I'm very certain that the Br is not a contaminant. Also, I } should point out that if this is a chemical degradation of the resin } then the physical characteristics of the cured resin appear to be normal. I } would appreciate any thoughts on the matter. Regards } } Martin Roe } Macaulay Land Use Research Institute } Aberdeen } AB15 8QH } Scotland } United Kingdom } } }
} I am a school counselor at the elementary level. Our leadership team is } spending some of our grant money on microscopes for a Science lab which } we are developing. I would like to know basic information on purchasing } microscopes for the elementary student. We hope to purchase five-six } scopes depending on the price. Can you help with this information? } Please e-mail me at diorado-at-stc.net Thanks! Diane
I'm happy to help; that's what Project MICRO is all about. You'll find detailed general purchase and evaluation advice on the MICRO web page (URL below), plus a list of possible sources. For elementary science, I strongly favor monocular "dissecting" scopes; you can get good ones for under $80. Plus perhaps one conventional 3-objective (4, 10, 40x) compound scope, for $120. Where are you located? I may be able to get you some advice from a local MSA member.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Martin J. Roe 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 fellow listservers, } I've recently discovered a problem with a batch of a particular } type of cold setting epoxy resin. I have been using this batch of } resin on an infrequent basis for about 2 years (and is probably } about 4 years old). When I analysed some of the cured resin by } microanalysis last week appreciable bromine (Br) was detected as well } as the usual chlorine content (Br} Cl). The 'introduction' of the Br } is a recent thing because I also analysed some resin (from the same } batch) prepared approximately one year ago. The analysis of this } showed no Br with Cl the only detectable element present (with the Be } window of the EDS detector in place that is). Is it possible } therefore for the resin to in some way deteriorate with time and for } Br compounds to form? If so, what are the chemical reactions going on } here. I'm very certain that the Br is not a contaminant. Also, I } should point out that if this is a chemical degradation of the resin } then the physical characteristics of the cured resin are normal. I } would appreciate any thoughts on the matter. Regards } } Martin Roe } Macaulay Land Use Research Institute } Aberdeen } AB15 8QH } Scotland } United Kingdom
Martin, One possibility that comes to mind is the storage cabinet of the resin: if you store bromine in the same cabinet and the resin is packed in a polymer flask, I can think of some diffusion out of the bromine into the resin bottle.
An other possibiliy is that there is a solvent used with a part of the resin, which contains Br. If the resin is growing older, there may be decomposition/degradation of the solvent, leaving a non volatile Br conaining component behind. Check each single component of the resin.
Hope this helps -- Gerhard Frank UNIVERSITAET ERLANGEN-NUERNBERG Phone: +49 9131 85 28606 Institut fuer Werkstoffwissenschaften VII Fax: +49 9131 85 28602 Cauerstr. 6, D-91058 Erlangen, Germany frank-at-ww.uni.erlangen.de
I ordinarily wear bifocals with an astigmatism correction. My company has bought me distance-only safety glasses, which I use irregularly but satisfactorily in place of my bifocals at the LM. The main advantage is the lack of the bifocal line.
If you can't bear the glasses, you might propose writing a "job hazard analysis" in which you analyze the hazards involved in your specific operation and propose specific solutions other than safety glasses, e.g., a splash shield between you and other workers, use of safety glasses for certain operations but not for viewing, after you have assured that risk of splash etc. during viewing is minimal.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
Our laboratory is in the process of 'remodelling' and have found several items which we would be able to part with at this time. These items include: 1. A Polaron Critical Point Dryer 2. An Arkay CD-80 cabinet dryer 3. A Gatan model 673 mark 2 wide angle TV system with a Data Translation 3851 A/D converter board 4. A Gatan model 622 fiber optically coupled TV system
This equipment was 100% functional the last time it was used. If you are interested in any of this equipment please contact me off line.
Jon Charlesworth, Coordinator Electron Microscopy Core Facility Mayo Clinic 1426 Guggenheim Building Rochester, MN 55905 ph: (507) 284-3148 fax: (507) 284-9349 email: charlesworth.jon-at-mayo.edu
Is anyone familliar with specific fixation- and/or staining techniques for rat and/or chicken eyes. Someone on our laboratory wants to to do some research on eyes. He wants to stain specific membranes like Bowmann and Desmett membrane.
Hi, I am looking for a program to convert weight % to Atomic % and vise versa preferably on a Power Mac.. An old program we have works on pre '90 Mac's but not on the newer ones. Anyone have any ideas? Thanks- Mike
I am looking for a 8cm by 10cm (TEM film format) film holder for a Leafscan 45 negative scanner.
I've been told that the company which makes the Leafscan was bought out. Does anyone know by whom, or who might now deal in parts?
Thanks, Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
Hope that there is room on this list for a bit of pre-millenial philosophy.
Before I saw John White's prototype laser scanner in 1986, I had spent my time trying to improve instrumentation for high-resolution SEM and TEM. Though capable of "seeing molecules" and even labelling them, these instruments and have one tremendous disadvantage for the biologist: because they form their images using quanta that carry (much) more than 5 electron volts of energy, the act of imaging destroys molecules. Consequently, these instruments can only be used to image dead cells.
So, even when the confocal microscope was seen primarily as a method of making 3D images of biological structures that had been fixed and stained, my own focus was on using it to image living specimens.
Early confocal microscopes were so inefficient in their use of the light from the specimen that one could only image a "living" specimen at high magnification for a very sort time (one frame?) before it was well on the way to death. However, the wide-field/deconvolution contingent under Agard and Sedat showed that 3D imaging of living cells was possible, so we tried harder.
Even though instrumental improvements have now reduced the amount of damage produced per image by about 100x, and even though "the hidden agenda" of the Second edition of the Confocal Handbook was to give researchers the tools needed to view living specimens optimally, it seemed (seems?) to me that the large preponderance of confocal images is still made on dead cells.
There were several important reasons:
* Not clear which biological questions could be best approached in this way. * Hard to keep cells alive on microscope stage. * Results tedious to obtain and outcome uncertain. * Unclear which dyes were most suitable for live-cell operation in terms of cytotoxicity and bleaching * The choice of operational parameters was complicated by interactions between them. If pixel and raster-size, NA, sampling time and, laser power all have to be optimized to produce the best signal-to-noise and signal-to-damage ratios. * Single=photon or multi-photon. * Finally, there seemed to be a geography problem. While individual groups were making great strides, their isolation kept the practical knowledge that they gained by trial and error from reaching many of those who had good biological problems but lacked "a place to start from".
This Email Listserve was a response to the isolation and, I believe, it has been a major factor in the development of the field. Thanks Bob and now Steve!!
However, sometimes one needs a more personal type of interaction than is yet possible over email. In an attempt to fill this perceived gap, 5 years ago I decided to organize a 10-day course devoted to the 3D microscopy of living cells. Although the course would also cover advanced 2D techniques, the bulk of the time would be devoted to hands-on, laboratory sessions with no more than 3-4 students to a 3D Workstation. To focus attention, students were encouraged to design and carry out a 3D Live-cell Microscopy project during 5 evening sessions in the 3D part of the course and them present the results on the last morning.
Since that time, the course has served over 80 students from 18 countries, and will be offered again this year June 16 -27 (with a 3D Image Processing Workshop from June 29 - July 1. More info at http://www.cs.ubc.ca/spider/ladic/course/bulletin.html).
My question is this:
Now that the field has progressed and so many groups are actively publishing in the field (and helping out on the Email Net!), is there still need for such a course?
Although organizing 15 faculty members to come to beautiful Vancouver isn't so difficult, it costs the manufacturers an estimated $200,000 to send the equipment and personnel for the 11, 3D Workstations and you know who pays for that in the end!!
In addition, the controversy about who can, and cannot, do 2-photon excitation and under what conditions becomes ever more strident. Besides making organizing more onerous, this is particularly sad because it is clear that the main advantage that 2-photon imaging has over other methods is in imaging live cells, where conventional anti-bleach agents cannot be used, and where objects thicker than 20 micrometers must sometimes be viewed.
Last year, this controversy was almost certainly a factor in the last minute non-appearance of a multi-photon instrument (we did have 2 others) and this year it may lead to the total absence of one our major sponsors from earlier years (we will have at least 9 others).
I ask whether we have outlived our usefulness because I want to know. And I don't any more knowledgeable group to ask.
Although the student evaluations have been predominantly very positive, and the course itself has definitely improved as we found out what worked best, the trend in student numbers has not been encouraging: the first year we had over 50 applications for 28 places, the second about 35 applications and last year we had only 24 students after 4 dropped out at the last moment. Applications this year are about the same as last year but the March 1 deadline is still far away so this doesn't mean much.
I ask for your thoughts in planning beyond this year, because, given the "informal" nature of the founding of the course, there is no "institutional support" and tuition and expenses are handled through my personal account. So far, we are solvent but I personally can't afford to make good losses, should they occur.
And just to make sure there is no misunderstanding: I am not talking about this year's course (1999). I am asking for the future.
The Emory University Neurology Microscopy Laboratory, the University of Georgia Botany Department EM Laboratory, and Leica Microsystems, Inc.
Present a Cryo Techniques and Immunogold Workshop.
April 18-23, 1999 at the Campus of Emory University in Atlanta, GA.
Objectives
1. To provide researchers the opportunity to learn the theory and practice of cryo techniques for biological sample preparation and immunogold labeling.
2. To permit participants to process their own samples using these techniques under expert guidance.
Scitex America owned Leaf and its products. The address for Scitex was: One O'Hare Center 6250 River Rd. Rosemont, IL 60018 (847) 692-6000
The film holders were negative carriers made by Beseler for the Beseler 45M series and CB7 enlargers. Why not contact Beseler either directly or through a dealer to get a carrier for, say, 35 mm film. Then you can have your shop cut the appropriate opening in the carrier for your film.
Below is the text of a message I received from Joe Peng at Scitex: --------------------------------------------------------
Dear Joost,
Are you doing these eyes for TEM or LM? We do mice eyes for TEM (in plastic) and have gotten some really beautiful shots of Desmett's membrane. We have a whole fixation and embedding protocol worked out. The membranes are perfect. If this is for TEM, let me know and I'll send you our fixation and embedding protocol.
Regards!!
Lesley Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
I think Tracy and Jean would be a good start. There are some other TEM labs on campus and in the Vet College and Labs. They should be able to tell you about them, too. IThat's about as much as I know since I am over on the Engineering/Materials side of campus.
Name: PEPPER TRACEY M Phone(w): 515-294-3872 Phone(w): 515-769-2471 Fax: 515-294-3932 Email(w): tpepper-at-iastate.edu Office: Address: 001 BESSEY City/State: AMES, IA 50011-1020
Name: OLSEN JEAN ANN Phone(w): 515-294-1009 Phone(w): 515-233-2696 Fax: 515-294-1401 Email(w): jaolsen-at-iastate.edu Department: VETERINARY MEDICAL RESEARCH INSTITUTE Office: Address: VMRI BLDG 2 City/State: AMES, IA 50011-1240
At 03:36 PM 1/19/99 -0600, you wrote: } } Who would be a contact person at Iowa State Univ. for biological TEM ? } Possibly involving Immuno EM. Thanks. } } Rick L. Vaughn }
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Hi,
Does anyone out there have a vacuum evaporator that is taking up space? I = have lots of space but no evaporator. =
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/apw.htm
with Novell_GroupWise; Wed, 20 Jan 1999 11:28:14 +1100 Message-Id: {s6a5bdce.092-at-rsbs.anu.edu.au} X-Mailer: Novell GroupWise 5.2
We are just setting up TEM EDXA in a multidisciplinary unit, and need to = acquire standards covering a wide range of applications. Any recommendatio= ns, pitfalls etc? (Philips 430 TEM up to 300kV, Oxford ISIS ultra-thin = window detector).=20 thanks Sally
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475 ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525
This message is in MIME format. The first part should be readable text, while the remaining parts are likely unreadable without MIME-aware tools. Send mail to mime-at-docserver.cac.washington.edu for more info.
I have a series of se images of a serial sectioned material which I would like to display as a volume rendered image. I have aligned the images, using an nih-image macro, however the brightness and contrast differs between them. As I understand it I need to perform histgram equalisation to correct for these differences.
I've noticed photoshop allows the user to load a transfer function using the "curves" dialog box, but it doesn't give the desired result.
Is this because the function which you specify is not the cumulative transfer function based on the reference histogram area??
Is there anyone who has done this before? I am using nih-image and photoshop for these tasks. As for the reconstruction, well any suggestions there would be apreciated also.
Thanks
---------------------------- Travis Baroni (PhD Student) | tbaroni-at-cyllene.uwa.edu.au | The University of Western | Australia, Nedlands, WA. | 6907. | ----------------------------
As Curriculum & Technology Specialist for the Alliance for Education, I take a Personal Scanning Electron Microscope (manufactured by RJ Lee, Pittsburgh, PA) to schools and museums. It is easily operated by anyone who can use a joystick and a mouse. I generally have students work in teams. Since our major funder is the Air Force, I refer to the responsibilities in flying terms, such as pilot and copilot.
We coordinate with the teacher to select specimens appropriate to their curriculum. Since we are using 17 inch monitors, it's not a particularly flashy activity for a large group. With large screens and appropriate sound systems, it could be. The main thing, though, is to choose a system that can be operated hands-on by young people.
For more information, you can call me at (937) 222-2934 between 9 and 5 EST.
I have a series of se images of a serial sectioned material which I would like to display as a volume rendered image. I have aligned the images, using an nih-image macro, however the brightness and contrast differs between them. As I understand it I need to perform histgram equalisation to correct for these differences.
I've noticed photoshop allows the user to load a transfer function using the "curves" dialog box, but it doesn't give the desired result.
Is this because the function which you specify is not the cumulative transfer function based on the reference histogram area??
Is there anyone who has done this before? I am using nih-image and photoshop for these tasks. As for the reconstruction, well any suggestions there would be apreciated also.
Thanks
Travis Baroni
---------------------------- Travis Baroni (PhD Student) | tbaroni-at-cyllene.uwa.edu.au | The University of Western | Australia, Nedlands, WA. | 6907. | ----------------------------
Is anyone famillair with good fixation and staining techniques for rat and chicken eyes. Is Davidson's fluid THE best fixative? Someone in our lab wants to stain slides in order to examin Desmett's and Bowman's membrane. Is it possible, to stain these kind of membranes?
Try using the excellent piece of Software "Electron Flight Simulator" version 3.1-E. It is marketed by D, Chernoff of Small Woprld somewhere in the US. I am running it on a PC and I am not sure its works on a Mac. Contact David Joy at {djoy-at-utk.edu} as he has written much of the software and will give you an adress for Chernoff.
Patrick Echlin Director, Multi-Imaging Centre University of Cambridge UK
On Tue, 19 Jan 1999, by way of Nestor J. Zaluzec wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Hi, I am looking for a program to convert weight % to Atomic % and vise } versa preferably on a Power Mac.. An old program we have works on pre '90 } Mac's but not on the newer ones. Anyone have any ideas? Thanks- Mike } } } }
Is anyone familiar with the re-embedding methods of paraffin embedded tissues in resin? I was asked to re-embed human lung tumor specimens into resin and reclassify them on electron microscope level. I will appreciate if you could send me your reembedding protocol.
Thanks in advance,
Regards,
Ranan
Ranan Gulhan AKTAS, M. D. Trakya University, Faculty of Medicine Pathology Department Edirne 22030 TURKEY
Paul:=20 I have two rather old evaporators which still get 10-6 torr. I will give=20 then to you but you must pay the shipping costs !
Patrick E=A3chlin Multi-Imaging Centre University of Cambridge UK
On 19 Jan 1999, Paul Webster 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 } -----------------------------------------------------------------------. } =20 } =20 } Hi, } =20 } Does anyone out there have a vacuum evaporator that is taking up space? = I have lots of space but no evaporator. =20 } 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/apw.htm } =20 } =20 } =20
Martin Roe discovered some Br in epoxies and asked for the source. I gave him an answer off line. However I see some other answers and I think that I should add my two cents for everybody:
I have a good friend who worked a long time as microscopist for CIBA before it came Novartis=8A
He told me about 5 years ago that bromine is voluntarily added (for some reasons=8A) in certain types of epoxies. So it is not a contamination, nor a new thing! So he (or his supplier) has probably just changed his epoxy source.
We used some compound sample made of A alloysl bits in epoxy to show to our students how dangerous it can be to work only at low kV in EDS. You have a nice superposition of Al Ka and Br L lines. If the uncertainty on sulfur/molybdenum is quite well known, that on Al/Br confuses quite a lot of people because Br is often unexpected!
Yours
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, I am looking for a program to convert weight % to Atomic % and vise } versa preferably on a Power Mac.. An old program we have works on pre '90 } Mac's but not on the newer ones. Anyone have any ideas? Thanks- Mike
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
In a message dated 1/19/99 10:19:06 PM, tbaroni-at-cyllene.uwa.edu.au writes:
} As I understand it I need to perform histgram equalisation } to correct for these differences.
In your case what you should really do is apply a transfer function that is the cumulative histogram of ALL of the images in the stack to each image, not equalize them individually.
UNIVERSITY OF LEEDS School of Process, Environmental and Materials Engineering
POSTDOCTORAL RESEARCH FELLOW
Applications are invited for this post funded by the EPSRC (Waste Minimisation Managed Programme) to join an active research group working with Dr. Mark A. Keane (Department of Chemical Engineering) and Dr. Rik Brydson (Department of Materials) on the development of supported metal catalysts for the "environmentally friendly" treatment of chlorinated aromatics (see, for example Appl. Catal. B: Environmental 18 (1998) 241-250). The project is a fundamental study of catalytic hydrodechlorination and will involve the preparation, testing and characterisation of a range of catalyst systems with an emphasis on developing an understanding of reaction kinetics and mechanism. The successful applicant should have a PhD in Chemistry, Chemical Engineering or Materials with a strong background= in Heterogeneous Catalysis: some experience in catalyst characterisation is a decided advantage. The post is available from 1 April 1999, or as soon as possible thereafter, for a period of three years
Salary Range: RA1A3 =A3 15,462-=A317,226
Informal Enquiries may be made to Dr. Mark A. Keane , Tel: +44 113 233 2428, E-mail: chemaak-at-leeds.ac.uk
Interested candidates should apply in writing with a detailed CV and the names of two referees to Dr. Mark A. Keane, Department of Chemical Engineering, University of Leeds, Leeds LS2 9JT, UK
Closing Date: 1 March 1999 __________________________________________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
There is a good shareware product named GraphicWorkshop Pro from Alchemy Mindworks in Ontario, Canada. It is a very good batch converter for Windows XXX.XXX based operating systems. They have a friendly web site to down load a copy:www.midworkshop.com. We of course have no financial interest in this product. Consider the Canadian dollar and our American friends can get it for about 65% of cost which I think is $40.00 if you register it. And we all register shareware, don't we? Jon McGovern J. P. McGovern and Associates jon-at-microscopy.net
available immediately in Department of Neuroscience and Neurology, University of Kuopio.
We are seeking suitably qualified persons to join a progressive multidisciplinary group investigating Alzheimer=92s disease. The experimental projects will investigate the efficacy of estrogen replacement therapy in preventing structural and functional impairment in different animal models of Alzheimer=92s disease.
We are looking for applicants who have experience in at least one of the following specialities:
Histopathology/histochemistry, electron microscopy, autoradiography.
Salary and date of commencement are negotiable.
For further information please contact:
Research Director Paavo Riekkinen, Jr. M.D., PhD. Dept. Neuroscience and Neurology Univ. Kuopio FIN-70211 Kuopio Finland Tel. 358-17-162016 Fax. 358-17-162048 E-mail: paavojr.riekkinen-at-uku.fi
Riitta Miettinen, Ph.D. Dept. Neuroscience and Neurology Univ. Kuopio P.O.Box 1627 FIN-70211 Kuopio FINLAND E-mail: Riitta.Miettinen-at-uku.fi Tel. 358-17-162761 Fax: 358-17-162048
I join John Russ in saying that you do NOT want to apply histogram equalization to each image individually, although I am not quite sure that what he suggested is quite what you want.
I suppose that there may be some differences in exposure between the images for whatever reason. The challenge will be to get the same features to the same level of gray. That is, if you have some light features on a dark background, you always want those light features to be at the same light gray level and the background to be at the same darker gray level. Histogram equalization is definitely NOT the tool you want to use since it will move gray levels depending on their population in the image. It normally increases contrast between some pixels and decreases it between others and hardly gives you a uniform transfer function for all images.
I am not very familiar with Photoshop. I thought the tool you would look for was called something like "levels". I think you would want to primarily use the gamma adjustment to bring two reference features (e.g., light features and dark background) to the same gray levels in all images, then maybe use some tweaking of brightness and contrast to finish the match (per John Mackenzie's recipe). I will leave it to you to figure out which tool to use (histogram or a pixel cursor or ...) to determine when the images are matched. In my own work I do not need a quantitative match, but only a qualitative, visually apparent match. Therefore, I just eyeball a match between one reference image and all the others as I adjust them.
WES
At 10:37 AM 1/20/99 +0800, you wrote: } } Hi all, } } I have a series of se images of a serial sectioned material which I would } like to display as a volume rendered image. I have aligned the images, } using an nih-image macro, however the brightness and contrast differs } between them. As I understand it I need to perform histgram equalisation } to correct for these differences. } } I've noticed photoshop allows the user to load a transfer function using } the "curves" dialog box, but it doesn't give the desired result. } } Is this because the function which you specify is not the cumulative } transfer function based on the reference histogram area?? } } Is there anyone who has done this before? I am using nih-image and } photoshop for these tasks. As for the reconstruction, well any suggestions } there would be apreciated also. } } Thanks } ---------------------------- } Travis Baroni (PhD Student) | } tbaroni-at-cyllene.uwa.edu.au | } The University of Western | } Australia, Nedlands, WA. | } 6907. | } ----------------------------
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
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Hi, Dear all -
I have an AO Reichert Ultracut and a LKB knifemaker. They have been in a storage for a while and need to be cleaned and lubricated. Does anyone has such experience in lubricating them and can give me some suggestions about what kind of lubricator(s) that I should use and how to do that?
Thank you in advance.
Gang Ning EM Facility Medical College of Wisconsin
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We have two old Reichert-Jung OMu2 microtomes which are in good working order with one exception - we are in need of a "female" allen key part which is used to tighten the block into the chuck. Does anyone have any they no longer need? Or know where to get one? Or has found a substitute which is easily available?
Thanks in advance,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-med.mcgill.ca
Hi Joost- you could refer to an article published in Cornea 12(3): 255-260, 1993 (Rock et al.,) we used a modified Karnovsky's 2.5% glut, 2.0% paraform, in a 300-momol, in cacodylate bfr., post fixed in OsO4, dehydrated, embedded in epon.
then using a Mallory's Azure II, methylene-blue followed by a basic fuchsin counter stain protocol we developed a fat and effective method for identifying and differentiating various regions of the cornea. Descemet's membrane stains blue, Bowman's layer stains pink, the collagen of the stroma striated pink & blue, scar tissue blue, and keratocytes blue. staining differences in the various regions are presumably attributed to the proteoglycan content and/or the various collagen types
hope this helps -Mike Rock
On Wed, 20 Jan 1999 J.Bruyntjes-at-voeding.tno.nl-at-Sparc5.Microscopy.Com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi there } } Is anyone famillair with good fixation and staining techniques for rat and } chicken eyes. Is Davidson's fluid THE best fixative? } Someone in our lab wants to stain slides in order to examin Desmett's and } Bowman's membrane. Is it possible, to stain these kind of membranes? } } Thanks in advance } } Joost Bruyntjes } TNO Zeist } The Netherlands } }
Dear All, I need to obtain some TEM analytical standards for stainless steel (for example, a standard containing just Ni, Cr, and Fe, in a well characterized composition). I was hoping for something along the lines of Cr ~ 28%, Ni ~22% and Fe ~ 50%. It would be great if I could find a sample that was TEM "ready", so that I don't have to make a new sample. If anyone knows of a source for such a sample, I would be grateful for a reference. Thanks in advance, Dorrance
Does anyone have epifluorescence attachments for a Nikon Optiphot that they wish to sell? I'm looking for a complete epi set-up.
Thanks, Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
The box said "Requires Windows98 or better.", so I bought a Macintosh.
We have a Microm HM 500 0 sold by Zeiss. It has been very reliable for three years and gives excellent sections with minimal adjustment. The only caveat is the refrigeration system. A compressor was replaced under warranty and recently we experienced a leak of refrigerant. Local refrigeration people can do the work (Zeiss is not licensed to do it). I might add that refrigeration problems are common in this building due to various factors. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
I will be receiving a sample consisting of a culture grown on a polycarbonate membrane (Transwell permeable). I need to know any pitfalls working with the membrane! Is this membrane compatible with acetone dehydration? Is it compatible with a Spurrs resin? Does anyone have experience in this area?
Thanks - Sally Burns
Sally Burns Center for Electron Optics B5 Pesticide Research Center (517) 355-5004
I have processed many filter membranes for TEM and SEM. I follow standard procedures with the following changes;
-dehydrate in EtOH, not acetone (eats the polycarbonate) -embed in Epon 812 or equivalent (I use Ted Pella's Eponate 12) -times can be shortened conciderably (Fix 20 min, wash and dehydrate steps 5 min, resin changes around 30 min.) -do a couple more resin changes than normal
I carry out the entire process (including embedment) in a 24 well plate and cut the filters out with a jeweler's saw after polymerization.
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu University of Iowa Central Microscopy Research Facility http://www.uiowa.edu/~cemrf Views expressed are mine.
On Wed, 20 Jan 1999, Sally Burns 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 will be receiving a sample consisting of a culture grown on a } polycarbonate membrane (Transwell permeable). I need to know any pitfalls } working with the membrane! } Is this membrane compatible with acetone dehydration? Is it compatible with } a Spurrs resin? Does anyone have experience in this area? } } Thanks - } Sally Burns } } Sally Burns } Center for Electron Optics } B5 Pesticide Research Center } (517) 355-5004 } } burnssal-at-pilot.msu.edu } }
We have recently acquired a LEO 438 and have need, for safety reasons, to provide small particle (1 micron) filtration at the inlet side of both the roughing and turbo pumps. I believe we can use conventional filters in-line on the roughing pump, but probably cannot afford to restrict flow at the turbo pump inlet. Does anyone have advice or ideas on how we might provide small particle filtration, perhaps via a commercial or custom built free-flow filtering device placed just under the perforated screen at the floor of the chamber. Servicing is a consideration. Also, any advice on filtering the roughing pump would also be much appreciated.
Thank you very much for your consideration.
Bruce Cunningham High Explosives Application Facility (HEAF) Lawrence Livermore Laboratory
-------------------------------------- Bruce Cunningham Lawrence Livermore National Laboratory P.O. Box 808, L-282 Livermore, CA 94550 cunningham1-at-llnl.gov TEL: 510-423-0135 FAX: 510-424-3281
I am looking for the company that makes ACLAR. I need to contact them. I don't need vendors I need the manufacturer of this wonder substance. So if anybody out there knows who it is, please let me know.
Tired of changing my fingerprints with borken slides,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
} With a good camera for initial camera, the results can be better than any } mid range color camera!! Definitely worth it.
What filter sets would people recommend? And for light microscopy I assume the filters are usually placed out of the focal plane, maybe somewhere near the field diaphram?
Gary Radice 804-289-8107 Department of Biology 804-289-8233 (FAX) University of Richmond gradice-at-richmond.edu Richmond VA 23173
HI I am attempting to examine embrylogical samples of a marine bivalve using SEM. But any containers I have used during the processing proceedure have proved indequate. I need containers which will hold samples down to a size of 30um. I have used filters placed in processing capsules but they are very awkward, and the sample has more often than not been lost during the proceedure. Any ideas would be greatly appreciated. Jean Raleigh, Marine Zoology Dept., Natioal University Galway Ireland
COMPUTER VIRUS NOW INFECTING HUMANS! Trojan Horse Supervirus Could Be Worse Than AIDS, Warn Docs -- And TWICE As Deadly! by Kevin Creed/Weekly World News
CHICAGO, ILL. -- Concerned scientists say the dreaded "Trojan Horse" computer virus has made the jump to humans -- and the brain-eating bug may soon be sweeping through America, claiming even more human lives than the AIDS epidemic! An unidentified 38-year-old man known only as Patient Zero has been diagnosed with the virus that had been heretofore found only in PCs.
"We've been dreading this day," said noted virologist Dr. Frederick Attingale who made the terrifying diagnosis. "We knew it was only a matter of time. That's how these things work. "At some point, Acquired Immune Deficiency Syndrome -- AIDS -- made the jump from monkeys to man. Now, in a similar way, the Trojan Horse virus has worked its way into the human population. Both viral transmissions were bound to happen sooner or later."
Dr. Attingale will not name the Chicago-area hospital where Patient Zero is being held. But the prognosis is not good. "People whose PCs have been infected know the virus eats away the hard disks and 'nervous systems' before anyone is aware that their computers have been infected.
"I'm afraid the same thing is happening in this man's body."
The patient, a junior executive with a large investment firm, is suffering from nerve spasms, hearing and vision loss and severe deterioration of the parts of the brain that govern memory, reasoning, math and language. His brain and nerves are being literally eaten away.
"There's no known cure and the illness continues to worsen," Dr. Attingale said. "He can barely communicate with us now."
The alarming situation came to light early last month when the patient went to his family physician complaining of headaches and memory lapses.
The doctor, suspecting a virus, referred him to Dr. Attingale.
For more information on this deadly virus, including how it enters the body and interviews with other doctors, pick up the current edition of Weekly World News (1/26/99), at your local supermarket or newsstand! Or better yet, why not subscribe and have it mailed to your home or office!
I have no experience with the Ultracut but I am aware that the surface between the cutting head and the main structure of the LKB Knifemaker (LKB 7801A) must NOT be lubricated. The metals are supposed to provide the correct level of friction to allow glass to be clamped at the correct pressure. So perhaps my message is don't lubricate anything until you are certain.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: Gang Ning To: microscopy-at-sparc5.microscopy.co
Aclar is made by and can be purchased in bulk from :
Pro Plastics Inc 1190 Sylvan Street P.O. Box 1489 Linden NJ 07036
201-925-5555
Area code may have changed At 04:48 PM 01/20/1999 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} With a good camera for initial camera, the results can be better than any } mid range color camera!! Definitely worth it.
What filter sets would people recommend? And for light microscopy I assume the filters are usually placed out of the focal plane, maybe somewhere near the field diaphram?
Gary Radice 804-289-8107 Department of Biology 804-289-8233 (FAX) University of Richmond gradice-at-richmond.edu Richmond VA 23173
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would appreciate hearing from anyone with an ESEM-FEG. Please respond to me personally as I would like to talk to you re: this instrument.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
by imo29.mx.aol.com (IMOv18.1) id DMMGa01223; Thu, 21 Jan 1999 09:38:42 -0500 (EST) Message-ID: {cb9380fd.36a73bf2-at-aol.com}
Not only should the knifemaker not be lubricated but all sliding surfaces between the clamping head and pedestal should be periodically cleaned with acetone or alcohol. Any lubrication will attract dirt which will affect the gravity drop of the pins onto the glass,it will also cause the clamping head to move up when pressure is applied to make the break. Many times poor knives can be eliminated by this simple cleaning procedure. It will necessitate removing the clamping head from the pedestal Good luck, Norm Woodside former LKB product specialist
Out of curiosity I mentioned this to a chemist colleague. He indicated that bromine is sometimes added to resins as a flame retardant. In some cases antimony is also added to boost the effect of the bromine. Everett Ramer Federal Energy Technology Center
} } } "Martin J. Roe" {mi596-at-mluri.sari.ac.uk} 01/18/99 07:11am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello fellow listservers, I've recently discovered a problem with a batch of a particular type of cold setting epoxy resin. I have been using this batch of resin on an infrequent basis for about 2 years (and is probably about 4 years old). When I analysed some of the cured resin by microanalysis last week appreciable bromine (Br) was detected as well as the usual chlorine content (Br} Cl). The 'introduction' of the Br is a recent thing because I also analysed some resin (from the same batch) prepared approximately one year ago. The analysis of this showed no Br with Cl the only detectable element present (with the Be window of the EDS detector in place that is). Is it possible therefore for the resin to in some way deteriorate with time and for Br compounds to form? If so, what are the chemical reactions going on here. I'm very certain that the Br is not a contaminant. Also, I should point out that if this is a chemical degradation of the resin then the physical characteristics of the cured resin are normal. I would appreciate any thoughts on the matter. Regards
Martin Roe Macaulay Land Use Research Institute Aberdeen AB15 8QH Scotland United Kingdom
I have a question about the KEVEX 7700 XRF system. We have been trying = for some time to get the output of the unit into a PC through the = printer port. Apparently it can be done and we would like to know if = anyone out there has tried this and what sort of parameters and software = have they used to connect to the 7700 unit. Any help would be greatly = appreciated. Currently all we get from the printer output to the PC via = a hyperterminal link is just garbeld. Thanks in advance.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD} {META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {STYLE} {/STYLE}
{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D2} I have a question about the KEVEX 7700 XRF system. = We have=20 been trying for some time to get the output of the unit into a PC = through the=20 printer port. Apparently it can be done and we would like to know if = anyone out=20 there has tried this and what sort of parameters and software have they = used to=20 connect to the 7700 unit. Any help would be greatly appreciated. = Currently all=20 we get from the printer output to the PC via a hyperterminal link is = just=20 garbeld. Thanks in advance. {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D2} ______________________ {BR} Roberto Garcia {BR} Senior = Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC 27695-7531 {BR} {A=20 href=3D"mailto:rgarcia-at-unity.ncsu.com"} rgarcia-at-unity.ncsu.com {/A} {/FONT} {= /DIV} {/BODY} {/HTML}
You can get ACLAR from Allied Signal, Inc. Westwood Road. Pottsville, PA. 17901. The phone number that we have appears to no longer be the correct one but I am sure you can find the number using information.
Good Luck.
--------------------------------------- David McKemy Dept. of Pharmacology/318 University of Nevada School of Medicine Howard Bldg. Rm.#214 Reno, Nevada 89557 Phone: (775) 784-4537 Fax: (775) 784-1620 Email: ddmckemy-at-med.unr.edu
On Wed, 20 Jan 1999, Paula Sicurello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hey Boarders, } } I am looking for the company that makes ACLAR. I need to contact } them. I don't need vendors I need the manufacturer of this wonder } substance. So if anybody out there knows who it is, please let me know. } } } Tired of changing my fingerprints with borken slides, } } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207 } http://biology.berkeley.edu/EML } } }
I routinely refer to two articles regarding this type of SEM prep.
Preparation of Microbiological Specimens for Scanning Electron Microscopy, L.P. Watson, A.E. Mckee, and B.R. Merrell. Scanning Electron Microscopy/1980/II, pages 45-56.
Specimen Preparation Techniques for Aquatic Organisms, T.K. Maugel, D.B. Bonar, W.J. Creegan and E.B. Small. Scanning Electron Microscopy/1980/II, pages 57-77.
Hope this helps, Louie
At 11:16 AM +0000 1/21/99, Jean Raleigh wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
Soon I may need to study an optical fiber with a periodic refractive index (doped). Since I am just beginning TEM studies, I am hoping that someone may be able to suggest the typical method of observing optical fibers with a TEM (these optical fibers are typically 100-125 microns in diameter). I suspect that thinning the fibers lengthwise may be a possible means, however, I would like to see a reference where optical fibers have been studied...or perhaps similar geometries like biological hair samples (though optical fibers are generally much thinner).
cross-sectional thinning (circular samples) is useless since we need to study period structures along the length of the fiber.
Thanks
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3392 [x4] `___/ //\./=/~\====\ (704)509-6622 [Hm] \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
OK, so we should not lube the knifemaker at the clamping head, but what should we do with ours that squeeks so bad it makes your teeth hurt? Our LKB 7800 has some serious squeeking when you increase the upward pressure for the break (and release it). Can we lube the arm that causes the pressing up motion (the one on the front of the machine)? It's driving us all crazier than we already are. So any suggestions will be greatly appreciated.
I was thinking of taking the cover off the knifebreaker and see if I could find any lubrication points, is that a bad thing to do?
Squeeking loudly in Berkeley,
Paula :-)
} Not only should the knifemaker not be lubricated but all sliding surfaces } between the clamping head and pedestal should be periodically cleaned with } acetone or alcohol. Any lubrication will attract dirt which will affect the } gravity drop of the pins onto the glass,it will also cause the clamping head } to move up when pressure is applied to make the break. Many times poor knives } can be eliminated by this simple cleaning procedure. It will necessitate } removing the clamping head from the pedestal } Good luck, } Norm Woodside } former LKB product specialist
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Does anyone knows where can I get high temperature conductive glue for TEM sample preparation? I need to glue a ceramic sample to Cu grid and Ion-mill it then heat treat to 1000 C in air before TEM observation. Thank you in advance.
Maoxu Qian
**************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * Seattle, WA 98195 * * mxq-at-u.washington.edu * * (206)616-3973(phone) * * (206)543-3100(fax) * ****************************
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I would think that it would be a simple enough issue. I have done similar things in the past. In fact you are probably pretty close to the solution.
It sounds like you probably have a mismatch in the baud rate between the two computers - it would give you garbage. The PC side is fairly easy to change for experimentation while the Kevex side is harder to change. I don't think the old DEC serial cards supported more than 19,200 baud. We had some old printers that required the rate set down to 2400. You could check your printer or Kevex manuals to see if there is any info in there. Otherwise, you could just keep setting the baud rate down on the PC side and reprinting until something sensible comes through.
The specs on the port are usually 8-bit, no parity, 1 stop bit. That should be the same as the default PC setting. Most DEC stuff was setup to use XON/XOFF software handshaking as oppsed to DTR or other hardware handshaking.
Good luck, and please let me know how it works.
WS
At 10:48 AM 1/21/99 -0500, you wrote: } } I have a question about the KEVEX 7700 XRF system. We have been trying for } some time to get the output of the unit into a PC through the printer port. } Apparently it can be done and we would like to know if anyone out there has } tried this and what sort of parameters and software have they used to connect } to the 7700 unit. Any help would be greatly appreciated. Currently all we get } from the printer output to the PC via a hyperterminal link is just garbeld. } Thanks in advance. } } ______________________ } Roberto Garcia } Senior Analyst, Metallography } North Carolina State University } Analytical Instrumentation Facility } Box 7531, Room 303 EGRC } Raleigh, NC 27695-7531 } {mailto:rgarcia-at-unity.ncsu.com} rgarcia-at-unity.ncsu.com
Hello, everyone Is there anyone that has "PC-PDF" JCPDS-ICDD PDF-2 DATABASE files of RuO4 and RuO2 or knows from where we can get these files? We have a very old PC-PDF file that has not been upgraded for several years. I would appreciate any information from anyone.
With best regards, Pipat Prayoonthong Graduate student Materials Science & Engineering Department. Stevens Institute of Technology Hoboken, NJ 07030
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Greetings Everyone, At one time or another, i have recieved an email from you and am just letting you know my email addy has changed from pclypool-at-sgi.net to claypool-at-serve.com
This email might concern the SX-50 Users group, contacts dealing with a Task8verC Microprobe, or you might just be a personal friend (or all the above hehe).
The following procedure may sound a little obvious, but I imagine you may be facing a rather limited access to all those books and journals, and I would feel really happy if this could help. I do wish you all the best in your EM lab-raising mission!
First, you will have to select smaller, "EM-size", portions of your paraffin-embedded specimens, cut them out and thoroughly deparaffinate. I would recommend at least 2 hours in xylene with frequent changes of xylene and some agitation; you should be able to see when it's all gone. Then several changes of 100% (anhydrous) ethanol, at least 1-1.5 h total time, and then gradually down the ethanol concentrations, like 95-85-70-50, 30 min or more at each step. Rinse 3x5 min in your choice of phosphate or cacodylate buffer, and then osmicate for 1-2 h, 1% OsO4 in the same buffer. Then dehydrate and embed for EM as you normally would.
Needless to say, even with the best original fixation the material will look pityful, but most of those diagnostically significant cell-to-cell junctions, filaments, etc. must still be there.
Best of luck! Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
I have a slip that was attached to a roll of ACLAR giving specifications and the following company name and address: Engineered Plastics Specialty Plastic Films Pottsville, PA 17901
I have no knowledge of whether this company still exists. I would guess the roll of ACLAR was 5 years old or more.
Missy Josephson
Eleanor Josephson, DVM, PhD University of Connecticut Health Center Department of Anatomy MC-3405 263 Farmington Ave. Farmington, CT 06030-3405 Ph.(860)679-2463 Fax (860)679-1274 ejosephs-at-neuron.uchc.edu
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I have not seen a reply to your posting so try this.
Astigmatism in an image of a modern TEM is always down to cleanlyness. I= f you have a gross change in astigmatism, if everything is working correctl= y, it has to be dirt. Lets look at typical symptoms for dirt in a system:-
1. An increase in astigmatism levels due to the charge on the dirt affecting the beam.
2. Beam movement, gradual in one direction as the charge grows then rapid in the return direction as the problem is discharged. When the charge reaches a level sufficient to touch earthed components within the system it discharges, the problem starts all over again.
3. The problem will vary depending upon the number of electrons hitting the area that is charging and their energy. The greater the numb= er of electrons you place in the charging area the faster it will charge. T= he higher the kV the more chance you have of the beam penetrating the contamination and finding an earth; less or no charge. The mode of operation in which you are working will also change the charge rate. In the normal imaging mode the setting of the diffraction lens does not plac= e the beam near the problem area of your coulmn - no noticable astigmatism change in this mode. In the diffraction mode the diffraction lens focal length is dramatically changed hence electrons in larger numbers strike t= he problem area and we see what happens; trouble.
There is only one solution as the problem will get worse - strip the column. I am sorry for the engineer because this is a rare move on a modern machine BUT IT MUST BE DONE!
How to make friends with service engineers this is not, but it is THE onl= y solution!
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Here is the method used by the pathology people around here.
- Remove tissue of interest from parafin embedded specimen, cut in 1-2 mm cubes. - Leave the specimens in fresh 1% OsO4 in XYLENE for 1,5 - 2 hrs. - Wash in xylene 2 times 10 min - One part resin(we use Epon-Araldite) + two parts xylene for 1 hr. - 1 part resin + 1 part xylene for 1 hr. - Resin for 30 min to 1 hour without a lid on the glass
All this steps on a carousel.
Embedd as usual, but keep the specimens in the resin over night before polymerization.
Good luck!
Best regards Randi Olsen
Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
HI I am attempting to examine embrylogical samples of a marine bivalve using SEM. But any containers I have used during the processing proceedure have proved indequate. I need containers which will hold samples down to a size of 30um. I have used filters placed in processing capsules but they are very awkward, and the sample has more often than not been lost during the proceedure. Any ideas would be greatly appreciated. Jean Raleigh, Marine Zoology Dept., Natioal University Galway Ireland
For the SEM preparations of spores and other small samples we put nets made of metal or nylon, meshs 20 =B5m, in our holder for CPD. For very small things, e.g. bacteria, zoospores, we introduce the samples with syringe and needle into LUMitainers. Lumitainers are little balls D=3D2,5-3,5 mm made of a kind of cellulose. They are useful for both SEM and TEM. In Germany you can buy them by Plano W. Planet GmbH, Marburgerstr. 90, 35043 Marburg, Fax 06425-51173. They are not cheap! I do not know where to get it abroad? Good luck! Anne Heller _____________________ Dr. Anne Heller Institut fuer Botanik (210) Universitaet Hohenheim Garbenstr.30 D-70593 Stuttgart Tel.0049-711-459-2180 Fax 0049-711-459-3355
Try folding small (10 x 10 mm) envelopes from lens tissue. Close with a small staple. These envelopes are permeable enough for processing. They work very well for pollen and erythrocyte samples.
Some types of lens tissue have rather large pores - I have seen holes up to 30 microns in some makes of tissue - check yours before using.
} I am attempting to examine embrylogical samples of a marine bivalve } using SEM. But any containers I have used during the processing } proceedure have proved indequate. I need containers which will hold } samples down to a size of 30um. I have used filters placed in processing } capsules but they are very awkward, and the sample has more often than not } been lost during the proceedure. Any ideas would be greatly appreciated. } Jean Raleigh, Marine Zoology Dept., Natioal University Galway Ireland
Prof Jan Coetzee Head: Lab for Microscopy and Microanalysis Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-362-5150 Pretoria 0002, South Africa http://www.up.ac.za/science/electron/emunit1.htm
I was interested in the use of osmium in xylene that you described. Does the tissue blacken as in a water-solution or does it simply become yellowish/brown? I am curious because the latter is what happens in the absence of water during freeze-substitution using e.g. 1% in acetone at -80 *C, although the colour change probably happens when warming up from low temperature.
Hello. I am looking for a company that is willing to build optical u-scope objectives with wavelength specific AR coatings. All leads will be greatly appreciated.
The Advanced Materials Processing and Analysis Center (AMPAC) at the University of Central Florida will be sponsoring two TEM specimen preparation short courses in conjunction with the FL AVS/FSM conference and vendor participation.
AMPAC's UCF/Cirent Materials Characterization Facility is offering two short courses in March.
"Tripod Polisher" will be offered March 12 and 13 instructor: Ron Anderson, IBM co-sponsored by UCF and South Bay Technology
"Focused Ion Beam Specimen Preparation" will be offered March 18 and 19. Instructors: Fred Stevie, Lucent Technologies and Lucille Giannuzzi, UCF co-sponsored by FEI and Micro Optics
Both courses will be held at the MCF located at 12443 Research Parkway, Suite 305, Orlando in Research Park {1 mile from the UCF campus. The registration fee is $750 per course or $1200 for both courses. Fees include course materials and meals (breakfast/lunch/breaks) all days. Registration deadline is March 1 and space is limited.
Additional information and registration forms for all of these events can be found at the AMPAC website: http://pegasus.cc.ucf.edu/~ampac
Please contact Dr. Giannuzzi below for any additional questions.
We look forward to seeing you in March!
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
The 2nd Annual AMPAC Golf Scholarship Invitational will be held on Sunday, March 14, at the Ekana Golf Club in Oviedo, Florida, in conjunction with the FL AVS/FSM conference. The tournament starts at 8:00 a.m. Entry fee is $135 per player or $500 per foursome. There will be lots of prizes and food! Hole sponsorships (both corporate and individual) are also available. A corporate donation of $1000 will also include 4 golfers. A corporate donation of $500 will include 2 golfers. Individual hole sponsors may donate $100. Deadline for entries is February 26. The tournament is limited to the first 80 golfers so register today! Proceeds will benefit graduate student fellowships in Materials Science and Engineering at UCF.
Additional information and registration forms for all of this event can be found at the AMPAC website: http://pegasus.cc.ucf.edu/~ampac
We look forward to seeing you in March!
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
This is easy and cheap to make: _ /_____\ _ | | | | | | | | {-- BEEM capsule |_|_____ |_| \ / ^ ^ |____|__ plankton netting
(I hope the ascii "art" survives the 'net...)
Cut out the center of the lid of a BEEM capsule, leaving a ring that still snaps onto the body of the capsule as usual. Close the lid, trapping a taut bit of plankton netting under it.
Cut off the lid from a second BEEM capsule and cut out its center as above. Cut of the pyramidal end from the first BEEM capsule, and snap the cut-off end over the new end of this capsule, again trapping a bit of plankton netting.
The netting comes in various sizes, so you can get one small enough for your needs, but use the largest size mesh you can to ease fluid flow.
Problem: when changing solutions before drying, the mesh will trap air. The best way to deal with this is to leave the 2nd end of the capsule open, and place the capsule mesh-end down in a 4mL Wheaton vial or similar. Change solutions by sucking out the old solution from the vial outside the capsule, and pipetting in the new solution into the open end of the capsule. Don't fill the capsule/vial completely, or the specimens might float out into the vial.
When ready to dry, snap on the 2nd lid with netting, suck some of the final fluid change into the capsule so there is no air bubble, then CPD.
I've also dried marine gastropod larvae from HMDS, no CPD with good results. These capsules work well for that also, and don't need the 2nd lid then.
Phil
} I am attempting to examine embrylogical samples of a marine bivalve } using SEM. But any containers I have used during the processing } proceedure have proved indequate. I need containers which will hold } samples down to a size of 30um. I have used filters placed in processing } capsules but they are very awkward, and the sample has more often than } not been lost during the proceedure. } Any ideas would be greatly appreciated. } Jean Raleigh, } Marine Zoology Dept., } Natioal University Galway } Ireland
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net
Jean Raleigh wrote: ============================================= I am attempting to examine embrylogical samples of a marine bivalve using SEM. But any containers I have used during the processing proceedure have proved indequate. I need containers which will hold samples down to a size of 30um. I have used filters placed in processing capsules but they are very awkward, and the sample has more often than not been lost during the proceedure. Any ideas would be greatly appreciated. ============================================= Have you tried what are called "Microporous Specimen Capsules"? They are available from SPI as well as several other of the main suppliers to the microscopy and histology market. They come in three pore sizes, the smallest being 30 um. They were designed and engineered just for your kind of application.
As the pores get smaller and smaller, the exchange times of fluids during the processing becomes longer. However, in this case patience can be a virtue since at 30 um, we believe these capsules to be far superior to the alternatives. And of course, 30 um entities are contained within the volume of the capsule.
Disclaimer: SPI Supplies is a supplier of these microporous capsules and would like to see their use increased! More information about these particular capsules are available on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
I am looking for a cpu-based aid for teaching crystallography at the undergraduate level.
Is anyone in ListLand familiar with CaRIne Crystallography software from France? This is popular in our chemistry dept., but I'm wondering whether other packages exist with comparable (or superior) libraries and algorithms to demonstrate 3D structures, rotate them, make substitutions, correlate structural data with XRD or CBED data, etc, etc.
You can reply to me off-list. If others are interested, I can summarize responses.
Thx, all--
Ann Hein Lehman, EM Facility Mgr Trinity College, Hartford, CT v 860-297-4289 f 860-297-2538 email: ann.lehman-at-exchange.cc.trincoll.edu
Hello to you all! Thanks to everyone for kindly responding to my recent enquiry about the Br in epoxy resins; I apologise for not responding sooner. Your answers were informative and on the basis of most of the suggestions, I favour the idea that the Br is from the resin itself and is not from an external i.e. contaminant source. It appears that Br is added by manufacturers to some types of epoxies. With regard to my particular resin, the company I got it from has still to get back to me. Regards
Hi everybody! I am looking for a used EDS, in good conditions, for installation on Jeol SEM. Any offer? Please inform price and I will estimate the transport & handling costs. Thanks in advance, Sincerely, Nelson Fava/Geosciences Institute/U. Brasilia, Brazil. MIcroprobe CAMECA/SX50#359 and SEM Lab.
3 D imaging of in vitro bio samples is a young technology with many new and exciting facets.
For example: The university of Linz Austria has made great strides in both 1- single molecule fluorescence and in 2- images characterizing the antibody antigen binding forces
Workshop beginning next Week end. See Web: http//:www.molec.com/linz
George
-----Original Message----- } From: James Pawley {jbpawley-at-facstaff.wisc.edu}
Hi Folks,
I was quite surprised that I had the only listing in answer to the call o= f Petra Wahlbring (with an EM912) who had problems of dirt in the TEM imagi= ng system!
Techniques for the identification and positioning of contamination within=
an EM column should be part of the general knowledge of every EM Unit. A= re there people who would like to take on a little more in terms of EM maintenance? Eight years ago we took the Royal Microscopical Society course "Monitoring and Maintaining the Electron Microscope" to Australia and it now runs each October at the University of Sydney, it has also tak= en place in South Africa and Hong Kong.
If there is a demand in the United States, and an organisation who feel they could host such a course, we would be pleased to help.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Thought I'd send out the responses to my post regarding aperture contamination in a VPSEM operated in the high vac mode.
I want to thank everyone who responded. Suggestions covered both improper sample preparation and microscope design problems due to using it in the high vac mode. I have passed the suggestions on and encouraged the SEM lab to contact me if they arrive at a solution so I could post it to the listserver.
Also if any one wants to contact an individual listed below, drop me an email and I can provide their address.
Lou
----- Harry Ekstrom wrote:
So, presumably, the signal is attenuated so much that simply by replacing the final apertures...the problem goes away? Is the inside of the chamber clean? ie Any contamination possibly from the diffusion pump? Other possibilities are outgassing samples causing contamination or a vacuum leak. How is the filament life? ---- Charles Garber wrote:
If these are semiconductor products, unless they are looking at photo resist , there should not be anything to really crud up their column or their apertures.
The only time I have seen this happen is when people were not cleaning the aperture holder and insertion rod properly and contamination was migrating down to the apertures and contaminating them.
The discover of this phenomenon was the basis of the Zaluzec patent and the plasma cleaning interest.
If the vacuum system is acceptable, then you might want to suggest an extra special cleaning of the aperture holder and insertion mechanism. I don't want you to think that I am suggesting this in order to sell them a plasma cleaner. Perhaps this might be of sufficient interest to Nestor Zaluzec at Argonne that your friend could pay him some fee to actually test out if doing a plasma cleaning of his insertion mechanism and aperture holder assembly actually would lead to a reduced contamination rate.
I could of course be wrong about this so you might want to ask Nestor what he thinks of my suggestion. But I think that at Argonne they do have some mechanism for helping industrial problems in that kind of a situation.
------ Steve Chapman wrote:
Dirty final apertures must be down to problems with the gas from the specimen or specimen chamber.
1. Could they use a bigger aperture without any problems? Most people do not push the instrument to its limit so the "normal apertures" could be too small for the application. A cure rather than a prevention!
2. If the specimen chamber has become highly contaminated try this. Pump down the microscope and use an industrial hot air (hair) dryer on ALL the metal areas. The vacuum will almost certainly fall back but as long as you are not trying to heat up an area right by an "O" ring this is what you should expect from the outgasing. Get the chamber hot is the secret.
3. Use an inflow of dry nitrogen gas to cut down the filth that may enter from the environment.
4. Check the rotary pump fluid and its efficiency?
5. What is the vacuum level, have all the LV inlet systems closed correctly?
---- Robert Foglia wtote:
I used to work for a company called High Yield Technology and they were selling an InSitu particle monitoring probe sensor, which could be used in high vacuum. I know they had developed an application for a Hitachi SEM which enabled the user to real time monitor the cleanliness of their system. They initially requested this sensor because they were seeing large amounts of contamination within their system, and the probe sensor was used to monitor while they cycled different valves and moved the robotics around (the robotic arm was scraping). The company is located in Silicon Valley and they might be an option to explore. By the way, they usually place the sensor on the exhaust port in order to get as close to the wafer as possible.
------
John Arnott wrote:
Based on your message concerning the contaminated aperture, my feeling is that the problem probably rests somewhere other than the aperture itself. But since there is the slightest possibility that there might be a problem with the aperture perhaps we can assist you. Ladd produces the vast majority of apertures/microholes used in the United States and the aperture you have may very will be a Ladd aperture. To make sure there is no residual contamination on an aperture we have a post production protocol that is designed to eliminate any contamination that results during production. In fact we have some apertures/microholes used in fluid and gas control that are required to be absolutely contamination free. We have a protocol to ensure that. It is also important that an aperture be shipped or stored in a glass or non-contaminating vial and that the aperture surface should not touch tissue or even lint free cloth. We would be glad to examine the aperture to see if the contamination was a result of packaging, handling or the production process. Please contact us if you if you wish us to do that.
----- Thomas C. Isabell wrote
Regarding the cleaning of SEM apertures, have you considered plasma cleaning?? We have had great success in removing the aperture strip from the microscope, plasma cleaning it in our Model 1020 Plasma Cleaner, and replacing it in the scope. Some of our results were published in a recent MRS proceedings - MRS Symp. Proc. Volume 523, pp 31-38. If you are interested in a copy, I could send one your way. Plasma cleaning returns the aperture strips to their "as new" state and eliminates the need to constantly replace them.
To take this one step further, if the apertures are indeed getting dirty because of dirty specimens, our plasma cleaner has also been shown to effectively clean specimens prior to insertion into the microscope. Some of these results are also shown in the aforementioned MRS Proceedings. Our instrument allows not only the cleaning of the specimen, but also the specimen stub, and if needed, the specimen stage.
If you have any further questions about the technique or our instruments, please do not hesitate to contact me. Additional information can be found on our website, at www.fischione.com .
------ Lisa Montanaro wrote:
There are several reasons why the final aperture may be getting dirty on a variable pressure SEM, and why this may be more noticeable on semiconductor samples. Several causes contribute to contamination within SEM chambers. If this system has been used for any other analysis other than semiconductor, at low vac modes, residues from prior analysis may be lining the chamber walls - these are difficult to remove even by baking the chamber out. Additionally, this system probably has oil-cooled pumps - the deleterious effect of backstreaming oils into chamber which "gums" onto the final aperture is well established, but can be diminished with a strategically located dry N2 purge port in the sample exchange chamber. Mounting media is also of concern - since most conductive paints outgas, these vapors can also contaminate the F.A. quickly, especially if the material is not exceedingly well dried. Simple cleanliness may also be the culprit - by handling a metal sample fixture with bare hands, oils and sweats are deposited on the fixture. These deposits love to attach to the F.A. (and cold tip of EDS detectors too!) As most semiconductor analysis requires fairly high magnification, and often lower KeV, these problems are exacerbated. I've delt with these and many other problems concerning the optimal performance of SEM's with semiconductor materials, and would be pleased to look over his system for performance improvements. Let me know if I may be of assistance!
----- Randy Nestor wrote:
I'll take a stab at the problem. Most of these types of SEM's have vacuum gradients from the chamber to the gun, even when run in the "normal" mode. The worst vacuum is in the chamber, the best in the gun. If your friend has any outgassing of the sample, the contaminants are proalby finding their way to the final aperature. It seems these apts are one way of controlling vacuum levels in different parts of the scope. We have seen similar problems with our Hitachi 2460N. What scope does your friend have?
-----
Ronald Vane wrote:
I have given the following advice to several users of variable pressure SEM who dirty microscopes and contamination, and they all said it cured the problem.
Don't leave it in high Vacuum mode all the time. Some variable pressure SEM are very dirty due to backstreaming at high vacuum. They are designed for low vacuum not high vacuum. By placing the system into low vacuum mode, the chamber is placed into viscous flow vacuum dynamics which stops backstreaming and purges out contaminants. Try leaving it in low vacuum mode overnight for a month and check the results. --- Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
To : scientist in MSA I'm a graduate student at Electronic Ceramics Lab. Dept. of Ceramic engineering Yonsei University. I've investigated electron diffraction of gas clusters in Korea Institute of Science and Technology. Please let me know electron-atomic scattering factor of In, Zn, and, Ar. Thank you for your effort.
Sung Han p.s. my e-mail address is pibhan-at-kistmail.kist.re.kr
At least for the specimens I happened to deal with so far (vertebrate tissues, cell culture, bacteria, microalgae), the material normally will not really blacken until you wash and start dehydrating it AFTER osmication in traditional aqueous OsO4 solutions. While in aqueous OsO4, it will remain brownish. (Unless, of course, you are using that simultaneous, single solution glutaraldehyde/OsO4 fixation, or a reducing buffer like PIPES, and have passed the time when it all turns black...)
The explanation used to be that, at the stage of osmication, while some of the OsO4, indeed, oxidizes nonsaturated lipids, etc., a lot of it simply dissolves in the lipids of the specimen as nonreduced OsO4. Then, say, the ethanol reduces that specifically accumulated OsO4, to form the so called "osmium black".
As for making FRESH solutions of OsO4 in xylene (or CCl4, etc.), I would just never have a spare OsO4 ampule (and enough paraffine blocks to EM-reevaluate) to try that! :-)
Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
I'm trying to adapt the TUNEL procedure of labeling cleaved DNA in apoptotic cells for TEM level observation. Basically I just want to rework it to utilize colloidal gold and localize the tagged 3'-OH ends of the degraded DNA. TUNEL has been done for several years at the LM level, but I've only come up with a handful of studies that successfuly nailed it with plastic embedded tissues and gold conjugates... and of those few if any seem to be A+ work (sometimes the specificity of the labelling looks a bit sketchy.) Anyone out there have any experience with this? I'm using a line of chronic myelogenous leukemia cells that has a well characterized apoptotic response to etoposide... previous ultrastructural studies show classic apoptotic changes. Any input will be appreciated.
I'm interested in finding out if there are any variable pressure SEMs available for paid use in the NY/NJ area. Specifically, I'm interested in doing some long-term imaging of large, uncoated specimens at very low mags (10X and below).
Many thanks in advance.
Best regards,
Angela
--------------------------------------------- Angela V. Klaus
Manager, Core Microscopy Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
www.coleoptera.org is working now, please be so kind and send me your comments. There is also large index of entomologist who worked or working on Tenebrionidae...
I apologise for crossposting.
Keep care and be of good cheer.
Regards
Vratislav Richard Eugene Maria John Baptiste of Bejsak (Bayshark)-Collorado-Mansfeld
Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
Temporally home address: 32 Girrawheen Ave. Kiama NSW 2533 Australia e-mail: 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.
I have received a letter asking if I knew of anyone who would donate two used light microscopes for use by clinics in northern Burma. The doctors there are currently using instruments with cracked lenses and no light source and have a desperate need for a better instrument.
If you have a microsacope you could donate please contact me privately and I can give you the details.
Probably the microscopes should have an oil objective and a built-in light source though the latter is not absolutely essential.
We are working with stereo microscope. We are more concern about the ope= rator health of fatigue after peeping into the microscope. The color temp= erature is about 3200K to 3950K. Anyone out there can suggest how long ca= n one (normal person) looks into the microscope without causing fatigue ?= We are now proposing for every 2 hours of looking into a stereo microsco= pe, an operator will rest for 10 minutes. Anyone make any research to fin= d out these ? We do not want our operator to experience epilepsy. Please help. Lim Hian Ho Senior Marketing And Application Engineer Wisma QES, No.6 Jalan USJ 9/5P, UEP Subang Jaya, 47620 Selangor, West Malaysia Tel : 603-7241188 ext 207 Fax: 603-7244488 e-mail : hhlim-at-qes.po.my
Hi everybody in microworld! I am looking for a second hand EDS, simple or equiped with an image soft, to attach to a JEOL SEM. Any offer? Thanks in advance, Sincerely, Nelson Fava. EPMA CAMECA SX50 Lab. Geosciences Institute/U. Brasilia/Brazil.
I am interested in using backscatter elelctron imaging to evaluate the distribution of iron in 1 um plastic sections of biological material. My intent is to provide an independent measure of the distribution of iron in my samples and compare this distribution with the distribution of iron as evidenced through use of the ferrocyanide reaction on adjacent sections. Any comments on the likelihood of success of such an endeavor and the pitfalls of using BEI for sectioned material would be appreciated.
Thank you very much,
WB Jaeckle
============================== Will Jaeckle Department of Biology Illinois Wesleyan University P.O. Box 2900 Bloomington, IL 61702-2900 TELE: 309-556-3779 FAX: 309-556-3864 EMAIL: wjaeckle-at-titan.iwu.edu =============================
Hi everybody in microworld! I am looking for a second hand EDS, simple or equiped with an image soft, to attach to a JEOL SEM. Any offer? Thanks in advance, Sincerely, Nelson Fava. EPMA CAMECA SX50 Lab. Geosciences Institute/U. Brasilia/Brazil.
Having been a subscriber to this list for half a year now, I'm convinced that someone can help me with the following problem. I want to use heparin conjugated with gold, without any "bridge" such as albumin in between, but I cannot find any commercial source. Since I'm a beginner in the gold business, is there any good recepy for the preparation of heparin-gold?
In a message dated 99-01-26 05:11:00 EST, hhlim-at-qes.po.my-at-sparc5.microscopy.com writes:
{ { Anyone out there can suggest how long can one (normal person) looks into the microscope without causing fatigue ? We are now proposing for every 2 hours of looking into a stereo microscope, an operator will rest for 10 minutes. } }
Lim,
I do not have any research on the subject, but the two most common causes of fatigue are: (1) eyestrain and (2) the physical strain on the neck, upper back and arms from staying in one position for such a long time. As far as eyestrain, I think your proposal for two hour blocks of time is realistic.
You don't mention the type of microscope, but if it is a fairly modern one you can probably obtain a tilting "ergotube" for viewing, so the operator can find the most comfortable position and adjust the viewing angle as needed. If this is not possible, you can also get an ergonomic table that can be raised and lowered either mechanically or via a motor. You can place the scope on the table and adjust the height to suit the operators.
You might also consider putting a video system on the microscope so the operator can choose to either look through the eyepieces or at a high resolution monitor. There are also video inspection stations that can take the place of the microscope entirely.
Good luck to you.
Best regards,
Bob ****************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel./Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Clinical, Research & Industrial Microscopy Cytology/Histology/Pathology/EM *******************************
I thought the "gold standard" for apoptosis is the ultrastructural changes that can be seen by TEM and that the DNA labeling was developed in order to view apoptosis/vs necrosis at the light level-over a larger area of tissue? At least that is the idea I've gotten from articles that I have read. Could someone please clarify this? Thanks in advance.
Karen Pawlowski Sr. Research. Assoc. UT Southwestern Med. Ctr. PhD candidate, UT Dallas
On Mon, 25 Jan 1999, Doug Matthews 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 everyone, } } I'm trying to adapt the TUNEL procedure of labeling cleaved DNA in } apoptotic cells for TEM level observation. Basically I just want to rework } it to utilize colloidal gold and localize the tagged 3'-OH ends of the } degraded DNA. TUNEL has been done for several years at the LM level, but } I've only come up with a handful of studies that successfuly nailed it with } plastic embedded tissues and gold conjugates... and of those few if any seem } to be A+ work (sometimes the specificity of the labelling looks a bit } sketchy.) } Anyone out there have any experience with this? I'm using a line of } chronic myelogenous leukemia cells that has a well characterized apoptotic } response to etoposide... previous ultrastructural studies show classic } apoptotic changes. Any input will be appreciated. } } Doug Matthews } } }
We offer the 1.4 nm Nanogold=AE gold cluster label with several reactivities for covalently labeling biomolecules. If you can selectively oxidise the terminal reducing sugar of heparin to give an aldehyde residue, it should react with Mono-amino-Nanogold=AE (you then reduce the resulting Schiff base and purify the conjugate by gel filtration). Alternatively, if you can selectively expose an amino- group, you can conjugate this with Mono-Sulfo-NHS-Nanogold=AE.
Protocols for using these reagents are posted on our web site (http://www.nanoprobes.com/Inf2021.html and http://www.nanoprobes.com/Inf2025.html). I hope this helps,
Rick Powell Nanoprobes, Incorporated
} } Dear all, } } Having been a subscriber to this list for half a year now, I'm convinced } that someone can help me with the following problem. } I want to use heparin conjugated with gold, without any "bridge" such as } albumin in between, but I cannot find any commercial source. Since I'm a } beginner in the gold business, is there any good recepy for the preparation } of heparin-gold? } } Thanks in advance! } } Anne
****************************************************************** * NANOPROBES, Incorporated | US Toll-free: (877) 808-2101 * * 25 East Loop Road, Suite 113 | Tel: (516) 444-8815 * * Stony Brook, NY 11790-3350, | Fax: (516) 444-8816 * * USA | rpowell-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
Hi everybody in microworld! I am looking for a second hand EDS, simple or equiped with an image soft, to attach to a JEOL SEM. Any offer? Thanks in advance, Sincerely, Nelson Fava. EPMA CAMECA SX50 Lab. Geosciences Institute/U. Brasilia/Brazil.
Hi everybody in microworld! I am looking for a second hand EDS, simple or equiped with an image soft, to attach to a JEOL SEM. Any offer? Thanks in advance, Sincerely, Nelson Fava. EPMA CAMECA SX50 Lab. Geosciences Institute/U. Brasilia/Brazil.
The Science of Biological Specimen Preparation for Microscopy
Edited by Marek Malecki and Godfried M. Roomans
Proceedings of the 14th Pfefferkorn Conference, Belleville, IL
Hardbound book with 31 papers; 466 + xii pages. Table of Contents available on request.
Published by and available from: Scanning Microscopy Intl., Box 66507, AMF O'Hare (Chicago), IL 60666-0507= , USA FAX: (847) 985-6698 / E.mail: 73211.647-at-compuserve.com
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BOOK REVIEW (published in January 1999 issue of the Journal of Biomedical=
Optics; vol 4 pages 191-192):
Advance in our knowledge of the molecular organization of living material=
rests on two kinds of technological development: (1) Microscope instrumentation (2) Specimen preparation. This includes new techniques=
appropriate for a specific mode of microscopy, and information on any structural alterations produced during such specimen preparation.
The present volume contains the proceedings of the 14th Pfefferkorn Conference organized by Scanning Microscopy International and dedicated t= o the late Prof. G.E. Pfefferkorn.
The organizors and editors of this conference brought together 31 leading=
scientists in this field from the USA, Europe and Japan, to present and discuss new techniques to prepare biological specimens for microscopy. =
Discussions with a panel of reviewers are added to each paper. There is not space for a detailed discussion of each paper. I shall try to indica= te the general substance of the contributions.
In general, three unique features of this book are worth stressing.
The first, it reflects current trends in science towards interdisciplinar= y approaches in solving important biological questions. An essential part = of incorporating microscopy into these interdisciplinary approaches is development of reporter molecules which will have features precisely determined in biochemistry and molecular biology labs and which will rema= in stable in changing environment of living cells being labeled for various modes of microscopy. Chapters by Heinfeld - nanogold, Kessels - boronate= d antibodies, Malecki - organo-metallic ligands, and Swartz - fluorescent derivatives of proteins are concerned with the development of such new probes which are based upon covalent bonds, therefore they can serve as reliable markers for structures of interest. Moreover, chemically define= d features can be translated into their functional architecture e.g., as shown for atomic force microscopy in the chapter by Woodward and Zasadzinski or for life time imaging by Lakowicz. The detailed protocols=
will allow an investigator to have them easily modified for a particular new application. This book also demonstrates how fruitful this interdisciplinary approach can be e.g., as demonstrated in chapters by DeBault and Gu, Malecki, and Nuovo, polymerase chain reaction developed f= or DNA amplification in molecular biology labs can be modified to determine morphological localization of selected sequences. The book clearly demonstrates, that in order to solve a biological question, it is nearly impossible to categorize approaches and restrain an investigator to traditional research techniques. To the contrary, the interdisciplinary approaches create backgrounds and precedents for breaking the boundaries = of traditional and specialized areas of science and opening new possibilitie= s.
The second, this book is also an attempt toward integrated microscopy. =
While many already published books were dedicated to very specialized are= as of one kind of microscopy, this one stands out as an eclectic (in a positive sense), but comprehensive review of the most recent developments=
in various areas of microscopy. This is particularly close to my scientific preferences, since I promoted this approach for many years. =
Integrated microscopy, allows us to overcome technical limitations of one=
kind of microscopy alone e.g., light microscopy of a living cell is limit= ed by resolution ~250 nm, while electron microscopy having atomic level of resolution can be pursued only on frozen or fixed cells, therefore studyi= ng the same cells with both types of microscopy creates an opportunity to study living cell phenomena at the molecular level. This approach is elegantly exemplified in the chapters by Malecki, Peachey et al., Ralston=
and Ploug. It also demonstrates how beneficial and cross-fertilizing thi= s integrating approach can be e.g., in the chapters by Lyubchenko where a technique of functional modifications of substrate surface used in TEM an= d SEM, now finds new applications in AFM. These chapters create not only a= n excellent starting point for a reader, but also collecting many other techniques in one book, opens for an investigator a compendium of choices= . =
The newest developments in specimen preparation for two-photon excitation=
fluorescence microscopy, atomic force microscopy, life-time imaging, ener= gy filtering transmission electron microscopy, etc. are all covered in this one volume. Therefore, scientists attempting to solve a life sciences problem with tools of modern microscopy can make a choice from the vast variety of techniques and examples presented in this book. The detailed hands-on specimen preparation protocols will guide them through. The discussions with the reviewers will provide them with the critical evaluation of choices.
The third, the book paves the road for future developments in microscopy.= =
The chapters and discussions with the reviewers, fairly define current technical limitations of various modes of microscopy and suggest possible=
ways towards overcoming these difficulties. In many cases, authors clear= ly state the directions of their future research. Therefore, not only it is=
updated information concerned with the status-quo, but also a proposal fo= r future research.
Specifically, the first group of papers deals with methods to study chromatin and nucleic acids. In the first paper M. Malecki describes new=
methods of gene transfer, which were developed based upon incorporation o= f reporter molecules allowing us imaging of cellular pathways in living cells, and in cryo-immobilized cells by energy filtered TEM from the cell=
surface to the chromatin. G.V. Childs describes methods to identify mRNA=
and proteins in the same cell, in tissue sections. L.E. De Bault and J. = Gu present detailed protocols for in situ hybridization, in situ transcripti= on and in situ polymerase chain reaction. M. Thiry developed the in situ terminal transferase - immunogold technique to pinpoint specific nucleic acid regions in thin sections. Scanning probe microscopy is represented = by five contributions. Hydration Scanning Tunneling Microscopy (Heim et al.= ) is based on conductivity of surface adsorbed water molecules and can imag= e hydrophilic insulators and biological specimens such as collagen IV molecules, TMV, and cryo-sectioned bovine tendon.
For atomic force microscopy imaging of macromolecules, a strong attachmen= t to the substrate is essential. Lyubchenko et al. show that treatment of mica with aminopropyltriethoxy- silane will hold DNA in place for imaging=
even in water. They also introduced other chemically reactive mica surfaces, hydrophobic or charged. Mueller-Reichert and Gross discuss DNA=
and DNA-protein assembly analyzed by TEM, Scanning Tunneling Microscopy (STM) and Atomic Force Microscopy (AFM). An interesting new application = of STM is imaging of freeze-fracture replicas (Woodward and Zasadzinski). I= t also can examine interior interfaces and provides quantitative informatio= n about the vertical dimension of interior structures.
Four contributions discuss light microscope techniques for the imaging of=
living cells. The first of these by N.S. Allen and M.N. Bennet use Alfal= fa root hairs before and after treatment with Nod factors (produced by Rhizobia) to study in live and fixed cells the role of actin and endoplasmic reticulum in growth form change. Imaging was with a confocal=
laser scanning microscope. The paper by Peachey et al. describes techniques to image cultured cells by phase, epifluorescence and confocal=
microscopy, and after fixation and critical point drying image the same cell as whole mount with the Jeol 400kV-EX intermediate voltage TEM, correlating structures seen in the living cell with the EM image. The paper by D.R. Swartz on covalent labeling of proteins with fluorescent compounds for imaging applications beautifully illustrates with alpha-actinin how a protein can be covalently linked to a fluorophor without interfering with its normal chemical interactions in the living cell. This paper is important for all who need fluorescently labeled cel= l proteins for imaging applications. L. Edelman and A. Ruf describe a simp= le treatment to stabilize freeze-dried cells during or after low temperature=
embedding in Lowicryl, to prevent loss of material from thin sections during wet cutting.
J.F. Hainfeld describes the techniques for labeling with Nanogold, Undecagold and FluoroNanogold. This paper is essential for anybody who requires gold labeling. The increasing availability of energy filtered TEMs will facilitate immuno-cytochemistry by providing electron spectroscopic imaging. Kessels et al. describe the development of organo-boron compounds which can be incorporated into organic molecules such as Fab'-boronated peptide conjugates for immunochemistry.
The last paper by H. Sitte contains a masterful critical review of cryofixation and the blueprints on how to build a cryomicrotome that can provide useful cryosections. I think that these seventy pages contain th= e most valuable part of this volume.
Finally, this volume obviously contains a wealth of stimulating and usefu= l information. It bears testimony that microscopy is alive and well.
Hans Ris, Zoology and Integrated Microscopy Resource, University of Wisconsin, Madison, WI
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Copies of several other reviews are available on request.
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Price: $95 (US delivery by uninsured mail) or $105 (elsewhere by uninsure= d mail); insured delivery, air mail, etc., are available for additional cos= t, please inquire.
REPRINTS or photo-copies of papers are available for (rates for delivery = by cheapest mail method); EACH: $5 (up to 8 pages); $10 (9-16 pages); $15 (17-24 pages) and $20 (25+ pages).
Recently we have been having problems with a resin mixture that we have been using for two years, until recently with good results. I am wondering if anyone else has any ideas about what might be the problem.
The resin is a mixture of Araldite 6005 (about 23% by wt.) SPIpon 812(about 23%) and DDSA (about 54%). We store the unopened resin bottles in the refrigerator, but warm them fully before opening. The opened, capped bottles are stored in a vented cabinet at room temperature. We typically make about 55 ml of resin at a time and store it without the accelerator (DMP-30) in the freezer. Again, we always make sure the bottle is completely warmed to room temp before opening it to use. To embed, we dehydrate through propylene oxide and then 50:50 PO:resin overnight. The next day the blocks are rolled for 4 hours or so in 100% resin + 1.5% DMP-30, then polymerized in fresh resin with accelerator for 24-48 hours at 60=B0.
Several months ago we began getting blocks which are difficult to trim smoothly; they seem to chip and crack. Although the resin sections well, when sections are picked up they tended to disintegrate, or at best, showed rifts and cracks throughout the resin. Increasing the curing time helps a bit, but does not eliminate the problem. We have tried new bottles of all the resins and the accelerator, without improvement. However, in some cases the new bottles may be from the same lots as the old (we haven't kept records of resin lot numbers).
Does this sound familiar? Specifically, we are wondering about storage and shelf life of the resins. Are we inviting problems by storing unopened bottles in the fridge (we have done so for years, but with the old Epon 812)? What about shelf life? The unopened bottles are not more than a year old. Do you store your opened resin bottles in desiccators? At what temperatures? How do you store resin mixtures? Finally, should DMP-30 be added at the 50:50 stage? We had not done this in the past, did for a year or so, then stopped doing it again and thought this wasn't a problem, but perhaps it is. Any ideas would be welcome. Thanks
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 =46ax: 860-4861936=20
Colleagues, can anyone help this person? Reply directly to him ...
Nestor Your Friendly Neighborhood SysOp -------------------------
Email: micnaut-at-aol.com Name: julius simon School: retired chemist school volunteer
State: florida
Question: i would like a description of a procedure for smear preparations of onion root tips to demonstrate the varios stages of mitosis-that could be quickly mastered by ahigh school bio student. I have a good scope with an automatic ca mera
Hello Doug, I have tried the TUNEL technique for EM. I got a very clean and specific labelling of heterochromatin in ALL nuclei, normal and apoptotic. My explanation is that colloidal gold labels only on the surface of the section. And there will be a lot DNA breaks at the surface due to sectioning and that is where the reaction will occur. As far as I can remeber, I could find only two references for EM. One of them (Thiery?) used this technique as a staining for all DNA. If you want more info, please contact me and I will dig deeper to find the references and the protocol I used (it worked beautifully on paraffin sections). Yours sincerely,
Sarka Lhotak
Hamilton Regional Cancer Centre Hamilton, Ontario, Canada lhotaks-at-mcmaster.ca
One of the most common causes of brittle tissue is exposure to propylene oxide (or a mixture containing it) in the presence of air, i.e. allowing the tissue to become exposed when in a propylene oxide (or PO:resin mixture) stage.
One or more of your ingredients going "off" could also be the cause of your problem, as you suggest. However, in over 30 years of TEM embedding with a whole variety of raw materials, many of which have stood on the shelf for years, we have very seldom experienced this sort of problem.
For a reliable protocol and resin mixture how about trying the one described in a paper I published several years ago? The reference is: Cross, R.H.M. (1989) A reliable epoxy resin mixture and its application in routine biological transmission electron microscopy. Micron & Microscopica Acta, 20(1), 1 - 7.
Good luck!
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za (or eurc-at-giraffe.ru.ac.za) tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
This is similar to a situation I was in many years ago. In my case it was self-induced because I was experimenting to find a mixture which would cut very large ultrathin sections. In that instance (as far as I can remember - it was late 1970's) I used a high anhydride:epoxy ratio combined with a partially hydrated anhydride. The latter was obtained by exposure to air over a long period, like the bottle was 10 years old before I got to it and it was pretty viscous.
Maybe your mixture already has the high A:E ratio and the anhydride has aged. Although I would not have thought that brittlenss would be the problem, DDSA is the long chain component whereas I was using both DDSA and MNA in my recipe and the MNA was the component which really gave hardness.
Brittleness was a sign of good cutability but it could be overdone! Then the sections would break up on cutting. One block would cut 3 mm square sections, and if you went down to grey in colour, they would simply dissolve on the water! They left bits of fixed tissue which were sort of de-embedded (I never loooked at these).
Brittleness was also induced by a long infiltration time - 48 hours at 35 Centigrade, then 24 hours at 45, followed by 24 hours at 60 C. This procedure induces sterical hindrance which catches the big molecules in certain configurations whereby they cannot form complete cross-linkage. A sign of this is that the blocks are thermoplastic - if you heat them in an oven, they get soft, you can impress the surface with tweezer tips etc. IS THIS TRUE OF YOURS?
Possibly you have aged components, or they have taken up water.
Alternatively, maybe your dehydration isn't complete. Water in the alcohol or acetone, or maybe propylene oxide (? don't know about this possibility, we haven't used it for 20 years or more because of the health aspect). Try adding molcular sieve to a sample of your dehydrating agent?
First of all it would be useful to clarify whether we are talking about a true stereo microscope or a binocular microscope. I personally find stereo microscopes a little more comfortable to work with - perhaps because you are looking at something with a more natural perspective and light than when examining a slide in transmitted light.
I would certainly think that a two hour working period may be possible but I would suspect that there might be problems with general fatigue, attention span, and even RSI (repetitive strain injury) if stage controls are being rotated very regularly. It would probably be necessary during that time to encourage 'micro-breaks' (meaning breaks of a few seconds every 10 or 15 minutes - rather than 'microscope breaks') as you should for display screen equipment/computers. The general rule of thumb with computer rest periods is little and often rather than saving time up over a couple of hours and I suspect that the problems are similar with microscopes.
Other considerations should include ergonomics (Microscope, work area, seating and the people using it - which of course is part of the ergonomics equation).
I hope this helps.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: "RCHIOVETTI-at-aol.com" To: hhlim; microscopy
Dear Marie,
Firstly, I 'm not quite sure how to store SPIPON-812, you should ask the manufacturer, but with my knowlege, all epoxy resins are stored at room temperature in a cool and away from direct sun light. Low temperature storage is not recommended for epoxy resins, especially araldites and anhydrides, such as Araldite 502, 6005, DDSA, NMA, NSA. DER... Meantime, DMP-30, DMAE, BDMA, they are contained amino group (-NH-), storing them in the refrigerator will increase its shelf life. Storing epoxy resins in the desiccator is the best. Secondly, you can store the resin mixture (without the addition of DMP-30) in the refrigerator for later use, but only 3 to 4 weeks is maximum, with well protected from contamination with moisture. After all, I think your sections problem is associated with the storage condition of your resins. For more general tips for embedding media, please refer to Electron Microscopy Sciences catalog XIII, page 48.
Sally Stowe wrote: } } We are just setting up TEM EDXA in a multidisciplinary unit, and need to acquire standards covering a wide range of applications. Any recommendations, pitfalls etc? (Philips 430 TEM up to 300kV, Oxford ISIS ultra-thin window detector). } thanks
Dear Sally, Chuck Fiori published an article about using lithium borate glass standards for biological work--the matrix is about the same as that for resin/organic material. There may be someone who sells these or similar standards. For frozen-hydrated work, one can dissolve a known amount of an appropriate compound in a sucrose solution which approximates the composition of tissue and cut cryo-sections. For materials work, there are some commercially available standards. It is important that the matrix of the standard match that of the unknown, so one type does not fit all. Good luck. Yours, Bill Tivol
Dear Anyone: I'm interested in finding out programs to simulate and to index electron diffraction patterns. If it is possible, for windows. Thaks in advance
Patricia Bozzano Comision Nacional de Energia Atomica Buenos Aires, Argentina.
Cytotechnologists who screen Pap smears, among other things, sit at a microscope up to 8 hours daily. Comfort is obviously important. Among the usual recommendations: sit upright, eyes forward (no bending of the neck), use elbow or forearm rest pads (e.g., "Wedgies"), feet flat on the floor (though some use a foot rest), break for 10 minutes hourly to stimulate circulation and minimize the vigilance decrement, maintain room light at a level comparable to the illumination intensity to minimize contrast differences and eyestrain, and look "through the eyepieces" (i.e., at infinity, relaxed accommodation), rather than "in the microscope".
I've coined a term to describe the collective response to the various ills (e.g., varicose veins, hemorrhoids, panorama butt, elbow calluses, repetitive strain injury, self-inflicted stab wounds from the dotting pens, eyepiece grooves under the eyes, and bruised orbits) that can beset a long sitting cytotechnologist: hyperchronokathistophobia, which translates to "fear of sitting for extended periods of time." Think we can claim workman's compensation and take early retirement?
Gary Gill
-----Original Message----- } From: "RCHIOVETTI-at-aol.com"-at-sparc5.microscopy.com [mailto:"RCHIOVETTI-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, January 26, 1999 11:32 AM To: hhlim-at-qes.po.mysparc5.microscopy.com; microscopy-at-sparc5.microscopy.com
In a message dated 99-01-26 05:11:00 EST, hhlim-at-qes.po.my-at-sparc5.microscopy.com writes:
{ { Anyone out there can suggest how long can one (normal person) looks into the microscope without causing fatigue ? We are now proposing for every 2 hours of looking into a stereo microscope, an operator will rest for 10 minutes. } }
Lim,
I do not have any research on the subject, but the two most common causes of fatigue are: (1) eyestrain and (2) the physical strain on the neck, upper back and arms from staying in one position for such a long time. As far as eyestrain, I think your proposal for two hour blocks of time is realistic.
You don't mention the type of microscope, but if it is a fairly modern one you can probably obtain a tilting "ergotube" for viewing, so the operator can find the most comfortable position and adjust the viewing angle as needed. If this is not possible, you can also get an ergonomic table that can be raised and lowered either mechanically or via a motor. You can place the scope on the table and adjust the height to suit the operators.
You might also consider putting a video system on the microscope so the operator can choose to either look through the eyepieces or at a high resolution monitor. There are also video inspection stations that can take the place of the microscope entirely.
Good luck to you.
Best regards,
Bob ****************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel./Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Clinical, Research & Industrial Microscopy Cytology/Histology/Pathology/EM *******************************
Marie, Our lab uses a variety of resins, ie, Spurrs, EMbed 812, Eponate 12, araldite, some very old Epon 812, and we store all of it at room temp. away from any water. We've had no problems with any of it. We also mix all ingredients including accelerator, for at least 15-20 min. on a stir plate. The more viscous resins need to be mixed first with a tongue depressor or applicator stick first for a minute or so. We do add the accelerator (BDMA, DMP-30, or DMAE)to the resin for the overnight 50:50 PO:resin step. The next morning we add 2 changes of fresh resin (again, with accelerator) for an hour each to make sure all the PO is gone and then embed at 60 degrees for 48 hrs. The only problem I've ever encountered similar to yours was from using PO that had been opened alot and was nearly empty. It evidently had gotten moisture in it. You might also check your absolute alcohol for moisture.
Mary Gail Engle Electron Microscopy & Imaging Facility] University of Kentucky
This may have nothing to do with your problem but we used to have seasonal changes in our block/resin quality before we got a proper air conditioning/heating system. Every summer when it was humid the resultant blocks would be soft and sticky, every winter hard and brittle. To help alleviate this problem our last step before actually embedding was to place the tissues in pure resin and let them sit in a vacuum for an hour or so. This might help if for some reason your humidity level in the lab has changed lately.
Pat Hales Dept. of Anatomy & Cell Biology McGill University hales-at-med.mcgill.ca
Institute of Physics EMAG '99 Conference - 25-27th August 1999, Sheffield= U.K. (Co-sponsored by RMS and IOM)
Electron and scanning probe microscopy and related techniques.
Scientific Content This biannual, three-day conference aims to bring together international S= cientists and Engineers both in Industry and Academia who employ electron and scanning p= robe microscopies, together with associated techniques such as surface science,= in both imaging and analytical applications. All aspects of Electron and Scanning Probe Micros= copy/ Spectroscopy will be discussed including:
o New Instrumentation (particularly Field Emission systems), Imaging and A= nalytical Techniques o High resolution Electron Microscopy and Electron Crystallography o Advanced SEM, Scanning Probe and Surface Science o Advances in Microanalysis and Elemental/Chemical Imaging o Microscopy of Catalysts, Sensors and Environmental Materials o Ferrous/ Non-ferrous metals and Intermetallics o Carbons, Ceramics, Electroceramics and Composites. o Semiconductors, Superconductors and Magnetic Materials. o Polymers o Microscopy of Interfaces and Surfaces
Sessions will be arranged by topic and through international keynote plena= ry lectures, invited and contributed papers, both oral and poster, current state-of-the-art iss= ues in the field will be highlighted and discussed. Papers from postgraduate students are particula= rly encouraged; correspondingly registration fees will be kept low and student bursaries w= ill be available through the Institute of Physics EMAG group.
o On Tuesday 24 August, prior to the conference, there will also be an Adv= anced School on HIGH RESOLUTION ELECTRON MICROSCOPY. Contact: c.j.hetherington-at-sheffield.ac.uk for further details
o A major trade exhibiton will be mounted in the purpose built Octagon Cen= tre which will run in parallel with the conference. Contact: i.m.reaney-at-sheffield.ac.uk for further details
SUBMISSION OF ABSTRACTS - DEADLINE MARCH 19 1999 Both oral and poster contributions are invited. Abstracts should be approx= . 300 words in length and may include figures and diagrams.
Please submit abstracts either by:
o WWW see http://www.iop.org/Confs/EMAG
o Email send blank email to confs-at-ioppublishing.com with "EMAG instructions" as th= e subject
o FTP download the files "readme.txt" and "EMAG.tem" from ftp.ioppublishing.com/outgoing/conferences/submisssions/EMAG/
In all cases please indicate whether you would prefer either ORAL or POSTE= R presentation and also which sessions you would prefer your contribution to be included = within.
Nominal Sessions o Interfaces and Surfaces (INT) o Electron Crystallography (ECR) o Catalysts/Sensors/Env. Materials (CAT) o High Resolution Electron Microscopy (HRM) o Semiconductors and Superconductors (SSC) o SEM/EBIC/CL (SEM) o Analytical Electron Microscopy (AEM) o Advanced Scanning Probe Techniques (SPT) o Ceramics/Carbons/Composites (CCC) o Metals/Intermetallics (MET) o Plenary (PLE)
STUDENT BURSARIES Students and young scholars are encouraged to apply for an EMAG bursary wh= ich will help meet costs incurred in attending the conference. Applications should = be sent to Dr R. Brydson, Department of Materials, School of Process, Environmental a= nd Materials Engineering, University of Leeds, Leeds LS2 9JT. email: mtlrmdb-at-leeds.ac.uk
GENERAL ENQUIRIES Contact: conferences-at-iop.org
_____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
The methods that I mention have been used for several years with undergraduate students and can produce quite good results. Success seems to depend, in part, on choosing the right sort of onion tips at the right time.
GROWING ROOT TIPS We have tried both seeds and bulbs and have only really had good results using root tips from onion bulbs which have been left to sprout over suitable beakers of water (the bulb should rest on the neck of the beaker and almost touch the water and after 1 to 2 weeks you should have a lot of tips - CAUTION supermarket onions may be incapable of sprouting roots you may have to try fresh ones). It is also possible that mitosis may vary at different times of day so you may have to experiment with 'harvesting times'.
STAINING METHODS These are not my methods but have been handed down since the dawn of time - the results are temporary mounts which should be examined soon after preparation. Caution the stains are harmful and acids are used to hydrolyse the cell walls/tissue. Also both methods use acetic acid solutions so the smell can be overpowering in an open lab. DO YOUR RISK ASSESSMENT FIRST. FEULGEN STAIN TECHNIQUE 1 Cut 1cm of root tip off onion, 2. fix in acetic alcohol (3:1) for at least 30 min 3. Hydrolyse in 1 molar/normal HCl at 60 deg C 10 - 15 min 4 Stain in Feulgen reagent for minimum of 60 min (IN THE DARK) - root tip will become purple in meristem 5. Without letting specimen dry out = place the end 2-3mm of root tip on a clean slide with a drop of 45% acetic acid 6. Cut root tip into 4 longitudinally then gently and carefully squash under a coverslip - protecting your thumb several layers of paper towel. 7. Examine for mitosis ACETIC ORCEIN TECHNIQUE 1, Cut 1cm of root tip off onion, 2. put into watch glass with concentrated HCl and absolute ethanol (1:1) for 10 min - NB take care when mixing this reagent 3. transfer root tips to watch glass with 45% acetic acid for 5 min 4. Without letting specimen dry out = place the end 2-3mm of root tip on a clean slide with a drop of ACETIC ORCEIN 5. Cut root tip into 4 longitudinally then gently and carefully squash under a coverslip - protecting your thumb several layers of paper towel. 6. Examine for mitosis
Good luck and take care.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
---------- } From: micnaut To: Microscopy
Colleagues, can anyone help this person? Reply directly to him ...
Nestor Your Friendly Neighborhood SysOp -------------------------
Email: micnaut-at-aol.com Name: julius simon School: retired chemist school volunteer
State: florida
Question: i would like a description of a procedure for smear preparations of onion root tips to demonstrate the varios stages of mitosis-that could be quickly mastered by ahigh school bio student. I have a good scope with an automatic ca mera
by smtp.uky.edu (8.8.8/8.8.8) with ESMTP id OAA12519 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 27 Jan 1999 14:52:35 -0500 (EST) Received: from mdc.novel.uky.edu ([128.163.67.41]) by pop.uky.edu (8.8.8/8.8.8) with SMTP id OAA13421 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 27 Jan 1999 14:52:35 -0500 (EST) Message-Id: {3.0.5.32.19990127145132.007f1700-at-pop.uky.edu} X-Sender: mgengle-at-pop.uky.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.5 (32)
Marie, Our lab uses a variety of resins, ie, Spurrs, EMbed 812, Eponate 12, araldite, some very old Epon 812, and we store all of it at room temp. away from any water. We've had no problems with any of it. We also mix all ingredients including accelerator, for at least 15-20 min. on a stir plate. The more viscous resins need to be mixed first with a tongue depressor or applicator stick first for a minute or so. We do add the accelerator (BDMA, DMP-30, or DMAE)to the resin for the overnight 50:50 PO:resin step. The next morning we add 2 changes of fresh resin (again, with accelerator) for an hour each to make sure all the PO is gone and then embed at 60 degrees for 48 hrs. The only problem I've ever encountered similar to yours was from using PO that had been opened alot and was nearly empty. It evidently had gotten moisture in it. You might also check your absolute alcohol for moisture. Good luck!
Mary Gail Engle Electron Microscopy & Imaging Facility] University of Kentucky mgengle-at-pop.uky.edu
Marie, Our lab uses a variety of resins, ie, Spurrs, EMbed 812, Eponate 12, araldite, some very old Epon 812, and we store all of it at room temp. away from any water. We've had no problems with any of it. We also mix all ingredients including accelerator, for at least 15-20 min. on a stir plate. The more viscous resins need to be mixed first with a tongue depressor or applicator stick first for a minute or so. We do add the accelerator (BDMA, DMP-30, or DMAE)to the resin for the overnight 50:50 PO:resin step. The next morning we add 2 changes of fresh resin (again, with accelerator) for an hour each to make sure all the PO is gone and then embed at 60 degrees for 48 hrs. The only problem I've ever encountered similar to yours was from using PO that had been opened alot and was nearly empty. It evidently had gotten moisture in it. You might also check your absolute alcohol for moisture. Good luck!
Mary Gail Engle Electron Microscopy & Imaging Facility] University of Kentucky mgengle-at-pop.uky.edu
1. I suggest that you not store your resins in the refrigerator. Room temp is best, tightly closed
2. Make sure that your propylene oxide is dry. Keep it strictly closed. Do not use small amounts which have been sitting in bottles.
3. Look up the dehydration schedule for your tissue in a textbook, or a paper. For instance, liver and tongue (same size blocks) need vastly differing dehydration and infiltration schedules.
4. Crucially important: Where did you get your formulation for your resin mixture? Does it meet the requirements of WPE weights of the components for a good TEM block? Have you contacted the manufacturer about any changes they might have made recently? Are you dealing with companies that are specialists for TEM supplies and carefully redistill all components once they get them from the manufacturer? (Resins bought after manufacture may contain many impurities at totally random times). Why are you using 6005? It is possible that your formulation is only marginally good, and is easily "derailed" by minor differences in environments.
5. Also crucially important: All resin coming into contact with tissue either during infiltration or during the final steps must be accelerated. Accelerating only part of the resin coming into contact with tissue used to be popular in the 60s. It is no longer considered good practice, because it yields uneven infiltration.
6. Is your accelerator dry? Your accelerator should not be older than 6 months. It should be kept strictly closed and at room temp.
We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of repair. They are leaking oil (suspect the oil seal is shot) but otherwise pump fine. Does anyone know where we could have them repaired? - Hitachi does not do this type of work and I hate to throw them away at $3,000 each.
Thanks,
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am hoping that somebody knows the answer to this;
I have been asked to look at some chromosomes in the TEM, and am wondering if someone knows of a good reference on stains that can be used for this work.
If you know of a guide or papers on the preparation tasks involved that would be of great help.
Many thanks
David
Dr. David C. Bell Room 13-1818 E-mail: dcb-at-MIT.EDU Center for Mat. Sci. and Eng. PH: (617) 253-3317 Massachusetts Institute of Technology FAX: (617) 258-6478 77 Massachusetts Ave, Cambridge, MA 02139
Postdoctoral positions at Karolinska Institutet, Stockholm
Membrane protein structure/Electron crystallography
Two postdoctoral positions are immediately available for work on structure determinations of membrane-bound enzymes using cryo electron microscopy. Two-dimensional crystals are available of proteins both in their native states and in the form of reaction intermediates. The work will initially be concentrated on data collection and processing. Previous experience in the field of electron crystallography is advantageous.
The Center for Structural Biochemistry (CSB) is a unit at the Department of Biosciences, Karolinska Institutet and comprises research groups with facilities for X-ray crystallography, NMR spectroscopy, electron microscopy and molecular modelling. CSB is located at the South Campus of Karolinska Institutet close to Stockholm, encompassing the Novum Research Center and Huddinge University Hospital. Excellent resources will be provided.
The postdoctoral positions/research associates are initially for two years with possibilities for extensions for up to four years.
Applications should be sent to Dr. Hans Hebert, Karolinska Institutet, Department of Biosciences at Novum, S-141 57 Huddinge, Sweden together with a CV and the names of two referees. Inquiries: Hans.Hebert-at-csb.ki.se or Philip.Koeck-at-csb.ki.se.
Qualifications (education, certification, language, etc.) and Experience required: A candidate with a BS or MS degree in polymer science, material science or chemistry is preferred with some 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 two professional level full time openings for Polymer Microscopists, one position at each of the Dow's facilities in Midland, MI and Freeport, TX. 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 ultramicrotomy and cryo-ultramicrotomy. 3. Operation of transmission electron microscope. 4. Interpretation of images. 5. Documentation of work. 6. Compliance with safety and quality programs. 7. Active member of project and SMX work teams.
Interested: 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, Workforce Planning 98-289, P. O. Box 150, Plaquemine, LA 70765. E-mail respondents must list Job 98-289 and their last name as the first and second items on the Subject line. Only those selected for an interview will be contacted. Only U.S. citizens or aliens who are authorized to work in the United States will be considered for employment.
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$20billion. Dow manufactures and supplies chemicals, plastics and agricultural products for customers in 164 countries and employs approx. 43,000 people worldwide. For more news and information about Dow, please visit our web site at www.dow.com.
Bob Cieslinski Microscopy & Microanalysis 1897 E Bldg. (517) 636-6875
John J. Bozzola 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 Vacuum-folks, } } We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of } repair. They are leaking oil (suspect the oil seal is shot) but otherwise } pump fine. Does anyone know where we could have them repaired? - Hitachi } does not do this type of work and I hate to throw them away at $3,000 each. } } Thanks, } } John } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } #################################################################### Dear John,
I have never had any luck with anyone rebuilding these pumps at an economical price. The companies that rebuild these pumps complain about the parts availability. Moreover, the rebuild doesn't last more than one year. At $600.00 to $1000.00 a rebuild it is cheaper in the long run to buy a replacement. I have used Alcatel Model 2010 with sucess.
There are several companies that offer rebuilding of vacuum pumps. One that I know of that is very reputable is Torr International in New Windsor, NY. You can reach them at ph: 914-565-4027 or visit their web sight at www.torr.com. I am sure they can help bring your pumps back up to spec. The contacts at Torr Int'l are Dr. Masud Naraghi and Mr. Jeff Terranova.
Good luck,
Steve Collins South Bay Technology East 4019 S. 16th St. Arlington, VA 22204
} } } } } Please visit us at http://www.southbaytech.com} } } } }
Celebrating our 35th year of Manufacturing Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy
John J. Bozzola 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 Vacuum-folks, } } We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of } repair. They are leaking oil (suspect the oil seal is shot) but otherwise } pump fine. Does anyone know where we could have them repaired? - Hitachi } does not do this type of work and I hate to throw them away at $3,000 each. } } Thanks, } } John } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
Hi !! One of my collagues is interested to found out a reprit of the paper: Identification of a new Zr-Nb- Fe Phase O.T.Woo and G.J.C.Carpenter. Proc. of the 12th Int. Congress for Electron Micoscopy. San Fransisco Press, 132(1990) Thank in advance
} } I was interested in the use of osmium in xylene that you described. Does the tissue blacken as } in a water-solution or does it simply become yellowish/brown? I am curious because the latter is } what happens in the absence of water during freeze-substitution using e.g. 1% in acetone at -80 } *C, although the colour change probably happens when warming up from low temperature.
Hello Keith.
According to the people here that most often works with this (Irene Lund) the blocks are not as dark as with standard methods, but light brown. We haven't done freeze-substitution with osmium, so it's not easy to compare directly. For this purpose we buy ampoulas with 0,1 gram OsO4.
Best regards Randi Olsen Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
Just made a trip to our Physics machine shop. They have sent pumps to Kurt J. Lesker in PA for repair. The cost to rebuild a 25 liter pump in 1997 was ~$600.
Kurt J. Lesker 1515 Worthington Ave. Clairton, PA 15025 1-800-245-1656
Hope this helps, Lou Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
Duniway Stockroom (http://www.duniway.com) lists a rebuild for a Hitachi VP-160 at $480. I assume that would come with shaft seals. Or you could order the shaft seals from Hitachi and rebuild the pumps yourself. The part number(s) ought to be in the CuteVac manual that came with the pump/microscope. I did this several years ago on a pump from a Hitachi S-570.
However, parts availability from Hitachi, in my experience, is not quick or easy (even with part numbers).
Carl
Earl Weltmer wrote: } } John J. Bozzola wrote: } } Hi Vacuum-folks, } } } } We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of } } repair. They are leaking oil (suspect the oil seal is shot) but otherwise } } pump fine. Does anyone know where we could have them repaired? - Hitachi } } does not do this type of work and I hate to throw them away at $3,000 each. } } } } Thanks, } } } } John } Dear John, } } I have never had any luck with anyone rebuilding these pumps at an } economical price. The companies that rebuild these pumps complain about } the parts availability. Moreover, the rebuild doesn't last more than one } year. At $600.00 to $1000.00 a rebuild it is cheaper in the long run to } buy a replacement. I have used Alcatel Model 2010 with sucess. } } Good Luck } } Earl Weltmer
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
Potential lead: Capital Equipment Co. (or Capital Vacuum) in the northern Virgina area offered repair for many brands of equipment. I cannot locate the address and do not know if they are still in business. A quick search did turn up a company in Herndon, VA (named Capital...), but I don't know if that is the correct one.
If the leak is not too fast, a tray under the pump is a cheap solution.
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Hi Vacuum-folks,
We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of repair. They are leaking oil (suspect the oil seal is shot) but otherwise pump fine. Does anyone know where we could have them repaired? - Hitachi does not do this type of work and I hate to throw them away at $3,000 each.
Thanks,
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Internet Mail Server 2.1); Thu, 28 Jan 1999 12:59:39 -0500 Mime-Version: 1.0 Content-Type: multipart/alternative; boundary="============_-1294576389==_ma============" Message-Id: {v04011727b2d654a02b0d-at-[128.206.162.35]}
Assistant or Associate Director Molecular Cytology Core Facility
The Molecular Biology Program at the University of Missouri is seeking an individual with experience in some or all of the following areas:
* confocal scanning laser microscopy * bright field microscopy * wide field fluorescence microscopy * low light video microscopy * image processing/analysis * deconvolution * all types of microtomy * immunocytochemistry * in situ hybridization * Adobe Photoshop
The Assistant/Associate Director will be responsible for training users, maintaining instruments and developing protocols for a campus-wide multi-user light micrscopy facility. Electron Microscopy is not a part of this facility. PhD desirable but not required for individuals with extensive experience. Although an ideal candidate would have experience in all of the areas listed above, candidates with extensive experience in selected areas and who have the desire and capacity to learn the additional areas will be considered. Excellent oral and written communication skills are essential. Experience in a multi-user core facility would be viewed positively. Women and minority candidates are especially encouraged to apply. Review of applications will begin immediately and continue until an appropriate candidate is hired.
Address applications (CV and 3 letters of reference) or inquires to:
Thomas E. Phillips, Ph.D. Division of Biological Sciences 3 Tucker Hall, University of Missouri Columbia, MO 65211-7400. 573-882-4712 PhillipsT-at-missouri.edu.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax) --============_-1294576389==_ma============ Content-Type: text/enriched; charset="us-ascii"
{bold} {bigger} {bigger} {bigger} Assistant or Associate Director
Molecular Cytology Core Facility
{/bigger} {/bigger} {/bigger} {/bold} The Molecular Biology Program at the University of Missouri is seeking an individual with experience in some or all of the following areas:
* confocal scanning laser microscopy
* bright field microscopy
* wide field fluorescence microscopy
* low light video microscopy
* image processing/analysis
* deconvolution
* all types of microtomy
* immunocytochemistry
* in situ hybridization
* Adobe Photoshop
The Assistant/Associate Director will be responsible for training users, maintaining instruments and developing protocols for a campus-wide multi-user light micrscopy facility. Electron Microscopy is not a part of this facility. PhD desirable but not required for individuals with extensive experience. Although an ideal candidate would have experience in all of the areas listed above, candidates with extensive experience in selected areas and who have the desire and capacity to learn the additional areas will be considered. Excellent oral and written communication skills are essential. Experience in a multi-user core facility would be viewed positively. Women and minority candidates are especially encouraged to apply. Review of applications will begin immediately and continue until an appropriate candidate is hired.
Address applications (CV and 3 letters of reference) or inquires to:
Folks: I am trying to get hold of a dual replica device for a Cressington freeze fracture machine. An alternate possibility would be a dual replica device from a Balzers 400 If anyone, anywhere, has one tucked in a drawer gathering dust etc. etc. and would like to part with it (for the purchase price, or whatever they feel is appropriate) please let me know as soon as possible Thanks Simon
---------------------------------------------------------------------------- -------- 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 Mobile: 412-607-3534 URL: http://sbic6.sbic.pitt.edu ---------------------------------------------------------------------------- -----
---------- } Van: HASWELL Malcolm {malcolm.haswell-at-sunderland.ac.uk} } Aan: Microscopy {microscopy-at-sparc5.microscopy.com} } Onderwerp: RE: retired chemist school volunteer ne
} GROWING ROOT TIPS ..CAUTION supermarket onions may be incapable of sprouting roots you } may have to try fresh ones).
This seems to become more and more a problem as onions are frequently treated with growth inhibitors. Instead of onions, garlic (Alium sativum) can be used! There was an article in MIKROKOSMOS some years ago, stating that garlic isn't treated that way, at least not in Germany (Europe?).
} STAINING METHODS
Some of my modifications:
} FEULGEN STAIN TECHNIQUE } 1 Cut 1cm of root tip off onion, } 2. fix in acetic alcohol (3:1) for at least 30 min
fix in ethylalcohol 95%-acetic acid 3/1 (v/v) for about 30 min
} 3. Hydrolyse in 1 molar/normal HCl at 60 deg C 10 - 15 min
when an incubator or a water bath isn't available: use HCl, 5N, 40 min at RT
} 4 Stain in Feulgen reagent for minimum of 60 min (IN THE DARK) - root tip
} will become purple in meristem } 5. Without letting specimen dry out = place the end 2-3mm of root tip on a } clean slide with a drop of 45% acetic acid } 6. Cut root tip into 4 longitudinally then gently and carefully squash under } a coverslip - protecting your thumb several layers of paper towel. } 7. Examine for mitosis
} ACETIC ORCEIN TECHNIQUE } 1, Cut 1cm of root tip off onion
fix in ethylalcohol 95%-acetic acid 3/1 (v/v) for about 30 min
} 2. put into watch glass with concentrated HCl and absolute ethanol (1:1) for } 10 min - NB take care when mixing this reagent
Wash in running water for at least 10 min
} 3. transfer root tips to watch glass with 45% acetic acid for 5 min
} 4. Without letting specimen dry out = place the end 2-3mm of root tip on a } clean slide with a drop of ACETIC ORCEIN
Slightly heat the slide untill steam apears, let cool, repeat this several times until the meristem comes dark red-brown.apply new staining solution when necesary. Don't let dry!
} 5. Cut root tip into 4 longitudinally then gently and carefully squash under } a coverslip - protecting your thumb several layers of paper towel. } 6. Examine for mitosis
Slides can be made permanent. There are lots of methods, I use these: For ACETIC ORCEIN-stained slides: use slides treated with Haupt's adhesive. Put the slides after examination in a jar of water, the cover slip down. Wait until the coverslip comes of. Wash slides and coverslips careful but toroughly in distilled water, dehydrate in ethylalcohol, 2-propanol. Treat with xylene and mount in DPX (or another resin). Most of the cells remain on the slide (coverslip).
For cells stained with Feulgen: Use slides treated with Haupt's adhesive. Put the slides after examination in a jar of water, the cover slip down. Wait until the coverslip comes of. Wash slides and coverslips careful but toroughly in distilled water, dehydrate in ethylalcohol 70%, stain in Light Green or Fast Green FCF in ethylalcohol 70% (concentration not critical, about .5% will do), dehydrate in ethylalcohol 90% and differentiate the green in it, 2-propanol. Treat with xylene and mount in DPX (or another resin). Most of the cells remain on the slide (coverslip). DNA: violet, RNA (nucleoli): green (Light Green) or blue-green (Fast Green FCF).
} Malcolm Haswell } Electron Microscopy } School of Sciences } Fleming Building } University of Sunderland } SUNDERLAND SR1 3SD } Tyne and Wear } UK } } Tel (0191) 515 2872 } e-mail: malcolm.haswell-at-sunderland.ac.uk
I am surprised that Hitachi will not repair the pumps for you. We had three pumps repaired ( Oil seals ) by Hitachi 1.5 years ago at minimal cost during our TEM move. The pumps are still working perfectly & have no leaks. It is not a major job to replace these seals yourself and in the past we have replaced a couple, with no difficulty getting parts from Hitachi (UK). The problem with replacement is seating the seals in evenly. It requires patience which is probably why Martin ( Hitachi, UK ) did a better job than us.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Carl Henderson {chender-at-umich.edu} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
I am still using Eric Shelden's Presentit {shelden-at-umich.edu} to do my digital presentations. It takes a folder full of PICT's and Quicktime's and place them in name order. No fancy transitions or anything, but I find that overkill. The main advantage is that it plays quicktimes cleanly and at maximum frame rate with easy access to the quicktime controls for frame by frame play. I keep a copy on my desktop so I can drag and drop a movie or image on it anytime I need to look at something. Doing quicktime in Powerpoint is tricky at best and requires a session with tech support to find out how to make it work correctly. For making text slides, and playing simple images Powerpoint is great although I still liked Persuasion better since you could apply multiple formats to a single presentation. The best of that lot may be Astound which is the only quicktime application that implements a speed control for quicktime movie playback rate. Dave
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Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
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REGARDING post-doc at EMAT, Belgium
Dear colleagues,
as of April 1, 1999, a 2-year TEM post-doc position will be available at = the EMAT group in Antwerp. The research will consist of using advanced = TEM techniques (HREM, EDX, EELS, low-dose, ...) for the study of new = photographic materials in close colaboration with the Agfa-Gevaert = company, one of the leading manufacturers of photographic material. The = position is open to EC citizens or equalized persons.
I look forward to your applications,
Nick Schryvers e-mail: schryver-at-ruca.ua.ac.be fax: 32-3-2180257
by upei.ca (PMDF V5.1-9 #20757) with ESMTP id {01J741DZDB3WA8CEC4-at-upei.ca} for microscopy-at-msa.microscopy.com; Fri, 29 Jan 1999 11:47:40 AST Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.43); Fri, 29 Jan 1999 11:51:38 -0400 Received: from SpoolDir by AVCN1 (Mercury 1.43); Fri, 29 Jan 1999 11:51:07 -0400
Hi! I have a question regarding the limits of Reichter-Jung Ultracut E ultramicrotome. One of the students wants to cut 1 micrometer sections of the blocks almost 5x5mm in diameter. So the sections would be 1 micrometer thick, 5 mm wide, 5 mm high. My questions is is what is the maximum size of the block face that can be safely (not destroying the machine) cut on that type of the microtome. I need to know numbers to back up my argument. Thanks in advance Dorota
Dr. Bozzola, I am not sure that my reply to your posting went through as I unknowingly did not follow protocol for this list server. I can rebuild these inexpensively, return them quickly, test ultimate vacuum, and provide a guarantee. I have rebuilt these pumps a couple hundred times. You are right the seals harden over time due to heat from the pump. Your also right in that it is silly to replace a perfectly good pump for $3K that can be rebuilt for a couple hundred. Give me a call. J. McClintock (606)257-1242 (606)277-6507 jcmcclin-at-pop.uky.edu
Dear John and all, In the USA the Hitachi service company Hitachi Instruments, Inc. (HII) used to have its Hitachi vacuum pumps rebuilt by Dunniway Stockroom (800) 446-8811 in Mountain View, CA. They (HII) found it uneconomic to do so and stopped. Now for warranty and service repairs of Hitachi pumps they replace them with Edwards pumps. The Hitachi pumps are thrown away! More of the new EMs from Japan are showing up with the Edwards pumps new. Dunniway has told me that the Hitachi pumps are hard to repair, and that there is no secondary market for them. You can buy used/rebuilt pump from Dunniway for a reasonable price. The Edwards replacement model for most Hitachi SEMs is E2M12 with a rebuilt price of $1400.
Disclaimer: I am a only a Customer of Dunniway with no financial interest.
Ron Vane XEI Scientific. -----Original Message----- } From: John J. Bozzola {bozzola-at-siu.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Dear fellow microscopists,
We are considering the purchase of a new semi-in-lens FESEM. Quality of service is of paramount importance to me in terms of the selection process. I would like to hear from others in the Pittsburgh area (say 500 mile radius) about the quality of their SEM service (not necessarily FESEM instruments) from the following vendors: LEO, JEOL, Hitachi, and Philips. I'm interested in the good, the bad, and the ugly kind of stories.
The types of things that I'm interested in is time it took for installation, timeliness (i.e. response time from the initial call), downtime, satisfaction with their work, your confidence in the service engineers, helpfulness in diagnosing problems and solving them over the phone without a service call, etc.
Please respond directly to me. I will keep your responses confidential.
Thanks in advance.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Hello All, Does anyone out there have any suggestions for getting good SEM cross= sectional images from dried wood? I have a student who would like to image= the microstructure of several types of wood. All the references I have= found so far suggest cutting fresh or moist wood with a new razor blade to= achieve good cross-sections. The student is studying the effects of two= different drying techniques, and therefore soaking the wood in water is out= of the question. Cutting the dried wood with razor blades has resulted in= crushed, damaged cross sections.=20
Thanks, Scott F. Scott Miller Electron Microscopy Lab smiller-at-umr.edu University of Missouri-Rolla =20 223 McNutt Hall voice: 573 341 4727 Rolla, MO 65409 USA fax: 573 341 6934
I am looking for a mounting medium with a high optical index (around 1.7). Once I used something called piperin (n=1.68). Could you please let me know where can one obtain such products?
Fellow Microscopists, I am soliciting your comments on what you consider the best approach to damp vibrations when using a light microscope. We are in the process of redesigning our light microscopy/image analysis work area and have a choice between a conventional marble table and a Newport BenchTop vibration isolation system (pneumatic). Which should it be? Are there other possibilities to consider?
Thanks for a moment of your time. Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
I would like to draw your attention to the materials science symposium being held as part of the Scanning 99 meeting in April this year. The abstract deadline is February 15th (the same as MSA) and the abstract format is the same as the MSA format. It is intended that there will be contributed presentations as part of the program, and depending on the number of submissions, the symposium may be extended for another day. Please forward this e-mail to anyone you think may be interested in attending the symposium.
{bold}
"Analyzing Materials Interfaces at Atomic Resolution" {/bold}
There will be a Materials Science symposium at Scanning 99 entitled "Analyzing Materials Interfaces at Atomic Resolution". Scanning 99 is being held at the Hyatt Regency O'Hare in Rosemont, IL from April 11-14 1999. The "Analyzing Materials Interfaces at Atomic Resolution" symposium is tentatively scheduled for Tuesday April 13th and will consist of invited and contributed presentations. The details of the conference and deadlines/forms for abstract submissions can be found at http://www.scanning.org or details can be requested from Mary Sullivan (e-mail: scanning-at-fams.org, tel:201-818-1010). Registration for members of the Midwest Microscopy and Microanalysis Society are at the reduced rates of $150 for the whole conference or $50 for a single day (all attendees of the MMMS symposium held at UIC last May are members of MMMS).
A preliminary list of invited speakers and the titles for their presentations is shown below.
{bold} Prof Ondrej Krivanek-University of Washington {/bold}
Title "Towards sub-Angstrom electron probes by Cs-corrected STEM."
{bold} Dr Max Haider-CEOS GmbH {/bold}
Title "Towards sub-Angstrom resolution by correction of spherical aberration"
{bold} Dr J. Murray Gibson -Argonne National Lab {/bold}
Title "Statistical Measurement of Electron Scattering Fluctuations in Amorphous
Materials - A new Structural Tool"
{bold} Prof Laurie Marks-Northwestern University {/bold}
Title "Picometer structure determination using Electron Diffraction"
{bold} Prof Marija Gajdardziska-Josifovska-University of Wisconsin at Milwaukee {/bold}
Title "Quantitative surface microscopy and diffraction over the length scales:
Morphology and crystallography of polar oxide surfaces. "
{bold} Dr David Muller-Lucent Technologies {/bold}
Title "The end of the Roadmap for Silicon Dioxide: Atomic resolution EELS of
of Hyper-Thin Gate Oxides"
{bold} Prof David Williams-Lehigh University {/bold}
Title "Atomic-Resolution X-ray Microanalysis in the TEM"
{bold} Dr Ed James-University of Illinois at Chicago {/bold}
Title "Atomic resolution scanning transmission electron microscopy on the 200kV
FEGTEM"
{bold} Dr Stephen Pennycook-Oak Ridge National Lab {/bold}
Title "Atomic scale analysis of interfaces by Z-contrast STEM, EELS and
If the wood is very dry, you might try simply fracturing a piece. A surface so prepared will have an extreme amount of relief, but it may provid you with a less deformed microstructure.
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Hello All, Does anyone out there have any suggestions for getting good SEM cross sectional images from dried wood? I have a student who would like to image the microstructure of several types of wood. All the references I have found so far suggest cutting fresh or moist wood with a new razor blade to achieve good cross-sections. The student is studying the effects of two different drying
techniques, and therefore soaking the wood in water is out of the question. Cutting the dried wood with razor blades has resulted in crushed, damaged cross sections.
Thanks, Scott F. Scott Miller Electron Microscopy Lab smiller-at-umr.edu University of Missouri-Rolla 223 McNutt Hall voice: 573 341 4727 Rolla, MO 65409 USA fax: 573 341 6934
It may well depend on the type of wood. Years ago I had some luck with Quercus blocks by cleaning up the top surface with a sliding microtome (e.g. AO 860) with a properly sharpened (and long) microtome knife set to a high shear angle and using a small advance. There were still some crumbs in the vessels but you could find good areas too.
I've never had much but frustration using razor blades on woody plants of any kind and lately it seems most brands of single edge blades we use around here for trimming blocks are softer and duller then they used to be.
cheers, John Heckman
} Hello All, } Does anyone out there have any suggestions for getting good SEM cross } sectional images from dried wood? I have a student who would like to image } the microstructure of several types of wood. All the references I have } found so far suggest cutting fresh or moist wood with a new razor blade to } achieve good cross-sections. The student is studying the effects of two } different drying techniques, and therefore soaking the wood in water is } out of the question. Cutting the dried wood with razor blades has } resulted in crushed, damaged cross sections. } } Thanks, } Scott } F. Scott Miller } Electron Microscopy Lab smiller-at-umr.edu } University of Missouri-Rolla } 223 McNutt Hall voice: 573 341 4727 } Rolla, MO 65409 USA fax: 573 341 6934
I am studying about Hf-Ta-V alloy. So, I want to make a TEM sample for that alloy(Hf and Ta of 1/3, V of 2/3) by jet-electropolishing technique using the apparatus made by Struers. I would like to know the experimental condition including solution, temperature and cathode. Please tell me about it who have experienced in that alloy.
won
Won-yong Kim Department of Materials Science and Engineering University of Pennsylvania, LRSM building 3231 Walnut St., Philadelphia, PA 19104
} redesigning our light microscopy/image analysis work area and have a choice } between a conventional marble table and a Newport BenchTop vibration } isolation system (pneumatic). Which should it be? Are there other } possibilities to consider? You are going to get lots of responses for cheap solutions using tennis balls or spaldeen balls or inner tubes from bicyles or wheelbarrows or layers of felt and sheets of concrete. Having no experience with these, I can't comment on their efficacy. However, the Newport or TMC table floating on air or N2 is going to be far superior to the marble table. Guarranteed. ******************************************************* * Michael Cammer Analytical Imaging Facility * * Albert Einstein College of Medicine * * Bronx, NY 10461 (718) 430-2890 * * work URL: http://www.ca.aecom.yu.edu/aif/ * * personal URL: http://cammer.home.mindspring.com/ * *******************************************************
Years ago, upon recommendation by folks out at Sharon Vacuum Company in Brockton MA, I bought a two component epoxy kit called A-12 Epoxy Adhesisve from the Armstrong Products Company on Argonne Road in Warsaw, Indiana, for repairing leaks in vacuum systems. It also proved useful for sealing cracked plastic parts and metal fittings on cryogenic systems, and for many other repairs. Great stuff, and it cured to a beautiful milk chocolate brown tone.
I'm just about out now, and I've written to Armstrong Products and they are no longer in existence,letter was not forwardable, etc.
Anybody out there know of their whereabouts? Or could you recommend any other suitable replacement product?
Thanks,
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.crl.umn.edu
I'm interested in doing some immuno work with the SEM. Basically tagging colloidal gold onto a surface antigen on cultured cells. I've got some old information and papers but as for up-to-date methodologies, I'm a little sketchy. Does anyone have any good review papers on the use of immuno surface markers w/ SEM (preferably colloidal gold?) Nothing too fancy. In case it helps, I'm specifically looking at phosphotidylserine exposed on the outer membrane leaflet during apoptosis in a cultured line of CML cells. Thanks in advance.
Gib Ahlstrand wrote: =============================================== Years ago, upon recommendation by folks out at Sharon Vacuum Company in Brockton MA, I bought a two component epoxy kit called A-12 Epoxy Adhesisve from the Armstrong Products Company on Argonne Road in Warsaw, Indiana, for repairing leaks in vacuum systems. It also proved useful for sealing cracked plastic parts and metal fittings on cryogenic systems, and for many other repairs. Great stuff, and it cured to a beautiful milk chocolate brown tone.
I'm just about out now, and I've written to Armstrong Products and they are no longer in existence,letter was not forwardable, etc.
Anybody out there know of their whereabouts? Or could you recommend any other suitable replacement product? ================================================= It is definitely not the same thing, but we (and a number of our customers) have had excellent results with VACSEAL® High Vacuum Leak Sealant. The product can withstand repeated temperature cycling from liquid helium temperatures to 450° C over long intervals of time.
It is described on URL http://www.2spi.com/catalog/vac/vacleak.html
Disclaimer: SPI Supplies offers this product for this kind of application so we would have a vested interest in seeing it used more widely.
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: www.2spi.com ############################ ==================================================
Hello Paul. Believe it or not 4 small inner tubes, inflated so that they have no real tension before having a table top put on them work very well. I have used this trick for Atomic force microscopy (AFM) & holography (on a wooden table). Check out how pneumatic legs are made, aside from the automatic leveling (option) they are not much different. An improvement is to build a base consisting of layered, carpet, plywood, 8x8x16" bricks (commonly called cinder blocks around here) up to the height needed. There are a lot of optical tables on this planet set up this way. You can get small tubes (6-8" dia) from cart & dolly vendors. Another trick is to to build a sand box. Check books on do it yourself holography Yea, I know these ideas don't look great but they do work & save lots of money.
HI Gib. Varian Vacuum Products has been selling "Torr Seal" for years. Don't know about it's cryo properties.
Bruce Brinson Rice U.
Gib Ahlstrand 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 Microscopy Listers, } } Years ago, upon recommendation by folks out at Sharon Vacuum Company in Brockton } MA, I bought a two component epoxy kit called A-12 Epoxy Adhesisve from the } Armstrong Products Company on Argonne Road in Warsaw, Indiana, for repairing } leaks in vacuum systems. It also proved useful for sealing cracked plastic parts } and metal fittings on cryogenic systems, and for many other repairs. Great } stuff, and it cured to a beautiful milk chocolate brown tone. } } I'm just about out now, and I've written to Armstrong Products and they are no } longer in existence,letter was not forwardable, etc. } } Anybody out there know of their whereabouts? Or could you recommend any other } suitable replacement product? } } Thanks, } } 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.crl.umn.edu
Scott - I never found razor blades satisfactory on fresh wood and rather suggest using a microtome with a permanent knife (not a disposable). I have not tried myself using a tungsten carbide knife but expect that this would give still better results. Small triangular TC knives which fit ultramicrotome chucks are an obvious and cheaper solution than the big TC knives.
Diamond knife sections would be too thin for SEM purposes but diamond knives can be used for facing a block. However, those knives are expensive and the operation is risky.
Another alternative is the use of a series of wet and dry papers on a lap. Use the paper wetted with kerosene or another slowly evaporating solvent and when finished use absolute alcohol to rinse. These papers are available to about 2000 gird, which is still not fine enough for SEM. For the final finish I suggest diamond lapping film. This material is expensive but the diamond particles are embedded with the plastic. The particles will rarely contaminate the specimen, the particle size is available down to 0.1 micrometre and the film is long lasting.
Using the grinding powder that is normally used for cutting of rock, makes a mess of a specimen like wood. Disclaimer PST is a supplier of some mentioned products.
Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 30, 1999 3:18 AM, Scott Miller [SMTP:smiller-at-umr.edu] wrote: } } Hello All, } Does anyone out there have any suggestions for getting } good SEM cross sectional images from dried wood? I have a } student who would like to image the microstructure of } several types of wood. All the references I have found } so far suggest cutting fresh or moist wood with a new } razor blade to achieve good cross-sections. The student } is studying the effects of two different drying } techniques, and therefore soaking the wood in water is } out of the question. Cutting the dried wood with razor } blades has resulted in crushed, damaged cross sections. } } Thanks, } Scott } F. Scott Miller } Electron Microscopy Lab smiller-at-umr.edu } University of Missouri-Rolla } 223 McNutt Hall voice: 573 341 4727 } Rolla, MO 65409 USA fax: 573 341 6934
These were once very popular, especialy for preparations of diatoms, but they seem very hard to find these days! (I have posted a question regarding mounting media with high RI on Histonet a few weeks ago: received only one answer...) The only source I know of is a Dutch company called Euromex Microscopes. They sell Naphrax (RI = 1.710) in 25 ml bottles. Catalog nr = PB0267. You can contact them trough email: info-at-euromex.nl Yvan Lindekens.
---------- } Van: SROUBEK {pasroube-at-mtu.edu} } Aan: Microscopy-at-sparc5.microscopy.com } Onderwerp: help on mounting medium } Datum: vrijdag 29 januari 1999 20:24 I am looking for a mounting medium with a high } optical index (around 1.7). Once I used something } called piperin (n=1.68). Could you please let me know } where can one obtain such products? } } Thanks } } Pavel Sroubek } } pasroube-at-mtu.edu }
Could the sample be planed at low temperature? Such as in a cryostat or cryoultramicrotome? The problem is that it would have to be returned to room temperature without condensation forming, maybe in a "sealed" container with dry nitrogen or air+dessicant.
Maybe it could be impregnated with a non-aqueus medium. I am wondering if 1-hexadecene (from Sigma) could be used. It is used as a mdium for filling gas spaces etc in leaves before high pressure freezing is carried out. It is a non-aquous, non-toxic, non-osmotically active type of paraffin. I am not sure about its removal afterwards.
Something that would sublime would be favoured! Maybe one of the media used instead of critical point drying? Hexamethyldisilazane? Combined with low temperature sectioning?
Another primitive approach would be to put it in liquid nitrogen in a styrofoam contained with a metal "plate" on the bottom and fracture the sample across with a cooled razor blade (held in tweezers) whacked with a hammer. Then it needs rewarming as above, in a container with dessicant (fresh phosphorus pentoxide, in fine powder form, is my favourite - this is a vicious chemical so beware of inhalation of the dust), preferably with dried nitrogen gas leaking through to prevent air plus atmosheric moisture ingressing.
At the recent resin cutting workshop in Seefeld (Austria), Prof. Sitte (who has had a lot to do with the design of Reichert/Leica ultramicrotomes) said that it is possible on modern instruments to cut 5 or 6 mm across by 12 mm in length. For TEM examination, ther grid size is the limiting factor! I have cut 2.5 x 2.5. mm ultrathinns for TEM. Someone else here has cut roughly the same size 0.5-1.0 micrometers thick, without a "boat"/water bath, of freeze-dried tissue, dry mounted, for x-ray microanalysis.
I would say that 5 x 5 mm, 1 micron thick should not be a problem for the instrument providing that the approach and trimming is done carefully and that the specimen is not very hard. The specimen should section easily, if it "sticks" on the knife and doesn't pass to cut a section then I would think again.
Hi Pavel: I don't know about Piperine, maybe somebody else can help with that. However, Meltmounts are available in a range of several refractive indices, including 1.68 and 1.704. The Meltmount quickstick is applied to a slide on a hotplate and after mounting the coverslip, the medium instantly solidifies at room temperature. The coverslip may be removed at anytime by re-heating the slide. Disclaimer: Meltmount-Quickstick are available from PST and other microscopy suppliers. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 30, 1999 5:24 AM, SROUBEK [SMTP:pasroube-at-mtu.edu] wrote: } } I am looking for a mounting medium with a high } optical index (around 1.7). Once I used something } called piperin (n=1.68). Could you please let me know } where can one obtain such products? } } Thanks } } Pavel Sroubek } } pasroube-at-mtu.edu
Dorota, I also have a Reichert ultracut and it does fine with very large thick sections. Knives tend to suffer however. Depending on your sample, you may find that you'll need a diamond histo knife. Glass however will also work . MG Engle Electron Microscopy & Imaging Facility University of Kentucky
Have you been experiencing trouble with vibration before now?
I'm on the 4th floor of my building, which has a lot of activity during the day. In my lab there are 4 sectioning stations with ultramicrotomes lined up in the same room, sited against the same wall. Three are on marble tables, one is on a 'floating' table.
Hands down, the N2-damped table beats out the marble tables.
For any critical work, the microtome on the floating table is usable at any hour of the day, while those on marble tables usually must be used after-hours, after traffic in the building has eased. The vibration in the floor is easily visible in the water's surface on any of the 3 marble tables; it is damped out in the N2-table. Well worth the investment.
Good luck! Ann Lehman EM Facility Mgr Trinity College Hartford, CT v 860-297-4289 f 860-297-2538 e ann.lehman-at-exchange.cc.trincoll.edu
-----Original Message----- } From: Gerroir, Paul J [mailto:Paul.Gerroir-at-crt.xerox.com] Sent: Friday, January 29, 1999 2:44 PM To: Microscopy-at-Sparc5.Microscopy.Com
January 29, 1999
Fellow Microscopists, I am soliciting your comments on what you consider the best approach to damp vibrations when using a light microscope. We are in the process of redesigning our light microscopy/image analysis work area and have a choice between a conventional marble table and a Newport BenchTop vibration isolation system (pneumatic). Which should it be? Are there other possibilities to consider?
Thanks for a moment of your time. Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
We do a lot of extremely sensitive micromanipulation work here with the LM and have a few very effective anti-vibration systems.
We placed a heavy slab (approx. 400lb) 2' x 3' of Boiler Plate Steel on top of about 150 tennis balls on a well supported bench. The steel is well-protected and finished to make a clean, safe work area. Total cost was about $250. Cdn.
If you'd like more details, you can contact me off-line.
Karen Rethoret Microscopy Lab York University Toronto, Ont. 416-736-2100 x33289
On Fri, 29 Jan 1999, Gerroir, Paul J wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } January 29, 1999 } } Fellow Microscopists, } I am soliciting your comments on what you consider the best approach to damp } vibrations when using a light microscope. We are in the process of } redesigning our light microscopy/image analysis work area and have a choice } between a conventional marble table and a Newport BenchTop vibration } isolation system (pneumatic). Which should it be? Are there other } possibilities to consider? } } Thanks for a moment of your time. } Paul } } Paul J. Gerroir } Microscopy } Materials Characterization } Xerox Research Centre of Canada } 2660 Speakman Drive } Mississauga, Ontario L5K 2L1 } } Phone: (905) 823-7091, ext. 216 } FAX: (905) 822-7022 } e-mail: paul.gerroir-at-crt.xerox.com } }
Research Opportunities Mount Sinai School of Medicine is a leader in medical research and education. The establishment of our new Microscopy Center has created opportunities for experienced Microscopists with a BS/BA or MS in Biology or Life Sciences. All applicants should have excellent organizational and communication skills, an understanding of basic laboratory procedures, and the ability to manage a large and varied workload.
Electron Microscopist The successful candidate will participate in ultrastructural studies of various biological systems. Qualifications include at least 2 years of experience in routine electron microscopy procedures (TEM, SEM), ultramicrotomy, immunogold labelling, specimen preparation, photographic darkroom work, and routine maintenance of equipment. Individuals with immunofluorescence and confocal microscopy experience are desirable. Code: EM
Light Microscopist The successful candidate will participate in biomedical studies that use a variety of advanced light microscopic techniques. Duties will include maintaining equipment, instructing users in equipment operation, and sample preparation. Qualifications include at least 2 years of experience in fluorescence and confocal microscopy, immunofluorescence labelling, in situ hybridization, digital imaging and analysis, cell culture, and histological techniques. Strong computer skills are essential. Code: LM
We offer a salary commensurate with experience and excellent benefits. For consideration, please mail your resume, which must indicate code for position of interest, to:
Scott Henderson, Ph.D., Director, Microscopy Center, Box 1007, Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029-6574
http://www.careermosaic.com/mountsinai
We are an equal opportunity employer fostering diversity in the workplace.
______________________________
Scott Henderson, Ph.D. Director of Microscopy, Mount Sinai School of Medicine, Dept. of Cell Biology & Anatomy, Box 1007, One Gustave L. Levy Place, New York, NY 10029-6574
Paul, we use a combination of marble tables and pneumatic shock absorbers on our microscopes. We have been very happy with the results. One of our microscopes has the bench top vibration table which is also very good.
At 02:44 PM 1/29/99 -0500, Gerroir, Paul J wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Doug: Try this paper. You will have to adapt the protocol to fit your needs. If you have any questions please feel free to call.
Coller, Barry S., Kutok, J.L., Scudder, L.E., Galanakis, D.K., West, S.M., Rudomen, G.S., Springer, K.T. . Studies of Activated GPIIb/IIIa Receptors on the Luminal Surface of Adherent Platelets: Paradoxical Loss of Luminal Receptors When Platelets Adhere to High Density Fibrinogen. J. Clin. Invest. (1993) Vol. 92, pp. 2796-2806.
Doug Matthews wrote:
} Hi everyone, } } I'm interested in doing some immuno work with the SEM. Basically } tagging colloidal gold onto a surface antigen on cultured cells. I've got } some old information and papers but as for up-to-date methodologies, I'm a } little sketchy. Does anyone have any good review papers on the use of immuno } surface markers w/ SEM (preferably colloidal gold?) Nothing too fancy. In } case it helps, I'm specifically looking at phosphotidylserine exposed on the } outer membrane leaflet during apoptosis in a cultured line of CML cells. } Thanks in advance. } } Doug Matthews } Providence College } }
-- Regards, Gregory Rudomen Technical Specialist University Microscopy Imaging Center State University of New York at Stony Brook 516-444-3126 Greg-at-umic.sunysb.edu *************************************** Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. ***************************************
Hi! Thanks to all of you who responded to my posting. All suggestions are very helpful. I forgot to add that the tissue (lung from rat) is embedded in Epon/Araldite. It is not cryosectioning. Thanks again Dorota
I am submitting a request for a faculty member who is not a member of the list. The researcher wishes to locate, for sale, antibodies and/or conjugated antibodies for Salmonella, Clostridium and Campylobacter.
Any replies may be directed to my e-mail address and not to the list.
With Best Wishes,
Bill Monroe
Bill Monroe EM Center Mississippi State University (601)-325-3019 Lab Fax 325-0246
Greetings, I'm looking for a conductive epoxy (for materials science use) in which I can embed metal samples, sand and polish, and then look at them in a field emission SEM. I have seen this used at other facilities, so I know it exists. I found something at Electron Microscopy Sciences that I thought might work, but the product has been discontinued. Does anyone know of an epoxy that can be used for the above application?
Any replies can be directed to me personally.
Thank you, Kevin Croat tkc-at-howdy.wustl.edu Dept. of Physics Washington University in St. Louis
Thanks to all those who responded to my question about resin problems. I got a number of good suggestions, mostly related to storage (most people thought storing resins in the fridge was unnecessary and probably a bad idea), use of accelerator (several people suggested that I switch to BDMA or add DMP-30 to all infiltrating steps) or water contamination in any of the components (several people suggested replacing all resins). We will definitely be following up. Many thanks for taking the time to reply.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-4861936
The Microscopy Society of America (MSA) and the MSA Technologists' Forum are the sponsors of the Professional Technical Staff Awards (PTSA) to provide assistance on a competitive basis to full-time professional staff who submit papers for presentation at Microscopy and Microanalysis '99 (M&M '99). The deadline (15 February 1999) is fast approaching! Eligible applicants: you are encouraged to submit an abstract and supporting documentation. Managers: you are encouraged to support eligible staff members in this effort. Please read the following, taken from the M&M '99 Registration Bulletin, for more information.
It is the intent of this award to stimulate attendance at M&M '99 for professional technical staff who ordinarily might not participate in the meeting, and to encourage employers to support their staff in professional activities. Awardees will be selected based on the quality of an abstract submitted for presentation at M&M '99. The applicant must be the first author of the submitted abstract. Applicants must be full paid-up members of MSA at the time of application. The awards consist of free full registration for the meeting, a copy of the Proceedings and the Sunday evening social event. MSA will reimburse awardees up to $600 for travel, lodging and other expenses. There will be a maximum of four awards given. Successful applicants must present their abstracts personally at M&M '99 in order to receive the award. They are expected to attend and participate in the entire meeting. Former winners will not be eligible for another award. Applications shall consist of: (1) a supporting letter from the applicant's employer, manager or supervisor, attesting to the applicant's status as a full-time professional staff member; (2) a scientific abstract (original and 4 photocopies) for presentation, as described in the Registration Bulletin, accompanied by a completed Data Form (available on-line at http://resolution.umn.edu:591/MandM/DataEntry.html, or if inaccessible, by calling the Meeting Manager at 708/361-6045; electronic submission of the Data Form is encouraged); (3) a copy of the abstract to be sent by 15 February 1999 to the Chair of the Technologists' Forum: Ms. Beverly E. Maleeff, SmithKline Beecham Pharmaceuticals, Safety Assessment-US, UE0462, 709 Swedeland Road, King of Prussia, PA 19406. Phone: 610/270-7987; Fax: 610/270-7202; e-mail: Beverly_E_Maleeff-at-sbphrd.com. In order to be considered, completed applications must be received by 15 February 1999. Abstracts will be judged by the MSA Technologists' Forum. All applicants will be notified of the outcome in early March. Applicants not receiving the award will have the opportunity to withdraw their abstract if necessary.
Regards, Bev Maleeff Chair, MSA Technologists' Forum
I'd like to thank all those who responsed to my question. It is clear now that we should use double glid (Ni) instead glue to hold the sammple. Attached are messages I've received.
} From zrdai-at-u.washington.edu Mon Feb 1 16:06:29 1999
You might consider the following, (a thick, rather crude adhesive). It is: Sauereisen Insa-Lute adhesive, No. 1 paste. We used it to hold U-Si slices for polishing mechanically. The as mounted slices were then ion irradiated in a UHV vacuum chamber at 350 C approximately. The glue held pretty well, but can't be dissolved-unless the company has a special solvent. It outperformed several "high temp" glues that I tried on hotplate tests. Source: Sauereisen 160 Gamma Drive Pittsburgh Pennsylvania, 15238-2989
Phone: (412) 963-0303 FAX: (412) 963-7620
Bernard Kestel Materials Science Div. 9700 So. Cass Ave. Argonne, Ill., 60439
} From stephan-at-gecko.biol.wits.ac.za Mon Feb 1 16:06:29 1999
I was wondering if anyone in the microscopic community could recommend an outfit that can polish petrographic thin sections for EMP work. I need to can the samples processed within a month timeframe and I am willing to pay some for this. Many thanks
The Emory University Neurology Microscopy Laboratory, the University of Georgia Botany Department EM Laboratory, and Leica Microsystems, Inc.
Present a Cryo Techniques and Immunogold Workshop.
April 18-23, 1999 at the Campus of Emory University in Atlanta, GA.
Objectives
1. To provide researchers the opportunity to learn the theory and practice of cryo techniques for biological sample preparation and immunogold labeling.
2. To permit participants to process their own samples using these techniques under expert guidance.
I was requested to identify a crystal phase of small (5-7 micron) alumina particles embedded into copper. There are only a few particles, and all of them are on the surface. Any ideas how it can be done?
Thanks,
Alexander Titkov Millennium Inorganic Chemicals PO Box 245 Bunbury WA 6231 Australia Ph 61 8 9780 8505 W FAX 61 8 9780 8500 E-mail: atitkov-at-micl.com.au
Do any of you probers out there have a spare TAP crystal for a JEOL 8600/733 probe that they may be willing to sell? ..we've had something of an accident..
Thanks,
Stu -------------------------------------------- Stuart Kearns Electron Microbeam Laboratories Department of Earth Sciences University of Bristol Queens Road Bristol BS8 1RJ UK tel: (0)117 954 5435 fax: (0)117 925 3385 e-mail: Stuart.Kearns-at-bristol.ac.uk http://eugf.gly.bris.ac.uk --------------------------------------------
When I was an undergraduate research assistant at Wash U, I worked in the McDonnell Center for the Space Sciences. They were using an epoxy called E-7 which I have been using ever since. It comes in two parts (A & B) which you mix in a 2 to 1 ratio. Curing time is 2 hours at 150 degrees F. If cured right, there is really no outgassing or beam damage, even at higher micrprobe currents. Contact Pat Swan on the 4th floor, he might still use it. You can buy it from:
Techkits PO Box 105 Demerest, NJ 07627 201-768-7334
The latest price was $17.25/set for {10 sets. Hope this helps, Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
American Lab will be running an extensive review of new equipment being displayed at the upcoming PITTCON meeting (March 7-11, Orlando). For the first time, microscopy and related imaging will have its own section, as part of the on-going FOCUS ON MICROSCOPY column. If you are: (1) showing products which have not been exhibited or presented at a prior PITTCON and/or (2) introducing new products at this PITTCON please send copies of press kits, press releases, slides/product shots, and any other information to me at the address below. Indicate on the envelope that this is for the PITTCON Microscopy/Imaging review.
This article will need to go to press shortly after the meeting, so if I can get a head start on your materials, I would greatly appreciate it. Also, I will be on the show floor, following through on any materials received, so please enclose your booth number, name of a pertinent contact, and, if possible, a selection of times when they might be in your booth.
This article will appear in the May issue of American Lab.
Please call/email if you have any questions. Thanks in advance for your assistance.
Best regards, Barbara Foster Microscopy/Marketing & Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Contributing editor to American Lab ("Focus on Microscopy" column)
"Jeff" wrote: ============================================== Good Day,
Does anyone know of a dry etch that has a high selectivity to etch SiO2 preferentially to Si? We need to image these samples in a SEM. ================================================= This is usually done in a plasma etcher, using CF4 or some other reactive F gas of the more expensive types. You can get more information about this on our website given below. Typically, in a 100 watt barrel etcher, you can remove 1 um of SiO2 in about 30 minutes. The process is completely dry and is used in failure analysis laboratories.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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We are beginning to use the Historesin plus embedding medium supplied by Leica for immunocytochemistry of the retina by light microscopy. We will be using the images to count photoreceptor and retinal pigment epithelial cells of animals fed special diets to examine the effects of nutrition on the retina. I would like to communicate with others who have used this product to learn the best ways of storing material to preserve immunoreactivity for long periods of time and to share information on other technical issues.
You can respond to me directly at the address below.
Max Snodderly, Ph.D. Schepens Eye Research Institute 20 Staniford Street Boston, MA 02114, USA
In a message dated 2/1/99 9:03:47 PM Eastern Standard Time, kjtobin-at-uic.edu writes:
{ { I was wondering if anyone in the microscopic community could recommend an outfit that can polish petrographic thin sections for EMP work. I need to can the samples processed within a month timeframe and I am willing to pay some for this. } }
We recommend:
Mineral Optics Laboratory 29 "A" Street, P. O. Box 828 Wilder, Vermont 05088 USA 802-295-9373 802-295-7540 (FAX)
Yours truly, Steve Stokowski Stone Products Consultants Concrete Petrographers 10 Clark Street, Suite A Ashland, Massachusetts, 01721 USA 508-881-6364 http://members.aol.com/CrushStone/petro.htm
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Hello to all!
I am looking for a set of negative cassettes (30 plates), a cassette magazine box, and a cassette receiver box for Hitachi-600 TEM. Please let me know if any of you or your friends have a Hitachi TEM being taken down apart and wants to give away or trade these things.
Thanks in advance.
Gang Ning EM Facility Medical College of Wisconsin 414-456-8344
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Hello to all!
I am looking for a set of negative cassettes (30 plates), a cassette magazine box, and a cassette receiver box for Hitachi-600 TEM. Please let me know if any of you or your friends has a Hitachi TEM being taken down apart and wants to give away or trade these things.
Thanks in advance.
Gang Ning EM Facility Medical College of Wisconsin 414-456-8344
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Colleagues will be growing Vero cells (Green Monkey Kidney Cells) and intestinal epithelial cells on cover slips for subsequent ultrathin sectioning and TEM examination. I am interested to know which cover slips are best to use for this purpose. Can they be purchased sterile? I have checked the Ted Pella and EMS catalogs, they both sell non sterile cellulose acetate cover slips. Are there compatibility issues with resins and solvents? I am interested in any tips or tricks to smooth the way. Thanks, Sally Burns
Sally Burns Center for Electron Optics B5 Pesticide Research Center Michigan State University East Lansing, MI 48823 (517) 355-5004 Phone (517) 353-5598 FAX
I need some microneedles to dislodge some very fine particles (micron size) on my sample but still flexible enough to bend. Any information is appreciated.
As an electron microscopist who has come from a neurophysiology background, I have used various fine needles for "dusting" off debris. You need to find the one that feels right.
Cat whiskers are long, pointy, strong, and flexible. They are particularly good for chasing tiny bubbles out of microelectrodes or capillary tubes.
Finely drawn glass. Heat a pipet or rod over a Bunsen burner and draw it out until it breaks. Find somewhere along the long string that has the right size and flexibility, but won't break and leave more debris!
Cactus spines. They come in many shapes and sizes. Also useful for pinning down things for dissection that can't come into contact with metal.
Find someone in neurobiology who does microelectrode recording and get them to make some electrodes, which are capillary tubes drawn to a very fine point. You can probably get some in the micron range. Beveled, even!
Insect Minutin pins mounted on a stick are very strong, but may scratch your substrate. They can be ground down for a finer point.
Eyelashes, beard hair, and other body hairs each have different properties. Have fun experimenting.
Good luck!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
We have used tungsten needles of about that dimension with our high- voltage electron microscope.. what we were looking for was a rather rigid "needle point" on which to mount micron sized objects. It was found that when we attempted to move these small specimens around and to get them to stick to the needles, the needles would bent very easily when they contacted the glass slides our specimens were prepared on. So you might try tungsten. We made them by electrolytically etching 5 mil. tungsten wires in NaNo3 solution, connecting the tungsten wire and a small graphite rod (1/8 inch dia.) to each of the terminals of a 6 volt AC transformer ( yes AC! ) and dipping both into the solution.The graphite rod was dipped well into the solution but the wtungsten would be dipped only an 1/8 inch or less below the surface. After a number minutes the wire would etch to a very small point, which could be examined under a light microscope until it was small enough for the application. Good luck.
For TEM of cultured cells, we grow the cultures on "Thermanox" tissue culture coverslips. From Nalge Nunc INternational, 50 sterile coverslips, 13 mm in diameter is catalog 174950. EMS also sells these thermanox cover slips in a variety of sizes (see page 143 of their cat XIII). The coverslips can be treated with all the same chemistry as tissue including propylene oxide and Spurrs epoxy, which are two components which solubilize polystyrene. These thermanox coverslips can be sunk cell side up in freshly made Spurrs, then following polymerization the coverslips can be removed by first sawing a small area of the epoxy/cell/substrate, then immersing in liquid nitrogen for a few seconds, then prying away the substrate. Now the embedded cells are immediate on the epoxy. Re-embed two fragments of the culture face to face for cross-sections, or cut the block parallel to the face for tangential sections. We particularly like the round 13 mm thermanox coverslips for immunocytochemistry of cultured cells since they can be floated cell side down in a drop of 100 microlitters antibody - gold conjugate, conserving reagents.
If you would like to grow a larger culture, you could also use "permanox" culture dishes, which are equally resistant to chemicals common in TEM processing. These are also available through EMS and other suppliers.
Good luck,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
Orlando in March!!!! To Register follow the local meetings in www.vacuum.org or http://www.msa.microscopy.com
Over 30 Invited Talks, 40 vendors, and over 40 student posters!
Golf Tourny on Sunday March 14 - www.pegasus.cc.ucf.edu/~ampac
AVS short courses and UCF/Vendor Sponsored Short Courses in Tripod Polisher and FIB TEM Specimen Preparation see www.pegasus.cc.ucf.edu/~ampac -----------------------------
Monday March 15, 1999
Opening and Keynote Address
8:00-8:15 am Welcome and Introduction, Lucille Giannuzzi, 1999 FL AVS/FSM Program Chair
8:15-9:00 am Keynote Address, Sam Durrance, Astronaut, Professor, University of Central Florida
Monday, March 15, 1999
Session I: Thin Films
Session Moderator: Maggie Puga-Lambers, University of Florida
9:00-9:25 am Tim Anderson, Chemical Eng. Dept., University of Florida
9:25-9:50 am CONCENTRATION DEPTH PROFILING OF IMPURITIES AND DOPANTS IN FLAT PANEL DISPLAYS AND GLASSES BY SIMS, Temel Buyuklimanli, Evans East
9:50-10:15 am MODIFICATION AND CHARACTERIZATION OF DEFECT STATES IN ZnO FILMS, Gregory J. Exarhos and Charles F. Windisch, Jr., Pacific Northwest National Laboratory
10:15-10:45 am Coffee Break
10:45-11:10 am FUNDAMENTALS OF TUNGSTEN CMP DURING CMOS DEVICE FABRICATION, Rajiv Singh, Engineering Research Center for Particle Science and Technology, University of Florida
11:10-11:35 am SURFACE ANALYSIS APPLICATIONS IN SEMICONDUCTOR TECHNOLOGY, Bridget Rogers, Vanderbilt University
11:35-12:00 pm THE INFLUENCE OF AIR ON THE PROPERTIES OF THIN FILMS DEPOSITED FROM BEAMS OF NANOPARTICLES USING A COPPER SOURCE, F. K. Urban, III, A. Khabari, A. Housseini-Tehrani, P. Griffiths, and G. Fernandez
12:00-1:00 pm Lunch Monday, March 15, 1999
Session II: Microscopy of Biological Samples
Session Moderator: Karl Muffly, University of South Florida Jo Ann Moore, University of South Florida
9:00-9:25 am DIGITAL MANIPULATION OF ACQUIRED IMAGES, WHAT IS POSSIBLE VS. WHAT IS ETHICAL, John Kinnamon, University of Denver and The Rocky Mountain Taste and Smell Center
9:25-9:50 am THE USE OF CORRELATIVE MICROSCOPY IN BIOLOGICAL PROBLEM SOLVING, Ralph Albrecht, University of Wisconsin
9:50-10:15 am APPLICATIONS OF A 300 KV, FIELD EMISSION ELECTRON MICROSCOPE IN STRUCTURAL BIOLOGY, Kenneth A. Taylor, Florida State University Institute for Molecular Biophysics
10:15-10:45 am Coffee Break
10:45-11:10 am DECONVOLUTION VS STANDARD FLUORESCENCE MICROSCOPY, Karl Muffly, University of South Florida College of Medicine
11:10-11:35 am SURFACE AND MICROSCOPIC ANALYSIS OF BIOMATERIALS AS THEY CHANGE IN VIVO: HOW FAR ARE WE FROM NEEDED DATA?, Chris Batich and Anthony Brennan, University of Florida
11:35-12:00 pm CARBOHYDRATE DEPOSITION PATTERNS IN ETIOLATED SOYBEAN SEEDLINGS GROWN IN MICROGRAVITY, E.C. Stryjewski, NASA Gravitational Biology Laboratory, Dynamac Corp., K.M. Johnson, National Research Council, NASA/KSC, W.C. Piastuch1, H.G. Levine1, and L.H. Levine, NASA Gravitational Biology Laboratory, Dynamac Corp., O. Martynenko3, and V. Prima,, Institute for Molecular Biology and Genetics, National Academy Of Sciences, Ukraine
12:00-1:00 pm Lunch
1:30-3:30 pm WORKSHOP ON MANIPULATING DIGITAL IMAGES, John Kinnamon, University of Denver and The Rocky Mountain Taste and Smell Center
3:30-4:00 pm Florida Society for Microscopy Annual Business Meeting
3:30-6:00 pm Competition Session and Student Competition (Session IV) Monday, March 15, 1999
Session III: Surface Science and Analysis
Session Moderators: Sudipta Seal, University of Central Florida
1:00-1:25 pm WETTABILITY AND INTERFACES IN METAL/NITRIDE SYSTEMS, Natalia Sobczak, Foundry Research Institute, Cracow, POLAND
1:25-1:50 pm SOFT X-RAY FLUORESCENCE SPECTROSCOPY IN MATERIALS SCIENCE, E. Joseph Nordgren, Uppsala University, Uppsala, Sweden
1:50-2:15 pm MAGNETIC PHASE DIAGRAMS OF ULTRA-THIN FILM BINARY ALLOYS FOR SPIN-VALVE APPLICATIONS, Brian Tonner, University of Central Florida
2:15-2:40 pm PRACTICAL ASPECTS OF HIGH RESOLUTION XPS WITH MONOCHROMATIC AlKA X-RAYS, A.C. Miller, Lehigh University
2:40-3:05 pm STUDIES OF OXIDATION AND CORROSION USING AN ANAEROBIC CELL APPROACH, Peter M.A. Sherwood, Kansas State University
3:05-3:30 pm MICRO-ESCA/NEXAFS AT A THIRD GENERATION SYNCHROTRON LIGHT SOURCE, J. H. Underwood, U. Kleineberg, S. Mrowka, P. J. Batson, S. B. Rekawa, M. S. Jones, R. C. C. Perera, P. N. Ross, University of California, Berkeley
3:30-4:00 pm Coffee Break
3:30-6:00 pm Competition Session and Student Competition (Session IV)
Monday, March 15, 1999
Session IV: Poster Session
Session Moderators: Larry Plew, Cirent Semiconductor
CHARACTERIZATION AND SURFACE ANALYSIS OF HYDROXOCARBONATE COMPOUNDS, B. B. Adhyaru, K. R. Williams, and V.Y. Young, University of Florida
INTERFACIAL REACTIONS BETWEEN METAL/P-GaN FOR FORMATION OF OHMIC CONTACTS, M. Ahonen, B. Liu, P.H. Holloway, University of Florida
SURFACE METASTABLE STRUCTURE OF KTa03 (001) BY HELIUM ATOM SCATTERING, E. A. Akhadov, T. W. Trelenberg, J. A. Li, J. G. Skofronick, S. A. Safron, Florida State University and L. A. Boatner, Oak Ridge National Laboratory
SIMS STUDY OF DIFFUSION PHENOMENA OF METAL ELEMENTS IMPLANTED INTO SILICON, Elvira V. Anoshkina,a,b) Hughes Francois-Saint-Cyr,a,b,c) Ashfaq Hussain,a,b) , Isaiah Oladeji,d) Fred A. Stevie,e) Lee Chow,b,d) Dan Zhou a-dUniversity of Central Florida, eCirent Semiconductor
FAILURE ANALYSIS : AN AES-SEM STUDY, K. R. Beaulieu, (UNDERGRADUATE) A. S. Kale, S. Seal, V. Desai, University of Central Florida
IDENTIFICATION OF SURFACE CHEMICAL FUNCTIONAL GROUPS IN POLYMER MEMBRANES: AN X-RAY PHOTOELECTRON SPECTROSCOPY STUDY, Sharon D. Beverly 1,2, Satyajit V. Shukla, 3,4 Seungkwan Hong, 2 and Sudipta Seal3, 4, 1NASA, 2-4University of Central Florida
STUDY OF CORROSION FAILURES UNDER ATMOSPHERIC CONDITIONS, L.A. Bracho, V. H. Desai, S. Seal, University of Central Florida
ANISOTROPIC PATTERN TRANSFER IN GaN BY PHOTO-ENHANCED WET ETCHING, Hyun Cho(1), S.M. Donovan(1), C.R. Abernathy(1), J. Han(2), R.J. Shul(2), F. Ren(3) and S.J. Pearton(1), (1) & (3) University of Florida (2) Sandia National Laboratories
NOVEL EMITTER BASE SELF-ALIGNED PROCESS FOR AlGaN/GaN HETEROJUCTION BIPOLAR TRANSISTORS, X. A. Cao1, H. Cho1, S. J. Pearton1, C. R. Abernathy1, F. Ren1, J. Han2, R. J. Shul2, and A. G. Baca2, 1 University of Florida, 2 Sandia National Laboratories
HELIUM ISOLATION IMPLANT FOR GALLIUM NITRIDE BASED FIELD EFFECT TRANSISTORS, G. Dang1, X. Cao1, F. Ren1, S.J. Pearton1, J. Hang2, and R. J. Shul2, 1University of Florida, 2Sandia National Labs
CAPILLARIZATION OF SKELETAL MUSCLE IN RATS UNDERGOING HEART FAILURE, Hans Degens, Rebecca K. Anderson, Karl E. Muffly and Stephen E. Always, University of South Florida
UN-ANNEALED AND ANNEALED PD ULTRA-THIN FILM ON SIC CHARACTERIZED BY ATOMIC FORCE MICROSCOPY, SCANNING TUNNELING MICROSCOPY, AND X-RAY PHOTOELECTRON SPECTROSCOPY, K. Elshot, Mechanical Materials and Aerospace Engineering, University of Central Florida, Orlando, FL 32826, W. Lu, D.T. Shi,E. Bryant, K. Lafate, H. Chen, A. Burger, W. E. Collins, Department of Physics, Fisk University, Nashville, TN 37208
FUNDAMENTAL PROPERTIES ON E-BEAM EVAPORATED ZnS:Mn AND Zn1-xMgxS:Mn ELECTROLUMINESCENT THIN FILMS, Tao Feng, Mark Davidson, Paul Holloway, University of Florida
DESIGN, DEVELOPMENT AND TESTING OF A COMPUTERIZED DATA ACQUISITION AND CONTROL SYSTEM FOR A NANOPARTICLE DEPOSITION SYSTEM, Frank K. Urban and G Fernandez, Florida International University, Miami, Florida
CHARACTERIZATION OF THE DIFFUSION PROPERTIES OF Mg, Cl, K, Ge, AND Mo IN SILICON BY SIMS, Hughes Francois-Saint-Cyr,a,b,c) Elvira V. Anoshkina,a,b) Ashfaq Hussain,a,b) ,Fred A. Stevie,e) Lee Chow,c,d) Kathleen Richardson, a,b,c) and Dan Zhou a,b), a-dUniversity of Central Florida, eCirent Semiconductor
A STUDY OF THE SURFACE COMPOSITION OF KTAO3 DOPED WITH CA, BA, SR, AND NB, P.W. Gresser Florida State University
NOZZLE DESIGN IN A DIFFERENTIALLY PUMPED NANO-PARTICLE INSTRUMENT, Peter D. Griffiths and Frank K. Urban, Florida International University
INTERFACIAL CHRACTERISTICS OF AlN TO Si, SiC AND GaN, K. Harris, B. P. Gila, F. Ren, J. Deroaches, K. N. Lee, J. D. MacKenzie, C. M. Zitterling+, M. Ostling+, S. N. G. Chu**, C. R. Abernathy, and S. J. Pearton, University of Florida, Gainesville, FL, +KTH, Royal Institute of Technology, Stockholm, **Bell Laboratories, Lucent Technologies
SELECTIVE DRY ETCHING USING INDUCTIVELY COUPLED PLASMAS: GaAs/AlGaAs AND GaAs/InGaP, D.C. Hays, H. Cho, K.B. Jung, Y.B. Hahn*, C.R. Abernathy, S.J. Pearton, and F. Ren, University of Florida, W.S. Hobson, Bell Laboratories, Lucent Technologies
HOT FILAMENT CVD OF DIAMOND THIN FILMS, Ashfaq Hussain, Lee Chow, and Dan Zhou, University of Central Florida
QUANTITATIVE COMPARISON OF VON WILLBRAND FACTOR (VWF) EXPRESSION IN HUMAN NON-MALIGNANT AND MALIGNANT TISSUE USING CONFOCAL LASER SCANNING MICROSCOPY (Undergraduate), D Janarious, J Biggerstaff, JL Francis, Walt Disney Memorial Cancer Institute
DIETARY MODIFICATION AND THE ALZHEIMER'S-LIKE PHENOTYPE IN mAPP/mPS1 TRANSGENIC MICE, P.T. Jantzen, M.N. Gordon and D.G. Morgan, University of South Florida
COMPARISON OF CHARACTERISTICS DRY ETCHING OF LaCaMnO3, NiMnSb AND NiFe THIN FILMS, K. B .Jung(1), Hyun Cho(1), J. J. Wang(1), J. Caballero(1), Tao Feng(1), J. R. Childress(2), K. H. Dahmen(3) and S. J. Pearton(1), (1)University of Florida, (2)IBM Almaden Research Center (3)Florida State University
FABRICATION OF DC-MAGNETRON SPUTTERED 70Ti-30Al THIN FILMS, A. S. Kale, K. R. Beaulieu, K. B. Sundaram, V. H. Desai, S. Seal, University of Central Florida
THE ATOMIC FORCE MICROSCOPY INVETIGATION OF THIN FILM DEPOSITED FROM NANOPARTICLE SOURSE, F.K. Urban, A. Khabari, Florida International University
COMPARISON OF ZnS:TbOF THIN FILMS DEPOSITED BY R.F MAGNETRON SPUTTERING USING ZnS, TbOF MIXED TARGET AND SEPERATED ZnS,TbOF TARGETS, Jongpyo Kim, Mark Davidson, Barbara Speck, David Moorehead, Qing Zhai, and Paul Holloway, University of Florida
ELECTRON BEAM DEGRADATION OF OXIDE AND SULFIDE THIN FILM PHOSPHORS FOR FIELD EMMISION DISPLAYS, Caroline A. Kondoleon, Billie Abrams, Philip Rack* and Paul Holloway, University of Florida, Rochester Institute of Technology
ELECTRON CYCLOTRON RESONANCE CHEMICAL VAPOR DEPOSITED SILICON NITRIDE FOR T-GATE PASSIVATION, J. LaRoche1, F. Ren1, S.J. Pearton1, J. R. Lothian2, J.W. Lee3, and D. Johnson3, 1University of Florida, 2Multiplex Inc, 3Plasma-Therm, Incorporated
DRY ETCHING OF BaSrTiO3 AND LaNiO3 THIN FILMS IN INDUCTIVELY COUPLED PLASMAS, K. P. Lee, K. B. Jung, A.Srivastava, D. Kumar, R. K. Singh and S. J. Pearton, University of Florida
EFFECTS OF Ni THICKNESS ON Ni/Ti/Ag OHMIC CONTACT TO p-GaN, B. Liu, M. H. Ahonen and P. H. Holloway, University of Florida
TITANIUM MAGNESIUM NICKEL ALLOY AND HYDROGEN STORAGE, Janice K. Lomness, Sudipta Seal, Michael D. Hampton, and Meredith Stowell (Undergraduate), University of Central Florida
(Undergradutate) CHARACTERIZATION OF BEAM PRODUCED BY PULSED ARC CLUSTER ION SOURCE, Samantha A. Moore and Anne J. Cox, Eckerd College
COMPARISON OF THE MICROSTRUCTURE AND ELECTROLUMINESCENT PROPERTIES OF ZnS:Mn DEPOSITED BY SPUTTERING AND ATOMIC LAYER EPITAXY, David J. Moorehead Karen E. Waldrip, M.R. Davidson*, J.H. Lee, B. Pathangey*, M.Puga-Lambers*, K.S. Jones, P.H. Holloway, University of Florida, and S.S. Sun and C.N. King, Planar Systems
TRI-LAYER ELECTRON BEAM RESIST FOR SUBMICRON T-GATE GaN BASED FIELD EFFECT TRANSISTORS, M. Mshewa, H. Hudspeth, F. Sharifi, S. J. Pearton, and F. Ren, University of Florida
A MODIFICATION OF THE VARIABLY DAMPED LEAST SQUARE ALGORITHM ASSISTED BY MEASUERED DATA POINTS AND DERIVATIVE PRESELECTION FOR IMPROVEMENT OF SOLUTIONS, J. Pontillo, F.K. Urban, Florida International University
PULSED CURRENT ELECTROMIGRATION, Cross Reardon and Rolf Hummel, University of Florida
ILLUSTRATION OF CAPILLARY PERFUSION IN HYPERTROPHIED CARDIAC MUSCLE USING THE FLUORESCENT DYE, THIOFLAVIN-S, Corey A. Schoder, Hans Degens, Don R. Hilbelink, Karl E. Muffly University of South Florida
FORMATION OF SILVER SULFIDE NANO-PARTICLES BY NOVEL SOL-GEL METHOD FOR INDUSTRIAL APPLICATIONS, S. Shukla and S. Seal, University of Central Florida
CO-LOCALIZATION OF GLIAL FIBRILLARY ACIDIC PROTEIN (GFAP), OX-42 AND OX-6 IN SPINAL CORD FOLLOWING SCIATIC NERVE DAMAGE, Stacy Sinibaldi, Edward Haller, and Samuel Saporta University of South Florida
CHARACTERIZATION OF THE CONSORTIUM BETWEEN THE RED TIDE CAUSING DINOFLAGELLATE GYMNODINIUM BREVE, AN UNIDENTIFIED BACTERIUM AND A PHAGE, Theresa R. Slifko, Lee Houchin, Anthony Greco, and John H. Paul, University of South Florida
INVESTIGATIONS INTO CHEMICAL MECHANICAL POLISHING OF TUNGSTEN USING VARIOUS ELECTROCHEMICAL AND SURFACE SCIENCE TECHNIQUES, Dnyanesh Tamboli, Sudipta Seal, Vimal Desai University of Central Florida
THE MINERAL CONTENT AND CELLULAR STRUCTURE OF HETEROTOPIC BONE, Gregory S. Taylor University of Florida College of Medicine, and Melinda K. Harman and W. Andrew Hodge, Orthopaedic Research Laboratory
EFFECTS OF FIB OPERATING CONDITIONS ON BEAM DAMAGE OCCURRING IN SILICON TEM SAMPLES, C.A. Urbanik, B.I. Prenitzer, L.A. Giannuzzi, S.R. Brown1, R.B. Irwin1, B. Rossi2 , T.L. Shofner2, and F.A. Stevie1, University of Central Florida, 1Cirent Semiconductor, 2The Bartech Group
IMPROVED PERFORMANCE IN THIN FILM ELECTROLUMINESCENT PHOSPHORS BY DONOR DOPING, K.E. Waldrip, J.S. Lewis, III, Q. Zhai, D. Moorehead, M.R. Davidson*, and P.H. Holloway, University of Florida, and S.S. Sun, Planar Systems, Inc.
EFFECTS OF FLUX AGENTS ON THE MICROSTRUCTURE AND ELECTROLUMINESCENCE OF SPUTTERED ZnS:Mn THIN FILM PHOSPHORS, Qing Zhai, Karen Waldrip, Jay Lewis, Jungpyo Kim, David Moorhead, Jinghong Li, Kevin Jones, and Paul Holloway, Maggie Puga-Lambers, Barbara Speck, and Mark Davidson, Microfebritech, University of Florida
ICP Cl/Ar PLASMA DAMAGE ON GaN SCHOTTKY, A. Zhang1, H. Cho, F. Ren1, S.J. Pearton1, J. M. Van Hove2, P. P. Chow2, R. Hickmand2, J. J. Klaasen2 and R. J. Shul3, 1University of Florida, 2SVT Associates, 3Sandia National Labs
MICROSTRUCTURE EVALUATION OF STRESS CORROSION CRACKING USING FIB TECHNIQUES, Hanlin Zhang, Brian Kempshall, Carrie Urbanik and Lucille A. Giannuzzi, University of Central Florida
Tuesday, March 16, 1999
Session V: Microscopy of Physical Samples
Session Moderator: Lucille A. Giannuzzi, University of Central Florida
8:30-8:55 am ANALYTICAL MICROSCOPY IN THE REAL SEMICONDUCTOR PROCESSING WORLD, Ron Anderson, IBM Analytical Services
8:55-9:20 am FIB APPLICATIONS IN MATERIAL SCIENCE PROBLEMS, M.W. Phaneuf, J. Li, G.A. Botton*, L. Weaver, Fibics Incorporated, *Materials Technology Laboratory
9:20-9:45 am FOCUSED ION BEAM IMAGING OF MICROELECTRONIC STRUCTURES, Ann N. Campbell, Sandia National Laboratories
9:45-10:10 am DETERMINATION OF THE COMPOSITION OF GRAIN BOUNDARIES AND INTERFACES BY X-RAY MICROANALYSIS, David B. Williams, Lehigh University
10:10-10:40 am Coffee Break
10:40-11:05 am MATERIALS APPLICATIONS OF ELECTRON HOLOGRAPHY, Altaf Carim, The Pennsylvania State Univeristy
11:05-11:30 am PHASE MAPPING AND PHASE IDENTIFICATION USING ELECTRON BACKSCATTER DIFFRACTION, David J Dingley and Stuart I Wright, TexSEM Laboratories
11:30-11:55 pm AFM, Phil Russell, North Carolina State University
12:00-1:00 pm Lunch and Vendor Talks (Session VI)
Tuesday, March 16, 1999
Session VI: Vendor Presentations
Session Moderators: Fred Stevie, Cirent Semiconductor
12:10-12:20 pm LOW ENERGY ION MILLING FOR TEM, David Henriks, South Bay Technologies
12:20-12:30 pm INNOVATIONS IN MASS FLOW CONTROLLERS FROM UNIT, Greg Vaughan, Schoonover, Inc.
12:30-12:40 pm RECENT DEVELOPMENTS IN ATOMIC FORCE MICROSCOPY, Matt Thompson, Digital Instruments
12:40-12:50 pm NEW PRODUCTS FROM SPECS: Er-LEED, ULTRA HIGH RESOLUTION EEES: DELTA0.5, AND PLASMA UV SOURCE UV300, Dietrich von Diemar, SPECS U.S.A.
Tuesday, March 16, 1999
Session VII: Electronic Materials
Session Moderator: Drew Hoff, University of South Florida
1:00-1:25 pm LOW FIELD ELECTRON EMISSION FROM UNDOPED NANO-STRUCTURED DIAMOND, W. Zhu, G. P. Kochanski and S. Jin, Bell Laboratories, Lucent Technologies
1:25-1:50 pm SYNTHESIS AND APPLICATIONS OF NANOCRYSTALLINE DIAMOND FILMS, Dieter M. Gruen, Argonne National Laboratory
1:50-2:15 pm CHARACTERIZATION OF SHALLOW JUNCTIONS USING SECONDARY ION MASS SPECTROMETRY, Charles W. Magee, I.M. Abdelrehim, T.H. Buyuklimanli, J.T. Marino and W. Ou, Evans East
2:15-2:40 pm NOVEL PROCESSING OF ELECTRONIC MATERIALS, W. V. Lampert, Air Force Research Laboratory, WPAFB, OH
2:40-3:05 pm GaN BASED ELECTRONICS FOR HIGH TMEPERATURE APPLICATION, F. Ren1, S. J. Pearton1, C. R. Abernathy1, J. M. Van Hove2, P. P. Chow2, R. Hickmand2, J. J. Klaasen2, J. Han3, A. G. Baca3, and R. J. Shul3, 1University of Florida, 2SVT Associates, 3Sandia National Labs
3:05-3:30 pm LOW ENERGY IMPLANTATION AND SHALLOW JUNCTIONS IN Si, Kevin Jones, University of Florida
3:30-4:00 pm Coffee Break, Student Awards and Door Prizes
Tripod Polisher, instructor: Ron Anderson March 12-13 FIB, instructors: Lucille Giannuzzi and Fred Stevie March 18-19
in conjunction with the FL AVS/FSM meeting week of March
for information see www.pegasus.cc.ucf.edu/~ampac or contact lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Tong - You might find that a glass needle will work for your purpose and you can easily make them yourself to almost any desired thinness. Simply take a thin 8 or 10 inch long glass rod (can be hollow too) and heat near the center with a bunsen burner or other source. After glowing pull the ends apart and you will draw the glass down to a suitable thickness. At the micron level glass is relatively strong for moving particles and as you will find quite flexible.
Dave Joswiak Dept. of Astronomy Univ. of Washington Seattle, WA 98195 (206)543-7702
On Tue, 2 Feb 1999, Tong Wang wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I need some microneedles to dislodge some very fine particles (micron size) } on my sample but still flexible enough to bend. } Any information is appreciated. } } Tong } } } } }
Postdoctoral Positions in Nuclear Materials Science - PD994571
The Materials Science and Processing Group (NMT-11) at Los Alamos National Laboratory is seeking qualified applicants to fill two postdoctoral positions in the science of nuclear materials. Successful candidates will work with technical staff members in the Materials Science and Technology (MST) and Nuclear Materials and Technology (NMT) Divisions. Both positions require the willingness to work on radioactive materials and the ability to obtain a DOE Q clearance (which usually requires US citizenship).
Position One: Electron Microscopy of Plutonium and Ceramic Actinide Waste Forms. The individual will have access to (a) the Electron Microscope Facility in MST for microstructural studies of unirradiated and surrogate materials and (b) the Materials Characterization Facility in NMT for analysis of radioactive materials. Techniques available for this research include SEM, OIM, TEM, HRTEM and STEM. This work will support research efforts in radiation effects in ceramics and plutonium characterization for weapons programs. Candidates must have prior experience in electron microscopy and microanalysis. Contact Jeremy Mitchell (505-665-3934, jeremy-at-lanl.gov) or Kurt Sickafus (505-665-3457 kurt-at-lanl.gov) for further technical information.
Position Two: Radiation Damage in Metals, Self-radiation Damage in Plutonium Metal, Alloys and Compounds. This work is basic research in support of plutonium metallurgy and chemistry for the weapons programs. Candidates must have prior experience in x-ray and/or neutron diffraction and the associated data analysis with GSAS, knowledge of mechanisms of radiation damage in metals and some knowledge of condensed matter physics. The individual will also have access to calorimeters, SEM, OIM, and TEM. Contact Luis Morales (505-665-7703 lmorales-at-lanl.gov) for additional technical information.
A Ph.D. in Materials Science, Metallurgical Engineering, Geosciences, or a related field completed within the last three years or soon to be completed is required. Candidates may compete for a Director's Fellowship and outstanding candidates may be considered for the prestigious J. Robert Oppenheimer, Richard P. Feynman or Frederick Reines Fellowships. Further details about the Postdoctoral Program may be found at: http://www.hr.lanl.gov/postdoc/ For consideration, submit a resume and publications list along with a cover letter outlining current research interests to postdoc-jobs-at-lanl.gov (no attachments, please!)
OR SUBMIT TWO COPIES to:
Postdoc Program Office, PD994571 MS P290 Los Alamos National Laboratory Los Alamos, NM 87545
NOTE: Advertisement #PD994571 must be referenced in the e-mail Subject line (or the address) and cover letter.
Affirmative Action/Equal Opportunity Employer. Individuals with disabilities needing reasonable accommodation should call (505) 667-8622. A Teletype Device for the Deaf (TDD) is available by calling (505) 665-5357. Los Alamos National Laboratory is operated by the University of California for the US Department of Energy. ============================ Jeremy N. Mitchell MS G730, NMT-11 Los Alamos National Laboratory Los Alamos, NM 87545 Phone: 505-665-3934 Fax: 505-665-4013
Dear Sally, The coverslips that everyone uses is my Thermanox plastic coverslps which are sterile and come in many different sizes depending on your need. They may be found wither in our hard copy catalog or on our website at www.emsdiasum.com. Go to our online catalog and click chapter 7. They are listed under Thermanox. In our hard copy catalog XIII they may be found on page 143. Please let me know if I may be of further assistance to you.
Sincerely, Electron Microscopy Sciences 215-646-1566
The best way I know of preparing pigments for TEM analysis is by blowing an aerosol of the sample dispersed in alcohol/water solution over a hot plate and collecting the sample on a grid placed at the far end of the hot plate. The company I work for is concerned about the safety issues surrounding this so does anybody know of any less hazardous sample preps. or a simple containment facility for the aerosol spray? I thought about building a glass box for containment but are there any commercially available containment/sample prep. units?
Thanks
Brian Manzor e-mail: brian.manzor-at-grmouth.zeneca.com
The best way I know of preparing pigments for TEM analysis is by blowing an aerosol of the sample dispersed in alcohol/water solution over a hot plate and collecting the sample on a grid placed at the far end of the hot plate. The company I work for is concerned about the safety issues surrounding this so does anybody know of any less hazardous sample preps. or a simple containment facility for the aerosol spray? I thought about building a glass box for containment but are there any commercially available containment/sample prep. units?
Thanks
Brian Manzor e-mail: brian.manzor-at-grmouth.zeneca.com
Hi Michael! To avoid tissue curling try 50% or 30% ETOH in the first dehydrating step. You can do dyhradation in cell-culture multiwells. You=B4ll have to remove the specimens if you=B4d like to use Propylene ox= ide because this interacts with the multi-well plastic. If you like to embed the sections flat, do it on silicated glass slides(o= r try a slide, coverd with slightly fatted aluminium foil) , put a drop of Epon/araldite on it, leave it at 45=B0C in the oven (12-24h) and then put= BEEM Capsules filled with Epon/araldit on the top(take care of bubbles!) , polymerize 2 days at 60=B0C. If you can=B4t remove the glass slide easily= , try it with fluid nitrogen. Good luck, Michael
MICHAEL DELANNOY schrieb:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Greetings, } I need assistance with flat embedding of rat cerebellum } (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically } we are worried about tissue curling during the dehydration. My first } inclination is argarose embed (before epon), but I would take any exper= t } advise. } Thank you, } } Mike D
Sprays can be contained to some extent within a box for spraying thin-layer chromatographic plates, 20 x 20 cm or a little larger. This has to be placed in a hood, since it won't catch and retain the smallest droplets. They come in cardboard and plastic versions. I can't find one in VWR, but any big Web catalog with a search capability should lead you to one.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
Hi all. We have identified a need for some hands on WDS training here in South = Africa. A number of our clients have either changed operators or = upgraded to new integrated systems. So we would like to know if there is = any one who would be able to help with presenting a WDS course here in = South Africa. No need to panic, South Africans do have money and are = willing to pay, not too much mind you.=20
We are open to the course content, however would like to cover a bit of = customer care operations, calibration and set up and then theory and = hands on operation.=20 We will have a Jeol 733 with an Advanced Microbeam / Edax integrated = system available by about June onwards.=20
If any one is interested please let us know so we can discuss details. Thanks
Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
Hello All, Thanks to all those who offered suggestions for the damping of vibrations sometimes experienced when using a light microscope. Once again you have proven to be an ingenious lot! Users have their microscope sitting on platforms supported by any of the following; tennis balls, "sandwiches" of bubble pack and aluminum plates or inner tubes all of which indicates that the pneumatic tables or benchtop isolation systems are preferable to the marble tables. An alternative approach is to support the microscope on a platform beneath which are sandwiched layers of Sorbothane (elastomer) and aluminum plates. One simple suggestion was to "plant" each leg of a conventional table; supporting your microscope, into its own little box of sand. I'm a little wary of this last suggestion as some of the laboratory wildlife might find the sand an attractive litter box!
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
To Dr. Naseem: TEM results on paraffin processed tissue will only be as good as the processing procedure used by the histology lab. In my experience, most histology labs use a very aggressive protocol, which causes extraction of cellular components to the point where only nuclei and possibly cell membranes might be recognizable. Laboratories that handle large volumes and are concerned with quick turnaround generally dehydrate and clear the heck out of specimens so they don't have to go back and reprocess if something was too big, not well fixed, etc. Our laboratory provides good fixation, gentle dehydration, clearing and infiltration of routine histology specimens so that when I have to do EM on one of these, there is usually enough cellular matrix left to make the effort worthwhile. I have been able to resolve Birbeck bodies, tonofilaments, junctions, microvilli and immune complex deposits (in glomeruli) in paraffin processed specimens. Good luck! Bob Santoianni Emory University Hospital Atlanta, Georgia robert_santoianni-at-emory.org
To process cell cultures for grown on round, 13 mm Thermanox tissue culture cover slips for TEM, we use the following hardware:
For fixation, we use porcelain multi-well dishes. These are what most people refer as "staining dishes". They are white, measure about 3.5 x 4.5" and have 12 depressions. We use these for glutaraldehyde, buffer rinses and OsO4. OsO4 can be completely rinsed from the dishes. Once in the last buffer after OsO4 and prior to ethanols and propylene oxide, we transfer the disks to 50 ml polypropylene culture tubes, such as Fisher 05-539-7 (these are sterile, but certainly it is not necessary to pay for sterile containers). There is plenty of room to enter a pipette for fluid exchange without touching the culture disks, and the culture surface will not touch the walls of the cylindrical tube so there is no worry that the cells will be rubbed off. Given the depth of the tube, we do not worry too much that the culture will dry out as fluids are exchanged, since the atmosphere within the tubes is fairly saturated with solvent vapor. Still, fluid exchange is done quickly. The polypropylene tubes are resistant to P.O. and Spurrs. Do not use tubes made from polystyrene as they will dissolve. Once infiltrated with the last change of epoxy, we fill a resin-resistant container with epoxy to a minimum depth of 5 mm, sink the disk so that the cells face up, then polymerize the epoxy. We steal the polypropylene lids from wheaten snap-cap vials (Fisher #0333520E) which are of the appropriate diameter for embedding 13 mm coverslips. Wearing a dust mask, We use a jewelers saw to free small blocks of embedded culture, loosen the cover slip with liquid nitrogen, then remove the disk which exposes the culture to the surface of the block. We then either re-embedded (in some of the same batch of media which was used to infiltrate the culture) face to face for cross sections, or cut the blocks parallel to the culture surface for tangential sections.
Good luck,
Doug Keene EM Facility Shriners Hospital for Children DRK-at-shcc.org
} } Hello All, } Does anyone out there have any suggestions for getting good SEM } cross sectional images from dried wood? I have a student who would } like to image the microstructure of several types of wood. All the
} } Thanks, } Scott
Hi Scott,
I think the classic paper you want is Exley et al. (1973 ?) J. Microscopy, vol 101, 21-30 "Preparation of wood specimens for the scanning microscope".
I don't have a copy handy to check, but I think this paper emanated from good ol' New Zealand! As I recall, the authors were able to cut fresh lignified wood satisfactorily using a fresh razor blade for each cut, and then cleaned the surfaces with hypochlorite solution before drying and coating. Most specimens were OK with just air drying but delicate features needed CPD. The authors often cut the one specimen in two planes to good effect, e.g. transverse and radial longitudinal.
If you want to look at long-dead wood it might be too hard to cut cleanly. You might need to soak it in something first. Exley et al. might have suggestions.
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
-----Original Message----- } From: Gerroir, Paul J {Paul.Gerroir-at-crt.xerox.com}
} One simple suggestion was to "plant" each leg of a } conventional table; supporting your microscope, into its own little box of } sand. I'm a little wary of this last suggestion as some of the laboratory } wildlife might find the sand an attractive litter box!
Mix red pepper with the sand that will keep the kitty out of the sand.
Gordon
Gordon Couger gcouger-at-couger.com Owner PRAG-L PRactical AGriculture List www.couger.com/prag-l Stillwater, OK 405 624-2855 GMT -6:00
I posted a notice several months back about a Siemens 101 Transmission Electron Microscope that was up for grabs. Although we had some interest, no one decided to take it. At this point, we have a major space crunch and need to dismantle and get rid of it. I've been told it contains 25 gms of liquid mercury which should be removed before disposal. Can anyone refer me to a technician familiar with this equiment in the NY area who could assist us in this project. Thanks in advance for your help. Please reply directly to me. I am also posting this notice on the safety list. ---------------------------------------- Gerry Griffin Environmental Services NYU Medical Center Email: Gerry.Griffin-at-med.nyu.edu
I'm studying the transport and localization of some proteins in the cytoplasm of cultured cells. The cells are fixed in paraformaldehyde and stained with a avidin-biotin-fluorescein system. I need to do double staining for this protein and the Golgi, so I intend to use the Bodipy-tr-ceramide probe, as suggested by Molecular Probes .
I would like to know if someone has experience or worked with this specific probe. Particularly I need to know if this probe may be used with fixed cells, that is, after the cells are fixed.
I will be very grateful for any information
Thanks in advance
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas Lyon - France
We are interested in buying a variety of used AFM and STM equipment. To see our wish list, go to our web page (or contact us directly) http://a1.com/asm/wantused.htm
thanks Don Chernoff
Advanced Surface Microscopy, Inc. E-Mail: asm-at-indy.net 6009 KNYGHTON RD. Voice: 317-251-1364 INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA) web: http://www.a1.com/asm Fax: 317-254-8690 (note: "a1"= letter "a", numeral "1")
I am looking for a postdoc/visiting scientist who will do developmental work on Direct Methods for structure determination using dynamical electron diffraction data. If you know of anyone who has: a) A good diffraction background b) A good crystallography background c) Good computer skills Please ask them to contact me. The position is available now.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
I just got an Amray 1600 with turbo and would like to correspond with others who are using this instrument. If there are any quirks about it, I would like to know what they are. If anyone has brought one up after several years of storage, I'd REALLY like to hear from you.
I'm hoping to be able to take 4x5" sheet film pix and 120 roll film pix.
Any info on the care and feeding of the turbo pump, roughing pump and inevitable column cleaning would be appreciated. I'm told that this SEM can take good pictures. I'm hoping to find out if this is true.
Having serviced Siemens microscopes I can tell you that the source of th= e mercury is a mercury diffusion pump. This pump is situated between the rotary pump and the "oil" diffusion pump in the vacuum circuit.
So that is where the mercury is now to dispose of it is another problem?
Stay safe.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
I am currently in the process of acquiring a TEM with accessory equipment. We are starting a new TEM-lab here at the Swedish Museum of Natural History.
One of the instruments I am considering is the LEO 912 AB. Anyone on the list with experience of that microscope? We will primarily be doing morphology including ESI imaging of thick sections, but not analytical work or cryo.
Also: any recommendations on which ultramicrotome to choose.
TIA
Ulf Jondelius
Ulf Jondelius, Invertebrate Zoology Swedish Museum of Natural History P.O.B. 50007, 104 05 Stockholm, Sweden phone: Int + 46 (0)8 666 5160 fax: Int + 46 (0)8 666 4125 e-mail: ulfj-at-nrm.se
Here's a few comments to follow from Doug Keene's reply to you celss on coverslips question.
You can process and section Thermanox quite easily with cells grown on them.
Orientation problems can be overcome if you cut the coverslip to fit a BEEM capsule before you innoculate with cells. I have seen good results with various cell types, and if you taper the cut coverslip to fit into the BEEM pot, trimming the block is pretty quick too. If you need, you can also bend up the coverslip at the non-tapered end to indicate which side the cells are growing.
Spurrs resin seems pretty good, but there can be some delamination between the coverslip and resin, but not always!
Hope this helps.
Plymouth Electron Microscopy Unit University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel: 01752 233092 Fax: 01752 233092 email: pbond-at-plymouth.ac.uk
I need to colorize some SEM pictures but I do not know which programm to use. If someone can help me, I use PC or Macintosh computer. Thanks a lot for your help Best regards
Didier
-------------------------------------------- Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
To save a long search can anyone remember which months archive of the listserver the discussion about preparing cd-roms for sem is in??
Chris
Chris Gilpin Experimental Officer Biological Sciences EM Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 0161 275 5170 Fax +44 0161 275 5171 Chris Gilpin http://www.empgu.man.ac.uk
I am now running a clinical path, diagnostic EM lab. but must reduce my storage. What are the clia and cap regs on how long to keep transmission em blocks, thick section slides, grids and phot mic's? I am being reduced from a 3 room lab to a room and a closet. thanks for you help. S. Drew
I would like to draw your attention to the materials science symposium being held as part of the Scanning 99 meeting in April this year. The abstract deadline is February 15th (the same as MSA) and the abstract format is the same as the MSA format. It is intended that there will be contributed presentations as part of the program, and depending on the number of submissions, the symposium may be extended for another day. Please forward this e-mail to anyone you think may be interested in attending the symposium.
{bold}
"Analyzing Materials Interfaces at Atomic Resolution" {/bold}
There will be a Materials Science symposium at Scanning 99 entitled "Analyzing Materials Interfaces at Atomic Resolution". Scanning 99 is being held at the Hyatt Regency O'Hare in Rosemont, IL from April 11-14 1999. The "Analyzing Materials Interfaces at Atomic Resolution" symposium is tentatively scheduled for Tuesday April 13th and will consist of invited and contributed presentations. The details of the conference and deadlines/forms for abstract submissions can be found at http://www.scanning.org or details can be requested from Mary Sullivan (e-mail: scanning-at-fams.org, tel:201-818-1010). Registration for members of the Midwest Microscopy and Microanalysis Society are at the reduced rates of $150 for the whole conference or $50 for a single day
The discussion was last September, maybe on into October. I will send you a copy of the summary that I have on hand.
Warren S.
At 02:18 PM 2/5/99 +0000, you wrote: } } Hi all } } To save a long search can anyone remember which months archive of the } listserver the discussion about preparing cd-roms for sem is in?? } } Chris Gilpin } Experimental Officer } Biological Sciences EM Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 0161 275 5170 } Fax +44 0161 275 5171 Chris Gilpin } http://www.empgu.man.ac.uk }
Just a reminder:ACS's Applied Optical Microscopy will be held in conjunction with PITTCON, March 5-7, in Orlando, Florida.
This is a three-day, hands-on total immersion workshop. Although this workshop is offered under the American Chemical Society Short Course programming, it is not just for chemists. The curriculum focuses on key techniques to help you be more effective in the lab. Topics range from alignment, care and cleaning to contrast techniques and basic measurement. There is also a special Saturday evening program on video and digital imaging and a full day on qualitative and quantitative Polarized Light Microscopy. Participants are encouraged to bring samples.
For details and registration information, visit the MME website {http://www.MME-Microscopy.com/education} or call the course coordinator, Barbara Foster
Barbara Foster Course Coordinator ACS Applied Optical Microscopy Course c/o Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 (413)746-6931 Fax: (413)746-9311 email: mme-at-map.com
According to some of the messages I saved, the dates are mid-August and September '98. Hope this helps.
Chris Gilpin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all } } To save a long search can anyone remember which months archive of the } listserver the discussion about preparing cd-roms for sem is in?? } } Chris } } Chris Gilpin } Experimental Officer } Biological Sciences EM Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 0161 275 5170 } Fax +44 0161 275 5171 Chris Gilpin } http://www.empgu.man.ac.uk
I was just asked to provide the equation for gamma adjustment to image contrast. I had always assumed that it was just
Output = (Input)^gamma i.e. a simple power law.
Thus for gamma = 0.5 you get a square root curve and for gamma = 2.0 you have a parabola. However, the contrast curves in various books I've checked do not appear to be a power law, but look more like circular arcs. Unfortunately these books only describe gamma but do not provide the equation. Are the curves just artist's license or is my understanding of the gamma correction wrong?
If you include Contrast and Brightness, I would expect the general equation to be
Output = B + C*(Input)^gamma
Where B = Brightness and C = Contrast.
Thanks, Henk Colijn Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true.
Following a very successful year, NORAN Instruments is pleased to announce employment opportunities within its Confocal Sales Division.
The position of District Sales Manager will be responsible for all confocal sales activities within a predefined region of the United States.
Functions will include: * Evaluation and qualification of customer needs * Demonstration of confocal systems, including configuration and setup * Coordination of installation and initial user training
Necessary qualifications include: * B.A. or B.S. degree in Natural Sciences. Advanced degree desirable. * One year's hands on experience with light microscopy techniques. * Experience and practical knowledge of Confocal microscopy applications. * Experience with computer image analysis and processing preferred. * Sales experience is highly desirable.
The position requires travel of up to 50% within designated sales region.
For consideration, please fax or e-mail your resume in confidence to:
Could some kind, knowledgable person explain why the addition of calcium = to fixatives (buffered aldehydes, biological material, resin embedding and = TEM) is important? =
Regards,
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
I am looking for any written materials concerning Reichert MeF = microscope. I have one in mint condition (probably never used), but = without any instructions. The microscope was made after 1954. It is now = disassembled and, although I know more or less how to put it together, I = would rather like to do it according to producer's advices.
I am not sure if my question fits to this mailing list, but I think that = history of microscopy would be of interest for some of us.
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-2 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.2016.0"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} I am looking for any = written=20 materials concerning Reichert MeF microscope. I have one in mint = condition=20 (probably never used), but without any instructions. The microscope was = made=20 after 1954. It is now disassembled and, although I know more or less how = to put=20 it together, I would rather like to do it according to producer's=20 advices. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} {/FONT} {/DIV} {DIV} {FONT face=3DArial size=3D2} I am not sure if my question fits to = this mailing=20 list, but I think that history of microscopy would be of interest = for some=20 of us. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} Thank you {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} Pawel = Karaszkiewicz {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} {A=20 href=3D"mailto:zekarasz-at-cyf-kr.edu.pl"} zekarasz-at-cyf-kr.edu.pl {/A} {/FONT} {= /DIV} {DIV} {FONT color=3D#000000 face=3DArial = size=3D2} =20 {/FONT} {/DIV} {/BODY} {/HTML}
There is a Win95/WinNT program called EIKONA3D for volumetric image processing, analysis and visualization, which provides special features and tools for working with image sequences originated from microscopy (e.g. 3D image processing, image alignment, 3D reconstruction, 3D visualization, volume rendering, surface rendering, 3D registration, etc.). One of the tools it provides is 3D segmentation of a 3D data set (image sequence) and labeling/colorization of 3D regions. See the site: http://www.alphatecltd.com/eikona3d.html
Best regards, Nikos Nikopoulos
At 11:10 AM 2/6/99 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have used Adobe PhotoDeluxe which is available (and pretty cheap) for both the Mac and PCs. The program is deceptively simple, but quite usefull if you look into it a bit.
By adding a transparent layer, you can colorize the image without actually modifying it. Once a colored layer is created the colors can be rapidly changed to fit your tastes.
Photoshop will do the same....but for a much higher price, and learning curve.
} } Dear all, } } } } I need to colorize some SEM pictures but I do not know which } } programm to use. If someone can help me, I use PC or Macintosh computer. } } Thanks a lot for your help } } Best regards } } } } Didier } }
-- _______________________________________________ / Michael J. Herron / / U of MN,Medicine/Infectious Diseases / / herro001-at-maroon.tc.umn.edu / / http://128.101.243.213 / /__________________________________________/
I have some samples of muscle tissue in which I would like to determine the distribution of mitochondria within each myocyte.
My life would be made easier, and the sample size bigger, if I could do this at the LM level rather than in the TEM. I figure that the mitochondria will be visible in the LM because they occur in large groups in this tissue, rather than singly. Also, I don't need to see every mitochondrion, just the general pattern.
The question is: does anyone know of a suitable specific stain? I am after a bright-field permanent stain that will work on semi-thin resin sections, not a fluorescent or antibody method. A nice old-fashioned stain!
So far the tissue has been fixed in glutaraldehyde but not processed further. Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
I'm studying the transport and localization of some proteins in the cytoplasm of cultured cells. The cells are fixed in paraformaldehyde and stained with a avidin-biotin-fluorescein system. I need to do double staining for this protein and the Golgi, so I intend to use the Bodipy-tr-ceramide probe, as suggested by Molecular Probes .
I would like to know if someone has experience or worked with this specific probe. Particularly I need to know if this probe may be used with fixed cells, that is, after the cells are fixed.
I will be very grateful for any information
Thanks in advance
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas Lyon - France
I have copies of the operations manuals for both the MeF3 and for its camera system and would be happy to provide you with copies. There would be a slight fee for copying and postage. Please let me know if you are interested.
Best regards,
Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118
MME is America's first national consortium providing
customized on-site workshops in all areas of
microscopy, sample preparation, and image analysis.
} } } }
{excerpt}
{fontfamily} {param} Arial {/param} {smaller} I am looking for any written materials concerning Reichert MeF microscope. I have one in mint condition (probably never used), but without any instructions. The microscope was made after 1954. It is now disassembled and, although I know more or less how to put it together, I would rather like to do it according to producer's advices.
{/smaller} {/fontfamily}
{fontfamily} {param} Arial {/param} {smaller} I am not sure if my question fits to this mailing list, but I think that history of microscopy would be of interest for some of us.
{/smaller} {/fontfamily}
{fontfamily} {param} Arial {/param} {smaller} Thank you
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I'm studying the transport and localization of some proteins in the cytoplasm of cultured cells. The cells are fixed in paraformaldehyde and stained with a avidin-biotin-fluorescein system. I need to do double staining for this protein and the Golgi, so I intend to use the Bodipy-tr-ceramide probe, as suggested by Molecular Probes .
I would like to know if someone has experience or worked with this specific probe. Particularly I need to know if this probe may be used with fixed cells, that is, after the cells are fixed.
I will be very grateful for any information
Thanks in advance
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas Lyon - France
How can you run a lab with the space you are getting? Everytime we are = CAP inspected we are cited (non binding) for lack of space. We have 5 = large rooms that are used for clinical purposes. Fight with the powers = that be on this CAP requirement as opposed to throwing out clinical = specimens.(ie blocks, slides, etc.)
If more room is not posible, how about off site storage? If I had to, I = could store blocks, prints, negs, thicks, etc in a 10 x 10 foot room. = (not much room to move in ) that we have collected in the lab since 1974. = =20
I do not actually now the real requirements that you are asking about, but = check with the histology lab and use those rules as guidelines.
Best of Luck,
Ed
} } } Sharon Drew {drewsh-at-smtpgw2.musc.edu} 02/05 9:45 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html=20=
I am now running a clinical path, diagnostic EM lab. but must reduce my storage. What are the clia and cap regs on how long to keep transmission em blocks, thick section slides, grids and phot mic's? I am being reduced from a 3 room lab to a room and a closet. thanks for you help. S. Drew = = = = = = = = = = =20
I have some samples of muscle tissue in which I would like to=20 determine the distribution of mitochondria within each myocyte.=20
My life would be made easier, and the sample size bigger, if I could=20 do this at the LM level rather than in the TEM. I figure that the=20 mitochondria will be visible in the LM because they occur in large=20 groups in this tissue, rather than singly. Also, I don't need to=20 see every mitochondrion, just the general pattern.
The question is: does anyone know of a suitable specific stain? I am=20 after a bright-field permanent stain that will work on semi-thin resin=20 sections, not a fluorescent or antibody method. A nice old-fashioned=20 stain!
So far the tissue has been fixed in glutaraldehyde but not processed=20 further. Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
Hi Paul, During aldehyde fixation the calcium ions tend to leach out, thus the addition of CaCl2 to the buffer solution. Of course there are other important factors to consider such as pH, osmolality, and choice of buffer (PIPES, MOPES, or Cacodylate instead of Phosphate which may result in some percipitation). MgCl2,and KCl are are also sometimes added; depending on what is important to preserve. (Gronblad,M., (1983) Cell Tissue Res. 229,627 and Glauert, A.M., 1975. Fixation, dehydration, and embedding of Biological specimens. Practical Methods in Electron Microscopy. Amer. Elsevier Pub. Co.,Inc., New York 207pp.
At 10:41 PM 2/5/99 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi all I am wondering if there is any source for SEM images that can be downloaded as "pdf" files. The images are for corroded metals. Regards Mayouf
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Hello all,
I would like to demonstrate the juxtaglomerular cells(=myoepitheloid cells) of kidney on semi-thin sections by using different staining methods. I remember reading a very nice method to show these cells. However, I can not find it now.
I will greatly appreciate if you could send me the suitable staining methods which you used before and also your suggestions about that.
Thanks in advance.
Best regards,
Ranan Gulhan AKTAS, M.D. Trakya University, Faculty of Medicine Pathology Department Edirne 22030 Turkey
Henk, I think that your equation is wrong. The gamma correction is Out = (In) ^(1/gamma). This gives a parabola (upwards turning) for gamma=1/2 which will expand the contrast in the bright region at the expense of the dark regions of the image. For gamma=2, it is a square root function (also a parabola on its side) that will expand the contrast in the dark regions at the expense of the light region. I assume that you would normalize the output range so that it was 0 to 255 with a suitable coefficient in the equation. The curves should go through 0 and 255, so there is no offset constant in the equation. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Hendrik O. Colijn To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hi all,
I was just asked to provide the equation for gamma adjustment to image contrast. I had always assumed that it was just
Output = (Input)^gamma i.e. a simple power law.
Thus for gamma = 0.5 you get a square root curve and for gamma = 2.0 you have a parabola. However, the contrast curves in various books I've checked do not appear to be a power law, but look more like circular arcs. Unfortunately these books only describe gamma but do not provide the equation. Are the curves just artist's license or is my understanding of the gamma correction wrong?
If you include Contrast and Brightness, I would expect the general equation to be
Output = B + C*(Input)^gamma
Where B = Brightness and C = Contrast.
Thanks, Henk Colijn Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true.
I am looking for public domain images from electron microscopes or anyone in the New York area that might be interested in collaborating with a design firm to produce a high profile television spot. Any help would be appreciated..
Jeff Linnell Liquid Design Group jeff-at-liquidesign.com
My laboratory is equipped with a LEO 912 Omega, the model before the AB if I'm not mistaken, which was purchased 4 years ago. This is an excellent instrument, powerful, versatile and dependable. I use it mostly for conventional and cryo-EM with excellent results. I would reccomend it without hesitation.
For microtome, I have been a satisfied Reichert Ultracut user for years, which is why I bought an Ultracut S for my lab 4 years ago. The fully motorized controls needed getting used to in the beginning but have proven remarkably reliable.
Usual disclaimer: I have no interest or relation to either company other than being a very satisfied customer.
If you have specific questions, don't hesitate to contact me.
Regards, Michel **************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
Never used one of those, so I don't know if these are possible after fixation in glut_de. At least some are possible, acc. to the text, after fixation in "formaldehyde-based fixatives" and "postfixation" in potasium dichromate...
These are all described in Langeron, M: "Precis de microscopie", Masson, Paris, 1934...
Can send you a copy of the protocols if you want (*.tif, *.jpg, whatever...it's in French).
Yvan Lindekens.
---------- } Van: Stephen Edgar {s.edgar-at-auckland.ac.nz} } Aan: Microscopy-at-sparc5.microscopy.com } Onderwerp: LM: A stain for mitochondria? } Datum: maandag 8 februari 1999 5:13
} I have some samples of muscle tissue in which I would like to } determine the distribution of mitochondria within each myocyte. } } The question is: does anyone know of a suitable specific stain? I am } after a bright-field permanent stain that will work on semi-thin resin } sections, not a fluorescent or antibody method. A nice old-fashioned } stain! } } So far the tissue has been fixed in glutaraldehyde but not processed } further. } Regards } } Stephen Edgar
Our group has inherited a Boston-Bradley Adjustable Blade manufactured by Gardener Laboratory of Bethesda, MD. During an on-line search we were able to determine that the company existed in 1952, but no other information was available.
We believe this to be a crude microtome. It has a stainless steel "clamp" and brass inserts that are labeled with exact thickness from 0.001 to 0.101mil. The clamp is a rectangular block with 3/4" on each end 5mm higher than the center section
Profile ____ ____ ____________
Along one side of this center section is an adjustable stainless steel slab with an arrow pointing to the center of one edge. The knurled knobs are on the other side of the center section. This slab may be adjusted such that it is above or below the center height. It may also be loosened from the center area so that it is a distance from the center.
It seems that the only way to cut would be to raise the slab, place a specimen on the center area (top down), hold the specimen in place and cut it off with a razor blade. This seems upside-down to every other microtome I have dealt with.
Has anyone ever used one of these? We need a crude sledge microtome and this may fit our needs if we could just figure out how to use it.
I had always been told that it helped to preserve the membranes. I've never tested that information, however.
Bob Underwood Derm Research Center U of Washington
On 5 Feb 1999, Webster 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, } } Could some kind, knowledgable person explain why the addition of calcium to fixatives (buffered aldehydes, biological material, resin embedding and TEM) is important? } Regards, } } 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 } } }
} I am looking for public domain images from electron microscopes or } anyone in the New York area that might be interested in collaborating } with a design firm to produce a high profile television spot. Any help } would be appreciated.. } } Jeff Linnell
Jeff -
You'll find a wide variety of images hotlinked at the end of the "Microscopy for Children" bibliography on the Project MICRO web page (URL below). Don't miss the secondary set of links available at "K-12 microscopy resources". Some may be public domain, some are not; you need to check after you've found what you want. There's a stock photo CD-ROM (colorized SEM) available from Corel; it's also in the bibliography .
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
MSA List Recipients, If anyone in possession of a spare Philips CM or EM400 series specimen holder (rod), either regular, low-background, or double-tilt, is willing for a nominal fee or goods and services to part company with said device, please contact me immediately at any of the numbers below. I will be more than appreciative and most remunerative. Thanks. Winston
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supervisor 2/9/99 11:25:46 AM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} Polymer Microscopist - Freeport, Texas and Midland, } Michigan } } Company: The Dow Chemical Company } } Location: Freeport, Texas and Midland, Michigan } } Qualifications (education, certification, language, etc.) and } Experience required: } A candidate with a BS or MS degree in polymer science, material } science or chemistry is preferred with some 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 two professional level full time } openings for Polymer Microscopists, one position at each of the Dow's } facilities in Midland, MI and Freeport, TX. 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 } ultramicrotomy and cryo-ultramicrotomy. } 3. Operation of transmission electron microscope. } 4. Interpretation of images. } 5. Documentation of work. } 6. Compliance with safety and quality programs. } 7. Active member of project and SMX work teams. } } Interested: } 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, Workforce } Planning 98-289, P. O. Box 150, Plaquemine, LA 70765. E-mail } respondents must list Job 98-289 and their last name as the first and } second items on the Subject line. Only those selected for an } interview will be contacted. Only U.S. citizens or aliens who are } authorized to work in the United States will be considered for } employment. } } 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$20billion. Dow manufactures and supplies chemicals, } plastics and agricultural products for customers in 164 countries and } employs approx. 43,000 people worldwide. For more news and } information about Dow, please visit our web site at www.dow.com. } } Bob Cieslinski } Microscopy & Microanalysis } 1897 E Bldg. } (517) 636-6875 }
} } MSA List Recipients, } If anyone in possession of a spare Philips CM or EM400 series } specimen holder (rod), either regular, low-background, or } double-tilt, is willing for a nominal fee or goods and services } to part company with said device, please contact me immediately } at any of the numbers below. I will be more than appreciative } and most remunerative. Thanks. } Winston } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Winston W Wiggins, Supervisor 2/9/99 11:25:46 AM } CRC-Electron Microscopy Lab. Ofc:704/355-1267 } Carolinas Medical Center Fax:704/355-7648 } P.O. Box 32861 Lab:704/355-7220 } Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ }
-----------------End of Original Message-----------------
can anyone tell me if people still use BSA or bacitracin as 'wetting agents' when negative staining? I cannot find reference to them in any of my texts but I have used them since the dawn of time, so I think I have just lost the references and/or they've gone out of fashion.
If anyone has advice on use, advantages of particular chemicals or references I would be grateful.
thanks
Malcolm
PS our glow discharge unit doesn't work - so I can't use that.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
} } MSA List Recipients, } If anyone in possession of a spare Philips CM or EM400 series } specimen holder (rod), either regular, low-background, or } double-tilt, is willing for a nominal fee or goods and services } to part company with said device, please contact me immediately } at any of the numbers below. I will be more than appreciative } and most remunerative. Thanks. } Winston } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Winston W Wiggins, Supervisor 2/9/99 11:25:46 AM } CRC-Electron Microscopy Lab. Ofc:704/355-1267 } Carolinas Medical Center Fax:704/355-7648 } P.O. Box 32861 Lab:704/355-7220 } Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ }
-----------------End of Original Message-----------------
We have an "Image Galleries" page on the WWW-Virtual Library for microscopy site where you may be able to find something suitable. http://www.ou.edu/research/electron/www-vl/image.shtml
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Jeff wrote:
} I am looking for public domain images from electron microscopes or } anyone in the New York area that might be interested in collaborating } with a design firm to produce a high profile television spot. Any help } would be appreciated.. } } Jeff Linnell
Jeff -
You'll find a wide variety of images hotlinked at the end of the "Microscopy for Children" bibliography on the Project MICRO web page (URL below). Don't miss the secondary set of links available at "K-12 microscopy resources". Some may be public domain, some are not; you need to check after you've found what you want. There's a stock photo CD-ROM
(colorized SEM) available from Corel; it's also in the bibliography .
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Hi folks, it has been a while but I am back and of course I need help.
I should submit a proposal to my Dean and Chancellor on a potential centralized morphology core (CMC). They want to see numbers for outlay-cost-income, and I am having trouble coming up with any cost about structure, maintenance.
I will very much appreciate either a brief response to the points below or someone pointing pointing me to a source for this information on the web (phone numbers or email addresses of members with such information will be appreciated). I am envisioning a CMC to provide under one roof: TEM, SEM, Digital Imaging, Confocal, Atomic Force Microscopy and ancillary services such as frozen, histo, immuno-histo and photography. I realize that information for all these may come from different places. Overall, what I need is just an approximate idea for:
1) Cost to operate existing facilities in academic centers?
2) What is the cost for maintaining equipment present at those facilities and to upgrade components?
3) What is the cost to internal and outside users? -Do you charge per case, number of blocks, number of samples, number of cuts?
-How many prints do you provide and at what magnifications for each case?
-Do you retain negatives and when you do not, do you charge extra?
-Charges for embedding cutting and staining frozens, paraffin?
-Charges for plastic processing (JB-4, historesin, etc), sectioning and staining?
-Charges for dupplicating slides, making slides, prints,
-Charges for preparing PhotoShop publication quality composites on color sublimation paper?
-Charges for using microscopy equipment
-Light without and with phase, Normaski, fluorescence, etc.
-Digital confocal optical sectioning, reconstruction, etc.
4) Exceptions.
a) Do junior faculty get service for less than senior and funded faculty?
b) Are there internal mechanisms for covering the costs of promising-emerging faculty, but without active support?
c) How many places have internal mechanisms such as the now extinct NIH BSRG to cover costs?
d) Are any of those costs derived directly or indirectly from grant overheads?
5) Space now housing the facility you describe?
6) How many of the facilities started with external funds?
7) How many of the facilities started with Dean or Chancellor funds?
8) Any other information I miss, but you consider important when considering a CMC?
9) In particular I want to show that most CMC DO NOT make a profit, because of the huge costs for maintenance contract on equipment???
10) Is there anyone out there getting internal support from grant's overhead, and is the money used for CMC considered an incentive-kick-back to funded invetigators?
11) How many of the existing CMC out there offer Cell Sorting (flow) as part of the morphology services?
-Arrangements?
-Maintenance?
-Support?
-Internal and external charges?
Thanks a lot. I will collate and post the results.
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
The Michigan Microscopy and Microanalysis (MMM) Society will hold its Spring Meeting on May 7, 1999 at the Genoa Woods Conference Center, 7707 Conference Center Drive, Brighton, MI. The program chair is still soliciting additional student papers for the meeting. If interest or you would like more details on the meeting contact the local affiliate president, Rob Eversole email at eversole-at-wmich.edu.
MSA (the Microscopy Society of America) is making a concerted effort to provide a forum for multiphoton imagers to gather for candid discussions concerning the new technique.
This year's event is condensed to a one day symposium,
#31 Multi-Photon Excitation Microscopy: the Next Generation 2 or 3 AUG 99 in Portland, OR
This year there will be far less time available; however, there are several important goals for this symposium: 1) invite all new speakers for the various applications presentations 2) presentations describing the commercial systems currently available 3) provide a question and answer session forum for technique education
Poster and abstract submissions are welcome, but please hurry since the deadline for abstract inclusion is 15 FEB 99. See http://www.msa.microscopy.com/
The morning session covers a variety of strong applications of (currently quite expensive) ultra-short pulse laser based fluorescence microscopy...
Karel Svoboda, Ph.D Cold Spring Harbor Laboratory "2PELSM for High-Resolution Imaging in Scattering Biological Tissues: Applications to Neuroscience"
Jayne Squirrell University of Wisconsin "2PEFM of Cytoplasmic Organization in Living Mammalian Embryos"
Mary Dickinson, Ph.D. Caltech "Biological Applications of Chromophores With Large Two-Photon Cross-Sections"
Christopher Navara, Ph.D Wayne Hughes Institute "Dynamic Drug Effects Monitoring"
Paul M W French, Ph.D Imperial College of Science, Technology and Medicine "Fluorescence Lifetime Imaging of Biological Tissue"
Afternoon question/answer session panel: to include the morning speakers, plus
Dave Piston, Ph.D Vanderbilt University Warren Zipfel, Ph.D Cornell University Rebbeca Williams Cornell University Vickie Centonze-Frohlich, Ph.D University of Wisconsin John White, Ph.D University of Wisconsin
In addition, we plan to have vendor presentations and handouts for the commercially available two (multi) photon systems.
==============================================
David L. Wokosin Instrumentation Development Engineer Integrated Microscopy Resource University of Wisconsin-Madison 1675 Observatory Drive Madison, WI 53706-1205
******************************************************************************** Victoria Centonze Frohlich, Ph.D. Deputy Director, IMR University of Wisconsin, Madison P(608) 263-6288 F(608) 265-4076 http://www.bocklabs.wisc.edu/imr/home.htm ******************************************************************************** --============_-1293529032==_ma============ Content-Type: text/enriched; charset="us-ascii"
MSA (the Microscopy Society of America)
is making a concerted effort to provide a forum for multiphoton
imagers to gather for candid discussions concerning the new technique.
This year's event is condensed to a one day symposium,
{bold} #31 Multi-Photon Excitation Microscopy: the Next Generation
{/bold} 2 or 3 AUG 99 in Portland, OR
This year there will be far less time available; however, there are several important goals for this symposium:
1) invite all new speakers for the various applications presentations
2) presentations describing the commercial systems currently available
3) provide a question and answer session forum for technique education
Poster and abstract submissions are welcome, but please hurry since the deadline for abstract inclusion is 15 FEB 99. See http://www.msa.microscopy.com/
The morning session covers a variety of strong applications of (currently quite expensive) ultra-short pulse laser based fluorescence microscopy...
Karel Svoboda, Ph.D Cold Spring Harbor Laboratory
"2PELSM for High-Resolution Imaging in Scattering Biological Tissues:
Applications to Neuroscience"
Jayne Squirrell University of Wisconsin
"2PEFM of Cytoplasmic Organization in Living Mammalian Embryos"
Mary Dickinson, Ph.D. Caltech
"Biological Applications of Chromophores With Large Two-Photon Cross-Sections"
Christopher Navara, Ph.D Wayne Hughes Institute
"Dynamic Drug Effects Monitoring"
Paul M W French, Ph.D Imperial College of Science, Technology and Medicine
"Fluorescence Lifetime Imaging of Biological Tissue"
Afternoon question/answer session panel:
to include the morning speakers, plus
Dave Piston, Ph.D Vanderbilt University
Warren Zipfel, Ph.D Cornell University
Rebbeca Williams Cornell University
Vickie Centonze-Frohlich, Ph.D University of Wisconsin
John White, Ph.D University of Wisconsin
In addition, we plan to have vendor presentations and handouts for
the commercially available two (multi) photon systems.
Cesar- you mention what you envision in a CMC what equipment do you currently have? what equipment will you be purchasing? I have used NIH microscopy centers, and I have managed institutional microscopy facilities. The more money you have for staff the better your facility will operate. It is best if you have one staff member per microscope. It gets crazy if your staff is over worked. You will find maintenance costs will be lower, the quality of results will improve, and overall costs will decrease. Maintenace contracts are a killer, we were once paying over $95,000 per year on service contracts for 4 EM scopes. It was driving prices through the roof, try to negotiate with the manfacturers. If you buy all new scopes, maybe buy from one vendor, and see if they will throw in a service engineer as part of the package, or hire a maintenance person yourself. Try to set costs in a per protocol fashion avoid itemizing. If you need any more specifics please feel free to contact me directly. -Mike
On Tue, 9 Feb 1999, Cesar D. Fermin Ph.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 } -----------------------------------------------------------------------. } } } Hi folks, it has been a while but I am back and of course I need help. } } I should submit a proposal to my Dean and Chancellor on a potential } centralized morphology core (CMC). They want to see numbers for } outlay-cost-income, and I am having trouble coming up with any cost about } structure, maintenance. } } I will very much appreciate either a brief response to the points below } or someone pointing pointing me to a source for this information on the } web (phone numbers or email addresses of members with such information } will be appreciated). I am envisioning a CMC to provide under one roof: } TEM, SEM, Digital Imaging, Confocal, Atomic Force Microscopy and } ancillary services such as frozen, histo, immuno-histo and photography. } I realize that information for all these may come from different places. } Overall, what I need is just an approximate idea for: } } 1) Cost to operate existing facilities in academic centers? } } 2) What is the cost for maintaining equipment present at those facilities } and to upgrade components? } } 3) What is the cost to internal and outside users? } -Do you charge per case, number of blocks, number of samples, number of } cuts? } } -How many prints do you provide and at what magnifications for each } case? } } -Do you retain negatives and when you do not, do you charge extra? } } -Charges for embedding cutting and staining frozens, paraffin? } } -Charges for plastic processing (JB-4, historesin, etc), sectioning and } staining? } } -Charges for dupplicating slides, making slides, prints, } } -Charges for preparing PhotoShop publication quality composites on } color sublimation paper? } } -Charges for using microscopy equipment } } -Light without and with phase, Normaski, fluorescence, etc. } } -Digital confocal optical sectioning, reconstruction, etc. } } 4) Exceptions. } } a) Do junior faculty get service for less than senior and funded } faculty? } } b) Are there internal mechanisms for covering the costs of } promising-emerging faculty, but } without active support? } } } c) How many places have internal mechanisms such as the now extinct NIH } BSRG to cover } costs? } } d) Are any of those costs derived directly or indirectly from grant } overheads? } } 5) Space now housing the facility you describe? } } } 6) How many of the facilities started with external funds? } } } 7) How many of the facilities started with Dean or Chancellor funds? } } } 8) Any other information I miss, but you consider important when } considering a CMC? } } } 9) In particular I want to show that most CMC DO NOT make a profit, } because of the huge costs for maintenance contract on equipment??? } } 10) Is there anyone out there getting internal support from grant's } overhead, and is the money used for CMC considered an incentive-kick-back } to funded invetigators? } } } 11) How many of the existing CMC out there offer Cell Sorting (flow) as } part of the morphology services? } } -Arrangements? } } -Maintenance? } } -Support? } } -Internal and external charges? } } } Thanks a lot. I will collate and post the results. } } *Disclaimer: Whatever... is not Tulane opinion! } Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology } web:[ http://www.tmc.tulane.edu/ferminlab/] or } [http://www.tmc.tulane.edu/imaging/] Internet: } [cfermin-at-mailhost.tcs.tulane.edu] } 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 } Fax 504 587-7389, Voice 584-2521, Main Office 588-5224 } } }
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Our group has inherited a Boston-Bradley Adjustable Blade manufactured by Gardener Laboratory of Bethesda, MD. During an on-line search we were able to determine that the company existed in 1952, but no other information was available. We believe this to be a crude microtome.
It has a stainless steel "clamp" and brass inserts that are labeled with exact thickness from 0.001 to 0.101mil. The clamp is a rectangular block with 3/4" on each end 5mm higher than the center section
Along one side of this center section is an adjustable stainless steel slab with an arrow pointing to the center of one edge. The knurled knobs are on the other side of the center section. This slab maybe adjusted such that it is above or below the center height. It may also be loosened from the center area so that it is a distance from the center. It seems that the only way to cut would be to raise the slab,place a specimen on the center area (top down), hold the specimen in place and cut it off with a razor blade. This seems upside-down to everyother microtome I have dealt with.
Has anyone ever used one of these? We need a crude sledge microtome and this may fit our needs if we could just figure out how to useit.
Thank-you for your help.
Maureen Hunt BP-Amoco p.l.c.
_________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
We just replaced the filament in our Jeol 2010, with a Denka filament can anyone tell me who the supplier is in australia, or elsewhere, I'd like to buy another. The filament is a Denka LaB6 Cathode LKSH M3 104051
Thanks George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 3394 Fax: +61 3 9925 5290
We just replaced the filament in our Jeol 2010, with a Denka filament can anyone tell me who the supplier is in australia, or elsewhere, I'd like to buy another. The filament is a Denka LaB6 Cathode LKSH M3 104051
Thanks George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 3394 Fax: +61 3 9925 5290
Leica Microsystems, Diatome US, and Electron Microscopy Sciences, announce another in a series of TEM Specimen Preparation workshops. This seminar will focus on the hands on participation of the following techniques:
Embedding of industrial samples Specimen trimming
Ultramicrotomy of hard materials Ultramicrotomy of polymers
Collection & handling of sections Staining of polymer sections
Low temperature ultramicrotomy
The format of our workshop is half day lecture and half day bench work in small working groups. Video attachments will be used on the ultramicrotomes in order to maximize the teaching experience for all involved. Samples will be supplied by the course instructors. Participants are encouraged to bring their own samples to work with as time allows.
Course Speakers & Instructors
Dr. Tom Malis Mr. Bob Vastenhout CANMET DOW Chemical Characterization Group Leader Polymer Microscopist Materials Technology Laboratory Analytical Science Department Ottawa, Ontario Terneuzen, The Netherlands
Mr. Helmut Gnagi Product Manager Microtomist Extaordinaire Diatome, Ltd., Switzerland
Hosted by: JoAn Hudson Clemson University Microscopy Facility Clemson, SC
When: June 9-11, 1999
Tuition: $1,400.00
Includes three nights lodging at the lakefront Conference Center & Inn at Clemson University, continental breakfast and lunch daily, one group dinner, course supplies, and lab charges.
Contact: Mike Boykin, Leica Microsystems, Inc. 800-248-0665 X5092
If you are or were using Accu-Scope (3001, 3002, 3018PH etc.) or Wang Biomed (3002, 3004) microscopes - please share your opinion about them and your experience with me.
What did you use it for? Did it prove suitable for the task? Did you have any problems with it? Was its optical/mechanical quality OK for you and for the job?
If you had to use technical support, how helpful/quick were they?
If you used other microscopes too - how do they compare?
Depending on how interesting the answers would be to the other list members, you may either post the replies to the list, or e-mail me directly.
Thank you! -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
Dear Listmembers, Does anybody use or have used ruthenium tetroxide as a fixative for biological material? Any references? Thanks in advance. Chris vd Merwe.
I wonder if anyone knows a good textbook for EDS of minerals? I used to have one in the lab, but it's gone missing, and I don't remember the title or author(s). It had the mineralogical characteristics of most or all of the common minerals, and included typical EDS spectra for each one. A very valuable book for a lab that does a lot of geological work, and I'd really like to replace it (and maybe chain it to the wall this time!)
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
Can anyone recommend a good text on EDS of minerals? We used to have a good one in the lab, which described the characteristics of various common minerals, and showed typical EDS spectra for them. Unfortunately, it's disappeared and I cannot remember the title or author, so I guess it's time for a new one.
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
Dear microscopists, Our facility needs to replace our print processor. I have looked at several models and wanted some feedback about what models work well. We are limited on the amount of space we can use. The processor can not be any larger then 40" X 38", needs to be easy to clean and durable. We are considering purchasing the ILFORD 2150, but heard it may have lots of problems.
Cheers Malcolm, Yes, the bacitracin wetting technique is still valid as ever. The reference you want is: D.W. Gregory and B.J.S. Pirie, Wetting agents for electron microscopy of biological specimens. Proc. Fifth European Congress on Electron Microscopy, (1972). 234-235. David Gregory recommended using a minimum concentration of 7.5 ug/cm3 for formvar coated grids and 10 ug/cm3 on carbon coated formvar or carbon grids as a wetting agent. All the best, Henry
----------------------------------------------------------------------------- Malcolm wrote: } } Dear all } } can anyone tell me if people still use BSA or bacitracin as 'wetting agents' } when negative staining? I cannot find reference to them in any of my texts } but I have used them since the dawn of time, so I think I have just lost the } references and/or they've gone out of fashion. } } If anyone has advice on use, advantages of particular chemicals or } references I would be grateful. } } thanks } } Malcolm } } PS our glow discharge unit doesn't work - so I can't use that. } } Malcolm Haswell } Electron Microscopy } School of Sciences } Fleming Building } University of Sunderland } SUNDERLAND SR1 3SD } Tyne and Wear } UK } } Tel (0191) 515 2872 } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } Henry Eichelberger, EM Facility Manager Department of Biological Sciences Binghamton, University
I am working with the Zeiss LSM 510 Confocal microscope, and am having trouble achieving 3D image reconstruction using the 3D for LSM program. Do you have any recommendations, or know of the resources available through which I can learn this technique? I'd appreciate any information you have on the topic!
----------------------------------------- Laurie Wallin Technician UCSD Department of Anesthesiology and Neuropathology 9500 Gilman Drive 0629 La Jolla, CA 92093 (619)534-1339 or 822-3271
} Are there any grants available to purchase microscopes for an elementary } school in California?
On a national level, no; but there may be an announcement from a major corporation soon that could help you. Your best current possibility is a local corporation that supports education. You'll need about $1000; you'll find the rationale for that figure and advice on what to buy on the Project MICRO web page (URL below). Although I can't supply the funding, I CAN help you write a proposal, and I can write a supporting letter (but I'll be on vacation 2/18-3/28). Where are you located? California is a big state!
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Dear Chris, Ruthenium tetroxide is slow penetrating, but some structures are better preserved by its use. Examples: Madison, K. C., et. al. J. Invest. Dermatology 88 (1987) 714. and Eichelberger, et. al., Proc. Microscopy & Microanalysis 1994, 270. Best regards, Henry ----------------------------------------------------------------------------- Chris vd Merwe wrote:
} From: "Elektronmikroskopie EM 2075 Lab1-5 NW11" {lab1-at-ccnet.up.ac.za} } Organization: University of Pretoria } To: Microscopy-at-Sparc5.Microscopy.Com } Date: Thu, 10 Feb 1999 12:59:24 CAT-2 } Subject: Ruthenium O4 in biology } Priority: normal } X-mailer: Pegasus Mail for Windows (v2.42a) } } Dear Listmembers, } Does anybody use or have used ruthenium tetroxide as a fixative for } biological material? Any references? } Thanks in advance. } Chris vd Merwe. } } Henry Eichelberger, EM Facility Manager Department of Biological Sciences Binghamton, University
} I wonder if anyone knows a good textbook for EDS of minerals? I used to } have one in the lab, but it's gone missing, and I don't remember the title } or author(s). It had the mineralogical characteristics of most or all of } the common minerals, and included typical EDS spectra for each one. A very } valuable book for a lab that does a lot of geological work, and I'd really } like to replace it (and maybe chain it to the wall this time!)
} F.C. Thomas } MicroAnalysis Facility } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada B2Y 4A2
Perhaps you're thinking of "SEM Petrology Atlas" by Joann E. Welton, Chevron Oil Field Research Company, published by the American Association of Petroleum Geologists, 1984. And no, you can't have my copy! Don't know if it's still in print, but amazon.com will hunt around for a used copy for you (for a small fee, of course).
Cheers, eh?
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Patricia, Of the many print processors I've used, I'd choose the Ilford above them all. The older versions have a problem blowing fuses but that is supposed to have been corrected with the new models. The only difficulty I had was that with heavy usage, it must be cleaned more often which, for me, was more of a nuisance than a problem. It's easy to use and durable, not to mention relatively inexpensive. Go for it. Winston
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supervisor 2/10/99 2:53:24 PM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Today I was realesed online free Metallographic Etch's Database. This database is designed to allow you to quick find etchant via powerfull keyword search engine written in Perl. Here are some key features:
1763 records in the database 75 etchants for macro etching 1359 etchants for micro etching 1103 etchants for chemical etching 218 electrolytes for electrolytic etching 75 etchants for physical etching 329 electrolytes for electropolishing
Keyword search engine Browse database by macroetching Browse database by microetching Browse database by chemical etching Browse database by electrolytic etching Browse database by physical etching Browse database by electropolishing Powerfull online help
For more information please see link at microscopy vendors database: http://www.kaker.com/mvd/vendors.html
Henrik Kaker -- SEM-EDS Laboratory Metal Ravne Slovenia http://www2.arnes.si/guest/sgszmera1/index.html
} Can anyone recommend a good text on EDS of minerals? We used to have a good } one in the lab, which described the characteristics of various common } minerals, and showed typical EDS spectra for them. Unfortunately, it's } disappeared and I cannot remember the title or author, so I guess it's time } for a new one. } } F.C. Thomas
Frank,
I think the book you are referring to is SEM Petrology Atlas by Joann E. Welton. It was published in 1984 by The American Association of Petroleum Geologists, Tulsa, OK 74101. ISBN 0-89181-653-4.
Hope this helps. Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
Patricia, Our facility has had an Ilford 2150 for 8 years. Though it has had breakdowns, repairs are fairly simple. I feel that the service contract is too expensive for what you get, which is someone walking you through a repair over the phone. I'm no mechanic but I've been able to deal with most of the problems myself and Iford will sell parts if you need to replace something. The one thing that has made the biggest difference in the trouble free operation of the machine is the addition of a dirt/rust filter on the water supply line. After we did that we really have had no problems except for an occassional plumbing leak (also fixable if you can operate a wrench). You'll also find that you don't need to change the chemicals as often as the manufacturer suggests, as they can be expensive. If you need any more information don't hesitate to contact me.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky Medical Center
--IMA.Boundary.2444868190 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
A colleague and I built a device for doing something similar when I was in graduate school. It consisted of an ultrasonic nebulizer to produce the aerosol at one end of a column, N2 carrier gas (low velocity) to move the aerosol and external heaters to heat the column (along with a few other bells and whistles). Samples were collected on grids at the exit end and the whole thing sat in a hood. The path length was about 2m.
What I'd like to know is if you take any care to get a more uniform aerosol size? Do you work at low enough concentrations to have mostly one (or fewer) particles per drop?
|---------------------------------------------------------------| | Opinions expressed are mine an not those of Rohm and Haas Co. | |---------------------------------------------------------------| | Dr. John R. Reffner | rsrj2r-at-rohmhaas.com | | Rohm and Haas Co. | | | 727 Norristown Road | voice:215-619-5283 | | Spring House, PA 19477 | Fax: 215-619-1607 | |---------------------------------------------------------------|
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Hi
The best way I know of preparing pigments for TEM analysis is by blowing an aerosol of the sample dispersed in alcohol/water solution over a hot plate and collecting the sample on a grid placed at the far end of the hot plate. The company I work for is concerned about the safety issues surrounding this so does anybody know of any less hazardous sample preps. or a simple containment facility for the aerosol spray? I thought about building a glass box for containment but are there any commercially available containment/sample prep. units?
Thanks
Brian Manzor e-mail: brian.manzor-at-grmouth.zeneca.com
Received: from nmho05u.rohmhaas.com ([136.141.252.23]) by ima1.rohmhaas.com with SMTP (IMA Internet Exchange 3.11) id 000E8E24; Wed, 3 Feb 1999 08:41:25 -0500 Received: by nmho05u.rohmhaas.com; id IAA09730; Wed, 3 Feb 1999 08:42:12 -0500 (EST) Received: from sparc5.microscopy.com(206.69.208.10) by nmho05u.rohmhaas.com via smap (3.2) id xma009705; Wed, 3 Feb 99 08:42:01 -0500 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id FAA06651 for dist-Microscopy; Wed, 3 Feb 1999 05:19:30 -0600 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id FAA06644 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 3 Feb 1999 05:18:59 -0600 Received: from apsn05.zeneca.com (apsn05.zeneca.com [193.132.159.6]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id FAA06637 for {microscopy-at-msa.microscopy.com} ; Wed, 3 Feb 1999 05:18:45 -0600 Received: from apsn15.ukap.zeneca.com by apsn05.zeneca.com (SMI-8.6/SMI-SVR4) id LAA01931; Wed, 3 Feb 1999 11:33:45 GMT Received: by apsn15.ukap.zeneca.com; id LAA29776; Wed, 3 Feb 1999 11:31:39 GMT Received: from ukapzcmsxhub01.ukap.zeneca.com by gbzhp02.ukap.zeneca.com with ESMTP (1.40.112.8/16.2) id AA200221316; Wed, 3 Feb 1999 11:28:36 GMT Received: by ukapzcmsxhub01.ukap.zeneca.com with Internet Mail Service (5.5.2232.9) id {1F6WTK3F} ; Wed, 3 Feb 1999 11:27:36 -0000 Message-Id: {21E55FE938B6D0119A7100805FFEAEC4D0177B-at-UKGR07} } From: Manzor Brian BP {brian.manzor-at-grmouth.zeneca.com} To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-Sparc5.Microscopy.Com}
I'm looking for a gold or graphite/carbon sputter coater for bio specimen preparation.
If you have one to sell, pls let me know details and price.
I'm considering using digital video (DV) for time-lapse/real-time LM. Broadcast/domestic DVcam magazines suggest that DV gives improved resolution over analogue video (S-VHS etc). Added to this, it also seems to give better quality frame by frame editing (+ stills output) when recorded on DV tape (mini or standard) and managed with software like Adobe Premier using Firewire ( aka i.link/IEEE1394) fast serial links.
If anyone has used this technology I'd be interested to hear more about the pros and cons.
TIA
Kevin Jennings -.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-. SmithKline Beecham Pharmaceuticals Analytical Sciences -Microscopy Group New Frontiers Science Park Harlow Essex UK -.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.
I am working on the light reflection of mainly squid iridophores. Are there any groups, preferably in the UK, working with polarising and/or interference microscopes who could give me some advice with this matter?
Thank you very much,
Lydia
PS. Hello Daniele ... back to work !!!
Keith Ryan Marine Biological Association of the UK Citadel Hill Plymouth Devon PL1 2PB England
Gary Gaugler, Ph.D. wrote: =============================================== I'm looking for a gold or graphite/carbon sputter coater for bio specimen preparation.
If you have one to sell, pls let me know details and price. ================================================ There are several very excellent data bases of information of who manufactures what in the microscopy and microanalysis market. You might want to consult some of those listings. Two that I myself make frequent use of are the following:
Microscopy Vendors Data Base http://207.137.96.185/mvd/vendors.html
Microworld Resources and New http://www.mwrn.com/
SPI Supplies is one of the several leading manufacturers of this equipment and all the information you would need our units can be found on our website below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
I can recommend looking at www.bookfinder.com and www.bibliophile.com if you are looking for a used copy of the text (assuming it isn't available new or you are too poor/cheap to pay for it). These are services where you do your own search over a number of used booksellers - I found a couple of technical books I wanted this way and for quite reasonable prices.
Janet Rice MCC Senior Member Technical Staff rice-at-mcc.com 512-338-3266
Yet another lab in search of microtome parts. We have four Leica Ultrotome III ultramicrotomes in very serviceable condition, but a couple need new belts. A supply of spare tubes would also be handy. Is anyone aware of a source for new or used parts for these venerable instruments?
Thanks in advance!
Randy Tindall Electron Microscope Core College of Veterinary Medicine University of Missouri - Columbia Phone: 573-882-8304
Gary: I have a sputter coater for sale. Trouble is I am in Cambridge UK
Patrick Echlin Director, Multi-Imaging Centre School of Biological Sciences Cambridge University
On Wed, 10 Feb 1999, Dr. Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm looking for a gold or graphite/carbon sputter coater for } bio specimen preparation. } } If you have one to sell, pls let me know details and price. } } } } } } Cheers, } Gary Gaugler, Ph.D. } } PGP Public key: } BDC9 8860 AA83 1FB2 66D3 01BB 9EB8 8E6D 671F BEB3 } }
Steve, We actually have an old LKB Microtome we are getting rid of--you are welcome to it, but we are in Boston. If you want to make arrangements to have it shipped to you--it's yours! Peggy Sherwood MGH-Wellman Labs of Photomedicine 224 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-3192 (fax)
Patricia, I didn't see your original e-mail request, but saw the responses! We have a Rapidoprint DD3700 Agfa-Gevaert in our lab for processing EM prints. It's very simple, but like some other processors, it has to be cleaned frequently, especially the water stations, depending upon it's use. Peggy Sherwood MGH-Wellman Labs of Photomedicine
Having worked with minerals of many types, I find that a freeware program for the Mac has been extremely useful in predicting spectra of any mineral. DTSA for either 68K or PPC can be obtained from NIST or possibly from the MSA web site (maybe someone else can confirm this). The program can generate synthetic spectra for just about any detector and will simulate thin or thick specimens. It's a wonderful teaching and research tool that I recommend highly.
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets Johns Hopkins University Baltimore, Maryland 21218 USA Phone: (410) 516-8342 Fax: (410) 516-7933 e-mail: klivi-at-jhu.edu
} Having worked with minerals of many types, I find that a freeware program } for the Mac has been extremely useful in predicting spectra of any mineral. } DTSA for either 68K or PPC can be obtained from NIST or possibly from the } MSA web site (maybe someone else can confirm this). The program can } generate synthetic spectra for just about any detector and will simulate } thin or thick specimens. It's a wonderful teaching and research tool that I } recommend highly. }
DTSA can be downloaded at http://micro.nist.gov/DTSA/dtsa.html .
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Summary and compilation: Tissue cultures and coverslips: Thanks to everyone for the tremendous response to my query. The majority of responses suggested the use of Thermonox coverslips. These may be purchased sterile. One side is prepared for cell adhesion, and is labeled. They are compatible with alcohol, acetone and propylene oxide as solvents, and with Epon, Spurr's and araldite mixtures for resin. These coverslips come in sizes which fit well into tissue culture plates. Other investigators have successfully used glass coverslips, membrane inserts and plastic petri plates. Permanox plates were also recommended, cells may be grown, fixed and embedded in the chambers. Most people use a dip into liquid nitrogen to facilitate removal of the coverslip after embedding. Other simply pull the coverslip off while still warm from the embedding oven. If the coverslip is not removed, it may still be sectioned with a diamond knife, however, the coverslip may separate from the resin under the electron beam. If the coverslip if removed, various methods are suggested to position the cells parallel or perpendicular to the bloc face. A compilation of the responses follows, MANY helpful tricks will be found on these pages. -----------------------------------------------------------------------
I use standard glass coverslips - round ones that fit in 24 and 12 well culture dishes. I sterilize them sitting on a clean filter paper in glass petri dishes then transfer them to culture dishes and seed with cells. I fix and process in 20 ml glass scintillation vials, embed in epon (put coverslip cell side up on a slide with a drop of epon, heat 4-8 hrs at 58 C (timing is important), cross-hatch with a razor blade, slowly lower into liquid nitrogen, and little squares of epon pop off with cells attached. re-embed squares in flat molds and section as usual. Thomas E. Phillips, Ph.D. tphillips-at-biosci.mbp.missouri.edu.
We have had good results with thermonox coverslips and even better with aclar provided the cells will grow on it. Process as usual for TEM and IEM. Heat can curl the aclar sometimes. Scott D. Whittaker sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks " Dear Sally, I too have used Thermanox coverslips for TEM of tissue culture cells, but I found they often pulled out from or wrinkled-up my sections. I had much better luck using filter-membrane inserts. The insert fits into the wells (I used 24-well plates but they come in many sizes). You can even polarize the cell line if necessary by placing media with serum in the well under the insert and without serum in the filter-membrane cup. The membrane then can be fixed and processed in the holder and just before embedding, separate the filter, cut it into pieces, and embed. No wrinkles, no pulling out of the resin (I've used both Spurrs and LR White). Granted, they are more expensive, but. . .
"Tina Schwach" tschwach-at-tc.umn.edu
----------------------------------------------------------- I have used Thermanox coverslips extensively for this purpose. (Check your supplier of Lux tissue culture products, as well as your EM suppliers.) They are resistant to acetone (not sure about propylene oxide) and I've used them with Epon, spurr's and an araldite/epon mixture. They are supplied sterile, and peel off easily from a warm polymerised block, leaving the cells embedded in the resin block. I used to UV sterilise them in a laminar flow hood overnight before use. A word of warning though, they are not suitable for light microscopy, and some cells don't adhere very well to them. As I recall, I have used vero cells and other kidney derived epithelial cell lines on them successfully. You can section thermanox, but I think a better result is obtained by peeling it off, and polymerising some fresh resin over the cells. Contact me with problems if you wish. I have no commercial interest in this product. I've just processed heaps of cultured cells. Good luck, Rosey van Driel {Rosemary.van.driel-at-baker.edu.au}
--------------------- We use the Thermanox coverslips that you can buy from EMS. They work really well you just have to be careful to keep the correct side up (Thermanox has a sidedness to it and the cells only grow right on it). ACLAR works well too, though lately the stuff we buy from Ted Pella seems to be thinner than it used to be & has a tendency to curl up when polymerizing. I you have any questions, please feel free to ask. Paula :-) Paula Sicurello UC Berkeley psic-at-uclink4.berkeley.edu http://biology.berkeley.edu/EML --------------------------------------------------------- Besides the Lux coverslips you can also use membrane inserts made by Falcon or Costar. If you would like have the exact protocol let me know and I can send it to you. I have used them both for number of years now without any problems though I prefer costar inserts. Neelima Shah.............. From: Neelima Shah shahn-at-mail.med.upenn.edu http://www.MED.upenn.edu/morphlab/
---------------------------------------------------------- Visit the lab manual on //con-sgi.microbio.emory.edu/eyes-of-the-eagle/index.html For over ten years our lab has floated monolayers of epithelial celllines from the surface of common plastic culture ware with propylene oxide. This is far easier than trying to use coverslips. Call me at 404 727 3508 if you need some coaching on the technology. Regards, Skip } From: L R MELSEN lmelsen-at-emory.edu http://www.MED.upenn.edu/morphlab/
----------------------------------------------------------------- We routinely attach cells to thermanox coverslips (available from EMS). These coverslips can be sterilized, are compatible with conventional embedding methods and can be thin sectioned with a diamond knife. We fix in GA/FA, osmium, dehydrate and embedd in Spurr's with no problem.
If you must section the coverslip itself, try to arrange it diagonally in the block and pick up the sections on formvar, because the coverslip and the resin tend to separate under the beam.
If you are sectioning parallel to the coverslip, embedd this way. Fill a beam capsule with resin and lay the coverslip, cell side down on top. Polymerize; them immediately upon removing the blocks from the oven, pull the coverslip off. If you do this quickly while the blocks are still warm, the cells will remain on the block and you don't have to worry about sectioning the coverslip. Good Luck, Michelle Peiffer 814-865-212 email:mlk101-at-psu.edu
--------------------------------------------------------------------------- For TEM of cultured cells, we grow the cultures on "Thermanox" tissue culture coverslips. From Nalge Nunc INternational, 50 sterile coverslips, 13 mm in diameter is catalog 174950. EMS also sells these thermanox cover slips in a variety of sizes (see page 143 of their cat XIII). The coverslips can be treated with all the same chemistry as tissue including propylene oxide and Spurrs epoxy, which are two components which solubilize polystyrene. These thermanox coverslips can be sunk cell side up in freshly made Spurrs, then following polymerization the coverslips can be removed by first sawing a small area of the epoxy/cell/substrate, then immersing in liquid nitrogen for a few seconds, then prying away the substrate. Now the embedded cells are immediate on the epoxy. Re-embed two fragments of the culture face to face for cross-sections, or cut the block parallel to the face for tangential sections. We particularly like the round 13 mm thermanox coverslips for immunocytochemistry of cultured cells since they can be floated cell side down in a drop of 100 microlitters antibody - gold conjugate, conserving reagents.
If you would like to grow a larger culture, you could also use "permanox" culture dishes, which are equally re